Preparation method of albumin oligopeptide and application of albumin oligopeptide in promotion of postoperative patient recovery

Abstract

The invention relates to the technical field of albumin oligopeptides, in particular to a preparation method of albumin oligopeptides and application of the albumin oligopeptides in promotion of postoperative patient recovery. The albumin oligopeptide provided by the invention can be taken before, during or after an operation, not only can quickly promote postoperative recovery and reduce the content of total bilirubin and direct bilirubin of a patient, but also can reduce the activity of glutamic oxalacetic transaminase and glutamic-pyruvic transaminase, and also can reduce the activity of carbohydrate antigen 199 and the content of alpha fetoprotein. In addition, by taking the albumin oligopeptide, the bitter throat dryness can be improved, nausea and vomiting symptoms can be eliminated, and the albumin oligopeptide has the effect of promoting postoperative liver function recovery of obstructive jaundice.

Description

Technical Field

[0001] This application relates to the technical field of albumin, and specifically relates to a preparation method of albumin oligopeptide and its application in promoting the recovery of postoperative patients. Background Art

[0002] Albumin, also known as serum albumin, is a type of protein that widely exists in organisms and has important physiological functions. Albumin is a single-chain protein composed of 585 amino acid residues, with a molecular weight of approximately 66.5 kDa. Its molecular structure is oval-shaped, with multiple structural domains that are interconnected by disulfide bonds to form a stable three-dimensional structure. The structure of albumin has a high degree of flexibility and plasticity, enabling it to bind to a variety of different substances, which is the basis for its diverse physiological functions.

[0003] Albumin oligopeptide is obtained by methods such as enzymatic hydrolysis or chemical hydrolysis of albumin. During the production process, specific proteases are usually selected to degrade large albumin molecules into oligopeptides with relatively small molecular weights under mild conditions. These oligopeptides generally consist of 2 - 10 amino acids and have good water solubility, low antigenicity, and high biological activity. Summary of the Invention

[0004] Obstructive Jaundice (OJ) refers to a disease caused by lesions in the biliary tract and around the biliary tract, such as choledocholithiasis, cholangiocarcinoma, gallbladder cancer, pancreatic cancer, etc., which cause biliary stricture, resulting in the inability of bile to enter the duodenum smoothly, thereby increasing the pressure in the biliary tract. Bile flows back into the sinusoidal capillaries through hepatocytes and bile canaliculi, causing an increase in conjugated bilirubin in the blood and jaundice of the body and sclera. The key to the treatment of this disease lies in relieving the obstruction, dredging the biliary tract, and controlling infection. Clinically, surgical treatment is the first choice. Compared with traditional open surgery, laparoscopic technology has the advantages of less trauma, easy recovery, and fewer complications, and is becoming increasingly mature in hepatobiliary surgery.

[0005] For patients with common bile duct stones and / or combined gallbladder stones, the common treatment procedure is laparoscopic cholecystectomy + choledochotomy with stone removal and T-tube drainage (LC + LCBDE). After the operation, liver-protecting and cholagogic drugs are used to promote the regression of jaundice, and at the same time, symptomatic treatments such as nutritional support, acid suppression, and fluid replacement are combined. The jaundice symptoms of most patients will gradually subside. However, for patients with severe biliary tract obstruction and severe liver function damage before the operation, the postoperative treatment period is long, the jaundice subsides slowly, and the liver function recovery time is long. To better solve this problem and accelerate the postoperative recovery of patients, this invention combines the application of albumin oligopeptide on the basis of symptomatic treatment and achieves satisfactory results.

[0006] In view of this, the present invention provides a method for preparing albumin oligopeptide and its application in promoting the recovery of postoperative patients. To achieve the above invention purpose, the present invention adopts the following technical solutions:

[0007] In the first aspect, the embodiment discloses a method for preparing chicken blood albumin oligopeptide, including the following steps:

[0008] S1: Prepare an aqueous solution containing 10-30% crude chicken blood albumin with water;

[0009] S2: After enzymatic hydrolysis of the crude product aqueous solution with alkaline protease, inactivate the enzyme;

[0010] S3: Centrifuge the enzyme-inactivated mixture, take the supernatant for filtration and retention treatment, and dry it to obtain albumin oligopeptide powder;

[0011] S4: Take an appropriate amount of albumin oligopeptide powder and perform peptide segment separation by reversed-phase high performance liquid chromatography;

[0012] S5: Lyophilize each collected peptide segment separately, then dissolve it with an appropriate amount of 0.1% formic acid aqueous solution to prepare a sample solution with a concentration of about 0.5 mg / mL, and perform identification by electrospray ionization technology to obtain albumin oligopeptide KQTALYYC.

