Composition comprising placenta extract for preventing or treating liver disease and improving liver function

Abstract

To provide a pharmaceutical composition for preventing or treating fatty liver disease, and also to provide a food composition for improving liver function.SOLUTION: The present invention relates to a composition comprising a placenta extract for preventing or treating fatty liver disease and improving liver function. More specifically, by comprising the placenta extract, the present invention reduces fat accumulation associated with fatty liver and reduces serum AST and ALT activities, and thus may exhibit a prophylactic or therapeutic effect on fatty liver disease, such as non-alcoholic fatty liver or alcoholic fatty liver, and may exhibit the effect of improving liver function.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a composition for preventing or treating liver diseases and improving liver function.

Background Art

[0002] Non-alcoholic fatty liver disease (NAFLD) is a condition in which triglycerides, a type of neutral fat, accumulate in liver cells without the ingestion of excessive alcohol or drugs that induce fatty liver. Non-alcoholic fatty liver disease continues to increase due to the nutritional excess caused by the high-fat and high-carbohydrate intake of modern people. It has been reported that 80% of adults with non-alcoholic fatty liver disease develop metabolic disorders such as insulin-resistant diabetes and heart disease.

[0003] Non-alcoholic fatty liver disease means a wide range of liver diseases including non-alcoholic simple steatosis, non-alcoholic steatohepatitis (NASH), and liver diseases that progress to non-alcoholic fatty liver-related cirrhosis. Pathologically, non-alcoholic simple steatosis, which is non-alcoholic fatty liver disease without inflammation, and non-alcoholic steatohepatitis with inflammation, if left untreated for a long time, may lead to severe liver diseases such as hepatitis, liver fibrosis, and cirrhosis. Non-alcoholic fatty liver disease is characterized by the accumulation of fat (fatty infiltration) in hepatocytes. Non-alcoholic simple steatosis may lead to non-alcoholic steatohepatitis. The accumulation of fat in non-alcoholic steatohepatitis is associated with various degrees of liver inflammation and scarring and is often associated with insulin resistance, dyslipidemia, and hypertension.

[0004] The onset of non-alcoholic steatohepatitis is explained by the two-hit hypothesis. First, fat accumulation occurs in the liver tissue. When the fat accumulation in the liver tissue becomes severe, an inflammatory reaction occurs, which causes lipid peroxidation and exacerbation of inflammation. While the number of patients with non-alcoholic steatohepatitis is increasing rapidly, there is still no excellent therapeutic drug for non-alcoholic steatohepatitis. With the increase in the obese population, the number of patients with non-alcoholic steatohepatitis is increasing, and the therapeutic drug market for non-alcoholic steatohepatitis has developed on a huge scale. Also, the causes and mechanisms of non-alcoholic steatohepatitis have been clarified, and much attention has been focused on the development of therapeutic drugs for non-alcoholic steatohepatitis. However, the development of safe and long-term administrable therapeutic drugs for non-alcoholic steatohepatitis is still at a very low level. Generally, for the treatment of non-alcoholic simple fatty liver or non-alcoholic steatohepatitis, anti-obesity drugs, insulin resistance treatment drugs, hyperlipidemia treatment drugs, hepatocyte protectants, antioxidants, etc. are used. However, these drugs are not essential therapeutic drugs for non-alcoholic simple fatty liver or non-alcoholic steatohepatitis but drugs used as symptom improvers and have side effects for long-term administration. Therefore, there is an increasing demand for the development of new therapeutic compositions that are safer, can be administered long-term, and are suitable for the treatment of non-alcoholic simple fatty liver or non-alcoholic steatohepatitis, which is a chronic disease.

[0005] In particular, Korea has a high prevalence of non-alcoholic simple fatty liver and non-alcoholic steatohepatitis, but the development level of drugs for the prevention, improvement, alleviation, or treatment of non-alcoholic simple fatty liver or non-alcoholic steatohepatitis is low. In view of this problem, research on therapeutic drugs for non-alcoholic simple fatty liver or non-alcoholic steatohepatitis using natural products has been conducted. In the case of therapeutic drugs for non-alcoholic simple fatty liver or non-alcoholic steatohepatitis using such natural products, since the content of active ingredients in the natural extracts is low, large amounts are used to obtain the therapeutic effect on non-alcoholic simple fatty liver or non-alcoholic steatohepatitis. is necessary. Moreover, many of these merely utilize natural product materials in marketing, and further scientific research is needed regarding the substantial therapeutic effects on non-alcoholic simple fatty liver or non-alcoholic steatohepatitis.

