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Short peptide, preparation method thereof, and applications of the short peptide in diagnosis or treatment of apoA-I related diseases

A technique for simulating peptides, ac-d-w-f-k-a-f-y-d-k-v-a-e-k-f-k-e-a-f-nh2, applied in the field of peptides, can solve problems such as increasing the risk of atherosclerosis, achieve the level of reducing and antioxidative PL, facilitate automatic operation, and have high application value

Inactive Publication Date: 2014-02-12
温州康成健康管理咨询有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

How lupus increases the risk of developing atherosclerosis is not fully understood, although disease duration and SLE damage index are independent predictors of promotion of atherosclerosis (carotid plaque)

Method used

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  • Short peptide, preparation method thereof, and applications of the short peptide in diagnosis or treatment of apoA-I related diseases
  • Short peptide, preparation method thereof, and applications of the short peptide in diagnosis or treatment of apoA-I related diseases
  • Short peptide, preparation method thereof, and applications of the short peptide in diagnosis or treatment of apoA-I related diseases

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The preparation of embodiment 1 L-4F

[0023] The L-4F of this embodiment is prepared by solid-phase synthesis, and its specific steps are as follows:

[0024] The short peptide was synthesized using a PS3 small-scale fully automatic peptide synthesizer (PS3 Protein Technologies, Woburn, MA). The L-form FMOC amino acid was coupled to an amide resin, condensed with HBTU and NMM (N-methylmorpholine), and acetylated at the N-terminus with acetic anhydride. Add 1% anisole (dichloromethane anisole), 0.1% mercaptoethanol and tryptophan (20% by weight of the peptide resin), the short peptide is eluted from the column, and then purified by reverse-phase high-performance liquid chromatography . The chromatographic column is a VYDAC C-4 column (22mm×25cm, 10μm), the eluent is acetonitrile containing 0.1% TFA (trifluoroacetic acid), the elution gradient is 0-66min, 25%-85%, and the flow rate is 4.8ml / min. The purity of the peptide was analyzed by reverse-phase high-performanc...

Embodiment 2

[0025] Example 2 Inhibitory effect of L-4F on lupus-like phenotype of autoimmune disease

[0026] 26-27 week administration: 1) pravastatin, 2) L-4F, 3) L-4F+pravastatin, 4) blank control. Compared with the mice in the blank control group, the L4F or L-4F+pravastatin group administration group showed that the appearance of lupus-like autoimmunity was improved, and the lymph nodes were significantly reduced (0.17±0.17g: 0.40±0.22g and 0.16±0.10g : 0.40±0.22g; p=0.0012, 0.0007). However, there was no significant change in spleen size and splenocyte number in the treatment group compared with the blank control group, in which B cells, CD4+, CD8+ T cells, NK, CD11c, and CD11b cells were determined by multicolor flow analysis (data not shown). listed).

[0027] In the test, the total IgG levels of the four groups were not much different, and it is generally believed that there is no inhibitory effect on immunity ( figure 1C). Serum IgG anti-double-stranded DNA antibody in the L...

Embodiment 3

[0029] Example 3 L-4F Treatment Little Alters Circulating Serum Levels of Inflammatory Chemokines and Cytokines

[0030] To explore potential biomarker changes among treatment groups, we analyzed female apoE using a Luminex-based beadarray - / - Fas - / - 68 inflammatory chemokines and cytokines in mice. The results showed that compared with the control group, the levels of tissue damage and inflammation in the L-4F group decreased: including CRP (C-reactive protein), fibrinogen, TNF-α (tumor necrosis factor α), CCL12 (mono nuclear cell chemoattractant protein 5 [MCP-5]). Multi-sample (54 detected antigens) Bonferroni correction showed that whether or not pravastatin was used, serum IL-10 (interleukin 10), CCL9 (macrophage inflammatory protein 1γ [MIP-1γ]), VCAM-1 (vascular The level of cell adhesion molecule 1) was significantly lower in the L-4F group than in the control group, and the level of circulating CCL19 (macrophage inflammatory protein 3γ [MIP-3γ]) was also significa...

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Abstract

The invention discloses a short peptide, a preparation method thereof, and applications of the short pepetide in diagnosis or treatment of apoA-I related diseases. The sequence of the short peptide is Ac-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH2, wherein the amino acids used are all L-amino acids. An L-4F provided by the invention has functions similar to the apoA-I. In an accelerated arteriosclerotic mice lupus model, the L-4F is capable of reducing significantly double-stranded DNA resistance and oxidized-type PL resistance of IgG, improving degrees of proteinuria, glomerulonephritis and sclerotin reduction, increasing the amount of smooth muscle cells, reducing infiltration of macrophages, and reducing the level of inflammation chemotactic factors causing atherosclerosis at the same time. Accordingly, the L-4F treatment is a protective mechanism for lupus diseases, also has certain functions for rebuilding and fixing latent patches, and has a broad application prospect and high application value. The L-4F can be used for diagnosis and treatment of the apoA-I related diseases, and has high medicinal value. The L-4F is obtained by a solid-phase synthetic method with simple operation, and therefore the L-4F has a broad market prospect.

Description

technical field [0001] The invention relates to a polypeptide with pharmacological effects, in particular to a polypeptide and its preparation method and its application in diagnosing or treating diseases related to apoA-I. Background technique [0002] apoA-I is a main apolipoprotein component of HDL, which can drive the reverse transport of cholesterol, accept excess cholesterol in peripheral cells and transport it back to liver HDL particles, and also has anti-inflammatory, anti-oxidation and anti-coronary atherosclerosis hardening effect. At present, the apoA-I mimetic peptides designed and synthesized according to the biological structure and characteristics of apoA-I include 18A small peptides (such as Ac-DWLKAFYDKVAEKLKEAF-NH2), which do not have apoA-I homologous sequences, but can form similar The A-level amphipathic helical structure of apoA-I has the biological characteristics of linking lipids. By adjusting the number and position of phenylalanine residues (F) ...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K1/06C07K1/04G01N33/68A61K38/10A61P9/10A61P13/12A61P37/02A61P29/00A61P3/06A61P3/10A61P3/04A61P19/04A61P9/12A61P7/02A61P35/00A61P1/16A61P31/18
Inventor 林灼锋林绍强潘薛波吴帆龚琦
Owner 温州康成健康管理咨询有限公司
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