[0013] Specifically, in step S1, the crude product aqueous solution contains 20% crude chicken blood albumin.

[0014] Specifically, in step S2, the dosage of alkaline protease is 1.0% (w / v) of the crude product aqueous solution, adjust the pH to 10.5, the temperature is 55 °C, after enzymatic hydrolysis at 50 °C for 20 h, inactivate the enzyme.

[0015] Specifically, in step S3, filter with a ceramic membrane with a molecular weight cut-off of 2×10 6 U, take the middle supernatant, ultrafilter with an ultrafiltration membrane with a molecular weight cut-off of 3000 U to obtain a filtrate with a molecular weight less than 3000 U, and spray dry to obtain peptide dry powder.

[0016] Specifically, in step S4, prepare a 1 mg / mL solution of albumin oligopeptide powder with 0.1% formic acid aqueous solution for reversed-phase high performance liquid chromatography.

[0017] In the specific step S4, the separation conditions of the reversed-phase high-performance liquid chromatography are as follows: the chromatographic column is a C18 column (4.6 mm × 250 mm, 5 μm), the mobile phase A is an aqueous solution of 0.1% formic acid, and the mobile phase B is an acetonitrile solution of 0.1% formic acid. The gradient elution program is as follows: 0 - 5 minutes, 5% B; 5 - 45 minutes, 5% - 60% B; 45 - 50 minutes, 60% - 100% B; 50 - 55 minutes, 100% B. The flow rate is 1.0 mL / min, and the detection wavelength is 214 nm. The peptide segments corresponding to each elution peak are collected.

[0018] In a second aspect, the present invention discloses the use of the albumin oligopeptide KQTALYYC prepared in the first aspect in the preparation of a preparation for promoting the recovery after obstructive jaundice surgery. The preparation for promoting the recovery after obstructive jaundice surgery can be a granule, tablet, capsule, pill, powder or liquid preparation; the dissolution medium in the liquid preparation can be a buffer solution, physiological saline or drinking water. The particle size of the above-mentioned granule is 18 - 22 mesh.

[0019] The granule or tablet of the albumin oligopeptide can be taken after surgery, twice a day, and the dosage per time (calculated as the crude drug) is 100 mg for every 60 Kg of body weight. It can not only rapidly promote the postoperative recovery, reduce the total bilirubin and direct bilirubin content of the patient, but also reduce the activities of glutamic-oxaloacetic transaminase and glutamic-pyruvic transaminase, and also reduce the activity of carbohydrate antigen 199 and the content of alpha-fetoprotein. In addition, by taking the albumin oligopeptide, it can improve the symptoms of dryness and bitterness in the throat, eliminate the symptoms of nausea and vomiting, and has the effect of promoting the recovery of liver function after obstructive jaundice surgery. Detailed implementation manners

[0020] In order to make the objectives, technical solutions and advantages of the present application clearer, the present application will be further described in detail below with reference to the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application. The reagents not specifically described in detail in the present application are all conventional reagents and can be obtained from commercial channels; the methods not specifically described in detail are all conventional experimental methods and can be known from the prior art.

[0021] Example 1: Preparation of albumin oligopeptide

[0022] The present invention provides a method for preparing a chicken blood albumin oligopeptide, comprising the following steps:

[0023] S1: Prepare an aqueous solution containing 20% crude chicken blood albumin with water;

[0024] S2: Add 1.0% (w / v) of alkaline protease (Nanning Dongheng Huadao Biotechnology) to the aqueous solution of the crude product and adjust the pH to 10.5. After enzymatic hydrolysis at 50 °C for 20 h, perform enzyme inactivation treatment;

[0025] S3: Centrifuge the mixture after enzyme inactivation, take the supernatant for ultrafiltration retention treatment, and dry it to obtain albumin oligopeptide powder.

