Application of glutamine transferase 2 as a therapeutic target in screening drugs for ischemic heart disease
By promoting the expression of the Tgm2 gene in cardiomyocytes through the small molecule composition 5SM, the problem of cardiomyocyte proliferation and apoptosis in ischemic heart disease was solved, and an effective treatment for cardiac damage repair was achieved.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- UNIVERSITY OF HEALTH & REHABILITATION SCIENCES
- Filing Date
- 2026-05-09
- Publication Date
- 2026-07-31
AI Technical Summary
There is a lack of effective drug targets in the current technology for the treatment of ischemic heart disease, especially for promoting cardiomyocyte proliferation and inhibiting apoptosis by regulating glutaminase 2 (Tgm2) to repair cardiac damage.
The small molecule composition 5SM (phenylephrine hydrochloride, baricitinib, dehydrocamellia alkaloid, VO-Ohpic trihydrate, AZD3965) was used to promote the expression of the Tgm2 gene in cardiomyocytes, and to promote cardiomyocyte proliferation and inhibit apoptosis by regulating Tgm2 activity.
It significantly increased the expression level of the Tgm2 gene in cardiomyocytes, promoted cardiomyocyte proliferation, inhibited apoptosis, and improved cardiac function, providing a new drug intervention strategy for the treatment of ischemic heart disease.
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of medicine, specifically relating to the application of glutamine transferase 2 as a therapeutic target in screening drugs for ischemic heart disease. Background Technology
[0002] Ischemic heart disease is one of the leading causes of death worldwide. Myocardial ischemia is a pathological condition characterized by insufficient coronary blood supply to meet the metabolic needs of the myocardium. For many years, the main treatment strategy to reduce ischemic injury has been reperfusion therapy. However, timely restoration of blood flow to the ischemic area can also lead to myocardial injury, known as I / R injury. This injury is mainly caused by reactive oxygen species (ROS) generated during reperfusion, which can lead to cell death in the infarcted area and its adjacent regions through apoptosis, pyroptosis, autophagy, and necrosis.
[0003] While reperfusion strategies help reduce mortality, reduced left ventricular ejection fraction (EF) due to reperfusion injury is one of the leading causes of chronic heart failure. Compared to expensive equipment and heart transplantation, pharmacological strategies using small molecule drugs offer significant advantages in terms of affordability, ease of use, and manageable intervention.
[0004] The dynamic regulation of cardiomyocyte proliferation and apoptosis is a core element of cardiac injury repair. Recent data have revealed that multiple signaling pathways are closely related to cardiomyocyte apoptosis and proliferation.
[0005] (1) Key signaling pathways regulating cardiomyocyte apoptosis: The core mechanism of apoptosis depends on the activation of caspase, which gradually amplifies apoptotic signals. Hypotheses about the mechanism of cardiomyocyte apoptosis include the oxygen free radical theory and the calcium overload theory. These mechanisms mainly induce cardiomyocyte apoptosis by changing mitochondrial membrane permeability, releasing apoptosis factors.
[0006] (2) Key signaling pathways for the regulation of cardiomyocyte proliferation: In the Hippo-YAP pathway, core myokinase MST1 / 2 inhibits the nuclear entry of YAP protein by phosphorylating it and blocks the expression of proliferative genes. The Notch signaling pathway participates in the regeneration process by regulating cardiomyocyte dedifferentiation. Activation of the PI3K / Akt-mTOR pathway can promote cardiomyocyte proliferation.
[0007] Transglutaminase 2 (Tgm2) belongs to the transglutaminase family, which is widely expressed in various cell types and participates in many key cellular processes, including cell growth, proliferation, differentiation, migration, and apoptosis. Tgm2 is a multifunctional protein whose activities can be divided into calcium-dependent (such as transaminases and deaminases) and calcium-independent (such as GTPases, ATPases, protein kinases, and dithiothreonose isomerases).
[0008] Currently, there are no reports on using glutaminase 2 as a therapeutic target in the screening of drugs for ischemic heart disease. Summary of the Invention
[0009] The purpose of this invention is to provide the application of transglutaminase 2 (Tgm2) as a target in the treatment of ischemic heart disease.
