Use of lanthanum carbonate in the preparation of a medicament for the treatment of hepatic encephalopathy

Abstract

The application relates to the biomedical application field of inorganic materials, and particularly relates to application of lanthanum carbonate in preparation of a medicine for treating hepatic encephalopathy. The application relates to application of lanthanum carbonate in preparation of a medicine for treating hepatic encephalopathy, wherein the medicine comprises lanthanum carbonate and pharmaceutically acceptable adjuvants, and the lanthanum carbonate comprises any one or more of lanthanum carbonate hydrate and basic lanthanum carbonate. The application is based on the fact that glutamine enzyme can reduce blood ammonia, and the hepatic encephalopathy is reversed by reducing blood ammonia and systemic inflammation, the liver function is recovered by regulating inflammation, thereby helping to cure the hepatic encephalopathy, and good treatment effect is shown, so that a new candidate medicine for clinically treating the hepatic encephalopathy is provided.

Description

TECHNICAL FIELD

[0001] The present application relates to the biomedical application field of inorganic materials, and particularly to application of lanthanum carbonate in preparation of a medicine for treating hepatic encephalopathy. BACKGROUND

[0002] Hepatic encephalopathy (HE) refers to a central nervous system dysfunction syndrome with metabolic disorder as the main feature caused by severe liver function disorder or disorder. Patients with liver function disorder or disorder have neurological and mental symptoms, and after excluding other brain diseases, they can be diagnosed as HE. The main symptoms are disturbance of consciousness, behavioral disorders, and even coma.

[0003] The ammonia intoxication theory has been accepted since the 20th century and is in the central position of the pathogenesis of HE. The metabolism of ammonia in the human body is mainly participated by the intestinal tract, liver, kidney, and skeletal muscle. In patients with cirrhosis, the increase of ammonia produced in any of the links may lead to HE. At present, the purpose of treating HE in clinic is to reduce blood ammonia through various ways, including the application of antibiotics such as rifaximin to reduce the production of intestinal ammonia, acidification of the intestinal tract to reduce the absorption of ammonia, etc. However, the mechanism of ammonia leading to the onset of HE is not completely clear. On the other hand, a large number of pathological research results suggest that in HE caused by acute liver failure, patients are mainly with brain edema and intracranial hypertension, while in HE patients caused by chronic liver damage, the transformation of astrocytes to Alzheimer Ⅱ cells is mainly. On this basis, scholars at home and abroad explain the relationship between ammonia and brain edema and astrocyte edema through a large number of animal experiments and clinical studies to clarify the pathogenesis of HE. The permeability of the blood-brain barrier increases in liver failure, and ammonia is easy to enter the brain tissue. High levels of ammonia in the brain have great toxicity to the central nervous system. On the one hand, it can directly change the ratio of inhibitory neurotransmitters and excitatory neurotransmitters, and change the expression of some important brain genes, such as astrocyte structural proteins, glutamate transporters, glial fibrillary acidic protein, and aquaporin 4 (AQP4). AQP4 may be related to astrocyte edema in high blood ammonia state, and then damage the autoregulation function of intracranial blood flow. On the other hand, the clearance of ammonia in the brain mainly depends on the role of glutamine synthetase in astrocytes and the synthesis of glutamine from glutamate. Because glutamine has an osmotic effect, when glutamine increases significantly, it can cause intracellular water accumulation and cell edema. In addition, glutamine can also induce mitochondrial permeability transition (MPT) in astrocytes, leading to mitochondrial dysfunction and energy metabolism disorder in brain cells, damaging intracellular signaling pathways, promoting the occurrence of neuronal apoptosis cascade, and causing central nervous system disorders in HE patients. In addition, some scholars believe that ammonia can also affect energy supply by interfering with the tricarboxylic acid cycle to cause cell edema. However, clinical practice has proved that even if the patient's blood ammonia level is very high, HE may not necessarily occur, and there is no certain correlation between blood ammonia level and the severity of hepatic encephalopathy, therefore, the ammonia intoxication theory is not the only mechanism of HE.

