Targeted ileum endotoxin chelating agent delivery medicine as well as preparation method and application thereof

Abstract

The invention relates to a targeted ileum delivery endotoxin chelating agent drug and a preparation method and application thereof, and belongs to the technical field of pharmaceutical preparations, the drug comprises a specific high-molecular copolymer, and the high-molecular copolymer comprises a three-dimensional framework formed by organic carbon-carbon chemical bonds and comprises amino groups. The invention aims to deliver a macromolecular chelating agent to the far end of the small intestine to be released so as to chelate pathogen-related molecular patterns generated by intestinal microorganisms, including endotoxin and CpG-DNA. And a compound formed by the macromolecular chelating agent released in the ileum segment and a pathogen molecule mode can be discharged from the digestive tract, so that the inflammation of the organism is reduced, and various metabolic diseases are relieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparations, and relates to targeted ileum delivery of endotoxin chelator drugs, a preparation method thereof, and application in drugs for treating metabolic diseases. The present invention can deliver specific high-molecular polymer chelators to the end of the small intestine for release, so as to chelate and remove pathogenic molecular patterns produced by intestinal microorganisms, including bacterial endotoxins and CpG-DNA, so as to excrete them from the body, and is used to alleviate and treat various metabolic diseases. Background Art

[0002] Metabolic disorders in the form of obesity, fatty liver disease, and type 2 diabetes have brought huge health challenges and heavy economic burdens to societies around the world (The metabolic syndrome: prevalence in worldwide populations, Endocrinol Metab Clin North Am 2004; 33: 351-75) (The global burden of metabolic disease: Data from 2000 to 2019. Cell Metab 2023; 35: 414-428). Studies have shown that metabolic diseases affect a large proportion of the adult population. According to large-scale surveys, the prevalence ranges from about 10% to more than 30%, depending on the region and population studied. In the United States, metabolic syndrome is a major public health problem (Trends in Metabolic Syndrome Among US Youth, From 1999 to 2018, JAMA 2022; 176: 043-1045). The high prevalence is at least partly attributed to factors such as a diet rich in processed foods, high fat, high sugar intake, and a relatively sedentary lifestyle. Additionally, the high obesity rates in the United States greatly contribute to the development of metabolic syndrome and diabetes.

[0004] Drinking alcohol is associated with a wide range of diseases and health problems (Global, regional, and national comparative risk assessment of 84 behavioral, environmental and occupational, and metabolic risks or clusters of risks for 195 countries and territories, 1990-2017: a systematic analysis for the Global Burden of Disease Study 2017, Lancet 2018; 392: 1923-1994). First, excessive drinking leads to fat accumulation in the liver. Heavy drinkers may develop alcoholic hepatitis (AH), which is characterized by jaundice, abdominal pain, and fatigue. Persistent alcoholic hepatitis may progress to liver inflammation and cirrhosis, which may further develop into liver failure, cancer, and other complications such as bleeding from enlarged blood vessels in the esophagus. Drinking alcohol may also lead to cardiovascular disease (CVD), arrhythmias (irregular heartbeat), heart muscle damage, esophagitis and gastritis, peptic ulcers, pancreatitis, and even cancer.

[0005] Blood test indicators of alcoholic liver disease include elevated levels of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transpeptidase (GGT), total bilirubin (TBil), prothrombin time (PT), mean corpuscular volume (MCV), and glucose-deficient transferrin (CDT).

[0006] Under normal physiological conditions, a large number (1-2 kg) of intestinal microorganisms are mostly confined to the large intestine, while the upper small intestine is almost sterile. This gating mechanism is partly mediated by α-defensins 5 / 6 secreted by Paneth cells located in the microvilli crypts (Paneth cell alpha-defensins: peptide mediators of innate immunity in the small intestine. Springer Semin Immunopathol, 2005; 27: 133-46). Loss of Paneth cell function and reduced expression of α-defensins are often associated with many metabolic disorders. In addition, intestinal damage is often associated with aging, as well as intestinal leakage caused by loss of tight junctions of intestinal epithelial cells, which is common in patients with metabolic disorders (The ageing gastrointestinal tract, Curr Opin Clin Nutr Metab Care 2016; 19: 12-8).

[0007] Previous studies have shown that vitamin D signaling controls the function of Paneth cells to some extent by promoting the expression of α-defensins, and vitamin D deficiency, which is often associated with metabolic disorders, can lead to downregulation of defensin expression and small intestinal bacterial overgrowth (SIGO), which may in turn promote endotoxemia, leading to insulin resistance, metabolic disorders and hepatic steatosis (Vitamin D Signaling through Induction of Paneth Cell Defensins Maintains Gut Microbiota and Improves Metabolic Disorders and Hepatic Steatosis in Animal Models, Front Physiol 2016; 7: 498).

[0008] Once pathogen-associated molecular patterns (PAMPs) originating from intestinal microorganisms enter the human body, they can promote metabolic disorders through multiple mechanisms. For example, PAMPs can activate innate immunity through pattern recognition receptors (such as Toll-like receptors) on immune cells (PAMPs and DAMPs: signal 0s that spur autophagy and immunity. Immunol Rev 2012; 249: 158-75). The interaction between ligands and receptors triggers a series of signaling events that may lead to the release of proinflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β), thereby initiating an inflammatory response.

[0009] From a biochemical point of view, endotoxins are lipopolysaccharides (LPS) derived from the cell walls of Gram-negative bacteria. LPS shed from dead bacteria and the cell walls of organisms leads to elevated levels of circulating endotoxins, known as metabolic endotoxins (Gut-derived low-grade endotoxaemia, atherothrombosis and cardiovascular disease, Nat Rev Cardiol 2023; 20: 24-37). LPS levels in the peripheral circulation usually increase after eating. LPS is incorporated into chylomicrons, crosses the intestinal barrier into the lymphatic system, and then enters the blood. In the blood, endotoxins are transported by LPS-binding proteins called LBPs and others such as high-density lipoprotein cholesterol (Chylomicrons promote intestinal absorption of lipopolysaccharides, J Lipid Res 2009; 50: 90-7).

[0010] Under the physiological state of healthy subjects, the above-mentioned HDL-cholesterol or LBP-bound endotoxin is transported to the liver, where it is degraded by specific liver enzymes (such as acyloxyacyl hydroxylases and alkaline phosphatases) or excreted into the bile through scavenger receptors. Kupffer cells in the liver can detoxify endotoxins by phagocytosis. However, with aging or various forms of liver damage, hepatocytes are unable to clear circulating endotoxins, and the ensuing endotoxins overflow into the systemic circulation, leading to mild endotoxemia, which further promotes insulin resistance in type 2 diabetes (T2D) and non-alcoholic fatty liver disease (NAFLD). The pro-inflammatory signal exerted by LPS is generated by binding of its lipid moiety lipid A to TLR4 and binding to the membrane-bound co-receptor CD14. LPS binding to TLR4 leads to the recruitment of the adaptor protein MyD88 to the cytoplasmic domain of TLR4, thereby activating transcription factors such as NF-κB, triggering transcriptional cascades and inflammatory responses.

[0011] Endotoxins in the small intestine can also be absorbed into the liver through the portal vein, especially in cases of aging and intestinal damage caused by various reasons.

[0012] A large number of clinical studies have found that endotoxemia is strongly associated with a variety of diseases, especially liver disease. Long-term drinking, aging, viral infection, and vitamin D deficiency may lead to impaired intestinal integrity and intestinal leakage. In these cases, bacteria in the large intestine can migrate into the small intestine, known as small intestinal bacterial overgrowth (SIBO), which leads to the enrichment of toxins and other pathogen-related molecular patterns in the distal region of the small intestine.

[0013] A large number of studies have shown that the plasma endotoxin level in patients with alcoholic liver disease is several times that of healthy subjects (Endotoxemia in patients with alcoholic and non-alcoholic cirrhosis and in subjects with no evidence of chronic liver disease following acute alcohol excess. J Hepatol 1987; 4: 8-14). The plasma endotoxin level of normal subjects is between 0.3-10.4 pg / ml, while the plasma endotoxin level of patients with alcoholic liver disease is between 8.5-206 pg / ml (Plasma endotoxin and serum cytokine levels in patients with alcoholic hepatitis: relation to severeness of liver disturbance. Alcohol Clin Exp Res 2000; 24: 48S-54S). Although endotoxin varies greatly between different individuals, the plasma endotoxin level of patients with alcoholic liver disease is always 5-20 times that of normal subjects. Animal experimental models of alcoholic liver injury have confirmed endotoxemia in alcoholic liver disease. Based on a male rat model and through binge drinking, one report showed that acute or chronic ethanol administration increases plasma endotoxin levels and that this level is closely associated with the development of liver damage (Exacerbation of alcoholic liver injury by enteralendotoxin in rats. Hepatology 2000; 32: 1008-17). Another study showed that rats fed ethanol showed that increased endotoxemia and lipid peroxidation in female rats stimulated NF-κB activation and chemokine production, enhancing liver damage (Increased severity of alcoholic liver injury in female rats: role of oxidative stress, endotoxin, and chemokines. Am J Physiol Gastrointest Liver Physiol 2001; 281: G1348-56). The alleviation of alcohol-induced endotoxemia and liver damage by antibiotics suggests that endotoxin plays a key causal role in alcoholic liver damage.In fact, oral administration of large amounts of polymyxin B, sterilizing the intestinal microbiota and reducing endotoxins, can improve alcoholic hepatitis in Wistar rats (Antibiotics prevent liver injury in rats following long-term exposure to ethanol. Gastroenterology 1995; 108: 218-24). Emerging evidence suggests that LPS and ethanol have synergistic effects on hepatocytes. LPS alone cannot mimic ethanol-induced steatosis or hepatitis, but ethanol and LPS together can effectively induce liver injury. Ethanol feeding sensitizes the liver of experimental animals to LPS-induced cell damage (Chronic ethanol feeding increases activation of NADPH oxidase by lipopolysaccharide in rat Kupffer cells: role of increased reactive oxygen in LPS-stimulated ERK1 / 2 activation and TNF-alpha production. J Leukoc Biol 2006; 79: 1348-56) (Ethanol feeding enhances inflammatory cytokine expression in lipopolysaccharide-induced hepatitis. J Gastroenterol Hepatol 1997; 12: 305-13).

