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Anti-NgR and anti-NG2 mixed polypeptide vaccine, and application thereof in spinal cord injury repair

A technology for spinal cord injury and polypeptide vaccines, which can be used in the fields of medicine and bioengineering to solve problems such as limited regeneration ability and hindered axon regeneration.

Pending Publication Date: 2020-04-24
SHANGHAI CHANGZHENG HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The central nervous system of adult mammals can regenerate after injury, but its regenerative ability is limited, mainly due to inhibitory factors in the local microenvironment and glial scars that hinder axon regeneration

Method used

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  • Anti-NgR and anti-NG2 mixed polypeptide vaccine, and application thereof in spinal cord injury repair
  • Anti-NgR and anti-NG2 mixed polypeptide vaccine, and application thereof in spinal cord injury repair
  • Anti-NgR and anti-NG2 mixed polypeptide vaccine, and application thereof in spinal cord injury repair

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: NGR+NG2 mixed vaccine preparation

[0029] 1. First design the target protein polypeptide, the design results are as follows:

[0030] Three NGR protein target polypeptides:

[0031] 1, PGCSRKNRTRSHCR (SEQ ID NO. 1);

[0032] 2, Cys-GNGSGPRHINDSP (SEQ ID NO.2);

[0033] 3, Cys-TGRATDEEPLGLPK (SEQ ID NO.3);

[0034] There are 43 amino acids in total.

[0035] Four NG2 protein target polypeptides:

[0036] 1, Cys-GKPESSTPTGEPGPM (SEQ ID NO.4);

[0037] 2, Cys-PEGRAPGPAGDSLT (SEQ ID NO.5);

[0038] 3, GLPYLRGTSRPLRGC (SEQ ID NO. 6);

[0039] 4, Cys-PVERRDQPGEPATE (SEQ ID NO.7);

[0040] There are 61 amino acids in total.

[0041] 2. NGR+NG2 protein peptide synthesis

[0042] a. Resin swelling: Weigh the Fmoc-Arg(pbf)-wangResin resin and pour it into the reaction column, add DCM to soak for 30 minutes, and drain.

[0043] b. Deprotection: add an appropriate amount of deprotection solution to the reaction column, stir and agitate for 30 minutes with ...

Embodiment 2

[0056] Embodiment 2: animal modeling and administration

[0057]1. Modeling method: SD rats, male sex, mass 200-300g, 8 weeks old, 18 rats were adaptively fed for one week, anesthetized by intraperitoneal injection of 10% chloral hydrate (1mL / 100g), after successful anesthesia The rats were fixed on the animal operating table in the prone position, the hair on the chest and back was cut and the skin was prepared, and disinfected. Determine the sequence of the vertebral body according to the ribs, make a longitudinal incision at T10 in the center of the back centered on the T10 spinous process, make an incision about 2.0 cm long along the spine, cut the skin and muscles, separate and expose the spinous process, and carefully knock out T10 lamina. Open the dura mater to expose the spinal cord, cut the left spinal cord with the ophthalmic surgery sharp knife along the midline, and remove the residual fibrous tissue. After the incision, the rat tail convulsively swayed and the l...

Embodiment 3

[0068] Embodiment 3: experimental detection and statistical analysis

[0069] 1. Behavioral scoring

[0070] The BBB motor function score was performed on the animals in each group at 0, 1, 3, 7, 14, and 28 days after operation. The hindlimb movements of rats were divided into 22 grades. 0 is total paralysis of the hind limbs, 21 is completely normal. The basic content is: the number and range of joint activities, the degree of weight bearing and the coordination of the front and rear limbs. Raw data processing, statistical analysis, drawing graphs. See the BBB Sports Score data sheet for details.

[0071] Conclusion: if figure 1 As shown, as the time points progressed, group B (subcutaneous administration group 1) recovered the best, and group A (control group) recovered the worst.

[0072] 2. ELISA detection

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Abstract

The invention relates to the technical field of medicine and bioengineering, and especially relates to an anti-NgR and anti-NG2 mixed polypeptide vaccine. The anti-NgR and anti-NG2 mixed polypeptide vaccine is composed of three NgR protein target polypeptides and four NG2 protein target polypeptides, and the amino acid sequences of the three NgR protein target polypeptides and the four NG2 proteintarget polypeptides are represented by SEQ ID NO.1 to SEQ ID NO.7 respectively. The invention also provides an application of the polypeptide vaccine in the preparation of spinal cord injury repair drugs or reagents. The mixed polypeptide vaccine can inhibit expression of inflammatory factors IL6 and IL-1 beta in a spinal cord injury area, reduce formation of glial scars in the spinal cord injuryarea, promote axon growth in the spinal cord injury area and promote neurological function recovery.

Description

technical field [0001] The invention relates to the technical fields of medicine and bioengineering, in particular to the development of an anti-NgR+NG2 mixed polypeptide vaccine and its application in repairing spinal cord injuries. Background technique [0002] The central nervous system of adult mammals can regenerate after injury, but its regenerative ability is limited, mainly because inhibitory factors in the local microenvironment and glial scars hinder axon regeneration. Nogo receptor (NgR) is a high-affinity co-receptor of three myelin-related axon growth inhibitory factors (Nogo, MAG, and Omgp), and inactivation of NgR can block the signal transmission of axonal growth inhibitory factors and function. The glial scar formation in the spinal cord injury area is mainly regulated by NG2 cells and the NG2 proteoglycan secreted by them. In 2004, the research team of the present invention began to use NgR and NG2 genes as targets to conduct research on gene therapy for ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61P25/28
CPCA61K39/00A61P25/28A61K2039/54Y02A50/30
Inventor 吕碧涛
Owner SHANGHAI CHANGZHENG HOSPITAL
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