Use of 7586-3507 in the preparation of antitumor drugs
The small molecule compound 7586-3507 targets NAT10 to inhibit esophageal cancer cell invasion and metastasis, offering a safer and more effective treatment option for esophageal cancer with minimal side effects.
Patent Information
- Application Number
- JP2025546930
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- Filing Date
- 2023-02-13
- Publication Date
- 2026-02-20
- Estimated Expiration
- 2043-02-13
AI Technical Summary
Current treatments for esophageal cancer, particularly esophageal squamous cell carcinoma, have low efficacy and high toxicity, with a 5-year survival rate of only 25-30% and limited improvement in surgical resection efficiency, necessitating the development of more stable and effective antitumor drugs.
The use of the small molecule compound 7586-3507, targeting NAT10, to inhibit the invasion and metastasis of esophageal cancer cells, formulated into an anti-esophageal cancer drug with concentrations ranging from 1.25 to 5 μM, potentially combined with pharmaceutically acceptable auxiliary materials.
7586-3507 effectively inhibits esophageal cancer cell invasion and metastasis in a concentration-dependent manner, demonstrating safety and efficacy as a promising antitumor drug with minimal toxicity.
Smart Images
Figure 2026506057000001_ABST
Abstract
Description
[Technical Field]
[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to the use of 7586-3507 in the preparation of antitumor drugs. [Background technology]
[0002] Esophageal cancer is a relatively common gastrointestinal tumor in China and one of the most common malignant tumors, with high morbidity, mortality, and low overall survival rates. Esophageal cancer has two histological subtypes: esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EA). However, in the Asia-Pacific region, including China, the majority of cases are ESCC. Treatment methods for esophageal cancer mainly include surgical resection, chemotherapy, radiation therapy, and molecular targeted therapy. Existing treatment methods have not improved patient prognosis, and the 5-year survival rate for currently treated esophageal cancer patients is only 25–30% (PMID: 29635717). Statistics show that 90% of patients diagnosed with esophageal cancer in China progress to mid- and late-stage disease. Improvements in the efficiency of surgical resection have not been sufficient, and drug therapy remains an important treatment option for mid- and late-stage patients. Therefore, there is an urgent need to develop more stable, effective, and safe anticancer drugs.
[0003] With the continuous development of molecular biology mechanisms and the continuous promotion and support of the biotechnology industry, people's research into tumors has deepened, and more and more tumor-specific molecular targets have been identified. Accordingly, antitumor small molecule targeted drugs have also grown vigorously. Small molecule targeted drugs have become a hot topic in tumor treatment research due to their advantages such as high efficacy, low toxicity and side effects, and high specificity. Currently, small molecule targeted antitumor drugs widely used in clinical practice are mainly tyrosine kinase inhibitors such as gefitinib, erlotinib, imatinib, sunitinib, lapatinib, and sorafenib.
[0004] N-acetyltransferase 10 (NAT10) is a nuclear protein that functions as an acetyltransferase and can catalyze the acetylation modification of histones and nonhistones. Nonhistone acetylation modifications catalyzed by NAT10 play important roles in various biological activities, including regulating telomerase activity, rRNA synthesis, DNA damage repair, and maintaining mRNA stability. At the same time, research has shown that they are closely related to the initiation, development, and prognosis of cancer.
[0005] 7586-3507 (2-[(4-methylphenoxy)methyl]-5-({2-[(4-methylphenoxy)methyl]-1h-1,3-benzodiazol-5-yl}methyl)-1h-1,3-benzodiazole, 2-[(4-methylphenoxy)methyl]-5-({2-[(4-methylphenoxy)methyl]-1h-1,3-benzodiazol-5-yl}methyl)-1h-1,3-benzodiazole) is a small molecule compound with the structural formula shown below. [ka] Summary of the Invention [Problem to be solved by the invention]
[0006] The main object of the present invention is to overcome the drawbacks and deficiencies of the prior art and to provide the use of 7586-3507 in the preparation of anti-esophageal cancer drugs. [Means for solving the problem]
[0007] The object of the present invention is achieved by the following technical solutions:
[0008] The use of 7586-3507 in the preparation of an anti-esophageal cancer drug, wherein the structural formula of said 7586-3507 is represented by formula (I): [ka]
[0009] Furthermore, the effective concentration of the 7586-3507 is 1.25 to 5 μM.