[0006] On the other hand, human placenta extract (HPE) contains various growth factors, cytokines, and other bioactive substances and is widely used for applications such as fatigue reduction and antioxidant effects (Lee KK, et al., Evid Based Complement Alternat. Med., vol. 2012,(2012) p.130875). However, despite much interest in human placenta extract, a complete study on its functions has not yet been conducted.

Summary of the Invention

Problems to be Solved by the Invention

[0007] An object of the present invention is to provide a pharmaceutical composition for preventing or treating fatty liver disease. An object of the present invention is to provide a food composition for improving liver function.

Means for Solving the Problems

[0008] 1. A pharmaceutical composition for preventing or treating fatty liver disease, comprising placenta extract. 2. The pharmaceutical composition for preventing or treating fatty liver disease according to item 1, wherein the placenta extract is human placenta extract.

[0009] 3. The pharmaceutical composition for preventing or treating fatty liver disease according to item 1, wherein the placenta extract is obtained by treating the placenta with pepsin and hydrochloric acid.

[0010] 4. The pharmaceutical composition for preventing or treating fatty liver disease according to item 3, wherein the placenta is defatted by acetone treatment.

[0011] 5. In item 3 above, the placenta extract is obtained by purifying with anion exchange chromatography one or more times after treatment with the pepsin and hydrochloric acid, and is a pharmaceutical composition for preventing or treating fatty liver disease.

[0012] 6. In item 5 above, the functional group of the anion exchange chromatography resin is selected from the group consisting of Q, QAE, TEAE, and DEAE, and is a pharmaceutical composition for preventing or treating fatty liver disease.

[0013] 7. In item 6 above, the anion exchange chromatography is performed using a column selected from the group consisting of a cross-linked agarose column, a polymethacrylate resin column, a hydrogel column, and a cross-linked polymethacrylate resin column, and is a pharmaceutical composition for preventing or treating fatty liver disease.

[0014] 8. In item 1 above, the pharmaceutical composition is administered by any one or more of intravenous injection (IV), subcutaneous injection (SC), and intramuscular injection (IM), and is a pharmaceutical composition for preventing or treating fatty liver disease.

[0015] 9. In item 1 above, the fatty liver disease is non-alcoholic fatty liver disease or alcoholic fatty liver disease, and is a pharmaceutical composition for preventing or treating fatty liver disease.

Advantages of the Invention

[0016] The pharmaceutical composition of the present invention shows a preventive or therapeutic effect on fatty liver disease, such as non-alcoholic fatty liver or alcoholic fatty liver, and can show an effect of improving liver function by reducing fat accumulation related to fatty liver and decreasing the AST and ALT levels in serum.

Brief Description of the Drawings

[0017]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

DETAILED DESCRIPTION OF THE INVENTION

[0018] The present invention provides a pharmaceutical composition for preventing or treating non-alcoholic fatty liver disease containing a placenta extract.

[0019] The term "placenta extract" is an extract obtained from the placenta of a human or an animal. The extract can be obtained by treating the placenta obtained from the placenta of a human or an animal with an acid and / or an enzyme. Here, the animal may be derived from a mammal, specifically a human, a cow, a horse, a sheep or a pig.

[0020] The term "placenta" is an organ formed during pregnancy, and functions to supply nutrients and enzymes from the mother to the fetus, discharge waste products and carbon dioxide generated in the fetus to the mother, inhibit the transfer of pathogens and drugs, which are foreign substances in the body, to the fetus, and regulate the endocrine of the fetus.

[0021] The placenta contains amino acids, proteins, sugars, nucleic acids, lipids, inorganic substances, enzymes, hormones, etc. Amino acids include aspartic acid, glutamic acid, leucine, lysine, glycine, alanine, serine, threonine, phenylalanine, tyrosine, methionine, histidine, etc. Proteins and enzymes include albumin, globulin, acid and alkaline phosphatase, hyaluronidase, etc. Sugars include glucose, galactose, ribose, etc. Nucleic acids include uracil, xanthine, hypoxanthine, etc., and lipids include lauric acid, palmitic acid, linoleic acid, etc. Inorganics include Na, K, Ca, P, Fe, Cl, etc., and hormones include gonadothrombin, lactogen, steroid hormones, etc.

