A kind of alcohol-detoxifying polypeptide

Abstract

The present invention belongs to the field of biomedicine technology, and specifically provides a new active alcohol-resolving peptide, the sequence of which is shown in SEQ ID NO.1; the present invention also provides a composition containing the peptide, which can be a medicine, a health product or a food. The peptide of the present invention has a small molecular weight, is easy to synthesize, is safe and has no toxic side effects, and has good safety and practicality.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to a new active alcohol-detoxifying peptide. Background Art

[0002] Drunkenness is the common name for alcohol poisoning, which refers to the abnormal state of body function that occurs after a patient drinks a large amount of alcohol (ethanol) at one time. It is divided into acute and chronic drunkenness.

[0003] As we all know, ethanol, the main component of alcohol, is mainly absorbed into tissues and organs through the mouth, esophagus, stomach, intestinal mucosa, etc., and then mainly metabolized by the liver. Ethanol is metabolized in the liver mainly through three enzyme systems: alcohol dehydrogenase (ADH) system, cytochrome enzyme (P4502E1) system and peroxidase (CAT) system. When drinking moderate or small amounts of alcohol, it is mainly metabolized through the ADH system.

[0004] Based on the formation mechanism of alcohol metabolism and alcoholic liver damage, a large number of studies have shown that the accumulation of alcohol and its harmful metabolites in the body is the main cause of alcoholic liver damage. Therefore, hangover, accelerating alcohol metabolism, and reducing the retention of harmful metabolites in the body are of great significance for maintaining human health. At present, many people have claimed to have obtained hangover methods, but innovation based on biotechnology is still needed. Summary of the invention

[0005] One purpose of the present invention is to provide a new active alcohol-detoxifying peptide, the amino acid sequence of which is Phe-Ala-Pro-Glu-Gly;

[0006] Another object of the present invention is to provide the use of the pentapeptide for sobering up;

[0007] Accordingly, the present invention also provides a composition containing the pentapeptide, such as a product for sobering up, which can be a drug for sobering up.

[0008] Furthermore, any of the alcohol sobering methods, including preventing and treating acute and chronic alcohol poisoning and / or alcoholic liver damage, can produce at least any of the following effects:

[0009] (1) It can accelerate the metabolism of alcohol in the body;

[0010] (2) It can shorten the residence time of ethanol and its harmful metabolites in the body;

[0011] (3) increase the activity of ADH, ALDH and / or CYP2E1 enzymes in the liver;

[0012] (4) increasing the levels of ADH, T-AOC, GSH and / or CAT in the blood;

[0013] (5) increasing the levels of T-AOC, GSH and / or CAT in the liver;

[0014] (6) reduce the levels of AST and / or ALT enzymes in the blood;

[0015] (7) Reduce the level of TG in the liver;

[0016] Preferably, the composition is an alcohol sobering drug, which can be in the form of granules, capsules, oral liquid, etc.

[0017] Furthermore, the alcohol relieving medicine also contains pharmaceutical excipients known to those skilled in the art.

[0018] Compared with the prior art, the present invention has the following positive and beneficial effects:

[0019] The pentapeptide of the present invention has a small molecular weight, is easy to synthesize, is safe and has no toxic side effects, and has good safety and practicality. Compared with the existing alcohol sobering agents and other drugs in the prior art, the raw materials are simple and easy to prepare, and the pentapeptide can achieve a good alcohol sobering effect when injected alone, which is more convenient for application in medicines and more convenient for its wide application in alcohol sobering. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a schematic diagram of the results of the activation rate of ADH by the pentapeptide of the present invention;

[0021] Figure 2 This is a graph showing the effect of the pentapeptide of the present invention on the alcohol concentration in mice;

[0022] Figure 3 The results of the pentapeptide of the present invention on ethanol metabolic enzymes in mouse serum and liver (A: ADH activity in serum, B: ADH activity in liver, C: ALDH activity in liver, D: CYP2E1 activity in liver);

[0023] Figure 4 The results of the pentapeptide of the present invention on the antioxidant enzymes in mouse serum (A: T-AOC content in mouse serum, B: GSH content in mouse serum, C: CAT content in mouse serum, D: MDA content in mouse serum);