[0014] In the circulation system, lipopolysaccharide binding protein (LBP) presents bacterial toxins to cell receptors CD14 and TLR-4. Therefore, long-term feeding of ethanol will enhance the expression of LBP and CD14. In contrast, LBP, CD14 and TLR-4 gene knockout mice show resistance to ethanol stimulation, indicating that endotoxin plays a causal role in promoting liver damage (Role of lipopolysaccharide-binding protein in early alcohol-induced liver injury in mice. J Immunol 2002. 168; 6: 2963-9).

[0015] A variety of methods that can reduce endotoxins in the blood have been shown to be an effective method for treating severe liver diseases. To this end, polymyxin B-immobilized fiber columns (PMX) in extracorporeal circulation devices have been used clinically to remove endotoxins from patients with liver failure and septic shock (Efficacy of polymyxin B-immobilized fiber column direct hemolysis for non-endotoxin-associated severe septic shock. Pediatr Int 2016; 58: 1346-1347). Extracorporeal circulation or plasma exchange has been used to reduce hepatotoxins and endotoxins to treat various diseases, such as liver failure, multiple organ failure, and sepsis. However, high costs and surgical risks are major obstacles to extracorporeal circulation methods, including PMX. Importantly, these treatments cannot prevent the influx of intestinal endotoxins into the body. Therefore, the development of oral drugs that can chelate enteric pathogen-associated molecular patterns, including endotoxins, has far-reaching potential application value for the prevention and treatment of various metabolic disorders, including liver diseases. Summary of the invention

[0016] Bile acid sequestrants with a variety of different structures, including cholestyramine, colesevelam, and colestipol, are biocompatible resins. They are not absorbed by the body and can be completely excreted from the digestive tract after binding bile acids. These types of high molecular weight copolymers have been used clinically to reduce hypercholesterolemia by sequestering bile acids, during which cholesterol is converted to bile acids in a compensatory manner.

[0017] The present invention demonstrates that specific copolymers can be used as active pharmaceutical ingredients and can be used to chelate intestinal sources of endotoxins and other pathogen-associated molecular patterns (PAMPs). In addition, the present invention further demonstrates a targeted delivery system that can deliver the above-mentioned chelators to the distal region of the small intestine to achieve efficient chelation and therapeutic potential. The present invention not only improves the chelation efficiency of endotoxins and other PAMs in the ileum, but also significantly reduces the dosage, reduces the adsorption of hydrophobic nutrients, and improves the safety of treatment. These two factors therefore constitute the basis of a new treatment method called PASA (ppathogen adsorption and sequestration agents, pathogen adsorption and chelating agents) therapy. In the current application, we propose a method for preparing a targeted ileum delivery endotoxin chelator and related therapeutic applications.

[0018] The present invention relates to a method for preparing a drug and its potential therapeutic applications. Specifically, it involves the use of specific copolymers as active pharmaceutical ingredients and formulating the drug for targeted delivery to the ileum, thereby effectively sequestering endotoxins and other pathogen-associated molecular patterns (PAMPs) for therapeutic applications. This innovative method has obvious advantages in the pharmaceutical field, enabling precise and effective drug delivery to sequester endotoxins that are abundant in the distal region of the small intestine. The claimed method has great potential in treating a variety of conditions and represents a novel and valuable contribution to the art of drug preparation and therapeutic applications.

[0019] In one aspect, the present invention provides a chelating agent composition for treating a subject suffering from a metabolic disorder caused or promoted by a metabolic endotoxin or other PAMPs originating from the intestine. The pharmaceutical composition comprises a plurality of biocompatible particles and / or polymers as active pharmaceutical ingredients (APIs) that cannot be digested and absorbed by the digestive tract of the subject. The chelating agent composition binds to at least endotoxin or lipopolysaccharide (LPS) and other PAMPs (e.g., CpG-DNA derived from intestinal microorganisms) in the form of a chelating agent-PAMP complex, which may include covalent bonds or non-covalent bonds. Due to the formation of the chelating agent-PAMP complex, intestinal PAMPs including endotoxins are cleared from the digestive tract along with the chelating agent complex.

[0020] Tablets or capsules for sequestering endotoxins among other PAMPs consist of core particles containing specific copolymers as API, a barrier layer, and a pH-sensitive coating. The insoluble and indigestible copolymers in the formulation can be targeted for delivery to distal regions of the small intestine to sequester certain pathogen-associated molecular patterns, such as endotoxins.

[0021] On the other hand, the present invention discloses a method for treating a subject with a metabolic disorder that is closely related to persistent inflammation and the resulting insulin resistance, which is driven in part by metabolic endotoxins. Metabolic disorders are a prominent cause and basis for complications and liver diseases such as alcoholic hepatitis, metabolic-associated fatty hepatitis (MASH), liver fibrosis and cirrhosis, liver failure, and even liver cancer. In addition, metabolic disorders caused by intestinal endotoxins and other PAMPs can also promote complications such as cardiovascular disease (CVD), type 2 diabetes (T2D), and central obesity. Therefore, the principles of PASA therapy and the pH-sensitive and ileum-targeted delivery of endotoxin chelators including specific copolymers can be used to treat these metabolic diseases.

[0022] The following are the innovative points related to the present invention:

[0023] Existing treatments for metabolic diseases (such as non-alcoholic steatohepatitis / metabolism-related steatohepatitis, alcoholic hepatitis) and efforts to develop new drugs mostly target specific cellular targets, including key enzymes and receptors that control metabolic pathways. Metabolic diseases, including various alcoholic liver diseases and non-alcoholic liver diseases, have multiple genetic and epigenetic factors. Therefore, it is difficult for a single targeted drug to achieve broad efficacy. For example, obeticholic acid (OCA) is a new drug being developed for the treatment of NASH, which is a farnesoid X receptor (FXR) agonist. FXR is a nuclear receptor that plays a vital role in bile acid, lipid and glucose metabolism. By activating FXR, OCA should improve liver function and reduce inflammation and fibrosis in patients with NASH. Although OCA has shown some benefits in patients with NASH, the extent of improvement may not be sufficient to meet regulatory requirements for approval. Elafibranor is a dual peroxisome proliferator-activated receptor (PPAR) α / δ agonist designed to improve insulin sensitivity, lipid metabolism and reduce inflammation, but it failed in clinical trials. Different patients may respond differently due to genetic and environmental factors, which makes it difficult to achieve consistent results in different patient populations. Hundreds of genes or single nucleotide polymorphisms (SNPs) have been implicated in the pathogenesis of NASH. For example, genes involved in lipid metabolism, such as PNPLA3 (patatin-like phospholipase domain-containing protein 3), TM6SF2 (transmembrane 6 superfamily member 2), and MBOAT7 (membrane-bound O-acyltransferase domain-containing 7), are strongly associated with an increased risk of NASH. These genes play a role in processes such as triglyceride metabolism and lipid droplet formation. In addition, genes associated with inflammation and immune responses, such as those encoding cytokines and chemokines, also contribute to the pathogenesis of NASH. Indeed, genes involved in the tumor necrosis factor-α (TNF-α) pathway and interleukin-6 (IL-6) signaling can influence inflammation in the liver. Genes involved in oxidative stress and mitochondrial function, such as SOD2 (superoxide dismutase 2), may also be associated with NASH. Mitochondrial dysfunction and related genes can also lead to increased oxidative stress and contribute to liver damage. These small molecule drugs that target specific pathways are not very effective in addressing the broad diversity of NASH pathogenesis. This is also true for other metabolic diseases. Therefore, we need to think creatively and address the upstream sources or causes of liver disease. In this regard, this article proposes a novel treatment approach called PASA (pathogen adsorbent and chelator) therapy. This approach shows promise in the treatment of alcoholic hepatitis and related metabolic disorders. Through the targeted release of copolymer-based chelators in the distal region of the small intestine, the drug can effectively adsorb and chelate microbial-derived pathogens such as endotoxins, thereby blocking the upstream and intestinal sources of inflammation, providing a potential solution to address the root causes of metabolic diseases.

[0024] Gut-derived pathogenic molecular patterns are a common source of inflammation in multiple metabolic diseases. Endotoxins and other pathogen-associated molecular patterns (PAMPs) derived from bacteria translocated from the colon to the small intestine can enter the circulation and liver via two major pathways: absorption by the lymphatic system or via the portal vein. Such high levels of metabolic endotoxins from the gut are a major source of systemic and tissue inflammation. For example, metabolic endotoxins can promote insulin resistance by inactivating insulin-mediated signaling pathways. This underlies type 2 diabetes and other related metabolic disorders.

[0025] Our studies have found that bile acid sequestrants such as cholestyramine and colesevelam have the ability to effectively bind and sequester endotoxins and bacterial CpG-DNA. In addition, we found that oral administration of these chelators can reduce metabolic endotoxin levels in other pathological conditions such as metabolic syndrome, liver fibrosis, and pancreatic cancer. However, the high molecular weight chelators in these original formulations are designed to be released in the upper gastrointestinal tract and are less efficient in capturing endotoxins that are mainly enriched in the distal region of the small intestine. In addition, previous formulations of these chelators are dispersed in the upper small intestine and may also chelate hydrophobic nutrients such as vitamin D and vitamin K to produce some adverse reactions. In response to these problems, the present invention proposes a new method for the preparation, composition and formulation of pharmaceutical preparations for new medical applications and indications by targeting new pathogens.