[0004] Clinical statistics show that the inflammatory marker (CRP, WBC, TNF, IL-1β, IL-6, IL-10, etc.) level of patients with liver cirrhosis combined with hepatic encephalopathy is significantly higher than that of patients without hepatic encephalopathy. In the process of liver damage, viruses and toxins can stimulate mononuclear macrophage system to produce a large amount of TNF-α, IL-1β, IL-6, IL-10 and other inflammatory mediators or cytokines in addition to directly damaging hepatocytes. Under normal circumstances, the blood-brain barrier can prevent these cytokines and other macromolecules from entering the endothelial cells on the brain side. However, it has been confirmed in animal experiments and in vitro models that inflammatory factors such as TNF-α can promote the increase of blood-brain barrier permeability, which in turn makes it easier for inflammatory factors to enter the brain. On the other hand, the astrocytes in the brain can also produce inflammatory factors. In the early stage, astrocytes mainly produce TNF, which can in turn stimulate astrocytes to produce IL-1β and IL-6. Peripheral and central cytokines form a "cascade effect" of cytokines and systemic inflammatory response syndrome (SIRS) through a series of cascade reactions, which can induce or aggravate HE. Based on this principle, an artificial liver that can remove inflammatory factors is currently considered an effective means of treating end-stage liver disease, and it has been confirmed that the effect of combined use of plasma exchange and hemofiltration is better than that of plasma exchange alone. However, whether the mechanism of action of inflammatory factors in the body is consistent with animal experiments has not been confirmed.

[0005] Lanthanum carbonate, with the trade name FOSRENOL, is developed and marketed by Shire. The marketed dosage form is chewable tablet. The indication is that the product is suitable for the treatment of hyperphosphatemia in patients with end-stage renal disease. The product should be taken with meals or immediately after meals. The recommended initial dose is 750-1500 mg per day, and the dose should be increased every 2-3 weeks until the target level of serum phosphate is reached. The product dissociates in the acidic environment of the upper gastrointestinal tract, binds with phosphate in food to form insoluble lanthanum phosphate complex to inhibit the absorption of phosphate, thereby reducing the levels of serum phosphate and calcium phosphate in the body. Lanthanum carbonate has high biological safety, and in clinical studies, the maximum daily dose of patients can reach 3750 mg. SUMMARY

[0006] The purpose of the present application is to provide the use of lanthanum carbonate in the preparation of a drug for treating hepatic encephalopathy, in order to improve the problems existing in the prior art.

[0007] The present application provides the use of lanthanum carbonate in the preparation of a drug for treating hepatic encephalopathy, wherein the hepatic encephalopathy is type A hepatic encephalopathy and type C hepatic encephalopathy.

[0008] Further, the medicine comprises lanthanum carbonate and pharmaceutically acceptable excipients, and the pharmaceutically acceptable excipients comprise any one or more of lactose, starch, gelatin.

[0009] Further, the lanthanum carbonate comprises any one or more of lanthanum carbonate hydrate and basic lanthanum carbonate.

[0010] Compared with the prior art, the present application has the following advantages: the current drug for hepatic encephalopathy mainly includes ammonia reduction, but only ammonia reduction without recovery of liver function, and the patient will be repeatedly hospitalized, and the degree of deterioration is more and more serious. The present medicine is a breakthrough design based on glutamine enzyme inhibition to reduce blood ammonia, and helps to cure hepatic encephalopathy by regulating inflammation to recover liver function. BRIEF DESCRIPTION OF DRAWINGS

[0011] The accompanying drawings are included to provide a further understanding of the present application, and are incorporated in and constitute a part of this specification, illustrate embodiments of the present application, and together with the description serve to explain the principles of the present application, and should not be considered limiting of the present application in any way. In the drawings:

[0012] Figure 1 The graph of blood phosphorus and calcium concentration of the A-type and C-type hepatic encephalopathy rat model in Test Example 2.1 is shown.

[0013] Figure 2 The graph of glutamic-pyruvic transaminase and glutamic-oxaloacetic transaminase data of the A-type and C-type hepatic encephalopathy rat model in Test Example 2.1 is shown.

[0014] Figure 3 The graph of total protein and total bilirubin data of the A-type and C-type hepatic encephalopathy rat model in Test Example 2.1 is shown.

[0015] Figure 4 The graph of albumin and globulin data of the A-type and C-type hepatic encephalopathy rat model in Test Example 2.1 is shown.

[0016] Figure 5 The graph of albumin-globulin ratio and alkaline phosphatase data of the A-type and C-type hepatic encephalopathy rat model in Test Example 2.1 is shown.

[0017] Figure 6 The graph of creatinine and urea data of the A-type and C-type hepatic encephalopathy rat model in Test Example 2.1 is shown.