[0024] Figure 5 The results of the pentapeptide of the present invention on the antioxidant enzymes in the liver of mice (A: T-AOC content in the liver, B: GSH content in the liver, C: CAT content in the liver, D: MDA content in the liver)

[0025] Figure 6 The effect of the pentapeptide of the present invention on the marker enzyme of liver damage in mice (A: AST activity in mouse serum, B: ALT activity in mouse serum);

[0026] Figure 7 This is the effect of the pentapeptide described in the present invention on lipid metabolism (TG content in the liver). DETAILED DESCRIPTION

[0027] The pentapeptide Phe-Ala-Pro-Glu-Gly of the present invention is synthesized by solid phase chemical method. The technical scheme of the present invention is described more clearly and completely through specific implementation methods below to facilitate the understanding of the present invention by those skilled in the art, but it is not intended to limit the scope of protection of the present invention.

[0028] Example

[0029] The effect of pentapeptide (FAPEG) on ADH activity was determined in vitro. The determination method was as follows: 4 test tubes were prepared, and 150 μL of buffer (including 75 μL 0.1 mM sodium pyrophosphate buffer with pH 8.8, 25 μL 11.5% (v / v) ethanol, 50 μL 27 mM NAD + ), then add 50 μL of FAPEG solution of different concentrations (2mmol / L, 4mmol / L, 6mmol / L, 8mmol / L), and then incubate at 37°C for 5 minutes; immediately add 50 μL of 0.2U / mL ADH enzyme solution to each test tube after incubation and mix thoroughly, then read the absorbance change of each solution at a wavelength of 340nm, read once every 1min, and measure continuously for 5min. The absorption value of adding distilled water is used as the blank group. Calculate the relative enzyme activity, the results are as follows Figure 1 shown.

[0030] Figure 1 The results showed that pentapeptide FAPEG could significantly increase the activation rate of ADH in vitro.

[0031] Example

[0032] This example further demonstrates the effect of the pentapeptide of the present invention on alcohol metabolism in mice through animal experiments. The specific experiments are as follows:

[0033] 64 Kunming mice (20-22 g) were taken and fed adaptively in the animal room for 7 days, and then randomly divided into 4 groups: blank control group (Control), model group (Model), positive control group (Positive), and pentapeptide group (FAPEG).

[0034] Blank group: mice were gavaged with normal saline (30 mg / Kg) every day according to their body weight, and then gavaged with normal saline 12 ml / Kg once after gavage;

[0035] Model group: mice were gavaged with normal saline (30 mg / kg) every day according to their body weight, and then gavaged with 56% liquor (12 ml / kg) once after gavage with normal saline;

[0036] Positive group: mice were gavaged with bifendate emulsion (150mg / Kg) every day according to their body weight, and then gavaged with 56% liquor (12ml / Kg) at one time after gavage with bifendate emulsion alone;

[0037] Pentapeptide group: mice were gavaged with pentapeptide solution (30 mg / Kg) according to their body weight every day, and 12 ml / Kg of 56-proof liquor was gavaged once after gavage with pentapeptide.

[0038] Two hours after the experimental group was gavaged with liquor on the first day (the blank group was gavaged with saline), blood was collected from the eyeball and mixed in sodium citrate anticoagulant, stored at 4°C for later use. The ethanol concentration in the blood was detected by gas chromatography internal standard method. The results are as follows: Figure 2 As shown in the figure, alcohol intake caused the ethanol concentration in the model group mice to rise sharply to 4.85 mg / ml within 2 hours. Compared with the model group, 30 mg / Kg of pentapeptide gavage (pentapeptide group) can significantly reduce the ethanol content to 3.01 mg / mL, and the clearance rate reaches more than 38%, which directly reduces the toxic effect of alcohol on mice.

