Pharmaceutical composition with Na / K-ATPase ligand as main component
By combining the use of Na,K-ATPase agonists and antagonists to regulate the conformations of E1 and E2, the safety issues of cardiac glycosides in the treatment of cancer are resolved, achieving efficient treatment and low side effects in a variety of diseases.
Patent Information
- Application Number
- CN202410253567.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-03-06
- Publication Date
- 2025-09-09
AI Technical Summary
Existing cardiac glycosides have significant anti-tumor activity in the treatment of cancer but are accompanied by adverse cardiac reactions. In addition, single conformational target drugs interfere with ion transport or signal transduction, which may affect cell physiological functions and lead to safety issues.
The combined use of Na,K-ATPase agonists and antagonists can regulate the E1 and E2 conformations, synergistically inhibit the ion transport and signal transduction of Na,K-ATPase, and reduce adverse reactions.
It has achieved the goal of improving efficacy and reducing adverse drug reactions in the treatment of tumors, cardiovascular diseases, kidney diseases, etc., and enhancing the treatment effect through synergistic effects.
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Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of biomedicine. Background Art
[0002] Na,K-ATPase is widely present on cells throughout the body and plays a crucial role in maintaining electrolyte and fluid balance in cells, organs, and the entire body. Mammalian Na,K-ATPase not only functions as an ion pump but also interacts with various cell membrane and intracellular proteins, thereby acting as a signal transduction receptor. Src is a non-receptor tyrosine kinase that forms a receptor complex with Na,K-ATPase and regulates cell growth, differentiation, apoptosis, and fibrosis.
[0003] Cardiac glycosides (CGS) such as digoxin, digitoxin, and ouabain are a class of steroid derivatives extracted from plants. As specific ligands of Na,K-ATPase, they were used as cardiotonic drugs to treat heart failure more than 200 years ago and are still used clinically to treat heart failure and atrial fibrillation.
[0004] Cancer is the leading cause of death worldwide. According to the World Health Organization (WHO), cancer caused nearly 10 million deaths worldwide in 2020, accounting for one-sixth of all deaths. Cardiac glycosides not only inhibit Na,K-ATPase's ion transport function but also activate its conformational transition from E1 to E2. This interaction with Src leads to the assembly and activation of various pathways, including the ERK cascade, the PLC / PKC pathway, and the generation of ROS in mitochondria. This leads to secondary mitochondrial damage and dysfunction, inducing tumor cell apoptosis. CGS can also activate protein tyrosine kinases (PTKs) and ERK1 / 2 through c-Src activation, restoring the expression of cell adhesion molecules in tight junctions and adherens junctions, thereby inhibiting tumor cell proliferation and metastasis. There are also reports that they exert anti-tumor effects through multiple mechanisms, including inducing cell cycle arrest, inhibiting topoisomerase activity, and reducing intracellular glycolysis. An increasing number of studies confirm the anti-tumor effects of CGS. However, while higher concentrations of CGS have significant anti-tumor activity, they also have adverse cardiac reactions due to their cardiotonic effects. Therefore, safety issues have become the biggest obstacle to its clinical application in anti-cancer medicine.
[0005] Na,K-ATPase involves the conversion of two conformations, E1 and E2, in the process of ion transport or signal transduction (see attached Figure 1Na,K-ATPase agonists (such as cardiac glycosides) can bind to E2, while antagonists (such as hydroxyxanthones) can bind to E1. In fact, both cardiac glycosides and hydroxyxanthones have anti-tumor activity. The applicants have discovered that combinations of Na,K-ATPase agonists and antagonists have enhanced anti-tumor activity and exhibit a degree of synergism. This is speculated to be related to the inhibition of both E1 and E2 conformations of Na,K-ATPase, suggesting that Na,K-ATPase may participate in a wider range of molecular signaling in the body and regulate cell growth or metabolism. Furthermore, the combined use of antagonists and agonists can also mitigate the adverse reactions of CGS drugs.