[0018] Figure 7 The graph of behavior change of the A-type and C-type hepatic encephalopathy rat model observed by the open field experiment in Test Example 2.2 and the total walking distance statistics are shown.

[0019] Figure 8 The graph of central walking distance and central stay time data of the A-type and C-type hepatic encephalopathy rat model in Test Example 2.2 is shown.

[0020] Figure 9 Figure 2.2-1 is a graph showing the pathological detection of the cerebral cortex area of the rat model of type A and type C hepatic encephalopathy in Test Example 2.2.

[0021] Figure 10 Figure 2.2-2 is an electron microscope image of the cerebral cortex area of the rat model of type A and type C hepatic encephalopathy in Test Example 2.2 and a graph of the nuclear perimeter statistics thereof.

[0022] Figure 11 Figure 2.2-3 is a graph showing the concentration of blood ammonia in the serum of the rat model of type A and type C hepatic encephalopathy in Test Example 2.2.

[0023] Figure 12 Figure 2.2-4 is a graph showing the concentration of glutamine and glutamic acid in the serum of the rat model of type A and type C hepatic encephalopathy in Test Example 2.2.

[0024] Figure 13 Figure 2.2-5 is a graph showing the concentration of ammonia and glutamine in the cerebrospinal fluid of the rat model of type A and type C hepatic encephalopathy in Test Example 2.2.

[0025] Figure 14 Figure 2.2-6 is a graph showing the relative activity of glutamine enzyme in the small intestine and liver of the rat model of type A and type C hepatic encephalopathy in Test Example 2.2.

[0026] Figure 15 Figure 2.2-7 is a graph showing the data of interleukin 1β and interleukin 6 in the serum of the rat model of type A and type C hepatic encephalopathy in Test Example 2.2.

[0027] Figure 16 Figure 2.2-8 is a graph showing the data of tumor necrosis factor α in the serum of the rat model of type A and type C hepatic encephalopathy in Test Example 2.2.

[0028] Figure 17 Figure 2.3-1 is a flowchart showing the process of oral administration of the elastic cable to reduce blood ammonia and inflammation to reverse hepatic encephalopathy in the rat model. DETAILED DESCRIPTION

[0029] In order to make the purposes, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below. Obviously, the described embodiments are some but not all of the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative efforts should fall into the scope of the present application. Unless otherwise defined, the technical terms or scientific terms used herein should be understood as the common meanings of the terms to those of ordinary skill in the art to which the present application belongs. The terms such as "comprise" and the like used herein are intended to encompass the elements or components appearing before the terms as well as their equivalents and other elements or components.

[0030] The embodiment provides a preparation method of a medicine mixture containing lanthanum carbonate, and comprises the following steps: mixing medicine (lanthanum carbonate and pharmaceutically acceptable adjuvant) with animal food to obtain the medicine mixture.

[0031] Specifically, after the medicine mixture is prepared, the mass fraction of lanthanum carbonate in the medicine mixture is 1-10%.

[0032] In some embodiments, the lanthanum carbonate used in the medicine mixture comprises any one or more of lanthanum carbonate hydrate and basic lanthanum carbonate.

[0033] Specifically, the medicine is ground and then mixed with food to obtain the medicine mixture, and the animal model with hepatic encephalopathy is orally administered.

[0034] Specifically, the medicine mixture containing lanthanum carbonate can improve the symptoms of hepatic encephalopathy of the animal after continuous administration for four weeks.

[0035] Specifically, the mechanism of lanthanum carbonate for treating liver fibrosis is ① reducing blood ammonia; and ② reducing cell inflammatory factors.

[0036] In some embodiments, the therapeutic effect of lanthanum carbonate is comprehensively evaluated by rat behavior, microscopic analysis of cerebral cortical area, blood ammonia, blood inflammatory factors and total bile acid.

[0037] Specifically, the inflammatory factors include TNF-α, IL-1β and IL-6β.

[0038] In some embodiments, the safety of lanthanum carbonate is evaluated by detecting the levels of biochemical indicators in serum.

[0039] Embodiment 1, preparation of a medicine mixture:

[0040] S1, 20g of lanthanum carbonate is mixed with 380g of rat feed, and a grinder is used to grind for 3min to obtain uniform powder;

[0041] S2, 400 mL of pure water was added to the uniform powder obtained in step S1, and stirred until the feed became dough-like;

[0042] S3, the feed dough obtained in step S2 was shaped into equal-sized cubes and completely air-dried to obtain a drug mixture containing 5% lanthanum carbonate.