[0026] In the first part of the present invention, we describe a method for preparing a pH-sensitive and time-dependent formulation for targeted release of chelating agents in the distal region of the small intestine. This method can improve the limitations of existing drugs, improve the efficiency of chelation for eliminating endotoxins in the distal region of the small intestine, and minimize unnecessary chelation of hydrophobic nutrients.

[0027] In the second part of this invention, we have demonstrated for the first time that targeted release of chelators in the ileum can be a therapeutic approach for the treatment of relevant metabolic disorders such as alcoholic hepatitis. Significantly, we have also shown that ileum-targeted colesevelam or cholestyramine drugs can suppress systemic inflammation and liver inflammation, which are prominent factors in many diseases. These findings therefore form the basis for expanding the indication to alleviate other liver diseases.

[0028] In terms of molecular mechanism, we found that ileum-targeted endotoxin chelators can regulate many pathological pathways, including (1) hepatocellular carcinoma, (2) diabetic complications, (3) p53, (4) Hippo, (5) insulin resistance, (6) cell senescence, (7) rheumatoid arthritis, (8) drug metabolism, (9) chemokine signaling, and (10) Th1 and Th2 cell differentiation. This global regulation of liver pathways also lays the foundation for the widespread application of PASA therapy as a new treatment.

[0029] The purpose of the present invention is to provide a drug for treating metabolic disorders caused by intestinal pathogen-related molecular patterns, a preparation method and its application. The specific invention contents are as follows:

[0030] The drug comprises a specific high molecular copolymer, which comprises a three-dimensional framework composed of organic carbon-carbon chemical bonds and an amino group.

[0031] The dosage form of the medicine is pellets, tablets or capsules.

[0032] The micro-pellet or tablet is composed of a core particle, an isolation layer and a pH-sensitive coating; the core particle contains a high molecular copolymer as an active drug ingredient.

[0033] The high molecular copolymer comprises one or any combination of colesevelam, cholestyramine, colestipol and colesamide.

[0034] The pH sensitive coating comprises methyl methacrylate and methacrylic acid.

[0035] The pH sensitive coating ensures drug release within the pH range of 6.5 to 7.5.

[0036] A method for preparing a drug for treating metabolic disorders caused by intestinal pathogen-associated molecular patterns, comprising the following steps:

[0037] (1) Preparation of core particles:

[0038] Colesevelam or cholestyramine, microcrystalline cellulose, croscarmellose sodium and hypromellose were weighed and mixed for 15 minutes to obtain a premixed powder, and then magnesium stearate was added and mixed for 5 minutes to obtain a total mixed powder, and tableted;

[0039] (2) Coating with an isolation layer:

[0040] Pour anhydrous ethanol into a stainless steel barrel, turn on the stirrer and add methacrylic acid-methyl methacrylate copolymer and continue stirring until it is completely dissolved, then continue to add triethyl citrate while stirring, and continue stirring for 1 hour after the addition is complete to prepare an isolation layer coating solution;

[0041] The high-efficiency coating machine is turned on and preheated. When the exhaust air temperature reaches above 30° C., the core particles prepared in step 1) are placed in the high-efficiency coating machine for coating so that the weight gain of the isolation layer is between 2% and 3%, thereby obtaining isolation-coated tablets.

[0042] (3) pH sensitive coating:

[0043] A. Put pure water into a stainless steel barrel, add polyethylene glycol 6000 into part of the pure water and stir continuously until dissolved to obtain an aqueous solution of polyethylene glycol 6000 for later use;

[0044] B. Take another portion of pure water in a stainless steel barrel, add talcum powder into the pure water, and stir for 30 seconds to obtain a talcum powder suspension;

[0045] C. Add the aqueous solution of polyethylene glycol 6000 to the cooled talc suspension and stir for 30 min, then continue to add triethyl citrate and stir for 30 min to obtain suspension 3 for later use;

[0046] D. Weigh the methacrylic acid-ethyl acrylate copolymer aqueous dispersion and put it into a stainless steel bucket, add purified water and stir for 30 minutes to obtain a methacrylic acid-ethyl acrylate copolymer aqueous dispersion suspension;

[0047] E. Adding suspension 3 to the suspension of methacrylic acid-ethyl acrylate copolymer aqueous dispersion and stirring continuously for 60 min to obtain an enteric coating solution with a concentration of 20.0% (w / w);

[0048] F is to coat the enteric layer, start the high-efficiency coating machine, preheat the empty machine, add the isolation coating tablets and place them in the high-efficiency coating machine, adjust the atomization and fan-shaped pressure, and coat the enteric layer so that the weight gain is 8.0-10.0% to obtain the drug.

[0049] A drug for treating metabolic disorders caused by intestinal pathogen-associated molecular patterns in drugs for treating metabolic diseases.

[0050] The present invention also provides a method for preparing a drug for treating metabolic disorders caused by intestinal pathogen-associated molecular patterns (PAMPs), which comprises administering a specially formulated drug to a subject, wherein the drug is intended to deliver a specific high-molecular polymer chelator to the distal region of the small intestine to chelate PAMPs of intestinal microorganisms, including endotoxins and CpG-DNA, and the complex bound to the chelator and PAMPs can be excreted from the digestive tract.

[0051] The specific high molecular weight polymer chelator drug is targetedly released from the pH sensitive coating in the ileum to effectively neutralize the pathogenic molecular pattern at the end of the small intestine, so as to be used for treating the corresponding disease.

[0052] Metabolic diseases include alcoholic hepatitis, metabolic-associated steatohepatitis (MASH), liver fibrosis and cirrhosis, liver failure, liver cancer, cardiovascular disease (CVD), type 2 diabetes (T2D) and central obesity.

[0053] The beneficial effects of the present invention are:

[0054] The drugs of the present invention can effectively adsorb and chelate microbial-derived pathogens, such as endotoxins, thereby blocking the upstream and intestinal sources of inflammation, providing a potential solution to the root causes of metabolic diseases. The present invention can improve the limitations of existing drugs, improve the chelation efficiency of eliminating endotoxins in the distal area of ​​the small intestine, and minimize unnecessary chelation of hydrophobic nutrients. We have demonstrated for the first time that targeted release of chelators in the ileum can be used as a treatment for related metabolic disorders (such as alcoholic hepatitis). We also found that ileum-targeted colesevelam or cholestyramine drugs can inhibit systemic inflammation and liver inflammation.

[0055] The present invention delivers a specific high molecular weight polymer chelator to the distal end of the small intestine and releases it to chelate pathogen-related molecular patterns produced by intestinal microorganisms, including endotoxins and CpG-DNA. The complex formed by the specific high molecular weight polymer chelator released in the ileum segment and the pathogen molecular pattern can be discharged from the digestive tract, thereby reducing inflammation in the body and being used to alleviate various metabolic diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0056] Figure 1 A diagram of a high molecular weight polyamine drug enteric-coated tablet SQ1 prepared in an embodiment of the present invention.

[0057] Figure 2 Computed tomography-based X-ray radiography was used to determine the disintegration time and location profile for targeted ileal release of endotoxin chelators.

[0058] Figure 3 Figure 3. Ileal-releasing cholestyramine formulations have the ability to sequester lipopolysaccharide (LPS) and CpG-DNA fragments in vitro.

[0059] Figure 4 Schematic diagram of two pH-sensitive ileal release pellet prototypes and their in vitro disintegration.

[0060] Figure 5 In vitro disintegration profile of micropellet formulations recorded.

[0061] Figure 6 The in vivo disintegration of the two pH release pellets containing barium sulfate was recorded by X-ray radiography in Sprague Dawley rats at specific time points.

[0062] Figure 7 Acute alcoholic hepatitis in young rats and therapeutic effects of colesevelam and active pharmaceutical ingredient (API) powder suspension released at pH 6.5.

[0063] Figure 8 Ileal release of colesevelam reduces alcohol-induced endotoxemia.

[0064] Fig. 9 Ileal release of colesevelam reduces alcohol-induced systemic inflammation.

[0065] Fig.10 This is a diagram showing the effect of ileal release of colesevelam on reducing alcohol-induced systemic inflammation.

[0066] Fig.11 This is a diagram showing the effect of ileal release of colesevelam on reducing alcohol-induced fatty liver.

[0067] Fig.12 Figure 3. Ileal release of colesevelam reduces alcohol-induced inflammation.

[0068] Fig.13 This is a diagram showing the effect of ileal release of colesevelam on reducing alcohol-induced liver tissue inflammation.

[0069] Fig.14 Figure 1. Acute alcoholic hepatitis in middle-aged rats and the therapeutic efficacy of pH-sensitive, ileum-targeted colesevelam and cholestyramine.

[0070] Fig.15 Figure 3. Ileal-released colesevelam reduces acute alcohol-induced endotoxemia.

[0071] Fig.16 This is a diagram showing the effect of ileal release of colesevelam on reducing systemic inflammation induced by acute alcoholic hepatitis.

[0072] Fig.17 This is a diagram showing the effect of ileal-released colesevelam in reducing the elevation of transaminases induced by acute alcoholic hepatitis.

[0073] Fig.18 This is a diagram showing the effect of ileal release of colesevelam in reducing coagulopathy induced by acute alcoholic hepatitis.

[0074] Fig.19 This is a diagram showing the effect of ileal release of colesevelam on reducing pancreatic damage induced by acute alcoholic hepatitis.

[0075] Fig. 20 This is a diagram showing the effect of ileal release of colesevelam on reducing hyperlipidemia induced by acute alcoholic hepatitis.

[0076] Fig.21 Figure 1 is an analysis of gene expression in the liver of middle-aged rats in a state of acute alcoholic hepatitis compared with a control group.