[0026] S4: Take an appropriate amount of albumin oligopeptide powder, dissolve it with mobile phase A (0.1% formic acid aqueous solution) to prepare a solution with a concentration of 1 mg / mL. Peptide separation is carried out by reversed-phase high-performance liquid chromatography (RP-HPLC). The chromatographic column is a C18 column (4.6 mm × 250 mm, 5 μm), mobile phase A is 0.1% formic acid aqueous solution, and mobile phase B is 0.1% formic acid acetonitrile solution. The gradient elution program is as follows: 0 - 5 minutes, 5% B; 5 - 45 minutes, 5% - 60% B; 45 - 50 minutes, 60% - 100% B; 50 - 55 minutes, 100% B. The flow rate is 1.0 mL / min, and the detection wavelength is 214 nm. Collect the peptides corresponding to each elution peak.

[0027] S5: Mass spectrometry analysis:

[0028] Freeze-dry each of the collected peptides separately, then dissolve them with an appropriate amount of 0.1% formic acid aqueous solution to prepare a sample solution with a concentration of approximately 0.5 mg / mL. The electrospray ionization (ESI) technique is used. A Q-Exactive mass spectrometer is used, with the spray voltage set at 3.5 kV, the capillary temperature at 320 °C, the sheath gas flow rate at 40 arb, and the auxiliary gas flow rate at 10 arb. The mass scanning range is m / z 300 - 1800. Perform mass spectrometry analysis on each peptide sample to collect primary mass spectrometry and secondary mass spectrometry data. Use MaxQuant software to analyze the mass spectrometry data, compare the collected mass spectrometry data with the albumin sequence in the Swiss-Prot protein database, and determine the amino acid sequence of the peptide based on the precursor ion mass and fragment ion mass information of the peptide. If there are multiple peptides, according to the overlapping parts between the peptides, use bioinformatics software (such as DNAMAN) to splice the amino acid sequences of each peptide to obtain the complete amino acid sequence of albumin oligopeptide. Amino acid composition analysis and peptide mapping analysis are used to verify the identification results. Amino acid composition analysis is carried out by acid hydrolysis of albumin oligopeptide, and then the content of various amino acids in the hydrolysis product is determined using an amino acid analyzer and compared with the identified amino acid sequence. Peptide mapping analysis is to enzymatically hydrolyze albumin oligopeptide again with a specific protease (such as trypsin), and then analyze the peptide map of the enzymatic hydrolysis product by RP-HPLC and compare it with the theoretical peptide map to verify the correctness of the amino acid sequence. Refer to the amino acid sequence provided in "Application of Reversed-Phase High-Performance Liquid Chromatography in the Separation and Purification of Glycated Albumin Peptides, Progress in Biochemistry and Biophysics, 1999" for verification. As a result, a freeze-dried product of albumin oligopeptide with the amino acid sequence KQTALYYC was obtained from crude chicken blood albumin in the present invention.

[0029] Example 2: Albumin oligopeptide promotes the recovery of liver function after obstructive jaundice surgery

[0030] 1. Experimental materials

[0031] Inpatients in the Department of Integrated Traditional Chinese and Western Medicine Surgery of a affiliated hospital of a traditional Chinese medicine university, with a western diagnosis of obstructive jaundice.

[0032] Diagnostic criteria for obstructive jaundice: Refer to the "Expert Consensus on Percutaneous Transhepatic Biliary Drainage and Stent Implantation for Obstructive Jaundice (2018)" in 2018 and the "Diagnosis and Treatment Guidelines for Hepatocholangiolithiasis" in 2007. Clinical manifestations include: jaundice (icterus of the sclera, skin and mucous membranes, darkening of urine color), abdominal pain, abdominal distension, nausea, vomiting, fatigue, lightening of fecal color, skin pruritus, etc. Laboratory tests include blood routine, urine routine, stool routine, coagulation function, liver and kidney function, tumor markers and blood glucose. The results of liver function show an increase in the values of TBIL, DBIL, ALT, and AST. Common methods include abdominal ultrasound, CT, and MR, etc. Manifestations include bile duct dilation, choledocholithiasis shadow, or combined with intrahepatic choledocholithiasis, etc.

[0033] Inclusion criteria: The range of TBIL is 80 - 200 μmol / L; the age is 18 - 60 years old; the western diagnosis conforms to the diagnostic criteria of choledocholithiasis and obstructive jaundice; the traditional Chinese medicine diagnosis conforms to the diagnostic criteria of jaundice (syndrome of damp-heat in the liver and gallbladder); it meets the surgical indications, has no surgical contraindications, and all undergo laparoscopic cholecystectomy + choledocholithotomy under choledochoscope + T-tube drainage (LC + LCBDE combined with T-tube drainage). During the operation, the stones are completely removed and the drainage tube is unobstructed. There are no complications such as intra-abdominal infection, bile fistula, and biliary stricture after the operation; those whose treatment methods are consistent with this trial.