[0010] Furthermore, the effective concentration of 7586-3507 is 2.5-5 μM.
[0011] Furthermore, the anti-esophageal cancer drug refers to a drug that can inhibit the invasion and / or metastasis of esophageal cancer cells. 7586-3507 significantly inhibits the invasion and / or metastasis of esophageal cancer cells in a concentration- and time-dependent manner.
[0012] Furthermore, the anti-esophageal cancer drug is a drug for treating and / or preventing esophageal cancer.
[0013] Furthermore, 7586-3507 inhibits the invasion and / or metastasis of esophageal cancer by targeting NAT10. The nucleotide sequence encoding NAT10 is shown in the gene sequence of NCBI Gene bank database under accession number 55226.
[0014] Furthermore, the esophageal cancer is esophageal squamous cell carcinoma.
[0015] An anti-esophageal cancer drug, comprising 7586-3507, the structural formula of which is represented by formula (I). [ka]
[0016] Furthermore, the anti-esophageal cancer drug further comprises a pharmaceutically acceptable auxiliary material.
[0017] Furthermore, the pharmaceutically acceptable auxiliary material is preferably at least one of a sustained-release agent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption enhancer, an adsorption carrier, a surfactant, and a lubricant.
[0018] Furthermore, the administration method of the anti-esophageal cancer drug preferably includes at least one of oral administration, intragastric administration, and injection administration, but is not limited to these. [Effects of the Invention]
[0019] Compared with the prior art, the present invention has the following advantages and effects. The present invention has found through testing that the small molecule compound 7586-3507 can effectively inhibit the invasion and metastasis of esophageal cancer. The small molecule compound 7586-3507 of the present invention significantly inhibits the invasion and metastasis of esophageal cancer cells in a concentration- and time-dependent manner within the concentration range of 1.25 μM to 5 μM. The small molecule compound 7586-3507 of the present invention has a certain degree of safety as a promising antitumor drug, and furthermore, has significant advantages in terms of efficacy and price. [Brief explanation of the drawings]
[0020] [Figure 1] FIG. 1 shows the structural formula of 7586-3507. [Figure 2] FIG. 1 shows the results of surface plasmon resonance analysis of 7586-3507 and NAT10 protein. [Figure 3] FIG. 10 is a graph showing the effects of different concentrations of 7586-3507 on the invasive ability of esophageal cancer cells. [Figure 4] This figure shows the results of fluorescence detection of the effect of different concentrations of 7586-3507 on the lung metastatic potential of esophageal squamous cell carcinoma cells. [Figure 5] FIG. 10 is a graph showing the changes in body weight of nude mice treated with different concentrations of 7586-3507. [Figure 6] FIG. 10 is a graph showing morphological changes in the lungs, liver, kidneys, and spleen of nude mice treated with different concentrations of 7586-3507. DETAILED DESCRIPTION OF THE INVENTION
[0021] The present invention will be described in more detail below with reference to examples and drawings, but the embodiments of the present invention are not limited to these. Example 1: In vitro testing
[0022] (1) Experimental materials
[0023] Human esophageal squamous cell carcinoma cells KYSE150 and EC9706 were purchased from DSMZ. 7586-3507 was purchased from Shanghai Topscience Biochemical Technology Co., Ltd. (Shanghai Topscience Biochemical Technology Co., Ltd.), and its structural formula is shown in Figure 1. Female mice (NOD-SCID) were purchased from Beijing IDMO Biotechnology Co., Ltd. RPMI-1640 medium was purchased from Gibco (USA). DMSO was purchased from Sigma (USA). 7586-3507 was dissolved in DMSO and stored at a concentration of 10 mM in a -80°C refrigerator.
[0024] (2) Cell invasion experiment:
[0025] Esophageal squamous cell carcinoma cells were treated with different concentrations of 7586-3507, and then the invasive ability of the esophageal squamous cell carcinoma cells was detected to investigate the effects of different concentrations of 7586-3507 on the invasive ability of esophageal squamous cell carcinoma cells.
[0026] 1) Chamber treatment: 100 μL of 5% Matrigel was added to a clean invasion chamber and left in an incubator at 37° C. for 30 minutes.