[0077] Fig. 22 Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of liver gene expression in middle-aged rats in a state of acute alcoholic hepatitis compared with treatment with an ileum-targeted endotoxin chelator.

[0078] Fig.23 This is a diagram showing the effect of ileal release of colesevelam on reducing the expression of hepatic inflammatory genes induced by acute alcoholic hepatitis.

[0079] Fig.24 It is a diagram of liver fibrosis and pathological progression. DETAILED DESCRIPTION

[0080] The present invention is described in detail below in conjunction with the accompanying drawings and specific embodiments. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0081] Example 1

[0082] Preparation and formulation of pH-sensitive targeted ileal release formulations.

[0083] In the present invention, the selected specific high molecular polymer, such as cholestyramine, colesevelam, colestipol, etc., is used as an active pharmaceutical ingredient (API) to prepare a pH-sensitive ileum-targeted preparation or drug.

[0084] A key approach to pH-sensitive formulations is based on the Eudragit system. Specifically, Eudragit S100 and Eudragit L100 are two commonly used polymers, both of which belong to the Eudragit series, which are polymethacrylate copolymers used in drug delivery systems. These two polymers have different properties and applications in drug delivery. Eudragit S100 is an anionic polymer composed of methyl methacrylate (MMA) and methacrylic acid (MAA) in a molar ratio of approximately 1:2, and has a molecular weight of approximately 135 kDa. Eudragit S100 dissolves in environments with a pH greater than 7, making it suitable for colon-targeted drug delivery systems because the pH in the colon is generally above 7. Eudragit L100 is also an anionic polymer, but the molar ratio of MAA to MMA is approximately 1:1, and its molecular weight is also approximately 135 kDa. Eudragit L100 is generally a copolymer of methacrylic acid and methyl methacrylate. The chemical formula of methacrylic acid is C4H6O2. It consists of a carboxylic acid group (-COOH) attached to a carbon chain with a double bond. The chemical formula of methyl methacrylate is C5H8O2. It is the methyl ester of methacrylic acid. Eudragit L100 dissolves in an environment with a pH between 6 and 7.

[0085] Example 2

[0086] Protocols for the preparation of targeted colesevelam or cholestyramine at pH 6.5 and pH 7.5 for ileal release using the Eudragit system are shown in Tables 1 and 2.

[0087] Table 1 Core particle composition

[0088] Element gram API, polymer 70.0 Microcrystalline Cellulose 25 Sodium Carboxymethyl Cellulose 4.5 Magnesium Stearate 0.5 common 100

[0089] Table 2 pH sensitive ileal release

[0090]

[0091] This protocol uses the Eudragit system to prepare a colesevelam or cholestyramine formulation that is targeted for release in the ileum at a pH of approximately 6.5-7.5 as follows:

[0092] 1. Materials and Equipment:

[0093] (A) Colesevelam or cholestyramine powder as the active pharmaceutical ingredient.

[0094] (B) Eudragit polymers suitable for pH-dependent release (e.g., Eudragit L100 for pH 6.5, and Eudragit S100 for pH 7.5 versions).

[0095] (C) Organic solvents (eg, ethanol, isopropanol).

[0096] (D) High shear mixer.

[0097] (E) Spray dryer.

[0098] (F) pH buffer solution used for testing.

[0099] 2. Formula composition:

[0100] (A) Colesevelam or cholestyramine, the active ingredient intended to be released in the ileum.

[0101] (B) Eudragit polymer: Select a Eudragit polymer that is insoluble in acidic conditions and starts to dissolve near pH 6.5, which will ensure targeted release in the ileum.

[0102] (C) Additives: Optional additives such as plasticizers, stabilizers or glidants may be added to improve the properties of the formulation.

[0103] 3. Preparation steps:

[0104] (A) Dissolve Eudragit L00 or Eudragit S100 in a suitable organic solvent, such as ethanol, isopropanol, or acetone, to form a polymer solution.

[0105] (B) Colesevelam powder was added to the solution and mixed thoroughly using a high shear mixer to ensure that colesevelam was evenly dispersed in the polymer solution.

[0106] (C) Adjust the concentration and viscosity of the mixture as necessary to achieve the desired coating properties.

[0107] (D) The colesevelam or cholestyramine particles are coated with the Eudragit polymer using a spray dryer. The spray drying process will result in the formation of coated particles with controlled release properties.

[0108] (E) Optionally, the coated tablets can be further processed by sieving or grinding to obtain a uniform particle size distribution.

[0109] (F) Coated colesevelam formulations were tested in acidic to basic pH buffer solutions to confirm targeted release at pH 6.5.

[0110] 4.Quality Control:

[0111] (A) Particle size analysis: The particle size distribution of the coated colesevelam particles is determined using techniques such as laser diffraction or microscopy.

[0112] (B) Encapsulation efficiency: The amount of colesevelam encapsulated within the Eudragit coating was assessed by appropriate analytical methods.

[0113] (C) In vitro release studies: Release studies were conducted in pH buffered solutions simulating gastrointestinal conditions. The release of colesevelam was monitored over time and targeted release was ensured at pH 6.5.

[0114] (D) Stability testing: The formulations were stored under different conditions (e.g., temperature, humidity) and changes in particle size, encapsulation efficiency, and release characteristics were monitored over time.

[0115] 5. Conclusion: By following this protocol, targeted colesevelam formulations at pH 6.5 and pH 7.5 can be prepared using the Eudragit system for release in the ileum. This formulation has the potential to enhance the therapeutic efficacy of colesevelam or other polymers by targeting their release to specific sites of action in the gastrointestinal tract.

[0116] Example 3

[0117] Another protocol for the preparation of pH-sensitive, ileum-targeted colesevelam and cholestyramine mini-tablets using the Eudragit system is shown in Tables 3 and 4.

[0118] Table 3 Formulation example of SQ6.5, designed to disintegrate colesevelam at pH 6.5

[0119]

[0120] Table 4 Formulation examples of SQ7.5, colesevelam or cholestyramine designed to disintegrate at pH 7.5

[0121]

[0122] (1) Ingredients:

[0123] The core granules contain a bile acid sequestrant as the active pharmaceutical ingredient, along with other ingredients including lactose, microcrystalline cellulose, pregelatinized starch, starch, sodium carboxymethylcellulose, hypromellose, and magnesium stearate.

[0124] The pH sensitive enteric layer contains Eudragit S100 (methacrylic acid-ethyl acrylate) copolymer, hypromellose acetate succinate, carboxylated agarose, carboxymethyl yellow lemon calcium chloride hydrogel, anionic copolymer of methacrylic acid and methyl methacrylate, cellulose acetate phthalate, hypromellose phthalate, polyvinyl acetate phthalate, hypromellose acetate succinate, carboxymethyl cellulose, cellulose acetate, and copolymers of maleic acid and phthalic acid derivatives.

[0125] The coating comprises Eudragit L100 (or Eudragit S100) (5% to 7%), triethyl citrate (0.5% to 0.7%), talc (4% to 5%), ethanol (83% to 87%), and purified water (4% to 6%).

[0126] (2) Process: The specified amounts of colesevelam hydrochloride or cholestyramine, microcrystalline cellulose and sodium carboxymethylcellulose are mixed. The container is rotated at 15 rpm for 15 minutes. After uniform mixing, 5% hypromellose is slowly added for wet granulation, followed by drying in a forced air drying oven at 60°C, followed by further mixing by the addition of magnesium stearate. To prepare microtablets for animal testing, granules are compressed using a 2 mm punch. The core granules are then enteric coated with Eudragit L100 or Eudragit S100 and other ingredients.

[0127] (3) Measurement of disintegration time and pH: The mini-tablets were named SQ6.5, containing colesevelam as the active pharmaceutical ingredient, which disintegrated at pH 6.5, and SQ7.5, which disintegrated at pH 7.5. As shown in Table 5, both tablets could withstand up to 4 hours in 0.1N hydrochloric acid. In contrast, the SQ6.5 mini-tablets disintegrated in a pH 6.5 solution (50 mM Tris at pH 6.5), while the SQ7.5 mini-tablets were stable at pH 6.5 but completely disintegrated in less than 30 minutes at pH 7.5. We also evaluated the hardness of the tablets, including tablets containing barium sulfate for animal experimental tracking (Table 6). The prototype mini-tablets of SQ6.5 and SQ7.5 are shown in Table 5. Figure 4 As shown in A, SQ6.5 represents colesevelam that disintegrates at pH 6.5, and SQ7.5 represents colesevelam that is released at pH 7.5.

[0128] Table 5 Disintegration conditions of ileum-targeted tablets

[0129]

[0130] Table 6 Hardness of pH-sensitive ileum-targeted micropellets

[0131] Name, API Hardness, Newton SQ6.5 disintegrating colesevelam 25.04-32.23 SQ7.5 disintegrating colesevelam 27.12-33.09 SQ6.5 disintegrated barium sulfate 100.9-112.3 SQ6.5 disintegrated barium sulfate 99.8-110.6

[0132] Example 4

[0133] Hydroxypropyl methylcellulose phthalate (T-HPMCP, T-type) is commonly used in pH-sensitive formulations for drug delivery. T-HPMCP is a polymer that exhibits pH-dependent solubility properties. It is insoluble in acidic environments but becomes soluble at higher pH values. This property makes it well suited to protect drugs from degradation in the acidic environment of the stomach and ensure targeted release in higher pH regions of the intestine.

[0134] (1) T-component: For copolymer drugs that are pH-sensitive and targeted to the ileum, T-HPMCP formulations can be designed to release the drug at the appropriate site, improve drug bioavailability, and reduce side effects. Hydroxypropyl methylcellulose phthalate (HPMCP) is used as an enteric polymer to control release in the ileum. Other excipients may include fillers (e.g., microcrystalline cellulose), binders (e.g., polyvinyl pyrrolidone), lubricants (e.g., magnesium stearate), and disintegrants (e.g., cross-linked sodium carboxymethylcellulose).