[0034] Exclusion criteria: Those with a history of pancreatic mass, cholangiocarcinoma, ampullary cancer, liver malignant tumor, hepatitis, cirrhosis, hepatocellular jaundice, biliary ascariasis, or accompanied by intrahepatic choledocholithiasis, intrahepatic choledocholithiasis with jaundice, etc.; those with mental diseases who cannot cooperate with examinations and treatments; those whose various indicators cannot be detected according to requirements; those who do not meet the above inclusion criteria

[0035] Dropout criteria: Those who do not treat according to requirements and are misclassified as those who do not meet the inclusion criteria; those who change treatment methods due to changes in the condition; those who automatically withdraw from this treatment.

[0036] 2. Experimental methods

[0037] (1) Inclusion of cases

[0038] 63 patients who met the inclusion and exclusion criteria were collected. The control group was treated with conventional western medicine combined with liver-protecting drugs, and the experimental group was treated with the freeze-dried product of albumin oligopeptide with the amino acid sequence of KQTALYYC provided by the present invention.

[0039] (2) Treatment plan

[0040] The patients were fasted from food and water and received routine Western medicine treatment. Electrocardiogram, MRCP, blood and urine routine tests, liver and kidney function tests, coagulation tests, and other biochemical examinations were completed.

[0041] (3) Surgical methods

[0042] All patients in each group underwent LC + LCBDE combined with T-tube drainage.

[0043] (4) Postoperative treatment

[0044] Control group: Routine anti-infection treatment; acid suppression; nutritional support; liver protection treatment was carried out with magnesium isoglycyrrhizinate injection, once a day, 0.2 g each time, for a course of 2 weeks or until jaundice completely subsided and liver function returned to normal and the drug was discontinued.

[0045] Experimental group: On the basis of the treatment in the control group, 100 mg of freeze-dried albumin oligopeptide was taken orally on the first day after surgery, twice a day, for a course of 2 weeks or until jaundice completely subsided and liver function returned to normal and the drug was discontinued.

[0046] (5) Observation indicators

[0047] Observe the syndromes of bitter taste in the mouth, dryness in the throat, nausea, and vomiting.

[0048] 1) Clinical cure: The syndrome score decreased by ≥95%, and clinical symptoms and signs disappeared or basically disappeared.

[0049] 2) Marked effect: The syndrome score decreased by ≥70%, and clinical symptoms and signs were significantly improved.

[0050] 3) Effective: The syndrome score decreased by ≥30%, and clinical symptoms and signs were improved.

[0051] 4) Ineffective: The syndrome score decreased by <30%, and clinical symptoms and signs were not significantly improved or even worsened.

[0052] Note: The total syndrome score = the sum of the scores of each symptom; the syndrome curative effect index = (pre-treatment score - post-treatment score) / pre-treatment total score × 100%.

[0053] (6) Efficacy indicators:

[0054] Peripheral venous blood was collected, serum was taken, and ALT, AST, TBIL, and DBIL values of the two groups were detected and recorded on the 3rd, 5th, 8th, and 14th days before and after surgery using an automatic biochemical analyzer (Beckman Coulter). Peripheral venous blood was collected, serum was taken, and the contents of CA199 and AFP were measured using the same automatic biochemical analyzer. The body temperature, pulse, respiratory rate, and blood pressure of the patients; blood, urine, and stool routine tests; liver and kidney function, electrocardiogram.

[0055] 3. Experimental results

[0056] Comparison of total bilirubin (μmol / L) values in Table 1

[0057] Grouping Before surgery The 3rd day after surgery The 7th day after surgery The 14th day after surgery Control group 132.09±13.54 108.21±29.81 56.80±8.43 21.53±4.62 Experimental group 134.52±32.61 106.09±24.55 37.21±6.25** 12.72±1.35**

[0058] Table 1 shows the detection of serum total bilirubin in the control group and the experimental group before surgery, on the 3rd day after surgery, on the 7th day after surgery, and on the 14th day after surgery. Among them, "**" indicates a significant difference from the control group, P < 0.01. As can be seen from Table 1, the serum total bilirubin content in the experimental group on the 7th day and the 14th day after surgery was significantly lower than that in the control group. And the normal reference range of human serum total bilirubin is 3.4 - 17.1 μmol / L. On the 14th day after surgery, the experimental group was within this normal range, while the control group still had a high value for this index. This shows that using the freeze-dried product of albumin oligopeptide with the amino acid sequence of KQTALYYC provided by the present invention to care for patients after LC + LCBDE combined with T-tube drainage surgery can significantly reduce their serum total bilirubin and has the effect of promoting the recovery of liver function after obstructive jaundice surgery.