[0043] Test Example 2.1: Construction of a rat model of hepatic encephalopathy and verification of the safety of oral lanthanum carbonate;

[0044] Type A hepatic encephalopathy in a thioacetamide-induced acute liver failure model: 12 male SPF SD rats, 8 weeks old and weighing 230 ± 20 g, were randomly selected, 6 of which were normally fed, and the other 6 were fed with 3% thioacetamide (mass ratio) in their diet, and 300 mg / kg was injected intraperitoneally every other day, and the modeling was successful after two consecutive doses. Subsequently, the diet containing 5% lanthanum carbonate was fed for four consecutive weeks. Blood was taken from the eyeball before and at the end of the administration, and the rat heart, liver, spleen, lung, and kidney were taken at the end of the administration (four weeks) for use;

[0045] Type C hepatic encephalopathy in a carbon tetrachloride-induced acute liver failure model: 12 male SPF SD rats, 8 weeks old and weighing 230 ± 20 g, were randomly selected, 6 of which were normally fed, and the other 6 were given 30% carbon tetrachloride (dissolved in olive oil) by intraperitoneal injection, with a dose of 2 mL / kg twice a week for eight weeks, and the modeling was successful. Subsequently, the diet containing 5% lanthanum carbonate was fed for four consecutive weeks. Blood was taken from the eyeball before and at the end of the administration, and the rat heart, liver, spleen, lung, and kidney were taken at the end of the administration (four weeks) for use;

[0046] During blood collection, a safe dose of 5% chloral hydrate was injected intraperitoneally for anesthesia; then the eyeball site was disinfected and a capillary was used to puncture the lower part of the eyeball to collect 1 mL of blood; the collected blood was placed at 4°C for 12 h, and after standing, the serum and tissues were collected by centrifugation;

[0047] The serum biochemical indicators for evaluating liver function were determined using a biochemical analyzer, including: alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), total protein (TP), albumin (ALB), globulin (GLB), and alkaline phosphatase (ALP). The heart, liver, spleen, lung, and kidney tissues obtained were subjected to H&E staining.

[0048] Test Example 2.2: Verification of the therapeutic effect of lanthanum carbonate on hepatic encephalopathy;

[0049] 2.2.1 Construction of TAA and CCl4-induced hepatic encephalopathy rat models and grouping:

[0050] Eight-week-old male SPF-grade SD rats weighing 230±20g were used to model hepatic encephalopathy. Following the modeling of hepatic encephalopathy of types A and C as described above, the rats were divided into eight groups, including: normal group A and normal group C, hepatic encephalopathy group A and hepatic encephalopathy group C, lanthanum carbonate treatment group A and lanthanum carbonate treatment group C, and ornithine treatment group A and ornithine treatment group C.

[0051] The control groups A and C each consisted of 6 normal rats; the hepatic encephalopathy group A, lanthanum carbonate treatment group A, and ornithine treatment group A each consisted of 6 rats with successfully modeled hepatic encephalopathy type A; the hepatic encephalopathy group C, lanthanum carbonate treatment group C, and ornithine treatment group C each consisted of 6 rats with successfully modeled hepatic encephalopathy type C; the control groups A, C, A, and C were fed a normal diet; the lanthanum carbonate treatment groups A and C were given an oral mixture containing 5% lanthanum carbonate, with an average daily intake of 1.25-2.25 g of lanthanum carbonate; the ornithine treatment groups A and C were treated with ornithine aspartate according to dosage conversion, specifically, ornithine aspartate was added to drinking water at a rate of 2 g / kg / day, for four weeks, either by administration or a normal diet.

[0052] In some embodiments, for ease of description, normal group A and normal group C are collectively referred to as the normal group, hepatic encephalopathy group A and hepatic encephalopathy group C are collectively referred to as the hepatic encephalopathy group, lanthanum carbonate treatment group A and lanthanum carbonate treatment group C are collectively referred to as the lanthanum carbonate treatment group, and ornithine treatment group A and ornithine treatment group C are collectively referred to as the ornithine treatment group.

[0053] 2.2.2 Validation of the therapeutic effect of oral drug mixture on a rat model of hepatic encephalopathy;

[0054] Rat behavior was assessed using an open field test; the total walking distance, central walking distance, and central dwell time of the rats were recorded during the open field test.

