CN118903111A - Application of indole in preparation of medicine for treating hepatic fibrosis - Google Patents

Abstract

The application discloses application of indole in preparing a medicine for treating hepatic fibrosis. Experiments prove that the indole can effectively improve liver function indexes, improve liver cell structures, effectively reduce collagen deposition levels, provide a new thought for clinically treating liver fibrosis, and is expected to become a new therapeutic drug in the aspect of liver injury diseases.

Description

Use of indole in preparing drugs for treating liver fibrosis

Technical Field

The present application relates to the field of medical technology, and in particular to the use of indole in the preparation of drugs for treating liver fibrosis.

Background Art

The liver is an indispensable organ for maintaining human life activities, playing a key role in vitamin metabolism, protein metabolism, lipid metabolism and hormone metabolism. Liver fibrosis is a pathological syndrome characterized by extracellular matrix deposition and tissue scar formation caused by factors such as viruses, drugs, high-fat diet and alcohol. Any liver damage will undergo a process of liver fibrosis during the process of liver repair and healing. If the damaging factors cannot be removed for a long time, the fibrosis process will continue for a long time and will develop into cirrhosis or even liver cancer, which seriously threatens human health.

So far, there are no approved anti-liver fibrosis drugs. The main treatment method used in clinical practice is to remove the cause of the disease. Commonly used liver-protecting drugs such as silymarin and phosphatidylcholine have adverse reactions such as diarrhea. Currently, once cirrhosis develops, liver transplantation is the only treatment method. Therefore, it is particularly important to deeply explore the pathogenesis of liver fibrosis and develop anti-fibrosis drugs with good effects and low toxicity, which has great clinical value and social significance.

Summary of the invention

In view of the problems existing in the prior art, the present application provides the use of indole in preparing a drug for treating liver fibrosis.

Specifically, this application involves the following aspects:

1. Use of indole in the preparation of a medicament for treating and/or preventing diseases caused by liver damage.

2. Use of indole in the preparation of a medicament for treating and/or preventing a disease associated with a disease caused by liver damage.

3. The use described in item 1 or 2, wherein the disease caused by liver damage is selected from one of liver fibrosis, acute liver damage, chronic liver damage, cirrhosis, and liver cancer.

4. The use according to item 1 or 2, wherein the cause of liver damage is selected from one of trauma, drugs, alcohol, chemical toxic substances, viruses or autoimmunity.

5. The use according to item 1 or 2, wherein the dosage of indole is 6-13 mg/kg of the subject 1-2 times a day.

6. The use according to item 5, wherein the subject is a human.

7. The use according to item 1 or 2, wherein a single dose of the medicament contains 50-500 mg of indole, preferably 300-400 mg of indole.

8. A composition for treating and/or preventing a disease caused by liver damage or a disease associated with a disease caused by liver damage, wherein the composition comprises indole as the only active ingredient in the composition.

9. The composition according to item 8, wherein the disease caused by liver damage is selected from one of liver fibrosis, acute liver damage, chronic liver damage, cirrhosis, and liver cancer.

10. The composition according to item 8, wherein the cause of liver damage is selected from one of trauma, drugs, alcohol, chemical toxic substances, viruses or autoimmunity.

11. A composition according to any one of items 8 to 10, wherein the composition further comprises one or more pharmaceutically acceptable carriers or excipients.

This application provides a new use of indole. Experiments have shown that indole can effectively improve liver function indicators, improve liver cell structure, and effectively reduce collagen deposition levels, providing new ideas for the clinical treatment of liver fibrosis and is expected to become a new therapeutic drug for liver damage diseases.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings are used to better understand the present application and do not constitute an improper limitation on the present application.

FIG1 is the H&E staining observation results of liver tissue pathology of each group of mice in the present application example;

FIG2 is the Masson staining observation results of liver tissue pathology of each group of mice in the present application example;

FIG3 is the Sirius Red staining observation results of liver tissue pathology of each group of mice in the present application example;

FIG4 is a graph showing changes in alanine aminotransferase (ALT), an indicator of liver function in each group of mice in the present application example;

FIG. 5 shows the changes in aspartate aminotransferase (AST), an indicator of liver function, in each group of mice in the examples of the present application.