[0006] In addition to naturally occurring cardiac glycoside ligands, cardiotonic steroids also exist in the human body as agonists of Na,K-ATPase. Activation of the Na,K-ATPase / Src receptor complex by cardiotonic steroids at physiological concentrations plays a crucial role in regulating renal and cardiovascular function. Sustained activation of the Src receptor complex leads to ROS stress, contributing to the pathogenesis of various diseases, including cancer, hypertension, preeclampsia, end-stage renal disease, congestive heart failure, and diabetes. Because agonists and antagonists of the Na,K-ATPase / Src receptor complex act on different conformations of Na,K-ATPase, their combined use can modulate Na,K-ATPase-mediated signaling in vivo, broadening the safety threshold and potentially enabling the diagnosis or treatment of various diseases caused by dysregulated Na,K-ATPase signaling.
[0007] Na,K-ATPase is widely expressed in cells in the body and is crucial for maintaining the transmembrane ion gradient. + Pumps out of the cell and releases K + Pumped into the cell. Figure 1The Post-Albers ion transport cycle shown involves the activation and transition of two conformations, E1 and E2. Interfering with ion transport by drugs targeting a single conformation of Na,K-ATPase may only affect the transport of one ion, leading to the accumulation of the other ion-bound state, thereby impacting cellular physiological functions or signal transduction. However, the combined use of agonists and antagonists binding to both E1 and E2 conformations, i.e., agonists and antagonists, can regulate ion transport while reducing the accumulation of a single ion state, stabilizing the intracellular and extracellular ion gradients and mitigating the effects on cellular physiological functions and signal transduction. This can exert therapeutic effects while minimizing adverse drug reactions. The combination of two inhibitors with different conformations can produce synergistic therapeutic effects in certain areas. For example, dual-conformation inhibition can synergistically enhance positive inotropic and anti-tumor effects (see the relevant examples). Based on the above reasoning and experimental data, different small molecule compound formulations can be developed for the diagnosis or treatment of related diseases.
[0008] Modern herbal medicines or traditional Chinese medicines have the advantages of mild effects, few adverse reactions, and "treating both the symptoms and the root causes" in the diagnosis and treatment of tumors, cardiovascular diseases, kidney diseases, etc. Traditional Chinese medicines or active ingredients containing Na,K-ATPase / Src receptor complex agonists and antagonists can be developed into modern Chinese patent medicines or herbal medicines based on this principle, which will play an important role in the treatment of the above-mentioned diseases. Summary of the Invention
[0009] The present invention relates to a pharmaceutical composition, characterized in that it comprises two different types of ligands of Na,K-ATPase: (1) Agonist substances of the receptor Na,K-ATPase / Src complex; (2) Antagonists of the receptor Na,K-ATPase / Src complex.
[0010] The combined use of the two ligand-like substances will be able to exert a synergistic effect, improve efficacy, and reduce adverse reactions. It can be developed for the treatment of various tumor, cardiovascular, kidney, lung, liver, orthopedic and other related diseases caused by dysfunction of Na,K-ATPase ion transport or signal transduction function.
[0011] Example 1. Comparison of the inhibitory effects of ouabain and MB5 (3,4,5-trihydroxyxanthone) alone and in combination on DU145 and MCF-7 tumor cells. Cells were plated in 6-well culture plates, replaced with fresh medium overnight, and the test compound was added to a concentration of 1µM. After continued incubation, the number of viable cells was measured 48 hours later. Figure 2 It can be seen that the two compounds have a certain synergistic anti-tumor effect.