[0059] Comparison of direct bilirubin (μmol / L) values in Table 2

[0060] Grouping Before surgery The 3rd day after surgery The 7th day after surgery The 14th day after surgery Control group 61.22±9.61 37.08±7.64 26.22±4.15 9.21±1.21 Experimental group 63.22±8.57 35.65±6.23 8.51±0.82** 5.25±0.73**

[0061] Table 2 shows the detection of direct bilirubin in the control group and the experimental group before surgery, on the 3rd day after surgery, on the 7th day after surgery, and on the 14th day after surgery. Among them, "**" indicates a significant difference from the control group, P < 0.01. And the normal reference range of serum direct bilirubin in adults is 0 - 6.8 μmol / L. On the 14th day after surgery, the experimental group was within this normal range, while the control group still had a high value for this index.

[0062] As can be seen from Table 2, the serum direct bilirubin content in the experimental group on the 7th day and the 14th day after surgery was significantly lower than that in the control group. This shows that using the freeze-dried product of albumin oligopeptide with the amino acid sequence of KQTALYYC provided by the present invention to care for patients after LC + LCBDE combined with T-tube drainage surgery can significantly reduce their serum direct bilirubin and has the effect of promoting the recovery of liver function after obstructive jaundice surgery.

[0063] Comparison of aspartate aminotransferase (U / L) values in Table 3

[0064] Grouping Before surgery The 3rd day after surgery The 7th day after surgery The 14th day after surgery Control group 184.09±24.43 67.25±12.38 42.36±5.61 31.09±3.62 Experimental group 186.51±19.74 48.72±8.41 24.72±4.33** 7.37±1.54**

[0065] Table 3 shows the detection of glutamic-oxaloacetic transaminase (GOT) activity in the control group and the experimental group before surgery, on the 3rd day after surgery, on the 7th day after surgery, and on the 14th day after surgery. Among them, "**" indicates a significant difference from the control group, P < 0.01. As can be seen from Table 3, the GOT activity in the experimental group on the 7th day and the 14th day after surgery was significantly lower than that in the control group. This shows that using the freeze-dried product of albumin oligopeptide with the amino acid sequence KQTALYYC provided by the present invention to care for patients after LC + LCBDE combined with T-tube drainage surgery can significantly reduce the GOT activity, and has the effect of promoting the recovery of liver function after obstructive jaundice surgery.

[0066] Comparison of alanine aminotransferase (ALT) values (U / L) in Table 4

[0067] Grouping Before surgery The 3rd day after surgery The 7th day after surgery The 14th day after surgery Control group 107.82±38.54 58.11±14.87 49.25±3.82 29.32±6.52 Experimental group 106.49±27.35 51.63±8.41 26.15±3.18** 21.09±4.36**

[0068] Table 4 shows the detection of alanine aminotransferase (ALT) activity in the control group and the experimental group before surgery, on the 3rd day after surgery, on the 7th day after surgery, and on the 14th day after surgery. Among them, "**" indicates a significant difference from the control group, P < 0.01. As can be seen from Table 4, the ALT activity in the experimental group on the 7th day and the 14th day after surgery was significantly lower than that in the control group. This shows that using the freeze-dried product of albumin oligopeptide with the amino acid sequence KQTALYYC provided by the present invention to care for patients after LC + LCBDE combined with T-tube drainage surgery can significantly reduce the ALT activity, and has the effect of promoting the recovery of liver function after obstructive jaundice surgery.

[0069] Jaundice subsidence time in Table 5

[0070] Grouping Number of cases Jaundice remission time (days) Control group 50 14.36±2.52 Experimental group 50 8.35±1.68*

[0071] Table 5 shows the jaundice subsidence time of the control group and the experimental group. Among them, "*" indicates a significant difference from the control group, P < 0.05. As can be seen from Table 5, the jaundice subsidence time of the experimental group was significantly lower than that of the control group. This shows that using the freeze-dried product of albumin oligopeptide with the amino acid sequence KQTALYYC provided by the present invention to care for patients after LC + LCBDE combined with T-tube drainage surgery can significantly reduce the jaundice subsidence time, and has the effect of promoting the recovery of liver function after obstructive jaundice surgery.

