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CN-122005544-A - Method for preparing VA-ECMO lung injury treatment drug by using ginkgolide A to regulate YARS1

CN122005544ACN 122005544 ACN122005544 ACN 122005544ACN-122005544-A

Abstract

The invention discloses a method for preparing a VA-ECMO lung injury treatment drug by using ginkgolide A to regulate YARS, which relates to the technical field of biological medicines, and comprises the steps of preparing a drug for treating the VA-ECMO lung injury by using ginkgolide A as a YARS protein regulator through virtual screening, binding affinity confirmation and cell drug effect confirmation, wherein the virtual screening is based on a YARS1 protein structure model, and screening ginkgolide A with high affinity binding energy with YARS by molecular butt joint. The invention proves that the ginkgolide A can effectively improve the pulmonary ventilation function, repair the alveolar epithelial barrier structure and inhibit inflammatory oxidative stress reaction by regulating YARS a, and achieves the aim of improving the lung injury treatment effect and the clinical transformation safety under the support of VA-ECMO.

Inventors

  • JIANG MIN
  • ZHANG RAN
  • XIE LIXIN
  • LIU HUIYING
  • SHAO JUNJIE
  • Han fan

Assignees

  • 中国人民解放军总医院第八医学中心

Dates

Publication Date
20260512
Application Date
20260409

Claims (10)

  1. 1. A method for preparing a medicine for treating VA-ECMO lung injury by using ginkgolide A to regulate YARS is characterized by comprising the steps of preparing a medicine for treating VA-ECMO lung injury by using ginkgolide A as a regulator of YARS protein through virtual screening, binding affinity confirmation and cell medicine effect confirmation, wherein the virtual screening is based on a YARS1 protein structure model, and screening out ginkgolide A with high affinity binding energy with YARS1 through molecular docking.
  2. 2. The method for preparing the VA-ECMO lung injury therapeutic agent by using the ginkgolide A control YARS as set forth in claim 1, wherein the specific steps of the virtual screening include: constructing a library of compounds comprising natural products; potential binding pockets were determined based on the crystal structure of YARS a and blind screen docking was performed using AutoDock Vina software; candidate molecules are screened for binding energy scoring threshold and bilobalide a is preferred by conformational visualization.
  3. 3. The method for preparing the VA-ECMO lung injury therapeutic drug by using the ginkgolide A regulation YARS1 as claimed in claim 2, wherein the method is characterized in that in the preparation process, the combination relation of ginkgolide A and YARS1 is subjected to biophysical confirmation by using a surface plasmon resonance technology, specifically, a recombinant rat YARS1 protein is fixed on the surface of a CM5 sensor chip, and the combination kinetic parameters and equilibrium dissociation constants of ginkgolide A and YARS1 are measured by using a Biacore 8K instrument.
  4. 4. The method for preparing the medicine for treating the VA-ECMO lung injury by using the ginkgolide A regulation YARS as set forth in claim 3, wherein the cell efficacy confirmation is to simulate an ischemia and hypoxia environment by using an oxygen glucose deprivation model, and verify the protection effect of ginkgolide A on alveolar epithelial cells, specifically, an A549 cell is placed in a sugar-free culture medium and a 1% O 2 hypoxia environment to establish an OGD model, and the intervention is carried out by using ginkgolide A with the concentration of 5-20 mu M, and the preferable concentration is 15 mu M.
  5. 5. The method of preparing a medicine for treating VA-ECMO lung injury by using bilobalide A control YARS1 according to claim 4, wherein said VA-ECMO lung injury is pulmonary congestion and pulmonary edema caused by increased left ventricular afterload, and said medicine is administered prophylactically within 0-2 hours before or after VA-ECMO is started, or in parallel with a left ventricular unloading strategy.
  6. 6. The method for preparing the medicine for treating the VA-ECMO lung injury by using the bilobalide A control YARS1 according to claim 5, wherein the medicine is an injection preparation, the administration route is intravenous injection, the effective concentration of the administration dose in vitro is 1-30 mu M, and the dose in an animal body is 0.1-50 mg/kg, preferably 10 mg/kg.
  7. 7. The method for preparing the medicine for treating the VA-ECMO lung injury according to claim 6, wherein the preparation method further comprises the steps of constructing a VA-ECMO combined TAC (transverse aortic stenosis) rat model for in vivo efficacy verification, specifically, constructing a rat VA-ECMO system, and placing a 1.4mm ring constriction at an aortic arch to simulate the left ventricular afterload increase state under clinical VA-ECMO support, and then administering the medicine for treating the bilobalide A.
  8. 8. The method for preparing the VA-ECMO lung injury therapeutic agent by using the ginkgolide A control YARS according to claim 7, wherein in the in-vivo efficacy verification, a multi-dimensional efficacy evaluation system is used for evaluating the efficacy of the agent, comprising the following steps: Monitoring improvement of arterial oxygen partial pressure (PaO 2 ); Structural indexes are that the pathological component reduction of lung tissues is observed through H & E staining, and the ultrastructural recovery of alveolar II type epithelial cells is observed through TEM; Molecular index, namely detecting the level change of alveolar epithelial injury markers SP-D, inflammatory factors IL-1 beta and IL-6 and oxidative stress indexes MPO and MDA in plasma or lung tissues.
  9. 9. The method for preparing VA-ECMO lung injury therapeutic agent using bilobalide A control YARS1 of claim 8, wherein the reduction of SP-D level is used as a specific index for evaluating alveolar epithelial barrier structure repair in said multi-dimensional efficacy assessment system.
  10. 10. The method for preparing the medicine for treating the VA-ECMO lung injury by using the ginkgolide A regulation YARS1 according to claim 9, wherein the medicine prepared by the method is characterized in that YARS is established as an intervened molecular target of the VA-ECMO afterload-related lung injury in the preparation process, and the novel application of an aminoacyl-tRNA synthetase family in mechanical transduction and inflammation regulation is expanded.

