Pharmaceutical composition for preventing or treating cholestatic liver disease, comprising beta-lapachone and ursodeoxycholic acid as active ingredients

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating cholestatic liver disease, comprising beta-lapachone and ursodeoxycholic acid as active ingredients. Specifically, by orally administering beta-lapachone and ursodeoxycholic acid in combination, the composition is expected to contribute to the treatment of patients with primary sclerosing cholangitis or inflammatory bowel disease accompanied by primary sclerosing cholangitis, which are difficult to treat, by reducing the concentration of TNF-alpha in the blood of the patients and by also significantly reducing the severity of primary sclerosing cholangitis.

Description

Pharmaceutical composition for the prevention or treatment of cholestatic liver disease comprising beta-lapachone and ursodeoxycholic acid as active ingredients

[0001] The present invention relates to a pharmaceutical composition for the prevention or treatment of cholestatic liver disease comprising beta-lapachone (BL) and ursodeoxycholic acid (UDCA) as active ingredients.

[0002] The present invention relates to a pharmaceutical composition for treating primary sclerosing cholangitis or inflammatory bowel disease accompanied by primary sclerosing cholangitis, comprising beta-lapachone (BL) and ursodeoxycholic acid (UDCA) as active ingredients.

[0003] Cholestatic liver disease refers to a group of diseases in which bile abnormally accumulates within hepatocytes or bile ducts due to an obstruction in the flow of bile. Representative cholestatic diseases include primary biliary cholangitis (PBC) or primary sclerosing cholangitis (PSC), intrahepatic cholestasis in pregnant women, Progressive Familial Intrahepatic Cholestasis (PFIC), biliary atresia, cholelithiasis, infectious cholangitis, cholangitis associated with Langerhans cell histiocytosis, Alagille syndrome, nonsyndromic ductal paucity, cholestasis due to viral hepatitis, cholestasis due to alcoholic hepatitis, drug-induced cholestasis, and drug-induced neoplastic cholestasis. These include cholestasis after liver transplantation and cholestasis associated with total parenteral nutrition. Among these, Primary Sclerosing Cholangitis (PSC) is a representative progressive cholestatic liver disease characterized by chronic inflammation and fibrosis of the bile ducts; it not only progresses to cirrhosis and liver failure but also increases the risk of developing liver cancer and cholangiocarcinoma. To date, there is no effective treatment for PSC, and while liver transplantation is the most definitive treatment, there is a risk of recurrence even after transplantation. There are no treatments approved by the U.S. Food and Drug Administration (FDA) that target PSC. Some experts recommend ursodeoxycholic acid (UDCA), which has been proven effective in lowering elevated liver enzyme levels in PSC patients and in other cholestatic liver diseases.

[0004] However, the effect of UDCA on improving PSC symptoms and patient survival rates has not yet been clearly established. While guidelines from the American Association for the Study of Liver Diseases and the American College of Gastroenterology do not recommend the use of UDCA for the treatment of PSC, the European Association for the Study of the Liver recommends the use of intermediate doses (13-15 mg / kg) of UDCA. Therefore, liver transplantation is currently the only proven treatment for PSC, and since not all patients are eligible for a liver transplant, there is an urgent need to develop effective treatments for PSC.

[0005] Meanwhile, it is known that approximately 70–80% of PSC patients also suffer from Inflammatory Bowel Disease (IBD). Inflammatory Bowel Disease (IBD) is a chronic inflammation occurring in the intestines and is classified into Ulcerative Colitis (UC) and Crohn's Disease. In Crohn's Disease, inflammation can occur anywhere from the mouth to the anus, whereas in Ulcerative Colitis, inflammation occurs in the large intestine. Inflammatory bowel disease is prevalent in younger age groups and is characterized by recurring periods of symptom exacerbation and remission, placing a significant burden on patients' quality of life and socioeconomically.

