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Application of RXFP1/3 inhibitor in preparation of medicine for preventing or treating adolescent idiopathic scoliosis disease

一种脊柱侧凸、抑制剂的技术，应用在药物组合、骨骼疾病、疾病诊断等方向，能够解决病因不明确、不能完全解释AIS病因等问题

Inactive Publication Date: 2021-11-02
张莉
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The etiology of AIS is still unclear. Researchers have put forward many hypotheses, such as abnormal genetics hypothesis, neuroendocrine abnormality hypothesis, abnormal bone formation hypothesis, bone growth imbalance hypothesis, etc., but none of the above hypotheses can fully explain the cause of AIS. (Kikanloo SR, Tarpada SP, Cho W. Etiology of Adolescent Idiopathic Scoliosis: A Literature Review. Asian Spine J. 2019; 13(3):519-26.)

Method used

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  • Application of RXFP1/3 inhibitor in preparation of medicine for preventing or treating adolescent idiopathic scoliosis disease
  • Application of RXFP1/3 inhibitor in preparation of medicine for preventing or treating adolescent idiopathic scoliosis disease
  • Application of RXFP1/3 inhibitor in preparation of medicine for preventing or treating adolescent idiopathic scoliosis disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Determination of Relaxin Levels in AIS Patients

[0049] Enrollment of clinical subjects

[0050] The subjects of the study were female AIS patients in the outpatient and inpatient department of spine surgery of Changhai Hospital from May 2016 to April 2019. Diagnosis of AIS was made based on clinical history and X-ray examination. Non-idiopathic scoliosis such as connective tissue diseases (Marfan syndrome, Ehlers-Danlos syndrome, etc.) was excluded. Supraspinous ligament tissue was obtained from patients with AIS who underwent posterior orthopedic fixation. Serum samples of AIS patients were collected from AIS patients who underwent bracing or surgical treatment in the inpatient department and outpatient clinic. Age-matched female patients with lumbar disc herniation (without scoliosis) who were treated at the Spine Surgery Clinic of Changhai Hospital served as the control group. There were no statistically significant differences in body mass index, age...

Embodiment 2

[0064] Example 2 Determination of Relaxin Level in AIS Scoliosis Mouse Animal Model

[0065] 1. Preparation of scoliosis mouse model

[0066] As reported in the literature (Machida, M. et al. Experimental scoliosis in melatonin-deficient C57BL / 6J mice without pinealectomy. Journal of Pineal Research 41, 1-7 (2010)), C57BL / 6J mice can be resected without performing double forelimb The mouse scoliosis animal model was established by pinealectomy (incidence rate 60-90%). Three-week-old C57BL / 6J were obtained from Charles River Laboratory (n=100) and housed in groups in air-conditioned rooms (22±2°C) with controlled lighting (lighting from 07:00 to 19:00). :00). During the 4 months of modeling, the mice with the forelimbs removed were placed in a special cage, and the height of food and water was gradually increased to keep the mice in a standing position before eating. Mice with a Cobb angle > 10° by anteroposterior spinal X-ray were diagnosed with scoliosis. Vertebral rotati...

Embodiment 3

[0078] Example 3: Effect of Administration of Inhibitor of RLN3 Receptor RXFP3 on Growth of Ligament Fibroblasts in Scoliotic Mice

[0079] experimental method:

[0080] 1. Mouse ligament fibroblasts were isolated from normal ligament tissue in the control group by type I collagenase digestion. Digested cells were incubated in an incubator (37°C, 5% CO 2 ) in Dulbecco's modified Eagle's medium supplemented with fetal bovine serum (10%) and penicillin / streptomycin (1%). Ligament fibroblasts were identified by immunofluorescence of α-SMA. Subsequent cellular experiments were performed on ligament fibroblasts at passage 3-5. Ligament fibroblasts were stimulated by TGF-β1 (10ng / mL), RLN3 (100ng / mL) and R3(B1-22)R (0.1umol / L) for 48 hours.

[0081]2. Immunofluorescence assay: Immunofluorescence assay was performed on the ligament fibroblast α-SMA. 8 [mu]m cryosections of paraformaldehyde-fixed fibroblasts were incubated with 0.3% Triton X-100 in phosphate buffered saline for 2...

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Abstract

The application discloses that the level of RLN3 (Relaxin 3, Relaxin 3) in serum of an AIS patient is obviously higher than that of a normal control, and continuous injection of the RLN3 or administration of an RXFP3 (Relaxin Family Peptide Receptor 3) inhibitor can obviously change the scoliosis morbidity of a mouse animal model, which prompts that the RLN3 plays an important role in AIS morbidity. Therefore, the invention provides an application of the RXFP1 / 3 inhibitor in preparation of a medicine for preventing and / or treating adolescent idiopathic scoliosis, and a pharmaceutical composition which is used for preventing and / or treating adolescent idiopathic scoliosis and contains the RXFP1 / 3 inhibitor, an application of a reagent for detecting the RLN3 level in preparation of a diagnostic reagent for diagnosing the teenager idiopathic scoliosis level of a subject, and a kit for diagnosing teenager idiopathic scoliosis or the disease risk thereof.

Description

[0001] This application is a divisional case of Chinese patent application number 202110029072.9 with an application date of January 11, 2021 and an invention titled "Use of RXFP1 / 3 Inhibitors in the Preparation of Drugs for Preventing or Treating Juvenile Idiopathic Scoliosis" Apply. technical field [0002] The invention belongs to the field of biotechnology. More specifically, the present invention relates to methods and agents for preventing and treating juvenile idiopathic scoliosis. Background technique [0003] Adolescent idiopathic scoliosis (Adolescent Idiopathic Scoliosis, AIS) is an important clinical disease, and the molecular regulation mechanism of its occurrence and development has attracted extensive attention: scoliosis is an ancient disease. The lateral curvature of the coronal plane of the spine exceeds 10 degrees as measured by the Cobb method, and other secondary causes (such as congenital, degenerative, neuromuscular, etc.) are excluded. It is called i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K38/16A61K39/395A61K31/7088A61P19/08C12Q1/6883G01N33/68
CPCA61K45/00A61K38/16A61K39/3955A61K31/7088A61P19/08C12Q1/6883G01N33/6893C12Q2600/158G01N2800/10
Inventor 张莉
Owner 张莉
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