Application of digoxin in preparation of medicine for treating and / or preventing tumors

Abstract

The invention discloses an application of Digoxin in preparation of a medicine for treating and / or preventing tumors. It is found for the first time that Digoxin can enhance the esophageal cancer radiotherapy sensitivity. Digoxin is a medicine which is approved by FDA, small in toxic and side effects and moderate in price, can be prepared into capsules for oral administration, and is used for radiotherapy sensitization of esophageal cancer and inhibition of growth of esophageal cancer.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and specifically relates to the application of Digoxin in the preparation of drugs for the treatment and / or prevention of tumors. Background Technology

[0002] Esophageal cancer includes two subtypes: esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC). ESCC is the predominant type in Southeast Asia and Africa, while EAC is the predominant type in Europe and North America. Regardless of the subtype, current treatment outcomes are unsatisfactory, and patient prognosis is extremely poor.

[0003] Significant progress has been made in the treatment of esophageal cancer in recent years, but the outcomes remain unsatisfactory. Radiotherapy plays a crucial role in the comprehensive treatment of esophageal cancer. However, the sensitivity of esophageal cancer to radiotherapy varies considerably; some patients are not sensitive to radiotherapy and are prone to radioresistance, which negatively impacts the prognosis of severely affected patients. Screening for compounds that can effectively reverse radioresistance in esophageal cancer will help clinicians develop better treatment plans.

[0004] Digoxin is an organic compound with the chemical formula C63-C62 ... 41 H 64 O 14 It is a medium-potency cardiac glycoside, a white crystalline powder, odorless, and bitter in taste. During treatment, it exerts a positive inotropic effect on the heart, slowing the heart rate and inhibiting cardiac conduction. It is suitable for low-output congestive heart failure, atrial fibrillation, atrial flutter, and paroxysmal supraventricular tachycardia. Summary of the Invention

[0005] The primary objective of this invention is to overcome the shortcomings and deficiencies of the prior art and to provide the application of Digoxin in the preparation of medicaments for the treatment and / or prevention of esophageal cancer.

[0006] The above-mentioned objectives of the present invention are achieved through the following technical solutions: The use of Digoxin in the preparation of drugs for the treatment and / or prevention of esophageal cancer.

[0007] Furthermore, the effective dose of Digoxin is 2.5–5 mg / kg.

[0008] Application of Digoxin in the preparation of reagents that inhibit the growth of esophageal cancer cells.

[0009] Furthermore, the effective concentration of Digoxin is 25–50 nM.

[0010] Furthermore, the esophageal cancer mentioned includes, but is not limited to, esophageal squamous cell carcinoma.

[0011] Application of Digoxin in the preparation of drugs that enhance the radiosensitivity of esophageal cancer.

[0012] Furthermore, the effective dose of Digoxin is 2.5–5 mg / kg.

[0013] Application of Digoxin in the preparation of reagents that enhance the radiosensitivity of esophageal cancer cells.

[0014] Furthermore, the effective concentration of Digoxin is 25–50 nM.

[0015] Furthermore, the radiotherapy uses radiation selected from the following: X-rays, alpha rays, beta rays, gamma rays, neutrons, electron beams, proton beams, particle beams, or combinations thereof.

[0016] Preferably, the single irradiation dose of the radiotherapy is 2 Gy.

[0017] Application of Digoxin in combination with radiotherapy in the preparation of drugs for the treatment and / or prevention of esophageal cancer.

[0018] Furthermore, the structural formula of Digoxin is shown in formula (Ⅰ):

[0019] Equation (Ⅰ).

[0020] Furthermore, the drug also contains pharmaceutically acceptable excipients.

[0021] Furthermore, the pharmaceutically acceptable excipient is preferably at least one of sustained-release agents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, adsorbents, surfactants, and lubricants.

[0022] Furthermore, the administration methods of the drug include, but are not limited to, oral administration, injection administration, and gavage administration.

[0023] The present invention has the following advantages and effects compared with the prior art: This invention is the first to discover that Digoxin can enhance the radiosensitivity of esophageal cancer. Digoxin is an FDA-approved drug with few side effects and an affordable price. It can be formulated into capsules for oral administration to enhance radiosensitivity in esophageal cancer and inhibit its growth. Attached Figure Description

[0024] Figure 1 This is a structural diagram of Digoxin.

[0025] Figure 2The figure shows the effect of different concentrations of Digoxin on the radiosensitivity of esophageal squamous cell carcinoma cells.

[0026] Figure 3 Figure showing the effect of different concentrations of Digoxin on the radiosensitivity of esophageal squamous cell carcinoma cells and the resulting cell colony formation.

[0027] Figure 4 The graph shows the results of the radiosensitivity of Digoxin in tumor-bearing mice. Detailed Implementation

[0028] The present invention will be further described in detail below with reference to the embodiments and accompanying drawings, but the embodiments of the present invention are not limited thereto.

[0029] Human esophageal squamous cell carcinoma cells KYSE150 and KYSE270 were purchased from German Microbiology & Cell Culture GmbH; LB medium was purchased from Sangon Biotech (Shanghai) Co., Ltd.; and 0.25% trypsin was purchased from Gibco, USA. Experimental female mice (C57BL / 6) were purchased from Ruiye Model Animal Center.

[0030] Method for constructing radioresistant (DR) cells for human esophageal cancer: Human esophageal squamous cell carcinoma cells KYSE150 and KYSE270, in logarithmic growth phase, were seeded into T25 culture flasks. When the cell confluence reached approximately 60%, they were subjected to a single 2 Gy X-ray irradiation. After the cells recovered and grew to 90% confluence, they were passaged and subjected to another 2 Gy irradiation. This "irradiation-recovery-passaging" cycle was repeated until the cumulative radiation dose reached 60 Gy. After irradiation was stopped, the cells were passaged 2 to 3 more times to obtain a stable radioresistant cell line.