[0006] The pathological characteristics of inflammatory bowel disease can be explained by the excessive activation of pro-inflammatory pathways, dysregulation of inflammatory cytokines, and an autoimmune cascade. In particular, tumor necrosis factor-alpha (TNF-α) is one of the pro-inflammatory cytokines and a major mediator that triggers abnormal immune responses. Inflammatory responses caused by increased TNF-α in the intestinal mucosa and intestinal wall of patients with inflammatory bowel disease lead to the breakdown of the intestinal mucosa and epithelial wall barrier, resulting in edema, ulcers, tissue destruction, and granulomas. Consequently, TNF-α inhibitors are used to treat inflammatory bowel disease by suppressing the activity of TNF-α-mediated pro-inflammatory pathways and reducing chronic inflammation. However, due to the inconvenience of requiring continuous injections to maintain efficacy and the disadvantage of therapeutic efficacy being less than 40%, there is a need for the development of new treatments for inflammatory bowel disease.

[0007] When PSC and IBD coexist, this is classified not merely as a complication but as an independent clinical subtype known as PSC-IBD. Patients with PSC-IBD exhibit clinical features different from those with general IBD; while intestinal symptoms may be relatively mild or asymptomatic, the risk of developing colorectal and cholangiocarcinoma is very high, making early detection and long-term management essential.

[0008] Patients with both PSC and IBD require a complex therapeutic approach that simultaneously improves cholestasis and suppresses intestinal inflammation. However, to date, there are no established standard treatments for patients with PSC, IBD, or IBD accompanied by PSC, and treatments for PSC and IBD alone have limitations in sufficiently improving the prognosis of PSC-IBD patients. For example, UDCA is used to a limited extent for PSC by improving bile acid metabolism, but its effect on survival rate improvement has not been proven; meanwhile, IBD treatments such as anti-TNF agents present issues of increased infection risk and limited response rates in PSC-IBD patients. Therefore, developing new therapeutic strategies capable of addressing the specific pathophysiology of patients with IBD accompanied by PSC is an urgent medical task.

[0009] The object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of cholestatic liver disease comprising beta-lapachone and ursodeoxycholic acid as active ingredients.

[0010] In addition, the objective of the present invention is to provide a pharmaceutical composition for treating primary sclerosing cholangitis or inflammatory bowel disease accompanied by primary sclerosing cholangitis, comprising beta-lapachone and ursodeoxycholic acid as active ingredients.

[0011] The present invention relates to a cholestatic liver disease prevention or treatment method characterized by being one or more selected from the group consisting of primary biliary cholangitis (PBC), primary sclerosing cholangitis (PSC), intrahepatic cholestasis in pregnant women, progressive familial intrahepatic cholestasis (PFIC), biliary obstruction, cholelithiasis, infectious cholangitis, cholangitis associated with Langerhans cell histiocytosis, Alagille syndrome, nonsyndromic ductal paucity, cholestasis due to viral hepatitis, cholestasis due to alcoholic hepatitis, drug-induced cholestasis, drug-induced neoplastic cholestasis, cholestasis after liver transplantation, and total parenteral nutrition-associated cholestasis. Provides a pharmaceutical composition.

[0012] The present invention provides a pharmaceutical composition for treating primary sclerosing cholangitis or inflammatory bowel disease accompanied by primary sclerosing cholangitis, comprising beta-lapachone and ursodeoxycholic acid as active ingredients.

[0013] The above-mentioned Inflammatory Bowel Disease (IBD) is a group of diseases that cause chronic intestinal inflammation, including Crohn's disease or ulcerative colitis.

[0014] Crohn's disease is a chronic inflammatory bowel disease that can occur in any digestive organ, including the small and large intestines. It is a rare, intractable autoimmune disease characterized by chronic inflammation of the mucous membranes of the digestive tract. It is primarily accompanied by chronic abdominal pain (especially in the lower right abdomen), chronic diarrhea, weight loss, loss of appetite, and fever. Additionally, it may be accompanied by perianal pain, abscesses, or fistulas, as well as indigestion and growth disorders.