[0012] Example 2. Inhibitory effect of a traditional Chinese medicine formula composed of two herbs, Toad Venenum and Polygala tenuifolia, on tumor growth in a mouse tumor-bearing model. The Polygala tenuifolia herbs were air-dried, ground, sieved, and placed in a flat-bottomed flask. Ethanol was added and extracted twice with reflux in a water bath. The extracts were filtered, combined, and concentrated under reduced pressure to recover the ethanol to obtain an extract. The extract was then dried under reduced pressure and, if necessary, further purified using a macroporous resin to obtain a Polygala tenuifolia extract with a hydroxyxanthenone content of more than 30%. The test drugs were divided into three groups: a Polygala tenuifolia extract group, a Toad Venenum group, and a mixture of a Polygala tenuifolia extract and Toad Venenum group (2:1 weight ratio). All test samples were ground and mixed with a 0.5% sodium carboxymethylcellulose solution and administered orally. 5×10 6 DU145 cells were injected subcutaneously into the flank of NOD / SCID mice. 3 When the tumor tissue was transplanted, the tumor reached 100 mm. 3 The animals were given the drug by gavage at 3 o'clock and the size of the tumor was measured twice a week. Figure 3 , it can be seen that Polygala tenuifolia extract has a synergistic effect on tumor inhibition of Toad Venenum.
[0013] Example 3: Synergistic Effect of Deacetylated Scutellarin Injection and Astragalus Injection on an Acute Heart Failure Model in Rats. Wistar rats were randomly divided into a model control group (A), a deacetylated scutellarin injection group (B), an astragalus injection group (C), and a deacetylated scutellarin + astragalus injection group (D), with 8 rats in each group. Each group received an intravenous infusion of 0.4% propranolol (propranolol hydrochloride injection) to establish the model. After successful model establishment, 0.1 mg / kg of deacetylated scutellarin injection and 4 mg / kg of astragalus injection were administered. Baseline values were recorded for each group before model establishment. Pre-drug values were recorded 20 minutes after successful model establishment and stabilization. Cardiac function indicators, including heart rate (HR), left ventricular systolic pressure (LVSP), and maximum rate of rise of left ventricular pressure (+dp / dt), were recorded 5, 15, 30, 60, and 120 minutes after intravenous administration of deacetylated scutellarin injection or astragalus injection. max The recorded data showed that the combination of the two can better improve cardiac function indicators.
[0014] Figure 1 This is a Post-Albers cycle diagram of ion transport by Na / K-ATPase on the cell membrane. Figure 2 This figure shows the synergistic inhibitory effect of ouabain combined with MB5 (3,4,5-trihydroxyxanthone) on the growth of tumor cells DU145 cells and MCF-7 cells. Figure 3 This figure shows the synergistic inhibitory effect of Polygala tenuifolia extract combined with Toad Venenum on tumor growth in mice.
Claims
1. A pharmaceutical composition, characterized in that Two different types of ligands for Na / K-ATPase are included: (1) Agonist substances of the receptor Na / K-ATPase / Src complex; (2) Antagonists of the receptor Na / K-ATPase / Src complex.
2. The agonist substance of the receptor Na / K-ATPase / Src complex according to claim 1, comprising at least one of the following compounds:
3. The antagonist substance of the receptor Na / K-ATPase / Src complex according to claim 1, comprising at least one of the following compounds:
4. The Chinese medicinal materials or herbal extracts containing the agonist substances of the receptor Na / K-ATPase / Src complex according to claim 2 include but are not limited to Chinese medicinal materials of animal origin such as toad venom, dried toad, toad skin, toad skin, toad head, toad tongue, toad liver, toad gall, fish oil, etc. containing digitalis components, as well as Chinese medicinal materials of plant origin such as digitalis, oleander, and dieffenbachia.
5. Chinese medicinal materials or Chinese herbal medicine extracts containing the antagonist substance for the receptor Na / K-ATPase / Src complex according to claim 3, including but not limited to Codonopsis pilosula, Salvia miltiorrhiza, Radix Angelicae sinensis, Polygala tenuifolia, Astragalus membranaceus and other Chinese medicinal materials containing hydroxyketone components.
6. The pharmaceutical composition according to claim 1 is mainly used to treat various tumors, cardiovascular, kidney, lung, liver, orthopedic and other related diseases caused by dysfunction of Na / K-ATPase ion transport or signal transduction.
7. A pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is administered orally, by injection, by inhalation or external application.