[0072] Comparison of dryness and bitterness in the throat symptom score values in Table 6

[0073] Grouping Before surgery The 3rd day after surgery The 7th day after surgery The 14th day after surgery Control group 4.62±1.02 3.82±0.72 2.09±0.62 1.49±0.67 Experimental group 4.81±0.81 3.46±1.11 1.71±0.43** 0.97±0.43**

[0074] Table 6 shows the detection of the symptom integral values of dryness and bitterness in the throat for the control group and the experimental group before surgery, on the 3rd day after surgery, on the 7th day after surgery, and on the 14th day after surgery. Among them, "**" indicates a significant difference compared with the control group, P < 0.01. It can be seen from Table 6 that the symptom integral values of dryness and bitterness in the throat of the experimental group on the 7th day and the 14th day after surgery are significantly lower than those of the control group. This shows that using the freeze-dried product of albumin oligopeptide with the amino acid sequence of KQTALYYC provided by the present invention to care for the patients after LC + LCBDE combined with T-tube drainage can quickly relieve the symptom of dryness and bitterness in the throat, and has the effect of promoting the recovery of liver function after obstructive jaundice surgery.

[0075] Comparison of symptom integral values of nausea and vomiting

[0076] Grouping Before surgery The 3rd day after surgery The 7th day after surgery The 14th day after surgery Control group 2.62±0.61 1.88±0.45 0.94±0.41 1.49±0.67 Experimental group 2.81±0.48 1.69±0.61 0.00±0.00** 0.00±0.00**

[0077] Table 7 shows the detection of the symptom integral values of nausea and vomiting for the control group and the experimental group before surgery, on the 3rd day after surgery, on the 7th day after surgery, and on the 14th day after surgery. Among them, "**" indicates a significant difference compared with the control group, P < 0.01. It can be seen from Table 7 that the symptom integral values of nausea and vomiting of the experimental group on the 7th day and the 14th day after surgery are significantly lower than those of the control group. This shows that using the freeze-dried product of albumin oligopeptide with the amino acid sequence of KQTALYYC provided by the present invention to care for the patients after LC + LCBDE combined with T-tube drainage can quickly relieve the symptom of nausea and vomiting, and has the effect of promoting the recovery of liver function after obstructive jaundice surgery.

[0078] Comparison of CA199 (U / mL)

[0079] Grouping Before surgery The 3rd day after surgery The 7th day after surgery The 14th day after surgery Control group 631.47±59.47 576.35±53.82 485.85±63.52 392.52±42.15 Experimental group 648.04±45.35 525.28±63.35* 367.54±35.35** 324.36±29.52**

[0080] Table 8 shows the detection of the activity of carbohydrate antigen 199 for the control group and the experimental group before surgery, on the 3rd day after surgery, on the 7th day after surgery, and on the 14th day after surgery. Among them, "*" indicates a significant difference compared with the control group, P < 0.05; "**" indicates a significant difference compared with the control group, P < 0.01. It can be seen from Table 8 that the activity of carbohydrate antigen 199 of the experimental group on the 7th day and the 14th day after surgery is significantly lower than that of the control group. This shows that using the freeze-dried product of albumin oligopeptide with the amino acid sequence of KQTALYYC provided by the present invention to care for the patients after LC + LCBDE combined with T-tube drainage has the effect of promoting the recovery of liver function after obstructive jaundice surgery.

[0081] Comparison of AFP (ng / mL)

[0082] Grouping Before surgery The 3rd day after surgery The 7th day after surgery The 14th day after surgery Control group 9.15±0.35 8.69±0.45 6.58±0.52 5.87±0.48 Experimental group 9.31±0.52 7.12±0.61* 3.84±0.67** 2.91±0.67**

[0083] Table 9 shows the detection of alpha-fetoprotein content in the control group and the experimental group before surgery, on the 3rd day, 7th day, and 14th day after surgery. Among them, "*" indicates a significant difference from the control group, P < 0.05; "**" indicates a significant difference from the control group, P < 0.01. As can be seen from Table 9, the alpha-fetoprotein content in the experimental group on the 7th day and 14th day after surgery was significantly lower than that in the control group. This shows that using the freeze-dried product of albumin oligopeptide with the amino acid sequence of KQTALYYC provided by the present invention to care for patients after LC + LCBDE combined with T-tube drainage has the effect of promoting the recovery of liver function after obstructive jaundice surgery.