[0031] Example 1: In vitro test 1.1 Cell viability The constructed esophageal cancer DR cells were treated with different concentrations (0, 50 nM) of digoxin and simultaneously irradiated with 4 Gy of ionizing radiation. Cell viability was then assessed using the WST-1 assay to investigate the effect of digoxin on the radiosensitivity of esophageal cancer DR cells. The specific steps are as follows: (1) 3000 cells were seeded into a 96-well plate for cell culture. After adhesion, different concentrations (0, 50 nM) of digoxin were added to treat esophageal cancer DR cells for 24 h.

[0032] (2) Irradiate the cells with 4 Gy of ionizing radiation, add CCK-8 after 48 hours, and keep at 37°C in the dark for 1 hour.

[0033] (3) Detect cell absorbance using an enzyme-linked immunosorbent assay (ELISA) reader and calculate cell relative activity.

[0034] 1.2 Cloning The constructed esophageal cancer DR cells were treated with different concentrations (0, 50 nM) of digoxin and simultaneously irradiated with 4 Gy of ionizing radiation. Cell viability was assessed using a clonogenic assay to investigate the effect of digoxin on the radiosensitivity of esophageal cancer DR cells. The specific steps are as follows: (1) 3000 cells were seeded into a 6-well plate for cell culture. After adhesion, different concentrations (0, 50 nM) of digoxin were added for treatment.

[0035] (2) After 24 hours of Digoxin treatment, the cells were irradiated with 4 Gy of ionizing radiation, and then the cells were cultured in an incubator for 10 days, during which the medium was changed.

[0036] (3) The cells were fixed with methanol and stained with crystal violet to calculate their ability to form clones.

[0037] The results are as follows Figure 2 and Figure 3 As shown, the results indicate that digoxi can enhance the sensitivity of esophageal cancer DR cells to radiotherapy. From Figure 2 It can be seen that the proliferative activity of radiotherapy-resistant epithelial cells (KYSE150-DR and KYSE270-DR) in human esophageal cancer decreases with increasing digoxin levels under radiotherapy. From... Figure 2 It can be seen that the clonogenic ability of radiotherapy resistant cells (KYSE150-DR and KYSE270-DR) in human esophageal cancer epithelial cells weakens with the increase of digoxin under radiotherapy.

[0038] Example 2: In vivo experiment Twenty-four 6-week-old female C57BL / 6 mice (purchased from Ruiye Model Animal Center) were selected and divided into four groups to construct a subcutaneous tumorigenesis model: (1) Mouse esophageal cancer cells (AKR, ATCC) were resuspended in PBS buffer (pH=7.4, 0.01M) to obtain resuspended cells; (2) Before the experiment, the mice were anesthetized and the degree of anesthesia was assessed by painless and painful stimuli to determine whether the nude mice were in anesthetized state. (3) Mix the resuspended cells with matrix gel at a volume ratio of 1:1 to obtain a mixture. Take 100 μL of the mixture containing 2×10⁻⁶ cells using a microsyringe with a 25 G needle. 5 The mice were subcutaneously injected with a mixture of AKRs. (4) Digoxin treatment: Ten days after subcutaneous injection, the drug was administered via gavage. This experiment included four groups: a control group, a radiotherapy group (4 Gy), a digoxin group (2.5 mg / kg), and a digoxin (2.5 mg / kg) + radiotherapy group (4 Gy), with six mice in each group. Treatment was administered every two days for a total of four treatments. Digoxin was dissolved in 0.5% (m / v) sodium nitrocellulose, and 100 μL was administered to each mouse per dose. Tumor size and mouse weight were measured after each administration. The mice were then euthanized, and the tumors were removed. The results are as follows Figure 4 As shown in the figure. The results showed that digoxin combined with radiotherapy significantly inhibited tumor growth in mice, and digoxin enhanced the radiosensitivity of esophageal cancer.

[0039] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. The use of Digoxin in the preparation of drugs for the treatment and / or prevention of esophageal cancer.

2. Application of Digoxin in the preparation of reagents that inhibit the growth of esophageal cancer cells.

3. Application of Digoxin in the preparation of drugs that enhance the radiosensitivity of esophageal cancer.

4. Application of Digoxin in the preparation of reagents that enhance the radiosensitivity of esophageal cancer cells.

5. Application of Digoxin combined with radiotherapy in the preparation of drugs for the treatment and / or prevention of esophageal cancer.

6. The application according to any one of claims 3-5, characterized in that: The radiotherapy described herein uses radiation selected from the following: X-rays, alpha rays, beta rays, gamma rays, neutrons, electron beams, proton beams, particle beams, or combinations thereof.

7. The application according to any one of claims 2 or 4, characterized in that: The effective concentration of Digoxin is 25–50 nM.

8. The application according to any one of claims 1-5, characterized in that: The esophageal cancer mentioned is esophageal squamous cell carcinoma.

9. The application according to any one of claims 1-5, characterized in that: The structural formula of Digoxin is shown in formula (Ⅰ): Equation (Ⅰ).

10. The application according to claim 1, 3, or 5, characterized in that: The drug further comprises pharmaceutically acceptable excipients; the pharmaceutically acceptable excipients are at least one of sustained-release agents, excipients, fillers, binders, humectants, disintegrants, absorption enhancers, adsorbents, surfactants, and lubricants. The methods of administration of the drug include oral administration, injection administration, and gavage administration.

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