[0015] Ulcerative colitis refers to an inflammatory bowel disease that occurs in the large intestine. It involves congestion, swelling, and bleeding of the colonic mucosa, as well as the formation of multiple ulcers on the mucosa. It begins with proctitis and gradually spreads upward to invade the entire large intestine, characterized by a continuous connection of lesions.

[0016] The present invention provides a pharmaceutical composition for treating inflammatory bowel disease accompanied by primary sclerosing cholangitis (PSC), comprising beta-lapachone and ursodeoxycholic acid as active ingredients.

[0017] Primary sclerosing cholangitis is an inflammatory disease of the bile ducts in which chronic inflammation causes the walls of the bile ducts to thicken, leading to narrowing or stricture. When inflammatory bowel disease is accompanied by primary sclerosing cholangitis, treatment efficiency is significantly reduced and the patient's quality of life is significantly lowered because the activity of the inflammatory bowel disease and the management of bile duct diseases, such as strictures, must be performed simultaneously.

[0018] In this invention, the term beta-lapachone (BL) refers to a quinone-based compound that has long been obtained from the Lapacho tree and used for medicinal purposes. It is a natural compound found in high concentrations in plants such as the Lapacho tree and Salvia miltiorrhiza, and it receives two electrons from NADH through the intracellular NQO1 (NAD(P)H:quinone oxidoreductase) enzyme to form NAD +It is known to exhibit activity that promotes conversion.

[0019] The above beta-lapachone (BL) may be used in the form of a pharmaceutically acceptable salt, and the salt may be an acid addition salt formed by a pharmaceutically acceptable free acid or a pharmaceutically acceptable metal salt using a base, but is not limited thereto. For example, inorganic acids and organic acids may be used as free acids. Inorganic acids may include hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, etc., and organic acids may include citric acid, acetic acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, succinic acid, 4-toluenesulfonic acid, glutamic acid, or aspartic acid, etc.

[0020] In the present invention, the term ursodeoxycholic acid (UDCA) is a secondary bile acid, also known as ursodiol, that is produced through the metabolism of intestinal bacteria in humans and most other species.

[0021] In the present invention, beta-lapachone may be administered orally at a dose of 100 to 500 mg / day, preferably 100 to 400 mg / day, and ursodeoxycholic acid may be administered orally at a dose of 2 to 28 mg / kg / day, but is not limited thereto.

[0022] A pharmaceutical composition for treating inflammatory bowel disease comprising beta-lapachone and ursodeoxycholic acid as active ingredients according to the present invention can reduce the concentration of TNF-α in the blood of patients with inflammatory bowel disease.

[0023] A pharmaceutical composition for treating inflammatory bowel disease comprising beta-lapachone and ursodeoxycholic acid as active ingredients according to the present invention can improve primary sclerosing cholangitis in patients with inflammatory bowel disease accompanied by primary sclerosing cholangitis. The improvement of primary sclerosing cholangitis may be a decrease in blood alkaline phosphatase, an improvement in fibrosis indicators, or an improvement in bile duct stenosis.

[0024] The above beta-lapachone and ursodeoxycholic acid may be added in an amount preferably 0.001 to 50 weight%, more preferably 0.001 to 40 weight%, and most preferably 0.001 to 30 weight% based on the total weight of the pharmaceutical composition.

[0025] The above pharmaceutical composition may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, liquids, and aerosols, as well as external preparations, suppositories, and sterile injectable solutions, according to conventional methods. Carriers, excipients, and diluents that may be included in the above pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulating, the product is prepared using diluents or excipients such as commonly used fillers, fillers, binders, humectants, disintegrants, surfactants, sweeteners, and acidifiers. Solid dosage forms for oral administration include tablets, pills, powders, granules, and capsules, and these solid dosage forms are prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., with the beta-lapachone and ursodeoxycholic acid of the present invention. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid dosage forms for oral administration include suspensions, liquid formulations, emulsions, and syrups, and may include various excipients, for example, humectants, sweeteners, flavorings, preservatives, and acidifiers, in addition to commonly used simple diluents such as water and liquid paraffin. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. As non-aqueous solvents and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As bases for suppositories, witepsol, macrogol, tween-61, cacao oil, laurin oil, glycerogelatin, etc. may be used.