[0022] The placenta extract may be obtained by treating the placenta with an enzyme and / or an acid. According to one embodiment, the placenta extract may be obtained by treating placenta with pepsin and hydrochloric acid.

[0023] The placenta extract may be obtained by treating placenta with pepsin and hydrochloric acid, followed by one or more rounds of chromatographic purification.

[0024] When produced through a purification step by chromatography, a placenta extract containing fewer impurities can be effectively obtained compared to when the purification step by chromatography is not carried out.

[0025] The chromatography may be, but is not limited to, anion exchange chromatography or cation exchange chromatography.

[0026] According to one embodiment, the chromatography may be anion exchange chromatography.

[0027] The functional group of the anion exchange chromatography resin may be selected from the group consisting of Q, QAE, TEAE, and DEAE.

[0028] Anion exchange chromatography can be performed using a column selected from the group consisting of a cross-linked agarose column, a polymethacrylate resin column, a hydrogel column, and a cross-linked polymethacrylate resin column.

[0029] The placenta may be defatted. According to one embodiment, the placenta to be treated with an enzyme and / or an acid may be defatted by acetone treatment.

[0030] The placenta may be in a crushed form. The crushed placenta may be obtained by finely cutting or crushing the placenta obtained from a human or an animal by a method that can be adopted by an ordinary technician.

[0031] Fatty liver disease is a disease in which the degree of fat deposition in the liver is 5% or more of the liver weight, and may be non-alcoholic fatty liver disease in which the cause of fatty liver does not originate from alcohol, or alcoholic fatty liver disease in which the cause of fatty liver originates from alcohol.

[0032] Non-alcoholic fatty liver disease (NAFLD) is a disease that shows no significant alcohol intake, no taking of drugs that cause fatty liver, no liver diseases caused by other accompanying causes, and shows findings of intrahepatic fat deposition in imaging medical examinations and tissue examinations.

[0033] Non-alcoholic fatty liver disease may include, but is not limited to, non-alcoholic fatty liver, non-alcoholic simple steatosis, non-alcoholic nutritional fatty liver disease, non-alcoholic starvation fatty liver disease, non-alcoholic obesity-related fatty liver disease, non-alcoholic diabetes-related fatty liver disease, non-alcoholic steatohepatitis (NASH), non-alcoholic cirrhosis, non-alcoholic liver fibrosis, and non-alcoholic liver cirrhosis.

[0034] The term "treatment" means any act that improves or beneficially changes the symptoms of individuals suspected of having fatty liver and those with the disease.

[0035] The term "prevention" means any act that suppresses or delays fatty liver. In one embodiment of the present invention, the preventive and therapeutic effects of a placental extract on liver diseases in a liver disease model were observed. In one embodiment of the present invention, a fatty liver animal model was treated with a placental extract, and the effect of improving liver function was confirmed.

[0036] The pharmaceutical composition of the present invention can contain the placental extract in an amount of 0.01 to 1% (v / v) based on the total volume of the composition.

[0037] The pharmaceutical composition of the present invention can contain the placental extract in an amount of 0.01 to 1%, 0.01 to 0.9%, 0.01 to 0.8%, 0.01 to 0.7%, 0.01 to 0.6%, 0.01 to 0.5%, 0.01 to 0.4%, 0.01 to 0.3%, 0.01 to 0.2%, or 0.01 to 0.1% (v / v) based on the total volume of the composition.

[0038] If the pharmaceutical composition contains an excessive amount of the placental extract, cytotoxicity is shown, and there is a problem that it is difficult to use as a pharmaceutical composition. If the placental extract is contained in a small amount, the drug effect may not be achieved.

[0039] The dosage form of the pharmaceutical composition of the present invention may be in the form of an injection or a topical skin preparation, for example, in a form selected from the group consisting of injections, microneedles, rollers, oral tablets and capsules, granules, and combinations thereof.

[0040] The administration route of the pharmaceutical composition of the present invention may be oral, intravenous, intramuscular, intraarterial, intramedullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual or rectal, and is not limited thereto.

[0041] The pharmaceutical composition of the present invention can be administered orally or parenterally. As parenteral administration, an injection method such as topical skin application or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection can be selected.

[0042] The pharmaceutical composition of the present invention can be administered in an amount of 1 mL to 10 mL per day. For example, it can be administered into the body in an amount of 1 mL to 10 mL, 1 mL to 8 mL, 1 mL to 6 mL, 1 mL to 4 mL, or 1 mL to 2 mL per day.