[0135] (2) Methods: The copolymer as active pharmaceutical ingredient (e.g., colesevelam) and various excipients are weighed and mixed. Granulation is performed if necessary. The mixture is compressed into core granules. The granules are then coated with a T-HPMCP coating using techniques such as pan coating or fluidized bed coating. Coating parameters are adjusted to achieve targeted release of colesevelam in the ileum. Coating thickness and composition can be optimized to ensure that the formulation remains intact in the stomach and small intestine and releases the drug in the ileum.

[0136] Example 5

[0137] Protocol for preparation of a pH 6.5 targeted formulation for ileal release using a poly(lactic-co-glycolic acid) (PLGA) system.

[0138] (1) Introduction: This protocol describes the steps to create a formulation designed to release a copolymer as an active pharmaceutical ingredient in the ileum, which has a pH of approximately 6.5. Poly(lactic-co-glycolic acid) (PLGA) was used as a pH-sensitive polymer because of its ability to degrade and release the encapsulated substance in response to specific pH changes.

[0139] (2) Materials and equipment: poly(lactic-co-glycolic acid) with appropriate molecular weight and degradation characteristics; copolymers as active pharmaceutical ingredients (APIs), such as colesevelam and cholestyramine; organic solvents, such as dichloromethane or acetone; sonicator; rotary evaporator; freeze dryer; pH buffer solutions for testing.

[0140] (3) Formulation composition: PLGA is used as a pH-dependent release pellet encapsulation polymer. The selection of PLGA should be based on its degradation rate at pH 6.5-7.5 and its ability to release the active pharmaceutical ingredient. Different molecular weights and copolymer ratios can be considered to achieve the desired release profile. Colesevelam or cholestyramine is used as the active pharmaceutical ingredient delivered in the ileum, thereby effectively clearing PAMPs produced by intestinal microorganisms. Optional additives such as stabilizers, surfactants or excipients can also be added to the pellet core to enhance the stability and release characteristics of the formulation.

[0141] (4) Preparation steps:

[0142] (A) Dissolve PLGA in an appropriate organic solvent (such as acetone or ethyl acetate) to form a polymer solution. The concentration of PLGA should be optimized to ensure appropriate encapsulation and release properties.

[0143] (B) Add colesevelam or cholestyramine to the solution and mix thoroughly using a sonicator or other suitable mixing equipment. This step ensures that the active pharmaceutical ingredient is evenly dispersed in the polymer matrix.

[0144] (C) Using emulsification techniques, an emulsion of the polymer-API solution is created in the aqueous phase. This can be achieved by adding the polymer solution dropwise to a stirred aqueous solution containing a surfactant.

[0145] (D) The emulsion is allowed to stabilize and then the organic solvent is removed using a rotary evaporator. This step results in the formation of PLGA nanoparticles encapsulating the active pharmaceutical ingredient.

[0146] Example 6

[0147] Improved version of pH-sensitive targeted formulation (Table 7).

[0148] Table 7 Composition examples of cholestyramine (SQ1-T) or colesevelam (SQ1-C) that disintegrate at pH 6.8

[0149]

[0150] (1) Compressed core particles:

[0151] According to the specified amounts in Table 7, colesevelam or cholestyramine (active pharmaceutical ingredient), microcrystalline cellulose, croscarmellose sodium and hypromellose were weighed and mixed for 15 minutes to obtain a premixed powder. Then, magnesium stearate was added and mixed for 5 minutes to obtain a total mixed powder. The content of the intermediate product was tested. Based on the content of the intermediate product, the weight of the tablet to be compressed was calculated. The tablet weight variation was controlled within ±5.0%. A round deep concave punch and die with a diameter of 2 mm were used for compression. The hardness was controlled at 5-15 N / mm2.

[0152] (2) Coating with an isolation layer:

[0153] Turn on the high-efficiency coater and preheat without load. When the exhaust temperature reaches above 30°C, place the prepared core particles into the high-efficiency coater and randomly sample and weigh. Adjust the atomization and fan pressure within the range of 0.1-0.6 MPa. During the coating process, control the inlet temperature to 30±10°C, the exhaust temperature to 30±10°C, the peristaltic pump speed to 1-25 revolutions per minute, and the pot speed to 2-18 revolutions per minute. Ensure that the weight gain of the isolation layer is between 2-3%.

[0154] (3) Pre-preparation of coating:

[0155] (A) Purified water is placed in a stainless steel bucket, and polyethylene glycol 6000 is slowly added to part of the purified water and stirred continuously until dissolved to obtain an aqueous solution of polyethylene glycol 6000 (Solution 1) for later use.

[0156] (B) Take another portion of pure water in a stainless steel barrel, slowly add talcum powder into the pure water, and continue stirring for 30 minutes after addition. Use a high-speed homogenizer to homogenize the stirred suspension for 3 minutes (homogenizer speed: 8000 rpm) to obtain a talcum powder suspension (suspension 2).

[0157] (C) Solution 1 was added to the cooled suspension 2 and stirred for 30 minutes, and triethyl citrate was added and stirred for 30 minutes to obtain suspension 3 for later use.

[0158] (D) The methacrylic acid-ethyl acrylate copolymer aqueous dispersion was weighed and placed in a stainless steel bucket, and the remaining purified water was added and stirred for 30 minutes to obtain a methacrylic acid-ethyl acrylate copolymer aqueous dispersion suspension.

[0159] (E) Suspension 3 was slowly added to Suspension 4 and stirred continuously for 60 minutes to obtain an enteric coating solution with a concentration of about 20.0% (w / w).

[0160] (F) Enteric layer, turn on the high-efficiency coater and preheat without load. When the exhaust temperature reaches above 30°C, add the coated tablets (isolation layer) into the high-efficiency coater and preheat for 5 minutes. After preheating, randomly sample and weigh. Adjust the atomization and fan pressure within the range of 0.1-0.6 MPa. During the coating process, control the inlet temperature to 30±10°C, the exhaust temperature to 30±10°C, the peristaltic pump speed to 10-25 revolutions per minute, and the pot speed to 6-18 revolutions per minute. The weight gain of the enteric layer is between 8.0-10.0%.

[0161] The present invention describes pharmaceutical methods, compositions and formulations. Pharmaceutical formulations contain endotoxin chelators as active pharmaceutical ingredients, and these formulations are designed for targeted release in distal regions of the small intestine. The purpose of these formulations is to chelate and excrete endotoxins and other pathogen-associated molecular patterns. In addition, these formulations inhibit the entry of endotoxins into the circulation and are used for the medical prevention and treatment of metabolic disorders, including liver disease.

[0162] Example 7

[0163] The in vivo disintegration of colesevelam and cholestyramine for ileum-targeted release was investigated using an animal model.

[0164] like Figure 1 As shown, prototypes of three pH-sensitive formulations were prepared as mini-tablets. The blue tablets are for targeted barium sulfate at pH 6.8, which is used to track the time and location of dissolution in the body. The white tablets are for SQ1-C (colesevelam) and the red tablets are for SQ1-T (cholestyramine), both of which are designed to disintegrate at pH 6.5. The mini-tablets used for rat experiments are 2 mm in diameter, 5 mm in length, with a hardness between 5 and 15 Newtons / mm2, approximately 8-10 mg per tablet, and contain 5-7 mg of active pharmaceutical ingredient.

[0165] We measured the disintegration pH of mini tablets. First, it was demonstrated that the three types of tablets could withstand more than 60 min in 0.2 N hydrochloric acid. Subsequently, the tablets were transferred to buffers with increasing pH values ​​from 2.5 to 7.5, which contained 120 mM sodium chloride and 50 mM tris. The dissolution time was recorded, and the tablets were observed to disintegrate in the buffer with a pH of 6.5. Although the tablets disintegrated within 30 min in the buffer with a pH of 6.5, the actual in vivo dissolution time was determined by in vivo tracking using barium sulfate tablets.

[0166] To determine in vivo disintegration, adult SD rats were fasted overnight before oral gavage of 4-5 microtablets containing barium sulfate at pH 6.8. Standard chow and drinking water were then added to the cages. X-ray-based CT scans were performed on the anesthetized rats at the indicated time points. Figure 2As shown, the pH 6.8 tablets began to dissolve in 4-6 hours and were mostly disintegrated within 8 hours. No tablets were found in the gastrointestinal tract 24 hours after administration. To further confirm the disintegration in the distal area of ​​the small intestine, rats were sacrificed at different time points and dissected to determine the location of the tablets. SQ1-T is cholestyramine targeted for ileal release, and SQ1-C is colesevelam targeted for ileal release.

[0167] Example 8

[0168] In vitro chelation assay of endotoxin and CpG-DNA.

[0169] Endotoxin or LPS in solution was measured by the Limulus Amebocyte Lysate (LAL) assay, which is based on the activity of endotoxin. On the other hand, purified endotoxin has a specific adsorption peak at 258 nm, which was used in our in vitro binding assay for adsorption and chelation analysis. Figure 3 As shown in A, we determined the in vitro adsorption / neutralization capacity of SQ1-CT and the active pharmaceutical ingredient powder suspension. Specifically, LPS (B4, Sigma-Aldrich) was dissolved in pure water, and the specific adsorption peak was scanned and recorded at 258 nm. SQ1-CT and the active pharmaceutical ingredient powder were dissolved in a pH 6.5 buffer (120 mM sodium chloride, 50 mM tris, pH 6.5). The high molecular weight resin was washed twice with the same buffer, then centrifuged and collected. Next, a certain amount of LPS was mixed with a certain volume of resin or control buffer. After incubating the mixture for 1 hour, the system was centrifuged and the supernatant was collected for spectrophotometric analysis. The results showed that the adsorption / binding capacity of the SQ-1 formulation for LPS was very similar to that of the active pharmaceutical ingredient copolymer at pH 6.8. In particular, as shown in this test, 1 gram of SQ1 or colesevelam can adsorb or chelate approximately 0.1 gram of endotoxin in vitro. In a similar manner, bacterial genomic DNA was isolated and purified according to standard procedures and Figure 3 B shows the measurement of chelating capacity.