[0055] Four weeks after feeding, all rats were sacrificed, and the cerebral cortex was harvested for H&E staining to analyze the degenerated and necrotic neurons, particularly the swelling of neurons accompanied by Nissl body dissolution. Electron microscopy analysis of the cerebral cortex was performed to calculate the perimeter of astrocyte nuclei.

[0056] Four weeks after feeding, all rats were sacrificed, and cerebrospinal fluid was collected. Ammonia, glutamine, and total bile acids in the cerebrospinal fluid were quantitatively measured.

[0057] Four weeks after feeding, all rats were sacrificed, and whole blood was collected. After centrifugation, serum was obtained, and serum ammonia, calcium, phosphorus, glutamate, glutamine, and total bile acids were quantitatively measured.

[0058] Four weeks after feeding, all rats were sacrificed, and whole blood was collected. After centrifugation, serum was obtained, and liver and kidney function indicators in the serum were quantitatively measured.

[0059] After 4 weeks of feeding, all rats were sacrificed, whole blood was taken, serum was obtained after centrifugation, and inflammatory markers in serum were quantitatively measured.

[0060] After 4 weeks of feeding, all rats were sacrificed, small intestine and liver tissues were taken, and glutamine enzyme activity was detected using a kit;

[0061] Results analysis:

[0062] Data are expressed as mean ± standard deviation (SD), and statistical analysis was performed using SPSS 22.0 and Graphpad Prism 6.01. Differences between two groups were evaluated using t-test; mean values between 3 groups or more were compared using analysis of variance (ANOVA).

[0063] 1. Oral lanthanum carbonate biosafety verification in rats with type A and C hepatic encephalopathy in test example 2.1:

[0064] Reference Figures 1-5 After the end of administration, we detected the serological indicators of rats by serum biochemical analyzer. The results showed that the blood phosphorus and calcium of rats with hepatic encephalopathy did not change significantly after injury, but the blood phosphorus decreased by about 10-20% after taking lanthanum carbonate for four weeks, while the blood calcium did not change. Among them, the blood phosphorus level of rats: P 正常组A vs 肝性脑病组A =0.9993, P 正常组A vs 碳酸镧治疗组A =0.0036; P 正常组C vs 肝性脑病组C =0.9667, P 正常组C vs 碳酸镧治疗组C =0.018; Specifically, P represents the statistical difference value, when P value is less than 0.05, it is considered that there is a statistically significant difference; when P value is less than 0.01, it is considered that there is a statistically significant difference; when P value is less than 0.001, it is considered that there is a statistically significant difference; when P value is greater than 0.05, it is considered that there is no difference in statistics; it should be clear that P 正常组A vs 肝性脑病组A represents the statistical difference value between normal group A and hepatic encephalopathy group A, P 正常组C vs 肝性脑病组C represents the statistical difference value between normal group C and hepatic encephalopathy group C, and so on. Compared with the normal group, the liver function indicators of rats with type A and C hepatic encephalopathy, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), globulin (GLB), white and globulin ratio (A / G, i.e. albumin and globulin ratio), alkaline phosphatase (ALP) and total protein (TP), were significantly increased, indicating that the rat liver was severely damaged and the liver function was significantly decreased, but there was no difference in albumin (ALB). The liver function level of rats with type A hepatic encephalopathy: ALT (P 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A <0.0001, P 肝性脑病组Avs 碳酸镧治疗组A <0.0001), AST (P正常组A vs 肝性脑病组A =0.0050, P 肝性脑病组A vs 鸟氨酸治疗组A =0.9944, P 肝性脑病组Avs 碳酸镧治疗组A =0.2176), ALP (P 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A , P 肝性脑病组Avs 碳酸镧治疗组A <0.0001), ALB (no difference), GLB (P 正常组A vs 肝性脑病组A =0.0036, P 肝性脑病组A vs 鸟氨酸治疗组A =0.1140, P 肝性脑病组Avs 碳酸镧治疗组A =0.0437), A / G (P 正常组A vs 肝性脑病组A =0.0014, P 肝性脑病组A vs 鸟氨酸治疗组A , P 肝性脑病组Avs 碳酸镧治疗组A =0.0661), TP (P 正常组A vs 肝性脑病组A =0.0012, P 肝性脑病组A vs 鸟氨酸治疗组A =0.0201, P 肝性脑病组Avs 碳酸镧治疗组A =0.0731), TBIL (P 正常组A vs 肝性脑病组A =0.0002, P 肝性脑病组A vs 鸟氨酸治疗组A =0.0169, P 肝性脑病组Avs 碳酸镧治疗组A =0.0048). Hepatic function levels in rats with hepatic encephalopathy type C: ALT (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C <0.0001, P 肝性脑病组C vs 碳酸镧治疗组C <0.0001), AST (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C <0.0001, P 肝性脑病组C vs 碳酸镧治疗组C <0.0001), ALP (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C =0.0086, P 肝性脑病组C vs 碳酸镧治疗组C <0.0001), ALB (no difference), GLB (P 正常组C vs 肝性脑病组C <0.0001, 肝性脑病组C vs 鸟氨酸治疗组C =0.0017, P 肝性脑病组C vs 碳酸镧治疗组C =0.0047), A / G (P 正常组C vs 肝性脑病组C =0.0231, P 肝性脑病组C vs 鸟氨酸治疗组C =0.1867, P 肝性脑病组C vs 碳酸镧治疗组C =0.4497), TP (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C =0.0781, P 肝性脑病组C vs 碳酸镧治疗组C =0.0055), TBIL (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C =0.0074, P 肝性脑病组C vs 碳酸镧治疗组C =0.0006).