[0027] 2) Cell seeding: Human esophageal squamous cell carcinoma (ESCC) highly metastatic cell lines KYSE150-Luc-LM5 and EC9706-Luc-LM3 (PMID: 35507004) were digested with 0.25% trypsin to form single-cell suspensions. After cell counting, 500,000 KYSE150-Luc-LM5 cells and 200,000 EC9706-Luc-LM3 cells were added to the upper chamber of each chamber, and different concentrations of 7586-3507 (final concentrations: 0, 1.25 μM, 2.5 μM, and 5 μM, respectively) were added. 500 μL of medium containing 20% FBS was added to the lower chamber. The cells were cultured at 37°C and 5% CO2 for 24 hours.
[0028] 3) Crystal violet staining: The chamber was removed and washed twice with PBS buffer (pH 7.4, purchased from Sigma-Aldrich; the same applies below). 500 μL of methanol was added to the bottom of the chamber and fixed for 10 minutes. The methanol was then aspirated off, and 0.2% crystal violet was added and stained for 10 minutes. After aspirating off the crystal violet, the remaining crystal violet was removed with clean water, the chamber was dried, and photographed. The in vitro test results shown in Figure 3 indicate that the invasive ability of esophageal squamous cell carcinoma cells weakened with increasing 7586-3507 concentration.
[0029] (3) Biomolecular Interaction Analysis System SPR:
[0030] 1) Expression and purification of NAT10 protein
[0031] The NAT10 gene was constructed in a His-tagged pET-28b prokaryotic expression vector, and the NAT10 prokaryotic expression plasmid was obtained (commissioned from Shanghai Quanyang Biotechnology Co., Ltd.). The constructed plasmid was transformed into BL21 competent cells, plated, and then the expressed cells were picked up in 10 mL of LB medium and cultured overnight at 37°C and 250 rpm to obtain a bacterial suspension. One percent of the bacterial suspension was transferred to 1 L of LB medium and shaken at 37°C and 250 rpm for 3 hours. When the OD reached 0.6–0.8, IPTG (working concentration: 0.5 mM) was added and expression was induced for 4 hours. One L of NAT10-expressing cells was collected by centrifugation, resuspended in 100 mL of buffer, and sonicated in an ice bath (300 W, 10 seconds, 10-second intervals, 30 times). After centrifugation, the supernatants were collected and loaded onto an equilibrated NTA purification column. After loading, the column was washed with buffer. The column was eluted with 10 mM, 20 mM, 50 mM, 200 mM, and 500 mM imidazole buffer, and the eluates were collected. The eluates from each group were identified by Western blotting. Finally, the fractions containing the target protein were combined, ultrafiltered, and concentrated. The protein solution was stored at -80°C for future use.
[0032] 2) Surface plasmon resonance (SPR) experiments
[0033] A new CM7 chip was selected, and after the chip was locked, the solution was replaced with 0.4% (v / v) P20 in PBS buffer (pH 7.4, purchased from Sigma-Aldrich) to equilibrate the chip surface. NAT10 was diluted in 3M sodium acetate buffer (purchased from GE Healthcare Life Sciences) at different pH values (pH 4, 4.5, 5, and 5.5) and flowed onto a blank chip surface to test the effectiveness of electrostatic adsorption. The appropriate pH environment was selected for chip binding, and the binding volume and time were tested in manual mode to achieve optimal results. 7586-3507 was diluted to several concentrations using electrophoresis buffer (the diluted concentrations of 7586-3507 were 12.5 nM, 25 nM, 50 nM, 100 nM, 200 nM, and 400 nM, respectively), while ensuring that the DMSO content in the electrophoresis buffer was constant. Samples were manually loaded to test the response values of 7586-3507 at various concentrations. Based on the response values, appropriate concentrations of 7586-3507 were selected to measure the affinity.Finally, the analysis was performed using Biacore software.
[0034] The surface plasmon resonance results in Figure 2 show that 7586-3507 can bind to the NAT10 protein. Example 2: In vivo testing
[0035] A total of 27 6-week-old female mice (NCG), 9 in the control group and 18 in the experimental group, were selected to establish a tumor metastasis model.