DETAILED DESCRIPTION

The present application is further described below in conjunction with examples. It should be understood that the examples are only used to further describe and illustrate the present application and are not used to limit the present application.

Unless otherwise defined, the technical and scientific terms in this specification have the same meaning as those generally understood by those skilled in the art. Although similar or identical methods and materials as described herein may be used in experiments or practical applications, materials and methods are described herein below. In the event of a conflict, the present specification including the definitions therein shall prevail. In addition, materials, methods and examples are provided for illustration only and are not restrictive. The present application is further described below in conjunction with specific embodiments, but is not intended to limit the scope of the present application.

Indole is also known as benzopyrrole, azaindole, etc. As one of the metabolites of tryptophan, indole can activate the aromatic hydrocarbon receptor to produce anti-inflammatory effects. However, there have been no reports on the results of studies on its use in the treatment of liver fibrosis and related liver damage.

The present application innovatively discovered that indole has a therapeutic or preventive effect on diseases caused by liver damage.

Therefore, the present application provides the use of indole in a drug for treating and/or preventing diseases caused by liver damage.

The present application also provides use of indole in preparing a medicament for treating and/or preventing diseases caused by liver damage.

The indole of the present application can be obtained by commercial purchase or prepared by techniques known in the art.

Liver damage mainly refers to liver cell damage, liver structure and function abnormalities. Liver damage will stimulate the body to produce too many highly reactive oxygen free radicals, induce oxidative stress, trigger inflammatory reactions, lead to liver cell damage and apoptosis, and release too much transaminase into the blood, thus affecting normal liver function. If not prevented and treated in time, it may induce a series of complications or promote the development of other diseases.

There are many causes of liver damage, mainly drug poisoning, viral infection, excessive alcohol intake, immune response, contact with chemical toxic substances in the environment, radiation damage, etc. The Acorus calamus or its extract of the present application has a therapeutic or preventive effect on diseases caused by liver damage. The cause of liver damage is not limited. As long as the liver cells are damaged, the liver structure and function are abnormal, the liver damage is within the scope of the present application. In a specific embodiment, the liver damage is caused by alcohol, virus, drug and chemical toxic substances.

The specific types of diseases caused by liver damage are not limited in this application, as long as they are caused by liver damage, they are within the protection scope of this application, including but not limited to liver fibrosis, acute liver damage, chronic liver damage, cirrhosis, hepatitis, liver cancer, fatty liver, liver failure, etc., such as liver damage, fatty liver, hepatitis, cirrhosis, liver failure, etc. caused by factors such as alcohol. In a specific embodiment, the disease is liver fibrosis.

The diseases related to the diseases caused by liver damage refer to other diseases caused by liver damage. These diseases may appear due to liver damage, and may also be alleviated or disappear after the liver damage is treated.

As used herein, "treatment" refers to any improvement in any consequence of a disease, disorder, or condition, such as extended survival, lower morbidity, and/or mitigation of side effects caused by alternative treatment modalities. In some embodiments, treatment comprises delaying or ameliorating a disease, disorder, or condition (i.e., slowing or preventing or reducing the development of a disease or at least one clinical symptom thereof). In some embodiments, treatment comprises delaying, alleviating, or ameliorating at least one physical parameter of a disease, disorder, or condition, including physical parameters that may be indiscernible to the patient. In some embodiments, treatment comprises regulating a disease, disorder, or condition physically (e.g., stabilizing discernible symptoms), physiologically (e.g., stabilizing physical parameters), or both.

As used herein, the term "preventing" or "prevention" refers to prophylactic treatment of a disease or condition; or delaying the onset or progression of the disease, disorder or condition.

The dosage of the drug of the present application depends on many factors, such as the nature and severity of the disease to be prevented or treated, the sex, age, weight and individual response of the patient or animal, the route of administration and the number of administrations, etc. The above dosage can be administered in a single dosage form or divided into several, such as two, three or four dosage forms. The dosage level must be selected based on the specific route of administration, the severity of the condition to be treated, and the condition and previous medical history of the patient to be treated.