[0010] In a first aspect, the present invention claims the use of a substance that promotes the expression of the Tgm2 gene in cardiomyocytes in the preparation of a medicament for the prevention and / or treatment of ischemic heart disease.
[0011] Secondly, the present invention claims protection for the use of a substance that promotes the expression of the Tgm2 gene in cardiomyocytes in the preparation of a product for protecting the myocardium in ischemic heart disease, or for the protection of the myocardium in ischemic heart disease.
[0012] Thirdly, the present invention claims protection for the use of substances that promote the expression of the Tgm2 gene in cardiomyocytes in the preparation of products for promoting cardiomyocyte proliferation and / or inhibiting cardiomyocyte apoptosis in ischemic heart disease; or for use in promoting cardiomyocyte proliferation and / or inhibiting cardiomyocyte apoptosis in ischemic heart disease.
[0013] Fourthly, the present invention claims the use of transglutaminase 2 (Tgm2) as a target in screening drugs for the prevention and / or treatment of ischemic heart disease.
[0014] The screening method is as follows: the drugs or reagents to be screened are screened with glutaminase 2 (Tgm2) as the target, and the drugs or reagents that can increase the expression level of Tgm2 gene are used as candidate drugs for the prevention and / or treatment of ischemic heart disease.
[0015] Furthermore, the glutamine transferase 2 (Tgm2) is glutamine transferase 2 (Tgm2) in cardiomyocytes.
[0016] In the above aspects, the substance that can promote the expression of the Tgm2 gene in cardiomyocytes is a substance that directly targets the Tgm2 gene and can increase the expression of the Tgm2 gene.
[0017] Of the above aspects, the substance that promotes the expression of the Tgm2 gene in cardiomyocytes is a substance that specifically promotes the expression of the Tgm2 gene in cardiomyocytes.
[0018] The substance that promotes Tgm2 gene expression in cardiomyocytes is a small molecule combination 5SM, which consists of phenylephrine hydrochloride (α-adrenergic receptor agonist), baricitinib (JAK inhibitor), dehydrocamellidine (DRYK inhibitor), VO-Ohpic trihydrate (PTEN inhibitor), and AZD3965 (MCT1 inhibitor). The molar ratio of phenylephrine hydrochloride, baricitinib, dehydrocamellidine, VO-Ohpic trihydrate, and AZD3965 is 2:4:5:2:2.
[0019] Transglutaminase 2 (Tgm2) is a multifunctional enzyme that plays a crucial role in regulating cardiomyocyte proliferation and apoptosis. The regulatory role of Tgm2 in cardiac injury repair provides novel drug candidates and intervention targets for the treatment of ischemic heart disease. Attached Figure Description
[0020] Figure 1 To conduct a multi-omics analysis of changes in Tgm2 gene expression in cardiomyocytes treated with 5SM; Figure 2 The figure shows changes in Tgm2 expression and its effects on cardiomyocyte proliferation and apoptosis; in the figure, Sham represents the sham-operated group and MI represents the myocardial infarction model. Detailed Implementation
[0021] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.
[0022] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.
[0023] Example 1: Spatiotemporal Representation Characteristics of Tgm2 The inventors' previous research found that 5SM (five small-molecule chemical compounds) can significantly induce cardiomyocyte proliferation and improve impaired cardiac function [Reference: Du JY, Zheng LX, Gao P et al. A small-molecule cocktail promotes mammalian cardiomyocyte proliferation and heartregeneration. Cell Stem Cell 2022; 29: 545–558.e13.]. The small-molecule combination 5SM consists of phenylephrine hydrochloride (an α-adrenergic receptor agonist), baricitinib (a JAK inhibitor), dehydrocamipolamine (a DRYK inhibitor), VO-Ohpic trihydrate (a PTEN inhibitor), and AZD3965 (an MCT1 inhibitor). The molar ratio of phenylephrine hydrochloride, baricitinib, dehydrocamipolamine, VO-Ohpic trihydrate, and AZD3965 is 2:4:5:2:2.