[0036] 1) Human esophageal squamous cell carcinoma highly metastatic cell line KYSE150-Luc-LM5 cells were digested into single cells with 0.25% trypsin. 6 KYSE150-Luc-LM5 cells were resuspended in 100 μL of PBS buffer.
[0037] 2) Before the animal tail vein experiment, mice were anesthetized, and the degree of anesthesia was evaluated using painless and painful stimulation to confirm that the mice were in an anesthetized state.
[0038] 3) The cells were resuspended in a microsyringe with a 25G needle and injected into the tail vein of a total of 27 mice.
[0039] 4) 7586-3507 treatment: One week after tail vein injection of KYSE150-Luc-LM5 esophageal squamous cell carcinoma cells, mice were administered the drug via intragastric administration. 7586-3507 was dissolved in PBS buffer and administered weekly to nine mice in each group at concentrations of 0 mg / kg (i.e., control group), 2.5 mg / kg (i.e., experimental group 1), and 5 mg / kg (i.e., experimental group 2). D-luciferin (purchased from GOLDBIO) was administered intraperitoneally weekly for three consecutive weeks. In vivo fluorescence photographs were taken to observe cancer cell metastasis, and nude mice were weighed.
[0040] The results are shown in Figures 4 and 5. Bioluminescence in the lungs of mice treated with 7586-3507 was reduced. Bioluminescence is caused by an enzymatic reaction between luciferase in esophageal squamous cell carcinoma KYSE150-Luc-LM5 cells and D-luciferin in the body fluids, and the intensity of fluorescence reflects the density of KYSE150-Luc-LM5 cells. Therefore, the number of lung metastases of esophageal cancer cells in the 7586-3507-treated group (i.e., the experimental group) was significantly lower than that in the control group, indicating that the drug inhibited tumor cell metastasis in a concentration-dependent manner. Figure 5 shows that there was no significant difference in mouse weight, indicating that compound 7586-3507 has no obvious toxicity.
[0041] The liver, lung, kidney, and spleen of mice treated with 7586-3507 (5 mg / kg) were removed and subjected to immunohistochemical staining. The results (see Figure 6) showed that 7586-3507 did not affect the function of the liver, lung, or kidney of mice.
[0042] The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention are equivalent replacement methods and fall within the protection scope of the present invention.
Claims
1. 1. Use of 7586-3507 in the preparation of an anti-esophageal cancer drug, wherein the structural formula of 7586-3507 is represented by formula (I): 【Chemistry 1】
2. The use of 7586-3507 in preparing an anti-esophageal cancer drug according to claim 1, characterized in that the effective concentration of 7586-3507 is 1.25-5 μM.
3. The use of 7586-3507 in preparing an anti-esophageal cancer drug according to claim 1, characterized in that the effective concentration of 7586-3507 is 2.5-5 μM.
4. The use of 7586-3507 in the preparation of an anti-esophageal cancer drug according to any one of claims 1 to 3, characterized in that the anti-esophageal cancer drug refers to a drug that can inhibit the invasion and / or metastasis of esophageal cancer cells.
5. Use of 7586-3507 in the preparation of an anti-esophageal cancer drug according to any one of claims 1 to 3, characterized in that the anti-esophageal cancer drug is a drug for treating and / or preventing esophageal cancer.
6. The use of 7586-3507 in the preparation of an anti-esophageal cancer drug according to any one of claims 1 to 3, wherein the esophageal cancer is esophageal squamous cell carcinoma.
7. The use of 7586-3507 in the preparation of an anti-esophageal cancer drug according to any one of claims 1 to 3, characterized in that 7586-3507 inhibits the invasion and / or metastasis of esophageal cancer by targeting NAT10.
8. An anti-esophageal cancer drug comprising 7586-3507 having the structural formula (I). 【Chemistry 2】
9. The anti-esophageal cancer drug further comprises a pharmaceutically acceptable auxiliary material; The anti-esophageal cancer drug according to claim 8, wherein the pharmaceutically acceptable auxiliary material is at least one of a sustained-release agent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption enhancer, an adsorption carrier, a surfactant, and a lubricant.
10. The anti-esophageal cancer drug according to claim 8 or 9, wherein the method of administration of the anti-esophageal cancer drug is at least one of oral administration, intragastric administration, and injection administration.