However, it should be recognized that the total daily dosage of the drug of the present application must be determined by the attending physician within the scope of sound medical judgment. For any particular patient, the specific therapeutically effective dosage level must be determined based on a variety of factors, including the disorder being treated and the severity of the disorder; the specific composition used; the patient's age, weight, general health, sex and diet; administration time, route of administration and excretion rate; duration of treatment; drugs used in combination or concurrently; and similar factors known in the medical field. For example, the practice in the art is to start the administration of a dose lower than the level required to obtain the desired therapeutic effect and gradually increase the dose until the desired effect is obtained.

In a specific embodiment, the dosage of indole is 1-2 times a day, 6-13 mg/kg of the subject each time, for example, 6 mg/kg of the subject, 7 mg/kg of the subject, 8 mg/kg of the subject, 9 mg/kg of the subject, 10 mg/kg of the subject, 11 mg/kg of the subject, 12 mg/kg of the subject, 13 mg/kg of the subject each time.

The subject may be a mammal, including but not limited to humans, cows, horses, sheep, pigs, goats, rabbits, cats, dogs, mice, and any other mammals that have livers and can produce damage.

In a specific embodiment, the subject is a human.

Those skilled in the art will appreciate that the number and frequency of administration may be adjusted according to the specific condition of the subject or the severity of the disease caused by liver damage.

In a specific embodiment, the continuous administration is 3 to 10 days, such as 3 days, 5 days, 7 days, 10 days, etc., and any values between these values. In a specific embodiment, the dosage of indole is 6-13 mg/kg subject once a day, for 3 to 10 consecutive days.

In a specific embodiment, the drug is administered in a single dose form, i.e., a single dose form. In a specific embodiment, a single dose of the drug contains 50-500 mg of indole, for example, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, and any value between these values. It should be understood by those skilled in the art that, generally speaking, a person's weight is measured in 60 kg. If the person's weight is not 60 kg, it can be converted according to this standard.

In a specific embodiment, a single dose of the medicament contains 300-400 mg of indole.

The drug of the present application can be administered in a unit dosage form, and the administration route can be enteral or parenteral, such as oral, intramuscular, subcutaneous, nasal, oral mucosa, skin, peritoneum or rectum, etc. The dosage form is, for example, tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal agents, buccal tablets, suppositories, freeze-dried powder injections, etc. It can be a common preparation, a sustained-release preparation, a controlled-release preparation and various microparticle delivery systems.

In a specific embodiment, the medicine optionally contains a pharmaceutically acceptable carrier or excipient. Including but not limited to lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, gel, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talcum, magnesium stearate and mineral oil etc., but are not limited thereto. In the medicine of the present application, in addition to the above-mentioned components, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents and preservatives etc. may also be included.

In a specific embodiment, the drug of the present application may further contain one or more other active substances. The other active substances refer to substances other than indole that are active and produce certain functions. The active substances may be active substances for treating or preventing diseases caused by liver damage, or active substances for assisting in treating or preventing diseases caused by liver damage, or active substances that do not treat diseases caused by liver damage, but may play a positive effect or function in other aspects.

The present application also provides a composition for treating and/or preventing diseases caused by liver damage, wherein the composition contains indole, which is the only active ingredient in the composition. That is, indole is the only active ingredient of the composition for treating and/or preventing diseases caused by liver damage, and the composition does not contain other active ingredients that can treat and/or prevent diseases caused by liver damage.

The term "composition" is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.

In a specific embodiment, the composition further comprises one or more pharmaceutically acceptable carriers or excipients. The one or more pharmaceutically acceptable carriers or excipients are any of the carriers or excipients described above.

As mentioned above, the diseases caused by liver damage include but are not limited to liver fibrosis, acute liver damage, chronic liver damage, cirrhosis, hepatitis, liver cancer, fatty liver, liver failure, etc., such as liver damage, fatty liver, hepatitis, cirrhosis, liver failure, etc. caused by factors such as alcohol. In a specific embodiment, the disease is liver fibrosis.