[0026] The dosage of the pharmaceutical composition of the present invention will vary depending on the age, gender, and weight of the subject to treatment, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration, and the judgment of the prescriber. The determination of the dosage based on these factors is within the level of a person skilled in the art, and generally, the dosage ranges from 0.01 mg / kg / day to approximately 500 mg / kg / day. A preferred dosage is 0.1 mg / kg / day to 200 mg / kg / day, and a more preferred dosage is 1 mg / kg / day to 200 mg / kg / day. Administration may be performed once a day or divided into several doses. The above dosage does not limit the scope of the present invention in any way.

[0027] The pharmaceutical composition of the present invention can be administered to mammals, such as rats, livestock, and humans, via various routes. Any mode of administration is expected, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine or intrathecal injection, and topical application. Since the beta-lapachone and ursodeoxycholic acid of the present invention have minimal toxicity and side effects, they are drugs that can be safely used for long-term prophylactic purposes.

[0028] The present invention provides a health functional food for the prevention or improvement of inflammatory bowel disease accompanied by primary sclerosing cholangitis (PSC), comprising beta-lapachone and ursodeoxycholic acid as active ingredients.

[0029] The above health functional food may be in which beta-lapachone and ursodeoxycholic acid are added in an amount preferably 0.001 to 50 weight%, more preferably 0.001 to 30 weight%, and most preferably 0.001 to 10 weight% based on the total weight of the food.

[0030] The above-mentioned health functional food includes forms such as tablets, capsules, pills, or liquids, and foods to which the extract of the present invention can be added include, for example, various food products, beverages, chewing gum, tea, vitamin complexes, health functional food products, etc.

[0031] The present invention relates to a pharmaceutical composition for treating primary sclerosing cholangitis or inflammatory bowel disease accompanied by primary sclerosing cholangitis, comprising beta-lapachone and ursodeoxycholic acid as active ingredients. Specifically, by orally administering beta-lapachone and ursodeoxycholic acid in combination, the concentration of TNF-α in the blood of patients with primary sclerosing cholangitis or inflammatory bowel disease accompanied by primary sclerosing cholangitis is reduced, and at the same time, the severity of primary sclerosing cholangitis is significantly improved, thereby contributing to the treatment of patients with primary sclerosing cholangitis or inflammatory bowel disease accompanied by primary sclerosing cholangitis, which is difficult to treat.

[0032] Figure 1 shows the design of a Phase 2 clinical trial for PSC and PSC-associated IBD patients receiving combination therapy of beta-lapachone and UDCA according to the present invention.

[0033] Figure 2 shows the effect of combined administration of beta-lapachone and UDCA on ALP concentration in PSC patients.

[0034] Figure 3 shows the effect of combined administration of beta-lapachone and UDCA on reducing ALP in PSC patients.

[0035] Figure 4 shows the results of the analysis of the improvement rate in PSC severity through MRCP analysis of PSC patients with the combined administration of beta-lapachone and UDCA.

[0036] Figure 5 shows the effect of combined administration of beta-lapachone and UDCA on the improvement of PSC severity in PSC patients through MRCP analysis.

[0037] Figure 6 shows the effect of co-administration of beta-lapachone and UDCA on blood TNF-α concentration in patients with PSC and IBD accompanied by PSC.

[0038] Preferred embodiments of the present invention are described in detail below. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms, and the contents introduced herein are provided to sufficiently convey the concept of the present invention.

[0039] <Example 1. Therapeutic Effects of Beta-Lapacone and UDCA on PSC Patients>

[0040] Figure 1 shows the design of the Phase 2 clinical trial for PSC patients. The clinical trial was conducted as a Phase 2a trial to explore the safety and efficacy of beta-lapachone, and a total of 23 patients with PSC or IBD with PSC who were already receiving UDCA at a dose of <28 mg / kg / day were enrolled in this Phase 2a trial. The actual UDCA dosage for enrolled patients ranged from approximately 4.4 to 22.1 mg / kg / day depending on patient body weight.