[0170] Example 9

[0171] In vitro disintegration study of two pH-sensitive colesevelam formulations.

[0172] As described in the Examples section of this invention, two types of colesevelam were prepared using the Eudragit system, designated SQ6.5 and SQ7.5, for disintegration at pH 6.5 and pH 7.5, respectively. The mini tablets contained 4.8-5.6 mg of active pharmaceutical ingredient ( Figure 4 ). Figure 4A shows two pH-sensitive ileal release pellet prototypes, SQ6.5 represents colesevelam that disintegrates at pH 6.5, and SQ7.5 represents colesevelam that is released at pH 7.5. Figure 4 B shows the in vitro disintegration time and pH value of two pH-sensitive ileal pellets. SQ6.5 represents colesevelam disintegrated at pH 6.5, and SQ7.5 represents colesevelam released at pH 7.5.

[0173] First, we measured the disintegration pH of the mini-tablets in buffer at 37°C with shaking in vitro. The results showed that the tablets were tolerant and stable in 0.1N hydrochloric acid for up to 4 hours. Subsequently, the tablets were transferred to pH 6.5 and 7.5 buffers containing 120mM sodium chloride and 50mM tris. The dissolution time was recorded. As shown in the figure, the SQ6.5 tablet and the equivalent barium chloride dissolved in the pH 6.5 buffer within 2 hours, while the SQ7.5 tablet and barium chloride were stable at pH 6.5 but completely dissolved in the pH 7.5 buffer within 4 hours ( Figure 5 ). Figure 5 The in vitro disintegration profile of the micropellet formulations is shown, which were placed in a buffer solution and shaken at 37°C for 6 hours. The buffer solutions are: (a) 0.1 mol / L hydrochloric acid, (b) a buffer solution containing 50 mmol / L tris (hydroxymethylaminomethane) (Tris) and 120 mmol / L sodium chloride at a pH of 6.5, and (c) a buffer solution containing 50 mmol / L tris (hydroxymethylaminomethane) (Tris) and 120 mmol / L sodium chloride at a pH of 7.5.

[0174] Ileal targeted release polymer in vivo disintegration experiment. Adult SD rats were fasted overnight. Five SQ6.5 and SQ7.5 tablets containing barium sulfate were administered orally. Then, standard feed and drinking water were added to the cages. X-ray-based CT scans were performed on the anesthetized rats at designated time points. Figure 6 As shown, the pH-sensitive tablets began to dissolve at 8-10 hours, presumably in the distal region of the small intestine. We also confirmed this by feeding rats the mini-tablets and sacrificing and dissecting them at different time points.

[0175] Example 10

[0176] Experimental design for young rats that underwent three weeks of alcohol induction followed by three binge drinking treatments.

[0177] Experimental design, acute alcoholic hepatitis in young rats and evaluation of the efficacy of SQ1. It has been reported that after acute intragastric administration of ethanol (5 g / kg) to Sprague Dawley rats, endotoxin levels in portal blood can rise rapidly from 0 within 60 minutes to 10 pg / mL at 90 minutes. Here, we modified this method to determine endotoxin levels and the efficacy of treatment in rats with alcoholic hepatitis after three weeks. Figure 7 As shown, twenty 7-week-old SPF-grade SD rats were used. Among them, five rats were assigned to the normal control group, while the remaining fifteen rats were fed with Lieber-DeCarli standard alcohol liquid (containing 36% of the total calories from alcohol) for three weeks to establish the model. Subsequently, biochemical metabolic indicators were measured. The level of transaminase (ALT) in the blood increased from 25±10 units / ml in the normal control to 60±8 units / ml, indicating that alcohol caused liver damage. At this time, the model rats were further divided into three groups, each with five rats. While the model group (ASH) continued to be fed with an alcohol liquid diet for another three weeks, one group was treated with SQ1 (SQ1-CV, 7 mini tablets per day, the active drug ingredient dose was 25 mg per day, administered by gavage); the third group was treated with active drug ingredient powder (CV of colesevelam, 25 mg per day, administered by gavage). Finally, the three groups of rats induced by alcohol were treated with binge drinking at a dose of 5 grams of alcohol / kg (10 ml per kg of body weight). Rats were euthanized 24 h after the last binge, and blood, liver, and pancreatic tissues were collected for analysis.

[0178] Embodiment 11

[0179] Targeted ileal release of colesevelam reduces toxinemia in acute alcoholic hepatitis.

[0180] like Figure 8 As shown, we determined Figure 7 The blood endotoxin levels in young rats that were subjected to three binge drinking sessions after six weeks of alcohol induction. As shown in the figure, endotoxin levels increased sixfold from 0.3EU / ml in control rats to 2.0EU / ml in rats with alcoholic liver disease. It is worth noting that administration of SQ1-CV microtablets significantly reduced blood endotoxin levels by approximately 75%, while the active pharmaceutical ingredient powder (CV) also partially reduced blood endotoxin levels. Plasma endotoxin levels were suppressed after treatment with colesevelam (SQ1-C) with a pH of 6.5 and targeted to the ileum, as well as the corresponding colesevelam suspension (CV). The experimental design is as follows Figure 7 The plasma endotoxin level was measured by the Limulus amebocyte lysate (LAL) assay.

[0181] Example 12

[0182] Targeted ileal release of colesevelam can reduce systemic inflammation produced by acute alcoholic hepatitis.

[0183] Once bacterial endotoxin enters the blood, it can induce the expression of a series of inflammatory factors such as TNF-α and IL-1 by activating TLR-4 / CD14 cell membrane receptors and signaling pathways. Alcoholic liver disease (ALD) is characterized by steatosis and upregulation of proinflammatory cytokines including IL-1β. IL-1β, type I IL-1 receptor (IL-1R1), and IL-1 receptor antagonist (IL-1Ra) are important regulators of the IL-1 signaling complex and play a role in inflammation. Since IL-1 is significantly associated with key clinical symptoms of acute alcoholic hepatitis (such as fever, neutrophilia, and cachexia), interfering with the IL-1 pathway may be an attractive therapeutic strategy in the future. The important role of type I IL-1 cytokines and certain inflammasomes has also been confirmed in a mouse model of non-alcoholic fatty liver disease. Colesevelam, which targets the ileum, can effectively reduce blood endotoxin levels. Fig. 9 As shown in Figure 2, serum levels of interleukin-1β were measured to determine alcohol-induced systemic inflammation and its inhibitory effect by oral administration of ileal-targeted SQ1-C and colesevelam suspension (CV). Figure 7 The cytokine was detected by enzyme-linked immunosorbent assay (ELISA). Oral administration of colesevelam targeting the ileum reduced IL-1β levels in the blood by measuring the LPS / TLR4 / CD14 inflammatory signaling pathway in young mice that were induced with alcohol for six weeks and then binge-fed for three weeks, confirming the concept of the present invention.

[0184] Embodiment 13

[0185] Targeted ileal release of colesevelam reduces interleukin-6 levels in acute alcoholic hepatitis.

[0186] Interleukin-6 (IL-6) is a key inflammatory cytokine that plays a central role in the development and progression of liver disease. Serum IL-6 concentrations were significantly increased in patients with alcoholic or non-alcoholic cirrhosis and toxic hepatitis compared with controls. IL-6 can induce alcoholic liver disease (ALD) by activating signal transducer and activator of transcription 3 (STAT3), which subsequently induces the expression of various liver protective genes in hepatocytes. In addition, clinical studies have shown that serum IL-6 levels can be used as a prognostic factor for alcoholic liver disease. On the other hand, endotoxin can rapidly induce the expression of IL-6 in vivo. Studies have found that hepatocytes can express IL-6 under the stimulation of endotoxin. Therefore, liver damage can also reduce the ability to clear IL-6 and promote inflammatory pathogenesis. Fig.10As shown, SQ1-C (i.e., colesevelam with a pH of 6.5 and targeted to the ileum) and its corresponding active pharmaceutical ingredient (API) named CV were able to inhibit alcohol-induced systemic inflammation as measured by serum levels of interleukin-6. Figure 7 The cytokine was detected by enzyme-linked immunosorbent assay (ELISA). By measuring the LPS / TLR4 / CD14 inflammatory signaling pathway, oral ileum-targeted colesevelam can partially reduce the level of IL-6 inflammatory factor in the blood, confirming the concept of our disclosure.

[0187] Embodiment 14

[0188] Fatty liver was evaluated in young rats with alcoholic hepatitis.

[0189] In order to quantitatively evaluate clinical samples and animal experimental data, the International Association for the Study of the Liver divides the degree of fatty liver into mild, moderate and severe. Common evaluation indicators are the percentage of fat content in liver tissue or grading scores. Specifically as follows: F1 is mild fatty liver, with less than 30% fatty liver tissue. It is generally believed that this degree of fat deposition has minimal effect on liver function. F2 is moderate fatty liver, with a fat content in liver tissue between 30% and 60%, which is considered to be moderate fatty liver. Moderate fat deposition may cause inflammatory response and damage, affecting liver function to a certain extent. F3 is severe fatty liver, and when the fat content in liver tissue exceeds 60%, it is considered to be severe fatty liver. Severe fatty liver is usually accompanied by liver inflammation, fibrosis and cell damage, which has a significant impact on liver function. The evaluation of fatty liver is a qualitative and quantitative process, usually determined by medical imaging techniques such as ultrasound, computed tomography or magnetic resonance imaging. For example Figure 7 The liver tissues in the experiments were stained with hematoxylin and eosin and Fig.11 Slices selected according to the scoring criteria are shown in A.