The composition of the present application can be in the form of pills, capsules, tablets, powders, granules, injections or compound preparations.

The composition of the present application can be administered in a single dose form, and the administration route can be enteral or parenteral, such as oral, intramuscular, subcutaneous, nasal, oral mucosa, skin, peritoneum or rectum, etc. The dosage form is, for example, tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal agents, buccal tablets, suppositories, freeze-dried powder injections, etc. It can be a common preparation, a sustained-release preparation, a controlled-release preparation and various microparticle delivery systems.

The dosage of the composition of the present application can be converted with reference to the dosage of indole as described above.

In the present application, the terms "alanine aminotransferase (ALT)" and "aspartate aminotransferase (AST)" are enzymes whose blood levels increase when the liver is damaged, and are enzymes used as liver function indicators using this property.

The present application found that indole can effectively improve liver function indicators (including reducing ALT and AST levels), improve liver cell structure, and effectively reduce collagen deposition levels, providing new ideas for the clinical treatment of liver fibrosis and is expected to become a new therapeutic drug for liver damage diseases.

Example

Example 1

1. Materials

1.1 Experimental animals: SPF-grade 7-week-old male C57BL/6 mice, weighing approximately 22 g, were purchased from Zhuhai Baishitong Biological Co., Ltd., animal license number: [SCXK(粤)2022-0051]. The experiment was carried out after 7 days of adaptive feeding, and the mice were allowed to eat freely during the experiment.

1.2 Test sample: Indole, purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

1.3 Reagents: 95% ethanol (AR), Shanghai McLean Biochemical Technology Co., Ltd.; carbon tetrachloride (HPLC), Shanghai McLean Biochemical Technology Co., Ltd.; olive oil (pharmaceutical grade), Shanghai McLean Biochemical Technology Co., Ltd.; sodium carboxymethyl cellulose (viscosity: 800-1200mPa.s, USP grade), Shanghai Aladdin Biochemical Technology Co., Ltd. Liver function test kit, Nanjing Jiancheng Bioengineering Research Institute Co., Ltd.

2 Experimental methods

2.1 Animal grouping and drug administration

Thirty C57BL/6 mice were randomly divided into a blank control group (NC), a model group (CCl ₄ ), and an indole group (Indole) according to their body weight. Indole was administered by gavage 2 hours after the first modeling. Indole was administered with a 0.5% sodium carboxymethylcellulose solution as a solvent. The indole group was gavaged once a day at 50 mg/kg for 8 weeks, and the blank control group and the model group were gavaged with an equal amount of 0.5% sodium carboxymethylcellulose solution.

2.2 Establishment of liver fibrosis model

The model group and the indole-administered group were intraperitoneally injected with 5 mL/kg CCl ₄ solution (10% (v:v) dissolved in olive oil) twice a week for 8 weeks to induce liver fibrosis model; the blank control group was intraperitoneally injected with an equal volume of olive oil.

2.3 Sample collection and processing

Two hours after the last administration, the mice were anesthetized with sodium pentobarbital, and blood was collected from the eye sockets to kill the mice. The whole blood of the mice was allowed to stand at room temperature for 30 minutes, and then centrifuged at 4°C, 3000 rpm for 15 minutes. The supernatant was taken to measure liver function indicators (ALT, AST), etc. The left lobe of the mouse liver was taken for histopathological examination to determine the therapeutic effect of indole on the CCl _4- induced mouse liver fibrosis model.

2.4 Determination of serum liver function indicators

An appropriate amount of mouse serum was taken and serum alanine aminotransferase (ALT) was determined according to the kit method. It was purchased from Nanjing Jiancheng Bioengineering Research Institute Co., Ltd. with batch number 20240221; aspartate aminotransferase (AST) was purchased from Nanjing Jiancheng Bioengineering Research Institute Co., Ltd. with batch number 20240222.

2.5 Pathological observation of mice

The same part of the liver from each group of mice was fixed in 4% paraformaldehyde for 24 hours, washed, dehydrated, transparent, embedded, and sliced. H&E staining, Masson staining, and Sirius Red staining were performed, and the histopathological changes were observed under an optical microscope.

