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Application of combined use of RepSox and Y27632 in preparation of medicine for treating retinal related diseases

A technology of Y27632 and retinal pigment, which is applied in the field of biomedicine, can solve the problems of mesenchymal epithelial transition and other problems, and achieve the effect of improving visual function and maintaining shape

Pending Publication Date: 2020-12-11
上海优祺生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Indeed, most current studies focus on inhibiting EMT in cells in an early or partial EMT state, which predisposes to mesenchymal-epithelial transition (MET)

Method used

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  • Application of combined use of RepSox and Y27632 in preparation of medicine for treating retinal related diseases
  • Application of combined use of RepSox and Y27632 in preparation of medicine for treating retinal related diseases
  • Application of combined use of RepSox and Y27632 in preparation of medicine for treating retinal related diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: the EMT level of RPE cell has cell density dependence

[0027] In order to further clarify the relationship between cell culture density and EMT occurrence in RPE, in this example, RPE cells derived from hESCs were cultured at different densities of 50%-3.125% for 14 days. RPE cells can maintain epithelial morphology at starting densities of 50% and 25%, but abruptly change to mesenchymal morphology at starting densities ranging from 12.5% ​​to 3.125%, as figure 1As shown in A. It was confirmed by Q-PCR and immunoblot analysis that with the decrease of the starting density, the expression of RPE-related genes, such as MITF, OTX2, RPE65, CRALBP, TYR, TYRP1, PMEL, BEST1 and E-CAD were up-regulated, and EMT-related Gene expression, such as downregulation of PAI1, FN1, and a-SMA, as figure 1 B and figure 1 C shown. figure 1 D shows the immunofluorescence results of RPE cell tight junction protein ZO1, F-actin-labeled phalloidin and EMT markers a-SMA and PA...

Embodiment 2

[0028] Example 2: The EMT level of RPE cells is time-dependent

[0029] To test whether cell culture time is also involved in the activation of EMT, RPE cells were cultured at 50% and 12.5% ​​of the starting density for different days. With the prolongation of culture time, RPE cells with 50% initial density gradually formed epithelial morphology. 12.5% ​​of the initial density of RPE cells gradually formed a long spindle, showing a tendency of fibrosis, such as figure 2 As shown in A. Q-PCR analysis showed that there was little difference in the expression of RPE or EMT-related genes in cells with different days of 50% initial density, such as figure 2 B and figure 2 C shown. However, for RPE cells cultured at an initial density of 12.5%, as the culture time increased, the expression of RPE-related genes decreased and the expression of EMT-related genes gradually increased, as shown in figure 2 D and figure 2 As shown in E, it shows that under the low initial densi...

Embodiment 3

[0030] Example 3: In the RPE cell EMT model induced by low density, the combined use of RepSox and Y27632 inhibits the EMT level of RPE cells

[0031] Based on the results of implementation cases 1 and 2, a batch of small molecule drugs with potential effects were obtained through effective screening on the low-density induced RPE cell EMT model. In subsequent experiments, it was confirmed that although RepSox and Y27632 alone can inhibit RPE The combined use of 10 μM RepSox and 10 μM Y27632 has a better effect on the EMT level of cells, whether it is the morphology of RPE cells at different time points and the Q-PCR results show that the expressions of RPE-related genes and EMT-related genes are as follows Figure 3A and Figure 3B shown.

[0032] For the EMT level of relatively late RPE, RepSox and Y27632 were added to intervene after 3 and 6 days of culture at 12.5% ​​initial density, respectively, as Figure 4 A and Figure 4 As shown in B, the results of immunofluoresc...

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Abstract

The invention provides application of combined use of RepSox and Y27632 in preparation of a medicine for reversing epithelial-mesenchymal transition (EMT) of retinal pigment epithelium (RPE). According to the application of combined use of RepSox and Y27632 in preparation of the medicine for reversing epithelial-mesenchymal transition (EMT) of retinal pigment epithelium (RPE) provided by the invention, the EMT of the RPE can be effectively reversed; the form of the retina is maintained; and the visual function is improved. The strategy is expected to provide a new treatment medicine combination and treatment scheme for clinically treating diseases caused by the EMT of the RPE, such as PVR and AMD.

Description

technical field [0001] The invention relates to the combined use of RepSox and Y27632 in the preparation of retina-related disease medicines, belonging to the field of biomedicine. Background technique [0002] The retinal pigment epithelium (RPE), a single layer of cells located between the neural retina and the choroid, plays an important role in the survival and maintenance of function of the neural retina [1]. RPE cells are terminally differentiated and rarely divide throughout life. However, these quiescent adult RPE cells can re-enter the cell cycle and proliferate during retinal injury repair and disease progression. In lower organisms, such as amphibians, RPE cells have extraordinary regenerative capacity and can completely repair damaged RPE layers, even including the neurosensory retina. In contrast, in higher animals, such as humans, the damage caused by the proliferation of RPE cells is often accompanied by pathological EMT, often transforming into mesenchymal ...

Claims

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Application Information

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IPC IPC(8): A61K31/4375A61K31/44A61P27/02
CPCA61K31/4375A61K31/44A61K9/0048A61P27/02
Inventor 徐国彤高芙蓉吴彬欣陈羿
Owner 上海优祺生物医药科技有限公司
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