[0084] Comparison of syndrome efficacy indexes in Table 10

[0085] Grouping Syndrome efficacy index Control group 78.28±0.63 Experimental group 92.19±0.82**

[0086] Table 10 shows the syndrome efficacy indexes of the control group and the experimental group. Among them, "**" indicates a significant difference from the control group, P < 0.01. As can be seen from Table 10, the syndrome efficacy indexes in the experimental group on the 7th day and 14th day after surgery were significantly higher than those in the control group. This shows that using the freeze-dried product of albumin oligopeptide with the amino acid sequence of KQTALYYC provided by the present invention to care for patients after LC + LCBDE combined with T-tube drainage has the effect of promoting the recovery of liver function after obstructive jaundice surgery.

[0087] As described above, it is only a preferred specific embodiment of the present application, but the protection scope of the present application is not limited thereto. Any changes or substitutions that can be easily thought of by those skilled in the art within the technical scope disclosed by the present application should be covered by the protection scope of the present application.

Claims

1. A method for preparing chicken blood albumin oligopeptide, characterized in that: The following steps are involved: S1: preparing an aqueous solution containing 10-30% crude chicken blood albumin with water; S2: hydrolyzing the crude product aqueous solution with alkaline protease and then inactivating the enzyme; S3: centrifuging the enzyme-killed mixed solution, taking the supernatant, filtering and intercepting it, and drying it to obtain albumin oligopeptide powder; S4: Take an appropriate amount of albumin oligopeptide powder and separate the peptide segments using reversed-phase high performance liquid chromatography; S5: The collected peptide segments are freeze-dried separately, and then dissolved with an appropriate amount of 0.1% formic acid aqueous solution to prepare a sample solution with a concentration of about 0.5 mg / mL. The sample solution is identified using electrospray ionization technology to obtain the albumin oligopeptide KQTALYYC.

2. The preparation method according to claim 1, characterized in that: In step S1, the crude aqueous solution contains 20% crude chicken blood albumin.

3. The preparation method according to claim 1, characterized in that: In step S2, the amount of alkaline protease used is 1.0% (w / v) of the crude aqueous solution, the pH is adjusted to 10.5, the temperature is 55° C., and after enzymolysis at 50° C. for 20 h, the enzyme is inactivated.

4. The preparation method according to claim 1, characterized in that: In step S3, a molecular weight cut-off of 2×10 6 U ceramic membrane filtration, take the intermediate clear liquid, ultrafilter with ultrafiltration membrane with molecular weight cutoff of 3000U, obtain filtrate with molecular weight less than 3000U, spray dry to obtain peptide dry powder.

5. The preparation method according to claim 1, characterized in that: In step S4, the albumin oligopeptide powder is prepared into a 1 mg / mL solution with a 0.1% formic acid aqueous solution and subjected to reverse phase high performance liquid chromatography.

6. The preparation method according to claim 1, characterized in that: In step S4, the separation conditions of reversed-phase high performance liquid chromatography are as follows: the chromatographic column is a C18 column (4.6 mm×250 mm, 5 μm), the mobile phase A is a 0.1% formic acid aqueous solution, and the mobile phase B is a 0.1% formic acid acetonitrile solution. The gradient elution program is: 0-5 minutes, 5% B; 5-45 minutes, 5%-60% B; 45-50 minutes, 60%-100% B; 50-55 minutes, 100% B. The flow rate is 1.0 mL / min, and the detection wavelength is 214 nm. The peptides corresponding to each elution peak are collected.

7. Use of the albumin oligopeptide KQTALYYC prepared by the preparation method according to any one of claims 1 to 6 in the preparation of a preparation for promoting postoperative recovery of obstructive jaundice.

8. The use according to claim 7, characterized in that: The preparation for promoting postoperative recovery of obstructive jaundice can be a granule, tablet, capsule, pill, powder or liquid preparation.

9. The use according to claim 8, characterized in that: The dissolution medium in the liquid preparation can be a buffer solution, physiological saline or drinking water.

10. The use according to claim 8, characterized in that: The particle size of the granules is 18 to 22 meshes.