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Mimic epitope peptide and application thereof

A mimetic epitope, chronic technology, applied in the field of bioengineering, can solve the problems of lack of ductility, drug resistance, poor biological stability, etc., and achieve the effect of improving ductility and stability, improving specificity, and increasing antigenicity

Pending Publication Date: 2019-06-21
THE FIFTH PEOPLES HOSPITAL OF SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, CH12 monoclonal antibody drugs still have shortcomings: (1) some patients have poor response or resistance to monoclonal antibody drugs in clinical practice; (2) repeated long-term use is required; (3) Adverse events have affected the use of monoclonal antibody drugs; (4) Based on my country's national conditions, the price of monoclonal antibody drugs is an unavoidable problem in clinical application due to the great differences in the economic tolerance of individual patients. Monoclonal antibody drugs cannot be selected due to high treatment costs (Singh JA, SaagKG, Bridges SL Jr, Akl EA, Bannuru RR, Sullivan MC, Vaysbrot E, McNaughton C, OsaniM, Shmerling RH, Curtis JR, Furst DE, Parks D , Kavanaugh A, O'Dell J, King C, Leong A, Matteson EL, Schousboe JT, Drevlow B, Ginsberg S, Grober J, St Clair EW, Tindall E, Miller AS, McAlindon T; American College of Rheumatology. 2015 American College of Rheumatology Guidelines for the Treatment of Rheumatoid Arthritis.ArthritisCare Res(Hoboken).2016Jan;68(1):1-25; Wu Wei, He Dongchu
However, the mimetope peptide has the disadvantages of poor biological stability, easy degradation in vivo, lack of extensibility and antigenicity caused by short peptide chains, and the application of the mimetope peptide in the field of kidney diseases is not mentioned.

Method used

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  • Mimic epitope peptide and application thereof
  • Mimic epitope peptide and application thereof
  • Mimic epitope peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Synthesis of Example 1 Mimic Epitope Peptide, Animal Immunization

[0045] Synthesis of mimotope peptide: According to the amino acid sequence of SEQ NO.1, the background SBS company was commissioned to synthesize the mimotope peptide with a purity of >98%.

[0046] At the same time, the polypeptide whose sequence is SEQ NO.5 (AIPLVVPFYSHSGGGSGGGS) was used as a control group for the experiment.

[0047] The dose of each immunization was 100 μg / ml, and the adjuvants used were complete Freund's adjuvant (first immunization) and incomplete Freund's adjuvant (subsequent immunization). Dissolve the polypeptide in physiological saline (antigen solution with a concentration of 0.2 mg / mL), and then mix with an equal volume of complete Freund's adjuvant (or incomplete Freund's adjuvant) by double-push method (at this time, the antigen concentration is about 0.1 mg / mL), and then subcutaneously injected at multiple points in the abdomen or groin of mice, and noticed that subcuta...

Embodiment 2

[0048] The establishment of embodiment 2 renal fibrosis model

[0049] The renal fibrosis model was established by UUO method.

[0050] Healthy male C57 / B6 mice aged 10 weeks.

[0051] Anesthetized by intraperitoneal injection of 0.6% pentobarbital sodium solution (according to 100μl / 10g), the left lower abdomen was prepared for skin, and the skin, superficial and deep fascia, and peritoneum were incised in sequence, and the small intestine was opened with forceps to expose the left ureter. Starting from the middle and lower 1 / 3 of the left ureter, use straight forceps and hemostats to help pass the silk thread through the bottom of the left ureter, ligate the ureter at both ends, cut the ureter in the middle of the two ligatures, and reset the ureter carefully. , and then suture the incision continuously, and incubate at 37 degrees for half an hour to prevent the death of mice from hypothermia. The animals were sacrificed on the 14th day after UUO, 6 in each group, and the ...

Embodiment 3

[0054] Example 3 Kidney histological analysis (including MASSON staining, fibrosis-related molecular staining and macrophage staining)

[0055] 1. The MASSON staining method is:

[0056]1. Routinely dewax the slices to water, and stain with the prepared Weigert iron hematoxylin for 5-10 minutes.

[0057] 2. Differentiate with acidic ethanol differentiation solution and wash with water.

[0058] 3. Use Masson blue solution to return to blue, and wash with water.

[0059] 4. Wash with distilled water for 1 min.

[0060] 5. Stain with ponceau red fuchsin staining solution for 5-10 minutes.

[0061] 6. During the above operation process, prepare weak acid working solution according to the ratio of distilled water: weak acid solution = 2:1, and wash with weak acid working solution for 1 min.

[0062] 7. Wash with phosphomolybdic acid solution for 1-2 minutes.

[0063] 8. Wash with the prepared weak acid working solution for 1 min.

[0064] 9. Dye directly into the aniline blu...

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Abstract

The invention discloses a mimic epitope peptide and application thereof. The amino acid sequence of the mimic epitope peptide is as shown in SEQ No.1. The mimic epitope peptide can be used as the antigen to immunize animals to prepare a polyclonal antibodies and can be used for preparing drugs for preventing and / or treating chronic organ fibrotic diseases. After a mouse kidney fibrosis model builtby a unilateral ureteral obstruction method is processed by the mimic epitope peptide, MASSON shows that mouse kidney fibrosis degree is evidently relieved, immunohistochemistry shows that the expression of protein related to fibrosis in kidney tissue is lowered evidently, macrophage number in the kidney tissue and the spleen is lowered evidently, and mechanism researches show that the mouse signal channel is inhibited.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a mimetic epitope peptide and its application. Background technique [0002] Chronic kidney disease (CKD) refers to irreversible renal structure and dysfunction caused by many different diseases that last for more than 3 months, mainly including pathological damage to the kidney, abnormal blood or urine components, and abnormal imaging examinations , or other unexplained decrease in glomerular filtration rate (GFR) (<60ml / min·1.73m2). Diseases that cause CKD include primary and secondary glomerulonephritis, renal tubular and interstitial injuries, and renal vascular lesions. Renal pathology revealed glomerulosclerosis, tubular atrophy, and interstitial fibrosis, and biochemical examination revealed azotemia, anemia, and abnormal vitamin D, calcium, and phosphate metabolism. [0003] Although various CKDs have different etiologies, the increase of glomerular capillary ...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/11C07K16/28A61K39/00A61K39/395A61P13/12A61P1/16A61P11/00A61P9/00
Inventor 杨麟牛建英
Owner THE FIFTH PEOPLES HOSPITAL OF SHANGHAI
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