[0190] Embodiment 15

[0191] An ileum-targeted formulation of colesevelam improves alcoholic steatosis.

[0192] like Fig.11As shown in B, fatty liver developed in young mice fed Lieber-DeCarli alcohol plus three binge diets. However, there were significant individual differences within the groups. As shown in the figure, in the alcoholic liver disease model group, one subject showed F3, two showed F2, and two showed F1. After oral administration of SQ1-CV, two subjects showed F2 and the other three showed F1. Importantly, the partial improvement of fatty liver in subjects taking SQ1 was consistent with the reduction of systemic inflammation and expression of inflammatory factors in the liver, further verifying our working hypothesis that SQ1-CV can absorb endotoxins, reduce endotoxemia and systemic inflammation, and thus contribute to the improvement of fatty liver.

[0193] Example 16

[0194] Methods for assessing liver inflammation in experimental animal models of alcoholic hepatitis.

[0195] like Fig.12 As shown in Figure 2, based on the rat liver sections and hematoxylin-eosin staining results obtained in our experiments, we established the liver inflammation index as follows: grade 0, less than 2 lesions in 10 fields of view at 200x magnification; grade 1, exactly 2 lesions in 10 fields of view at 200x magnification; grade 2, 2 to 4 lesions in 10 fields of view at 200x magnification. Importantly, the ranking of the liver inflammation index among the groups was consistent with the levels of endotoxin and systemic inflammatory factors (IL-1), further illustrating the causal relationship of the pathogenesis, that is, the key role of endotoxemia caused by intestinal bacterial toxins in the occurrence of liver disease.

[0196] Embodiment 17

[0197] Targeted ileal release of colesevelam can inhibit the mRNA expression of inflammatory factors in liver tissue in alcoholic liver disease.

[0198] The mRNA expression of two inflammatory factors in liver tissue was measured. Fig.13 As shown, the increased liver inflammation determined by the messenger RNA (mRNA) levels of interleukin-1β (Panel A) in rats was alleviated by daily administration of the ileum-targeted colesevelam formulation SQ1-C at a dose of 91 mg / kg. The increased liver inflammation determined by the messenger RNA (mRNA) levels of interleukin-6 (Panel B) in rats was alleviated by daily administration of the ileum-targeted colesevelam formulation SQ1-C at a dose of 91 mg / kg.

[0199] Administration of 6 weeks of alcohol feeding plus three binge drinking sessions resulted in a 2-fold increase in the expression of the cytokine interleukin IL-1β and a 3-fold increase in the expression of IL-6 in liver tissue. Administration of colesevelam, targeted to the ileum, reduced the expression of both inflammatory factors in liver tissue. Importantly, the expression of the cytokine IL-1 is directly regulated by endotoxin / TLR4 / CD14. Here, we show that its expression in ALD livers follows the exact same trend as blood endotoxin concentrations, again demonstrating a causal pathological relationship. Furthermore, we demonstrate that SQ1 treatment reduces the expression of inflammatory genes in the liver. This suggests that chelating intestinal endotoxins could be used to attenuate the expression of inflammatory genes in the liver.

[0200] Embodiment 18

[0201] Experimental design of rats with acute alcoholic hepatitis and treatment with the ileum-targeted endotoxin chelators SQ1-C and SQ1-T.

[0202] like Fig.14 As shown, forty middle-aged rats (10-12 months old) were used in this experiment. Thirty of them were induced with alcohol for ten days on a Lieber-DeCarli diet (containing 5% alcohol, accounting for 36% of total calories). Then, the animals received a single binge dose of 5 g / kg, 10 ml / kg body weight. The experiment was terminated within 24 hours. During the experiment, pH 6.5 ileum-targeted mini-tablets SQ1-C and SQ1-T containing colesevelam and cholestyramine were administered, respectively, at an oral dose of 91 mg / kg, n=10 per group.

[0203] Embodiment 19

[0204] Inhibition of metabolic endotoxin levels in rats with acute alcoholic hepatitis using the ileum-targeted endotoxin chelators SQ1-C and SQ1-T.

[0205] Chronic alcohol consumption damages the intestinal barrier and increases its permeability. This allows bacterial endotoxins (lipopolysaccharides, LPS) to leak from the intestine into the blood. Once in the blood, LPS becomes a metabolic endotoxin. It binds to Toll-like receptor 4 (TLR4) on immune cells such as Kupffer cells in the liver. This activates the immune system and triggers the production of proinflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). A large amount of clinical research and animal experimental evidence shows that aging-related intestinal damage such as leaky gut further enhances the transport of endotoxins to the liver and systemic circulation. The release of these cytokines leads to liver inflammation and oxidative stress. If this process continues, it can lead to liver cell damage, steatosis (fat accumulation), inflammation (steatohepatitis), fibrosis (scarring), and ultimately cirrhosis. As Fig.15 As shown, for Fig.14 Using the methods mentioned in , we measured circulating levels of metabolized endotoxin in middle-aged rats with acute alcoholic hepatitis with or without drug treatment. Middle-aged rats were induced with alcohol (5% alcohol in the liquid diet, 36% of calories from alcohol) for ten days and then subjected to a binge diet treatment (5 g / kg body weight). This resulted in a significant increase in plasma endotoxin levels, i.e., metabolized endotoxin. Notably, copolymers SQ1-C and SQ1-T, designed at pH 6.5 and targeted to the ileum, could statistically inhibit plasma levels of endotoxin, measured by the Limulus amebocyte lysate (LAL) assay.

[0206] Embodiment 20

[0207] The ileum-targeted endotoxin chelators SQ1-C and SQ1-T with a pH of 6.5 can suppress systemic inflammation in acute alcoholic hepatitis.

[0208] It is well documented that endotoxins can directly induce systemic inflammation in various metabolic disorders including alcoholic hepatitis, manifested by elevations in interleukin-1β and tumor necrosis factor-α. In alcoholic liver disease, IL-1β is an important mediator of inflammation. It is upregulated in response to alcohol-induced liver damage. IL-1β can be induced by bacterial endotoxins that enter the blood and activate the immune system. IL-1β is significantly associated with key clinical symptoms of acute alcoholic hepatitis such as fever, neutrophilia (increased number of neutrophils in the blood), and cachexia (wasting syndrome). Tumor necrosis factor-α is also a key inflammatory cytokine involved in the inflammatory response in alcoholic diseases. Alcohol consumption can lead to increased production of tumor necrosis factor-α. High levels of tumor necrosis factor-α are associated with the progression of alcoholic liver disease from steatosis (fat accumulation in the liver) to more severe forms such as steatohepatitis (inflammation and damage in the liver with fat accumulation), fibrosis (scarring of the liver), and cirrhosis (advanced scarring of the liver). As Fig.16 As shown, for Fig.14 Methods mentioned in, middle-aged rats responded to acute alcoholic hepatitis with a rapid increase in circulating levels of interleukin-1β and tumor necrosis factor-α. In contrast, both colesevelam (SQ1-C) and cholestyramine (SQ1-T), which are pH-sensitive and targeted to the ileum, were able to inhibit systemic inflammation in acute alcoholic hepatitis. First, this result is consistent with a reduction in plasma endotoxin levels by intestinal sequestration of both copolymers, with targeted delivery via a pH-sensitive and time-dependent formulation designed for ileal release. Second, the results showing a reduction in systemic inflammation provide the basis for numerous therapeutic potentials. Cytokine levels in the blood of rats were measured by enzyme-linked immunosorbent assay. As Fig.14As described above, this pH-sensitive and ileum-targeted endotoxin chelator can inhibit the serum level of tumor necrosis factor-α (TNF-α) in acute alcoholic hepatitis and the serum level of interleukin-1β in acute alcoholic hepatitis through intestinal chelation.

[0209] Embodiment 21

[0210] Oral administration of the ileum-targeted endotoxin chelators SQ1-C and SQ1-T, with a pH of 6.5, attenuated elevated plasma transaminase levels in rats with acute alcoholic hepatitis.

[0211] like Fig.17 As shown, the pH-sensitive and ileum-targeted copolymers SQ1-C (with colesevelam as the active pharmaceutical ingredient) and SQ1-T (with cholestyramine as the active pharmaceutical ingredient) were able to suppress the increase in transaminase levels, reduce liver function and inhibit liver damage in middle-aged rats with acute alcoholic hepatitis. As shown in the figure, alcohol induction combined with a single binge drinking treatment in middle-aged rats resulted in statistically increased serum liver transaminases, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. Notably, SQ1-C and SQ1-T could improve liver damage, as shown by the suppression of transaminase levels.

[0212] Embodiment 22

[0213] Ileum-targeted endotoxin chelators SQ1-C and SQ1-T improve coagulopathy in acute alcoholic hepatitis.

[0214] like Fig.18 and Fig.14 As shown in the experimental conditions mentioned in , the coagulation system of middle-aged rats under acute alcoholic hepatitis is impaired, as manifested by increased prothrombin time (PT) and activated partial thromboplastin time (APTT). Importantly, oral administration of colesevelam (SQ1-C) with a pH of 6.5 and targeted ileal release, as well as the corresponding cholestyramine (SQ1-T), can restore coagulation homeostasis in rats with acute alcoholic hepatitis. Fig.14 In the rat model, the prolongation of activated partial thromboplastin time (APTT) caused by alcoholic hepatitis was detected by an automatic chemical analyzer and was found to be inhibited by oral administration of SQ1-C and SQ1-T.

[0215] Embodiment 23

[0216] Oral administration of the ileum-targeted endotoxin chelators SQ1-C and SQ1-T improves pancreatic injury in rats with acute alcoholic hepatitis.