[0041] The "beta-lapachone group (beta-lapachone and UDCA combined)" and the "placebo control group (placebo, UDCA only)" were randomized in a 2:1 ratio, and 100 mg tablets of beta-lapachone were administered orally twice daily. The study was designed with a treatment period of 12 weeks and 4 weeks of follow-up visits.

[0042] To determine the efficacy of the combined administration of beta-lapachone and UDCA, whole blood was collected from subjects before administration and 12 weeks after administration. Subsequently, the samples were centrifuged, and the supernatant was stored at -70°C for use in subsequent experiments.

[0043] ALP analysis

[0044] The therapeutic effect on PSC in patients with IBD accompanied by PSC was confirmed through the analysis of changes in ALP, magnetic resonance cholangiopancreatography (MRCP), and fibrosis markers using blood from PSC patients treated with the combination of beta-lapachone and UDCA in the Phase 2 clinical trial obtained above.

[0045] The full analysis set (FAset), for which efficacy could be evaluated, consisted of 2 patients who violated the criteria and 2 patients who did not take UDCA during the clinical trial period (1 patient per group), and these were excluded from the results, resulting in a group of 5 patients in the "placebo control group" and 14 patients in the "beta-lapachone treatment group." The protocol compliance set (PPset), which included only patients who fully complied with the clinical trial protocol until the end of the trial, consisted of 4 patients in the placebo control group (UDCA treatment group) and 10 patients in the beta-lapachone group (UDCA + beta-lapachone combination treatment group), and the analysis was conducted. As shown in Figure 2, the change in ALP after 12 weeks of treatment relative to baseline showed that the "beta-lapachone treatment group (beta-lapachone and UDCA combined)" had a significant reduction of more than 55 U / L in the FA set and more than 100 U / L in the PP set compared to the "placebo control group (Placebo, UDCA only)".

[0046] In Figure 3, the effect of beta-lapachone administration on ALP levels was evaluated in patients with primary sclerosing cholangitis (PSC) receiving UDCA. If the ALP level measured at baseline exceeds twice the Upper Limit of Normal (ULN) (bALP > 2XULN), it suggests cholestatic liver damage.

[0047] As a result, as shown in Figure 3, it was confirmed that the combined administration of beta-lapachone and UDCA significantly increased the proportion of patients with a decrease in ALP of 20% or more in PSC patients with cholestatic liver damage (bALP > 2XULN).

[0048] Enhanced Liver Fibrosis (ELF) score

[0049] Enhanced Liver Fibrosis (ELF), a single score related to the severity of liver fibrosis, was evaluated and quantified by combining the measurement of three biomarkers—blood hyaluronic acid (HA), procollagen III N-terminal peptide (PIIINP), and metalloproteinase-1 tissue inhibitor (TIMP-1)—in patients and is shown in Figure 4. As seen in Figure 4, none of the 5 patients who received only UDCA showed improvement in their ELF scores. However, 3 out of 14 patients (21.4%) who received beta-lapachone and UDCA in combination for 12 weeks showed improvement.

[0050] MRCP analysis

[0051] Changes in the severity of PSC patients were evaluated by referring the morphology, stenosis, and dilation of the bile ducts inside and outside the liver to three independent radiologists for MRCP imaging analysis of the patient's liver and bile ducts, and assigning a score (MRCP Anali Score). The MRCP (Magnetic Resonance Cholangiopancreatography) Anali Score is a radiological scoring system for primary sclerosing cholangitis (PSC) that evaluates bile duct dilation, liver deformation, and signs of portal hypertension.

[0052] Among the 5 patients who received only UDCA, none showed a decrease in MRCP Anali scores. However, among the 14 patients who received beta-lapachone and UDCA in combination for 12 weeks, 2 patients (14.3%) showed a decrease in MRCP Anali scores.

[0053] As shown in Figure 5, bile duct stenosis improved in 4 out of 14 patients (28.6%) who received beta-lapachone and UDCA in combination for 12 weeks.