[0065] In addition, as Figure 6As shown, we found that the changes of urea and creatinine concentrations in serum of rats showed injury, which also suggested that the kidney function of rats was damaged. The ornithine treatment group was slightly improved, but the recovery of liver and kidney function was weaker than that of the lanthanum carbonate treatment group. The concentrations of serum creatinine and urea in rats with C-type hepatic encephalopathy: creatinine (P 正常组C vs 肝性脑病组C =0.0086, P 肝性脑病组C vs 鸟氨酸治疗组C =0.0545, P 肝性脑病组C vs 碳酸镧治疗组C =0.9623), urea (P 正常组C vs 肝性脑病组C =0.0027, P 肝性脑病组C vs 鸟氨酸治疗组C =0.2813, P 肝性脑病组C vs 碳酸镧治疗组C =0.0005). The concentrations of serum creatinine and urea in rats with A-type hepatic encephalopathy: creatinine (P 正常组A vs 肝性脑病组A =0.0003, P 肝性脑病组A vs 鸟氨酸治疗组A =0.048, P 肝性脑病组Avs 碳酸镧治疗组A =0.0003), urea (P 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A =0.0148, P 肝性脑病组Avs 碳酸镧治疗组A =0.0002).

[0066] 2. Test the efficacy of lanthanum carbonate treatment of A and C type hepatic encephalopathy rats in Example 2.2.

[0067] (1) Preliminary treatment effect after four weeks of administration:

[0068] Figure 7 and Figure 8 The results showed that the total walking distance and central walking distance of rats with hepatic encephalopathy were significantly shortened, and the central residence time was significantly reduced, indicating that the rats had low exploration desire; similar to the behavior after brain injury of similar hepatic encephalopathy. The efficacy of the lanthanum carbonate treatment group was obvious, and the total walking distance and central walking distance were significantly increased, and the central residence time was significantly increased, which was close to the normal group and better than the ornithine treatment group. Rats with C-type hepatic encephalopathy: total walking distance (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A <0.0001, P 肝性脑病组C vs 碳酸镧治疗组C <0.0001), central walking distance (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C =0.0016, P 肝性脑病组C vs 碳酸镧治疗组C =0.0026), central residence time (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C <0.0001, P 肝性脑病组C vs 碳酸镧治疗组C <0.0001). Rats with A-type hepatic encephalopathy: total walking distance (P 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A =0.0002, P 肝性脑病组Avs 碳酸镧治疗组A=0.0091), the central walking distance of rats (P 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A =0.0006, P 肝性脑病组Avs 碳酸镧治疗组A =0.0489), rat center dwell time (P 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs鸟氨酸治疗组A =0.0007, P 肝性脑病组A vs 碳酸镧治疗组A =0.0005).

[0069] Figure 9 The results showed that the astrocytes in the cerebral cortex of rats with hepatic encephalopathy exhibited significant degeneration and necrosis, manifested as neuronal cell swelling accompanied by Nissl body dissolution. However, after treatment with lanthanum carbonate, such necrosis was rarely observed, and the results were similar to those in the normal group, which was superior to the ornithine treatment group.