[0217] like Fig.19As shown, alcohol induction for 10 days followed by a binge drinking episode resulted in pancreatic damage as evidenced by elevated plasma levels of the pancreatic markers alpha-amylase (AMY) and alkaline phosphatase (ALK). Notably, oral administration of SQ1-C and SQ1-T, two substances with high molecular weight copolymers in a pH 6.5 targeted ileum formulation as active pharmaceutical ingredients, completely inhibited pancreatic damage and reduced pathological markers. Furthermore, improved pancreatic function by chelation therapy was closely associated with reduced metabolic endotoxins and systemic inflammation, providing a potential causal relationship as a basis for new therapeutic approaches.

[0218] Embodiment 24

[0219] Oral ileum-targeted endotoxin chelators SQ1-C and SQ1-T attenuate hyperglycemia in alcoholic hepatitis.

[0220] like Fig. 20 As shown in the study, alcohol consumption plus overeating significantly increased blood glucose levels in middle-aged rats, i.e., hyperglycemia, an elevated blood glucose level or hyperglycemia, a common feature of alcoholic hepatitis. Notably, oral administration of colesevelam (SQ1-C) and cholestyramine (SQ1-T), which have a pH of 6.5 and are targeted to the ileum, can alleviate Fig.14 Plasma triglyceride and glucose levels were measured by automated chemical analyzers in the rat model of hyperglycemia described in the literature.

[0221] Embodiment 25

[0222] Gene ontology (GO) enrichment and KEGG enrichment analysis of rats with acute alcoholic hepatitis and PASA treatment.

[0223] like Fig.14 RNA extraction from liver tissues was performed for RNA-seq analysis as shown in the experimental design presented in . Rare data from RNA-seq were further subjected to gene ontology (GO) enrichment analysis, which can provide insights into the biological processes that are differentially regulated between the model group (SD rats in acute alcoholic hepatitis) and the control group ( Fig.21 ). The KEGG enrichment analysis results for “Model_vs_control_all_1” provide insights into the differences between animals that binge-drink alcohol (model) and controls in animal models. The results indicate that alcohol consumption mobilizes the following pathways: xenobiotic catabolic processes, sterol and steroid biosynthetic processes, cellular responses to glucocorticoid stimulation, and ketones.

[0224] in, Fig.21A shows gene ontology (GO) analysis of liver gene expression in middle-aged rats under acute alcoholic hepatitis compared with controls. Gene ontology (GO) enrichment analysis provides insight into biological processes that are differentially regulated between the model group (Sprague Dawley rats with alcoholic hepatitis) and the control group. Fig.21 B shows the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of liver gene expression in middle-aged rats under acute alcoholic hepatitis compared with the control group. The results of Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the alcohol model group and the control group help to gain a deeper understanding of the Fig.14 There were differences between the rats that drank excessive amounts of alcohol and the control rats in the experiment.

[0225] Embodiment 26

[0226] KEGG enrichment analysis of liver gene expression in rats with acute alcoholic hepatitis and treated with the ileum-targeted endotoxin chelators SQ1-C and SQ1-T.

[0227] KEGG enrichment analysis provided information on biological pathways involved in middle-aged rats under alcoholic hepatitis treated with SQ1-C or SQ1-T chelation. Fig.14 The analysis showed that binge drinking affects multiple pathways associated with disease and biological processes ( Fig. 22 ). Sequestration of endotoxin by SQ1-C and SQ1-T targeted to the ileum may act on these pathways to counteract the negative effects of alcohol and potentially prevent or treat a variety of diseases. Treatment with SQ1-C and SQ1-T appears to have effects on pathways associated with hepatocellular carcinoma, diabetic complications, p53 and Hippo, insulin resistance, cellular senescence, rheumatoid arthritis, as well as drug metabolism, chemokine signaling, and Th1 versus Th2 cell differentiation.

[0228] in, Fig. 22 A shows Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of liver gene expression in middle-aged rats in a state of acute alcoholic hepatitis compared with treatment with the ileum-targeted endotoxin chelator SQ1-C. Fig. 22 B shows another Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of liver gene expression in middle-aged rats in a state of acute alcoholic hepatitis compared with treatment with the ileum-targeted endotoxin chelator SQ1-T.

[0229] Embodiment 27

[0230] Oral administration of the ileum-targeted endotoxin chelators SQ1-C and SQ1-C inhibited the expression of hepatic TNF-α receptor and thrombospondin 1 (TSP-1) in rats with acute alcoholic hepatitis.

[0231] like Fig.14 As described, middle-aged rats were treated with alcohol for ten days and then binge-fed for one day, and gene expression in liver tissue was analyzed by RNA-seq. Fig.23 As shown, alcohol treatment significantly increased the expression of TNF-α and thrombospondin 1 in the liver, consistent with endotoxin levels and systemic inflammation caused by leukocytes in the body. Here we show that liver tissue has the intrinsic ability to express TNF-α receptors, indicating that the liver is a source of inflammation. TSP-1 can promote the fibrinolytic system, affect blood clot dissolution, and enhance systemic inflammation and disease severity in acute-chronic liver failure. LPS is known to induce the production of TSP-1. Notably, oral administration of two ileum-targeted high-molecular-weight copolymers, SQ1-C and SQ1-T, inhibited the expression of TNF-α (Figure A) and thrombospondin 1 (Figure B) in the liver.

[0232] Embodiment 28

[0233] Oral administration of an ileum-targeted enteric endotoxin sequestering agent reduces liver fibrosis in rats with alcoholic hepatitis.

[0234] Experimental and therapeutic Fig.14 As described in Fig.24 As shown, rats with acute alcoholic hepatitis exhibited liver fibrosis and pathological progression, with increased mRNA levels of lysine oxidase 2 (Lox 2, Figure A) and transforming growth factor beta-2 (Tgfb2, Figure B) in the liver. Strong evidence from clinical associations and genetic experiments suggests a key role for endotoxin signaling in liver fibrosis and hepatic stellate cell activation through Lox2 as well as TGF-beta signaling.

Claims

1. A drug for treating metabolic disorders caused by intestinal pathogen-associated molecular patterns, characterized in that: The drug comprises a specific high molecular copolymer, which comprises a three-dimensional framework composed of organic carbon-carbon chemical bonds and an amino group.

2. A drug for treating metabolic disorders caused by intestinal pathogen-associated molecular patterns according to claim 1, characterized in that: The dosage form of the medicine is pellets, tablets or capsules.

3. A drug for treating metabolic disorders caused by intestinal pathogen-associated molecular patterns according to claim 2, characterized in that: The micro-pellet or tablet is composed of a core particle, an isolation layer and a pH-sensitive coating; the core particle contains a specific high-molecular copolymer as an active drug ingredient.

4. A drug for treating metabolic disorders caused by intestinal pathogen-associated molecular patterns according to claim 3, characterized in that: The high molecular copolymer comprises one or any combination of colesevelam, cholestyramine, colestipol and colesamide.

5. A drug for treating metabolic disorders caused by intestinal pathogen-associated molecular patterns according to claim 3, characterized in that: The pH sensitive coating comprises methyl methacrylate and methacrylic acid.

6. A drug for treating metabolic disorders caused by intestinal pathogen-associated molecular patterns according to claim 3, characterized in that: The pH sensitive coating ensures drug release within the pH range of 6.5 to 7.

5.

7. A drug for treating metabolic disorders caused by intestinal pathogen-associated molecular patterns according to claim 3, characterized in that: The isolation layer contains methacrylic acid-methyl methacrylate copolymer, triethyl citrate and anhydrous ethanol.

8. A method for preparing a drug for treating metabolic disorders caused by intestinal pathogen-associated molecular patterns, characterized in that: The following steps are involved: (1) Preparation of core particles: Weigh colesevelam or cholestyramine, microcrystalline cellulose, croscarmellose sodium and hypromellose to obtain a premixed powder, then add magnesium stearate and mix to obtain a total mixed powder, and compress into tablets; (2) Coating with an isolation layer: Pour anhydrous ethanol into a stainless steel barrel, turn on the stirrer and add methacrylic acid-methyl methacrylate copolymer and continue stirring until it is completely dissolved, then continue to add triethyl citrate while stirring, and continue stirring for 1 hour after the addition is complete to prepare an isolation layer coating solution; The high-efficiency coating machine is turned on and preheated. When the exhaust air temperature reaches above 30° C., the core particles prepared in step 1) are placed in the high-efficiency coating machine for coating so that the weight gain of the isolation layer is between 2% and 3%, thereby obtaining isolation-coated tablets. (3) pH sensitive coating: A. Put pure water into a stainless steel barrel, add polyethylene glycol 6000 into part of the pure water and stir continuously until dissolved to obtain an aqueous solution of polyethylene glycol 6000 for later use; B. Take another portion of pure water in a stainless steel barrel, add talcum powder into the pure water, and stir for 30 seconds to obtain a talcum powder suspension; C. Add the aqueous solution of polyethylene glycol 6000 to the cooled talc suspension and stir for 30 min, then continue to add triethyl citrate and stir for 30 min to obtain suspension 3 for later use; D. Weigh the methacrylic acid-ethyl acrylate copolymer aqueous dispersion and put it into a stainless steel bucket, add purified water and stir for 30 minutes to obtain a methacrylic acid-ethyl acrylate copolymer aqueous dispersion suspension; E. Adding suspension 3 to the suspension of methacrylic acid-ethyl acrylate copolymer aqueous dispersion and stirring continuously for 60 min to obtain an enteric coating solution with a concentration of 20.0% (w / w); F is to coat the enteric layer, start the high-efficiency coating machine, preheat the empty machine, add the isolation coating tablets and place them in the high-efficiency coating machine, adjust the atomization and fan-shaped pressure, and coat the enteric layer so that the weight gain is 8.0-10.0% to obtain the drug.

9. Use of a drug for treating metabolic disorders caused by intestinal pathogen-associated molecular patterns in drugs for treating metabolic diseases.

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