[0024] Analysis of multi-omics data revealed a significant upregulation of the Tgm2 promoter region peak during myocardial proliferation induction, indicating that 5SM treatment significantly increased chromatin accessibility in this region and enhanced Tgm2 gene transcriptional activity. Simultaneously, bulk-RNASeq results showed a significant increase in Tgm2 expression levels after 5SM treatment, with statistically significant differences. Single-cell sequencing revealed the expression distribution of Tgm2 in cardiac cells, showing that Tgm2 is mainly expressed in cardiomyocytes. These results suggest that Tgm2 may be involved in the myocardial repair process following cardiac injury.
[0025] Example 2: The relationship between Tgm2 and myocardial proliferation / apoptosis The study found that Tgm2 expression was low in rat cardiomyocytes at 3 days (P3) and 7 days (P7) after birth, and significantly upregulated after 14 days (P14) (see [link to study]). Figure 2 In adult rats, the expression of Tgm2 was observed to change at 1, 3, and 7 days after myocardial infarction. The results showed that Tgm2 expression initially increased significantly and then decreased (see [link to original text]). Figure 2 (See Figure B). This suggests that Tgm2 may be involved in the repair and regeneration process after cardiac injury. Furthermore, Tgm2 expression was significantly upregulated in neonatal mice 1 day (P1) and 8 days (P8) after myocardial infarction, 1.5 days after the onset of myocardial infarction. In P1 mice, Tgm2 expression was upregulated at 3 and 7 days after myocardial infarction, but without statistical significance. In P8 mice, Tgm2 expression was also upregulated at 3 and 7 days after myocardial infarction, with statistical significance (see Figure B). Figure 2(C) The above results suggest that Tgm2 is significantly upregulated in both neonatal and adult rats after myocardial infarction and is involved in the process of cardiac injury repair. Knockdown of Tgm2 (Tgm2-siRNA) in neonatal rat cardiomyocytes significantly reduced the number of pH3-positive cardiomyocytes (see [reference needed]). Figure 2 In the middle D and E); overexpression of Tgm2 (Tgm2-OE) pH3-positive cardiomyocytes was significantly increased (see D, E); Figure 2 In mid-F and G mice, Tgm2 is overexpressed in neonatal rat cardiomyocytes (Tgm2-OE), and the expression of apoptosis genes Caspase, Bcl2, and BAX is decreased (see [link to relevant documentation]). Figure 2 The results (from the study of Tgm2 in the central nervous system and its associated apoptosis) suggest that Tgm2 may participate in cardioprotection by influencing cardiomyocyte apoptosis. These findings suggest that Tgm2 may affect cardiac repair after injury by influencing cardiomyocyte proliferation and apoptosis.
[0026] The present invention has been described in detail above. For those skilled in the art, the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. Although specific embodiments have been given, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. Some of the essential features can be applied within the scope of the following appended claims.
Claims
1. Application of substances that promote the expression of the Tgm2 gene in cardiomyocytes in the preparation of drugs for the prevention and / or treatment of ischemic heart disease.
2. The application of substances that promote the expression of the Tgm2 gene in cardiomyocytes in the preparation of products for protecting the myocardium in ischemic heart disease, or for the protection of the myocardium in ischemic heart disease.
3. The application of substances that promote the expression of the Tgm2 gene in cardiomyocytes in the preparation of products for promoting cardiomyocyte proliferation in ischemic heart disease; or for promoting cardiomyocyte proliferation in ischemic heart disease.
4. The application of substances that promote the expression of the Tgm2 gene in cardiomyocytes in the preparation of products for inhibiting cardiomyocyte apoptosis in ischemic heart disease; or for inhibiting cardiomyocyte apoptosis in ischemic heart disease.
5. Application of glutamine transferase 2 (Tgm2) as a target in screening drugs for the prevention and / or treatment of ischemic heart disease.
6. The application according to claim 5, characterized in that: The screening method is as follows: the drugs or reagents to be screened are screened with glutaminase 2 (Tgm2) as the target, and the drugs or reagents that can increase the expression level of Tgm2 gene are used as candidate drugs for the prevention and / or treatment of ischemic heart disease.
7. The application according to claim 5 or 6, characterized in that: The glutamine transferase 2 (Tgm2) mentioned is the glutamine transferase 2 (Tgm2) in cardiomyocytes.