[0070] Figure 10 The results showed that astrocytes in the cerebral cortex of rats with hepatic encephalopathy exhibited significant swelling, specifically characterized by increased nuclear volume, reduced and lighter-colored heterochromatin, indicating significant nuclear damage. After treatment with lanthanum carbonate, the nuclear perimeter changed little, and the heterochromatin color was darker than in the damaged group, approaching that of the normal group, which was superior to the ornithine treatment group. Astrocyte nuclear perimeter in the cerebral cortex: Type C hepatic encephalopathy rats: P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C =0.0072, P 肝性脑病组C vs 碳酸镧治疗组C =0.0014. Type A hepatic encephalopathy rats: P 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A =0.0142, P 肝性脑病组A vs 碳酸镧治疗组A =0.0028.

[0071] Figures 11-12 The results showed that serum ammonia levels were significantly increased, glutamine levels were decreased, and glutamate levels were increased in rats with hepatic encephalopathy after injury. After treatment with lanthanum carbonate, serum ammonia levels decreased to almost normal levels, glutamine levels increased, and glutamate levels decreased, approaching those of the normal group and showing better results than the ornithine treatment group. Type C hepatic encephalopathy rats: Serum ammonia levels (P...) 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C <0.0001, P 肝性脑病组C vs 碳酸镧治疗组C <0.0001), rat serum glutamate level (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C <0.0001, P 肝性脑病组C vs 碳酸镧治疗组C <0.0001), rat serum glutamine concentration (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C =0.0395, P 肝性脑病组C vs 碳酸镧治疗组C =0.0038). Type A hepatic encephalopathy rats: serum ammonia levels in rats (P 正常组A vs 肝性脑病组A<0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A <0.0001, P 肝性脑病组A vs 碳酸镧治疗组A <0.0001), rat serum glutamate level (P 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A <0.0001, P 肝性脑病组A vs 碳酸镧治疗组A <0.0001), rat serum glutamine concentration (P 正常组A vs 肝性脑病组A =0.0072, P 肝性脑病组A vs 鸟氨酸治疗组A =0.3213, P 肝性脑病组A vs 碳酸镧治疗组A =0.0078).

[0072] Figure 13 The results showed that, similar to serum results, ammonia and glutamine in cerebrospinal fluid increased significantly after the onset of the disease, which was consistent with the significant swelling of astrocytes. Figure 10 The results were consistent. After lanthanum carbonate treatment, ammonia levels decreased to almost normal, and glutamine levels decreased, approaching those of the normal group, which was superior to the ornithine treatment group. Glutamine levels in serum ( Figure 12 The difference between the changes in blood and cerebrospinal fluid (CSF) is that blood cannot cross the blood-brain barrier after injury, but ammonia can. In rats with hepatic encephalopathy type C: ammonia levels in the CSF of rats (P... 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C <0.0001, P 肝性脑病组C vs 碳酸镧治疗组C <0.0001), glutamine level in rat cerebrospinal fluid (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C =0.1686, P 肝性脑病组C vs 碳酸镧治疗组C =0.0138). Type A hepatic encephalopathy rats: ammonia level in rat cerebrospinal fluid (P 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A <0.0001, P 肝性脑病组A vs 碳酸镧治疗组A <0.0001), glutamine level in rat cerebrospinal fluid (P 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A =0.8623, P 肝性脑病组A vs 碳酸镧治疗组A =0.2712).

[0073] Figure 14 This indicates that, compared with the normal group, the activity of glutaminase in rats with hepatic encephalopathy was significantly increased in both the small intestine and liver. After oral treatment with lanthanum carbonate, the activity and expression of glutaminase were significantly inhibited, resulting in a significant decrease in its ability to break down glutamine to produce glutamate and ammonia, similar to the normal group and also superior to the ornithine treatment group. This result is consistent with... Figure 11 and Figure 12Consistent because the action of glutaminase GLS breaks down glutamine into glutamate and ammonia, the stronger the glutaminase (GLS) activity, the more glutamine is broken down, leading to a decrease in glutamine levels in the body, and correspondingly, a higher concentration of glutamate and ammonia. The GLS activity (P) in the small intestine of rats with hepatic encephalopathy type A (HEEA) is shown in the figure. 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A =0.459, P 肝性脑病组A vs 碳酸镧治疗组A =0.0186), liver GLS activity in rats with type A hepatic encephalopathy (P 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A =0.0001, P 肝性脑病组A vs 碳酸镧治疗组A <0.0001). Liver GLS activity in rats with hepatic encephalopathy type C (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C <0.0001, P 肝性脑病组C vs 碳酸镧治疗组C <0.0001), GLS activity in the small intestine of rats with hepatic encephalopathy type C (P 正常组C vs 肝性脑病组C =0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C =0.9095, P 肝性脑病组C vs 碳酸镧治疗组C =0.1142).