[0039] Then continue the gavage treatment, according to the above gavage method, for 7 consecutive days. 2 hours after the last gavage, collect the serum and liver tissue of the mice: take the blood sample of the mice and centrifuge it at a speed of 4000rpm and a temperature of 4℃ for 10min to prepare the serum; prepare the liver into a 10% liver homogenate with normal saline, and then centrifuge the liver homogenate sample at a speed of 3500rpm and a temperature of 4℃ for 10min, and then collect the supernatant of the liver homogenate. The serum and liver collected for 7 days after gavage were tested as follows:

[0040] The ADH enzyme activity in serum was detected using a kit (a method known to those skilled in the art), and the activities of ADH, ALDH and CYP2E1 in the liver were detected. The results were as follows: Figure 3 As shown; after mice continuously consumed ethanol for 7 days, the ADH and ALDH activities in the liver of the model group were significantly reduced, and the CYP2E1 activity was increased.

[0041] The test results show that continuous intragastric administration of the pentapeptide of the present invention for 7 days can significantly increase the activity of ADH (p < 0.05) in serum, significantly increase the activity of ADH (p < 0.05) and ALDH (p < 0.01) enzymes in the liver, and reduce the level of CYP2E1 (p < 0.01), thereby improving the alcohol metabolism ability of mice. The pentapeptide (FAPEG) accelerates ethanol metabolism by enhancing the activity of ethanol metabolic enzymes and reduces the damage to the liver caused by ethanol metabolites and ROS.

[0042] Based on the mouse serum and liver homogenate supernatant collected above, the antioxidant capacity (T-AOC), glutathione (GSH), catalase (CAT) and malondialdehyde (MDA) levels of mouse serum and liver tissue were determined using a kit. Figure 4 and Figure 5 As shown; Figure 4 Results Continuous oral administration of pentapeptide for 7 days significantly enhanced the levels of T-AOC (p<0.01), GSH (p<0.01), and CAT (p<0.05) antioxidant enzymes in the serum of mice, and significantly reduced the level of MDA (p<0.01) in the serum. Figure 5 The results also showed that continuous oral administration of pentapeptides for 7 days significantly enhanced the levels of antioxidant enzymes T-AOC (p < 0.001), GSH (p < 0.001), and CAT (p < 0.05) in the liver of mice, reduced the content of MDA (p < 0.05) in the liver, and reduced the oxidative stress damage caused by continuous and heavy drinking in mice.

[0043] Based on the mouse serum collected above, the activity of AST and ALT in the mouse serum was measured using a kit (a method known to those skilled in the art). The results are as follows: Figure 6 As shown. Figure 6 The results showed that compared with the model group, continuous oral administration of pentapeptide for 7 days significantly reduced the levels of AST (p < 0.01) and ALT (p < 0.05) in serum. Therefore, pentapeptide has a protective effect on alcohol-induced liver cell damage.

[0044] Based on the liver homogenate supernatant collected above, the TG content in the mouse liver was determined using a kit (a method known to those skilled in the art). The results are as follows: Figure 7 As shown. Figure 7 The results show that compared with the model group, continuous intragastric administration of the pentapeptide of the present invention for 7 days can reduce the content of TG in the liver (p < 0.05), so it has a good protective effect on liver lipid metabolism.

Claims

1. Application of the polypeptide shown in SEQ ID NO.1 in preparing a hangover-relieving composition.

2. The use according to claim 1, characterized in that: The hangover-resolving composition is used for preventing and treating acute and chronic drunkenness and / or alcoholic liver damage.

3. The use according to claim 1, characterized in that: The hangover relieving composition is used to produce at least one of the following effects: 1) Accelerate the metabolism of alcohol in the body; 2) Reduce the levels of malondialdehyde, alanine aminotransferase and / or aspartate aminotransferase in the blood; 3) Reduce the levels of malondialdehyde, ROS oxygen free radicals, total cholesterol and / or CYP2E1 enzyme in the liver; 4) Increase the levels of ADH, T-AOC, GSH and / or CAT in the blood; 5) Increase the levels of T-AOC, GSH and / or CAT in the liver; 6) Increase the activity of ADH and / or ALDH enzymes in the liver.

4. The use according to claim 1, characterized in that: The hangover-relief composition is a medicine for sobering up.

5. A hangover-relieving composition comprising the polypeptide shown in SEQ ID NO.

1.

6. The hangover-relieving composition according to claim 5, characterized in that: The hangover relieving composition is a medicine.

Images