[0043] The pharmaceutical composition of the present invention can be administered once a day, twice a day, three times a day or four times a day, or more frequently.

[0044] In addition, the present invention provides a food composition for improving liver function containing a placenta extract. Details of the placenta extract are as described above, so specific descriptions are omitted.

[0045] The food composition according to the present invention can have any dosage form selected from the group consisting of functional foods, nutritional supplements, health foods, food additives, feeds, and combinations thereof.

[0046] The food composition of the present invention can contain 0.01 to 1% by volume (v / v) of the placenta extract with respect to the total volume of the composition.

[0047] The food composition of the present invention contains the placenta extract in an amount of 0.01 to 1% by volume, 0.01 to 0.9% by volume, 0.01 to 0.8% by volume, 0.01 to 0.7% by volume, 0.01 to 0.6% by volume, 0.01 to 0.5% by volume, 0.01 to 0.4% by volume, 0.01 to 0.3% by volume, 0.01 to 0.2% by volume, or 0.01 to 0.1% by volume (v / v) with respect to the total volume of the composition. It can be included.

[0048] When the placenta extract is contained in an excessive amount in the food composition, cytotoxicity is shown, and there is a problem that it is difficult to use as a food composition. When the placenta extract is contained in a small amount, the effect of improving liver function may not be obtained.

Example

[0049] Hereinafter, the present invention will be described more specifically with reference to examples. However, these examples are merely illustrative of the present invention and do not limit the scope of the present invention.

[0050] Example 1. Production of placenta extract After degreasing the placenta by acetone treatment, the placenta extract was produced by sufficiently hydrolyzing it with pepsin and hydrochloric acid treatment so that an incomplete hydrolyzate was not generated (see Figure 1). Further, it was purified by anion exchange chromatography. This process is an example of a method for treating the placenta to obtain a placenta hydrolyzate and does not limit the method for obtaining a placenta hydrolyzate.

[0051] Experimental Example 1. Confirmation of the effect of reducing the size of fatty liver of placenta extract using a non-alcoholic fatty liver zebrafish model 1. Experimental method To confirm the effect of reducing fat accumulation in liver tissue of placental extracts, a tamoxifen-induced non-alcoholic fatty liver zebrafish model was used. Zebrafish embryos on the 5th day after fertilization were dispensed into 24-well plates at 5-7 per well, and 5 μM tamoxifen and placental extracts were diluted to appropriate concentrations in a 0.06% sea salt solution and co-treated. When the placental extract of the present invention was treated on zebrafish at a concentration of 1% or more, it was confirmed that the zebrafish individuals could not survive and died. Therefore, in this experiment, the substances of the present invention were each tested in concentration ranges of 0.05, 0.1, and 0.5%. After 24 hours of treatment (the 6th day after correction), the solution was replaced with 5 μM LipidGreen2 solution and reacted for 30 minutes under light-shielded conditions. After the zebrafish embryos were anesthetized with Tricaine, they were fixed in 3% methylcellulose, and the degree of staining of the liver part was photographed using a fluorescence microscope. Also, the area and fluorescence intensity of the fluorescently stained liver were quantitatively analyzed using the Gen5 (BioTek) program, and the heatmap image conversion was performed using the edit LUT function of the ImageJ (NIH) program.

[0052] 2. Experimental Results Figure 2 shows the liver region of zebrafish on the 6th day after fertilization after fat staining.

[0053] The upper row of images (black and white images) in Figure 2 are images obtained by a GFP filter (469, 525 nm), meaning that the higher the fat accumulation, the whiter it becomes. The dotted line part of each image means the liver part of the zebrafish. From the upper row of images in Figure 2, it can be confirmed that fatty liver accumulates due to tamoxifen (tamoxifen + placental extract 0% in Figure 2). The lower row of images in Figure 2 (images displayed from blue to red) are images converted into the form of a heatmap using the ImageJ program to make it easier to see the fluorescence degree of the upper row of images. In the liver part, the part showing a form close to red means fatty liver.

[0054] Figure 3 shows the results of Figure 2 in a graph, and the statistical significance of Figure 3 was indicated by *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001.