2.6 Data Analysis

The experimental data were statistically analyzed using GraphPad Prism 8.0 software. When the measurement data met the normal distribution, One-way ANOVA was used. If not, Tamhane’s T2(M) test was used. For data analysis, the number of each group was n=10. *P<0.05 indicated statistical significance, and **P<0.01 indicated significant statistical significance.

3. Results

3.1 Effects of indole on pathological conditions of mice in each group

As shown in Figure 1, H&E staining of mouse liver revealed that the liver cells in the blank control group were neatly arranged, uniform in size, uniformly distributed in the intercellular matrix, and had good nuclear morphology; compared with the blank control group, the liver cells in the model group were disordered, with cell vacuoles and inflammatory infiltration, nuclear condensation, and nuclear rupture and dissolution. Compared with the model group, the disorder of liver cells in the liver of mice in the indole administration group was reduced, cell vacuoles were reduced, and nuclear condensation and nuclear rupture were reduced, showing a certain improvement effect.

As shown in Figures 2 and 3, Masson staining (blue) and Sirus Red staining (red) of mouse livers revealed that there was no obvious collagen deposition and no pseudolobule formation in the livers of mice in the blank control group; compared with the blank control group, there was obvious collagen deposition and pseudolobule formation in the livers of mice in the model group; compared with the model group, there was significantly less blue collagen deposition and significantly less pseudolobule formation in the livers of mice in the indole-treated group, which suggests that indole can effectively improve liver damage in mice with _CCl4- induced liver fibrosis.

3.2 Effects of indole on serum liver function indices of mice in each group

As shown in Figures 4 and 5, compared with the blank control group, the ALT and AST activities in the serum of the model group increased significantly (P < 0.01). Compared with the model group, the ALT and AST activities of the indole group decreased to a certain extent (P < 0.05), indicating that indole can effectively improve the serum liver function of mice with liver fibrosis induced by CCl ₄ .

4. Summary

The present application proves through animal experiments that the indole shown in the present application has the effect of protecting liver fibrosis. It can effectively improve the liver pathology of mice with liver fibrosis induced by _CCl4 , reduce collagen deposition and lower liver function indicators, etc., and can be used as a new candidate drug with the function of protecting liver fibrosis.

Although the embodiments of the present application are described above, the present application is not limited to the above specific embodiments and application fields, and the above specific embodiments are merely illustrative and instructive, rather than restrictive. A person of ordinary skill in the art can make many forms under the guidance of this specification and without departing from the scope of protection of the claims of the present application, all of which belong to the protection of the present application.

Claims (11)

1. Use of indole for the preparation of a medicament for the treatment and/or prevention of a disease caused by liver damage.

2. Use of indole for the manufacture of a medicament for the treatment and/or prophylaxis of a disease associated with a disease caused by liver damage.

3. The use according to claim 1 or 2, wherein the disease caused by liver injury is selected from one of liver fibrosis, acute liver injury, chronic liver injury, cirrhosis, liver cancer.

4. The use of claim 1 or 2, wherein the cause of liver injury is selected from one of trauma, drug, alcohol, chemical toxic substance, virus or autoimmunity.

5. The use according to claim 1 or 2, wherein the indole is administered in a dose of 1-2 times daily, 6-13mg/kg subject.

6. The use of claim 5, wherein the subject is a human.

7. The use according to claim 1 or 2, wherein the single dose of the medicament comprises 50mg-500mg indole, preferably 300-400mg indole.

8. A composition for the treatment and/or prophylaxis of a disease caused by liver damage or a disease associated with a disease caused by liver damage, wherein the composition comprises indole as the sole active ingredient in the composition.

9. The composition of claim 8, wherein the disease caused by liver injury is selected from one of liver fibrosis, acute liver injury, chronic liver injury, cirrhosis, liver cancer.

10. The composition of claim 8, wherein the cause of liver injury is selected from one of trauma, drug, alcohol, chemical toxic substance, virus, or autoimmunity.

11. The composition of any one of claims 8-10, wherein the composition further comprises one or more pharmaceutically acceptable carriers or excipients.