Description

Method for preparing VA-ECMO lung injury treatment drug by using ginkgolide A to regulate YARS1 Technical Field The invention relates to the technical field of biological medicines, in particular to a method for preparing a VA-ECMO lung injury treatment drug by using ginkgolide A to regulate YARS. Background The device can provide effective gas exchange and simultaneously bear the function of heart pumping, and is widely applied to the treatment of cardiogenic shock, cardiac arrest and serious mixed respiratory cycle failure. With the advancement of medical technology, ECMO is increasingly used not only as a short-term "bridge" support means, but also in waiting for organ transplantation and waiting for recovery of cardiopulmonary function. However, the parallel circulation characteristic of VA-ECMO changes the inherent hemodynamic state of the organism, so that the left ventricular afterload is obviously increased, pulmonary circulation congestion and alveolar injury are further initiated or aggravated, and the final recovery prognosis of a patient is seriously restricted by the complication, so that the prevention and treatment of VA-ECMO-related pulmonary injury have become an important research direction in the field of in-vitro life support. Although strategies such as left ventricular unloading or drug assistance are often used clinically to alleviate hemodynamic disorders caused by VA-ECMO, specific drug intervention approaches to alveolar epithelial cell injury, inflammatory cascades, and oxidative stress caused by these strategies are relatively lacking. Traditional broad-spectrum anti-inflammatory or antioxidant therapies often lack precise molecular targets and it is difficult to achieve efficient lung protection in complex mechanical circulatory support environments. In the prior art, a medicine target point and a corresponding candidate molecule which can precisely intervene in the pathological process of lung injury caused by VA-ECMO are urgently needed to solve the problems of pulmonary congestion, edema, barrier function destruction and the like caused by post load increase. Therefore, establishing a controllable molecular target closely related to the pathological process and screening to obtain the regulator with high affinity and specificity is a key point for overcoming the bottleneck of the prior art and improving the safety of ECMO treatment. Disclosure of Invention The present invention has been made in view of the above-described problems occurring in the prior art. Therefore, the invention provides a method for preparing a VA-ECMO lung injury treatment drug by using ginkgolide A to regulate YARS, which solves the problems that in the prior art, because of lacking an accurate molecular target for the lung injury caused by VA-ECMO, the lung injury, inflammatory cascade reaction and oxidative stress are difficult to effectively relieve by specific drug intervention. In order to solve the technical problems, the invention provides the following technical scheme: in a first aspect, the present invention provides a method for preparing a VA-ECMO lung injury therapeutic agent by using bilobalide A control YARS1, which comprises, The preparation method comprises the steps of preparing a medicament for treating lung injury caused by VA-ECMO by using bilobalide A as a YARS protein regulator through virtual screening, binding affinity confirmation and cell efficacy confirmation, wherein the virtual screening is to screen out bilobalide A with high affinity binding energy with YARS by molecular butt joint based on a YARS1 protein structure model. As a preferable scheme of the method for preparing the VA-ECMO lung injury treatment drug by using the ginkgolide A regulation YARS, the method comprises the following specific steps of: constructing a library of compounds comprising natural products; potential binding pockets were determined based on the crystal structure of YARS a and blind screen docking was performed using AutoDock Vina software; candidate molecules are screened for binding energy scoring threshold and bilobalide a is preferred by conformational visualization. Furthermore, a natural product compound library with structural diversity and drug-like property is constructed, so that a rich chemical space is provided for subsequent discovery of the Miao compound. Secondly, based on the high-resolution crystal structure of YARS protein, the potential active binding pocket on the surface is determined by a computational chemistry method, and the blind screen docking (Blind Docking) of the whole protein surface is carried out by adopting AutoDock Vina molecular docking software, so that the interaction possibility of small molecules and target proteins is comprehensively explored in a mode of not presetting binding sites. Finally, a screening threshold is set according to Binding Affinity, candidate molecules with high Affinity are primarily screened out, and visual analysis is carried out