[0055] There was no difference in liver size between normal individuals (DMSO-treated) and fatty liver individuals (tamoxifen-treated), but the fluorescence intensity indicating fat accumulation was quantified to be approximately twice as high in fatty liver individuals (tamoxifen-treated) as in normal individuals (DMSO-treated) (see the left side of Figure 3).

[0056] When the fat-suppressing effect was confirmed in the zebrafish fatty liver model using the placental extract of the present invention, there was little difference in liver size, but the fluorescence intensity indicating fat accumulation decreased to approximately 60% compared to the fatty liver model (tamoxifen-treated) when the placental extract of the present invention was treated at a concentration of 0.05 - 0.1% (tamoxifen + placental extract 0.05% or 0.1% treatment). Therefore, it was confirmed that the placental extract at this concentration range exhibits the effect of inhibiting fat accumulation in the fatty liver model.

[0057] Experimental Example 2. Confirmation of the effect of decreasing ALT and AST using a high-fat diet mouse model 1. Experimental method In this experiment, 6-week-old male C57BL / 6J mice were bred using a normal-fat diet and a high-fat diet. After randomly separating the mice used in the test into groups of 6 per group, the high-fat diet group was divided into groups administered saline, placental extract (Laennec, 1.8 ml / kg), and metformin (300 mg / kg). In the saline and placental extract administration groups, intravenous injection (I.V.) was used, and in the metformin administration group, oral administration (P.O.) was used for administration for 21 days. Here, metformin is widely known as an anti-diabetic drug and was used as a positive control group that suppresses fat accumulation in liver tissue. After the experimental period ended, the mice were fasted for 16 hours, anesthetized with CO2 gas, and blood was collected from the abdominal aorta. After leaving the blood at room temperature for 1 hour, it was centrifuged at 3000 rpm at 4°C for 10 minutes to obtain serum. The collected serum was stored at -70°C until analysis, and using an automatic biochemical analyzer, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were analyzed. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) are useful biological indicators of liver damage and hepatotoxicity and were measured to confirm the effect of improving liver damage of the placental extract of the present invention.

[0058] 2. Experimental Results In mice fed a high-fat diet, the levels of AST and ALT were significantly increased. In contrast, when the placental extract of the present invention was administered to mice fed a high-fat diet, it was confirmed that the levels of AST and ALT were significantly inhibited. On the other hand, when metformin, which is a positive control group, was administered, it was confirmed that there was no difference in the value of blood AST from the high-fat diet group (saline administration), and only the value of ALT was significantly inhibited (see Figures 4 and 5). Therefore, as a result of observing the efficacy of the placental extract in a fatty liver model using blood biochemical evaluation, it was found that it is effective in suppressing liver damage caused by fatty liver, and it was confirmed that this is a higher level of efficacy than the metformin administration group, which is the positive control group.

Claims

1. A pharmaceutical composition for preventing or treating fatty liver disease, comprising a placenta extract.

2. The pharmaceutical composition for preventing or treating fatty liver disease according to Claim 1, wherein the placenta extract is obtained from a human placenta.

3. The pharmaceutical composition for preventing or treating fatty liver disease according to Claim 1, wherein the placenta extract is obtained by treating the placenta with pepsin and hydrochloric acid.

4. The pharmaceutical composition for preventing or treating fatty liver disease according to Claim 3, wherein the placenta is defatted by acetone treatment.

5. The pharmaceutical composition for preventing or treating fatty liver disease according to Claim 3, wherein the placenta extract is obtained by purifying the placenta extract treated with the pepsin and hydrochloric acid one or more times by anion exchange chromatography.

6. The pharmaceutical composition for preventing or treating fatty liver disease according to Claim 5, wherein the functional group of the anion exchange chromatography resin is selected from the group consisting of Q, QAE, TEAE, and DEAE.

7. The pharmaceutical composition for preventing or treating fatty liver disease according to Claim 6, wherein the anion exchange chromatography is performed using a column selected from the group consisting of a cross-linked agarose column, a polymethacrylate resin column, a hydrogel column, and a cross-linked polymethacrylate resin column.

8. The pharmaceutical composition for preventing or treating fatty liver disease according to Claim 1, wherein the pharmaceutical composition is administered by any one or more of intravenous injection (IV), subcutaneous injection (SC), and intramuscular injection (IM).

9. The pharmaceutical composition for preventing or treating fatty liver disease according to Claim 1, wherein the fatty liver disease is non-alcoholic fatty liver disease or alcoholic fatty liver disease.

Images