[0074] Figures 15-16 In Test Example 2.2, after 4 weeks of oral administration of the drug mixture, elevated levels of three cellular inflammatory factors in rats—tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6)—were directly associated with the progression and worsening of hepatic encephalopathy. However, after oral treatment with lanthanum carbonate, these levels almost returned to normal, while treatment with ornithine aspartate had almost no effect on inflammation levels. Type A hepatic encephalopathy rats: Serum TNF-α concentration (P...) 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A =0.9968, P 肝性脑病组A vs 碳酸镧治疗组A =0.0038), IL-1β concentration in rat serum (P) 正常组A vs 肝性脑病组A =0.0022, P 肝性脑病组A vs 鸟氨酸治疗组A =0.7168, P 肝性脑病组A vs 碳酸镧治疗组A =0.0091), rat serum IL-6 concentration (P) 正常组A vs 肝性脑病组A <0.0001, P 肝性脑病组A vs 鸟氨酸治疗组A =0.3543, P 肝性脑病组A vs 碳酸镧治疗组A =0.03). Type C hepatic encephalopathy rats: Serum TNF-α concentration in rats (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C =0.2246, P 肝性脑病组C vs 碳酸镧治疗组C =0.1114), IL-1β concentration in rat serum (P 正常组C vs 肝性脑病组C =0.0002, P 肝性脑病组C vs 鸟氨酸治疗组C =0.9987, P肝性脑病组C vs 碳酸镧治疗组C =0.1341), IL-6 concentration in rat serum (P 正常组C vs 肝性脑病组C <0.0001, P 肝性脑病组C vs 鸟氨酸治疗组C =0.5713, P 肝性脑病组C vs 碳酸镧治疗组C =0.0017).

[0075] Figure 17 Rat models of hepatic encephalopathy (HEE) of types A and C, which closely resemble clinical manifestations, were established using acute and chronic liver injury. Oral administration of lanthanum carbonate reversed HEE by reducing blood ammonia and systemic inflammation, demonstrating good therapeutic effects and providing a new candidate drug for the clinical treatment of HEE.

[0076] (2) Treatment summary:

[0077] Lanthanum carbonate, a phosphate binder, showed significant therapeutic effects on hepatic encephalopathy caused by both acute and chronic liver injury. We observed that astrocytes in the cerebral cortex of rats with hepatic encephalopathy exhibited marked swelling, specifically increased nuclear volume, reduced and lighter-colored heterochromatin, indicating significant nuclear damage. This may be due to elevated intracerebral ammonia levels, leading to increased glutamine synthesis, which can cause astrocyte swelling and consequently brain damage. After lanthanum carbonate treatment, the nuclear perimeter changed less, and the heterochromatin color was darker than in the injured group. While ornithine also has a restorative effect on nuclear damage, its efficacy is less than that of lanthanum carbonate.

[0078] While embodiments of the present invention have been described in detail above, it will be apparent to those skilled in the art that various modifications and variations can be made to these embodiments. However, it should be understood that such modifications and variations fall within the scope and spirit of the invention as defined in the claims. Furthermore, the invention described herein may have other embodiments and can be implemented or carried out in various ways.

Claims

1. The application of lanthanum carbonate in the preparation of a drug for treating hepatic encephalopathy, characterized in that, The hepatic encephalopathy mentioned refers to hepatic encephalopathy type A and hepatic encephalopathy type C.

2. The application according to claim 1, characterized in that, The drug comprises lanthanum carbonate and pharmaceutically acceptable excipients, wherein the pharmaceutically acceptable excipients include any one or more of lactose, starch, and gelatin.

3. The application according to claim 2, characterized in that, The lanthanum carbonate includes any one or more of lanthanum carbonate hydrate and basic lanthanum carbonate.

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