Composition for treating neonatal HIE

A composition, neonatal technology, applied in the directions of microorganisms, drug combinations, biochemical equipment and methods, etc., can solve the problems of rejection, high production cost, storage and transportation limitations, etc., to achieve improved therapeutic efficacy, excellent therapeutic effect, The effect of reducing production costs

Pending Publication Date: 2019-11-15
美迪因诺医疗创新科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this approach using stem cells themselves has the following limitations and side effects
[0005] First, basically, cell therapy agents may not rule out the possibility of tumorigenicity due to DNA transfer; second, because stem cells have a large size, vascular occlusion or myocardial infarction may occur; third, when using allogeneic cells ( such as umbilical cord blood) in transplants (allogeneic transplantation) have rejection problems due to cell surface antigens; fourth, cell therapeutics are often difficult to prepare and have many limitations in storage and transportation and high production costs
Therefore, due to the inherent limitations of stem cells, a method of improving efficacy using genetic manipulation has been developed as a method to reduce side effects and improve treatment effects (Korean Unexamined Patent Application No. 10-2017-0099382, etc.), but there is no clear alternative so far

Method used

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  • Composition for treating neonatal HIE
  • Composition for treating neonatal HIE
  • Composition for treating neonatal HIE

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1: Induction of enhanced efficacy by treatment of stem cells with thrombin

[0064] 1-1. Thrombin-induced exosome secretion

[0065] Human umbilical cord blood-derived mesenchymal stem cells (3×10 5 ) were inoculated on 60-mm Petri dishes (OrangeScientific cat#4450200) and cultured for 1 week. After confirming that the cells were saturated and proliferating in the culture dish, the medium was replaced with a serum-free medium (MEMα medium) (diluted 50 units / mL thrombin (REYON Pharmaceutical.Co., LTD) in a serum-free medium), and the cells were After culturing for another 6 hours, thrombin-treated stem cells were obtained.

[0066] Then, in order to confirm whether exosome secretion in mesenchymal stem cells was activated by thrombin treatment, the process of exosome secretion was confirmed by transmission electron microscope (TEM) images. The result is as figure 1 As shown, it can be seen that thrombin stimulation induces exosome secretion.

[0067] Alterna...

Embodiment 2

[0074] Example 2: Construction of an in vitro model of neonatal HIE

[0075] After 18.5 days of pregnancy, the lower abdomen of SD rats was dissected to remove the fetuses, and then the whole brains were carefully extracted. From the extracted brain tissue, only the cerebral cortex was carefully isolated, separated into individual cells using a pipette, and then cultured in neuronal cell culture solution. After 10 days of culture, cells were exposed to glucose-free medium under 1% hypoxic conditions (oxygen and glucose deprivation (OGD)) for 60 minutes to establish an in vitro model of neonatal HIE.

Embodiment 3

[0076] Example 3: In vitro inhibitory effect of thrombin-treated stem cells on neuronal cell death

[0077] The in vitro neonatal HIE model constructed in Example 2 was treated with the thrombin-treated stem cells obtained in Example 1, and then the nerve cell viability was evaluated by MTT assay.

[0078] The result is as Figure 4As shown, in the in vitro HIE model (OGD), neuronal cell death was significantly increased due to oxygen / glucose deprivation compared with normal neuronal cells (NC), while compared with non-thrombin-treated stem cell groups (OGD+naive MSC) or Neuronal cell death was considerably affected in the “thrombin-pretreated stem cell-treated in vitro neonatal HIE group (OGD+MSC_thrombin-pre)” compared to the hypoxic-preconditioned stem cell group (OGD+MSC_thrombin-pre) Large inhibition, proving that thrombin-treated stem cells exhibited the most excellent neuronal cell protection.

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Abstract

The present invention relates to a pharmaceutical composition for preventing or treating neonatal hypoxic ischemic encephalopathy (HIE), comprising a thrombin-treated stem cell or an exosome derived therefrom as an active ingredient, and a method for producing the same. According to the present invention, the thrombin-treated stem cell or the exosome derived therefrom has an increased expression of growth factors, immunoregulatory factors, antioxidant factors, or regeneration factors compared to a group not treated with thrombin and also enhances a neuronal apoptosis inhibitory effect, and thus has an advantage in that the therapeutic effect thereof on neonatal hypoxic ischemic encephalopathy (HIE) is excellent.

Description

technical field [0001] The present invention relates to a pharmaceutical composition for preventing or treating neonatal hypoxic-ischemic encephalopathy (HIE), which comprises thrombin-treated stem cells or exosomes derived therefrom as an active ingredient, and relates to its preparation method. Background technique [0002] Neonatal HIE has been reported to account for 23% of all neonatal deaths, in cases of severe injury, approximately 50% to 100% of patients die, and 65% to 75% of survivors continue to suffer from it during their lifetime Severe motor / cognitive impairment, convulsions, etc. The only current treatment for HIE is therapeutic hypothermia, but its effect is negligible, so about 44% of patients who received therapeutic hypothermia died or suffered from long-term neurological sequelae, and therapeutic hypothermia did not improve the effect, especially for Severe types do not improve the effect. Therefore, the development of effective and important treatment...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K35/545A61K35/51A61K35/50A61K38/48A61K9/00A61K9/12A61P9/10
CPCA61K9/00A61K9/12A61K35/28A61K35/51C12N5/0665C12N2501/998A61K38/4833A61K9/0019A61K35/50A61K35/545A61P25/28A61K9/0085C12N2501/734A61K35/12
Inventor 张允实朴元淳安沼玧成东庆
Owner 美迪因诺医疗创新科技有限公司
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