[0054] Based on the above results, it was confirmed that while there was no change in PSC symptoms in PSC patients with UDCA alone, liver fibrosis indicators and bile duct stenosis improved in patients who received beta-lapachone and UDCA in combination for 12 weeks.

[0055] <Example 2. Treatment of IBD Patients with PSC with Beta-Lapacone and UDCA>

[0056] We confirmed whether the combination of beta-lapachone and UDCA showed a therapeutic effect in patients with IBD accompanied by PSC. The concentrations of TNF-α in the blood of patients with PSC and patients with IBD accompanied by PSC obtained in Example 1 above were measured at the start of the experiment (0W) and after 12 weeks of combined administration of beta-lapachone and UDCA (12W), and the results are shown in Fig. 6. The concentration of TNF-α was measured according to the manufacturer's manual using a commercially produced Human TNF alpha ELISA kit (Thermo Fisher Invitrogen KHC3011), and the change from the start of the experiment is indicated.

[0057] As shown in Figure 6, an analysis of blood TNF-α levels in PSC patients and IBD patients with PSC participating in this clinical trial revealed that blood TNF-α levels decreased by an average of 0.06 pg / ml in the IBD patient group with PSC after 12 weeks of beta-lapachone administration, whereas they increased to 0.13 pg / ml in the PSC patient group without IBD. This indicates a superior TNF-α reduction effect in IBD patients with PSC, suggesting a selective therapeutic effect in the treatment of IBD with PSC.

Claims

1. A pharmaceutical composition for the prevention or treatment of cholestatic liver disease comprising beta-lapachone and ursodeoxycholic acid as active ingredients.

2. In Paragraph 1, A pharmaceutical composition characterized in that the above cholestatic liver disease is one or more selected from the group consisting of primary biliary cholangitis (PBC), primary sclerosing cholangitis (PSC), intrahepatic cholestasis in pregnant women, progressive familial intrahepatic cholestasis (PFIC), biliary obstruction, cholelithiasis, infectious cholangitis, cholangitis associated with Langerhans cell histiocytosis, Alagille syndrome, nonsyndromic ductal paucity, cholestasis due to viral hepatitis, cholestasis due to alcoholic hepatitis, drug-induced cholestasis, drug-induced neoplastic cholestasis, cholestasis after liver transplantation, and total parenteral nutrition-associated cholestasis.

3. In Paragraph 2, A pharmaceutical composition characterized in that the cholestatic liver disease is primary sclerosing cholangitis (PSC).

4. In Paragraph 3, A pharmaceutical composition characterized by the improvement of primary sclerosing cholangitis being a reduction in blood alkaline phosphatase, improvement of fibrosis indicators, or improvement of bile duct stenosis.

5. In any one of paragraphs 1 through 4, A pharmaceutical composition characterized by orally administering the above beta-lapachone and ursodeoxycholic acid at a dose of 100 to 400 mg / day of beta-lapachone and 2 to 28 mg / kg / day of ursodeoxycholic acid.

6. A pharmaceutical composition for treating inflammatory bowel disease accompanied by primary sclerosing cholangitis, comprising beta-lapachone and ursodeoxycholic acid as active ingredients.

7. In Paragraph 6, A pharmaceutical composition characterized in that the above-mentioned inflammatory bowel disease is Crohn's disease or ulcerative colitis.

8. In Paragraph 6, The above pharmaceutical composition is characterized by reducing the concentration of TNF-α in the blood of patients with inflammatory bowel disease.

9. In any one of paragraphs 6 through 8, A pharmaceutical composition characterized by orally administering the above beta-lapachone and ursodeoxycholic acid at a dose of 100 to 400 mg / day of beta-lapachone and 2 to 28 mg / kg / day of ursodeoxycholic acid.

10. A health functional food for the prevention or improvement of inflammatory bowel disease accompanied by primary sclerosing cholangitis (PSC), containing beta-lapachone and ursodeoxycholic acid as active ingredients.

Images