JQ1-ribociclib heterocomplex, synthetic method thereof and application of JQ1-ribociclib heterocomplex in preparation of anti-gastric cancer drugs

Abstract

The invention discloses a JQ1-ribociclib heterocomplex, a synthesis method thereof and application of the JQ1-ribociclib heterocomplex as an anti-gastric cancer drug. The structural formula of the JQ1-ribociclib heterocomplex is # imgabs0 #. The invention also specifically discloses a synthesis method of the JQ1-ribociclib heterocomplex and application of the JQ1-ribociclib heterocomplex in preparation of the anti-gastric cancer drug. The JQ1-ribociclib heterocomplex provided by the invention has relatively high gastric cancer inhibition activity, and a new way is provided for treatment of gastric cancer. The JQ1-ribociclib heterocomplex provided by the invention has the advantages of simple synthesis method, cheap and easily available raw materials, and easy industrial production and clinical conversion.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicinal chemistry, and particularly relates to a JQ1-ribociclib hybrid, a synthesis method thereof, and an application in preparing an anti-gastric cancer drug. Background Art

[0002] Cancer is an important disease threatening human life and health. Gastric cancer has the second highest fatality rate among all cancer types, and the five-year survival rate is less than 30%. In China, the incidence and death population of gastric cancer both exceed 40% of the global total. In addition to surgery, radiotherapy, and chemotherapy, targeted therapy is also clinically used for the treatment of gastric cancer. However, the targeted therapy for gastric cancer only has targets such as VEGFR-2, HER2, and PD-1. Therefore, it is of great significance to discover new effective therapeutic targets and inhibitors against these targets.

[0003] Bromodomain-containing protein 4 (BRD4) can recognize and bind to the acetylated lysine residues at the N-terminal tail of histones through two domains (BD1 and BD2), participate in the transmission of epigenetic histone post-translational modification acetylation signals, regulate downstream gene transcription, and play an important role in the processes of cell cycle, cell differentiation, and signal transduction. BRD4 inhibitors can competitively bind to the acetyl-lysine binding site of BRD4, block the binding of BRD4 to histone acetylated lysine, and antagonize the promoting effect of BRD4 on tumorigenesis and development, thereby inducing apoptosis of tumor cells and inhibiting the proliferation and metastasis of tumors. At present, many BRD4 inhibitors have been developed successively. Among them, a variety of BRD4 inhibitors are in the clinical trial stage, showing good application prospects. Compared with normal gastric tissue, the expression of BRD4 is significantly up-regulated in gastric cancer tissue. Overexpressed BRD4 in gastric cancer promotes tumor growth and development. Combining BRD4 inhibitors with other inhibitors can exert synergistic anti-gastric cancer activity and reduce the generation of drug resistance. These research results provide an important theoretical basis for the application of BRD4 inhibitors in the clinical treatment of gastric cancer.

[0004] Sustained proliferation is an important feature of tumor cells. Therefore, blocking the tumor cell cycle has become an important means to control tumor development. Cyclin-dependent kinases 4 / 6 (CDK4 / 6) are key regulators of the cell cycle. By binding to cyclin D (CyclinD), they phosphorylate the retinoblastoma protein (Rb), ultimately activating the E2F transcriptional program and promoting cells to enter the S phase of the cell cycle. In tumor cells, due to activating mutations, the protein kinases are always in an activated state, allowing cell division to be uncontrolled by proliferation and inhibitory signals. CDK4 / 6 inhibitors can bind to the ATP-binding site of the CDK4 / 6 protein to exert an inhibitory effect, blocking Rb phosphorylation and subsequent reactions, thereby preventing DNA synthesis and proliferation of tumor cells. Gastric cancer patients often exhibit abnormalities in cell cycle-related molecules, making it an attractive therapeutic target. Notably, in gastric cancer patients, the CDKN2A-CDK4 / CDK6 / CCND1 signaling axis is often in a hyperactive state. For example, CDKN2A silencing was found in 35% of GC patients, CDK6 amplification was observed in 9% of gastric cancer patients, and CCND1 amplification was observed in 7% of patients. These alterations accelerate the transition of tumor cells from the G1 phase to the S phase and lead to uncontrolled cell proliferation. Since CDK4 / 6 is a key mediator for cells to transition to the S phase, inhibiting the activity of CDK4 / 6 can effectively inhibit the occurrence and development of GC. Currently, in preclinical models, monotherapies and combination formulations based on CDK4 / 6 inhibitors have initially shown good efficacy and safety in gastric cancer patients.

[0005] Although CDK4 / 6 inhibitors have shown single-agent anti-tumor activity in gastric cancer patients, not all patients respond to these drugs, and most cancer patients develop acquired drug resistance after using CDK4 / 6 inhibitors. The clinical efficacy of using CDK4 / 6 inhibitors alone is limited. In 2022, Sun et al. (J Exp Clin Cancer Res. 2023, 42, 44) found that CDK4 / 6 inhibitors can induce chromatin remodeling in gastric cancer cells, significantly increasing the expression of H3K27ac and overactivating BRD4. Inhibiting BRD4 with small molecule inhibitors can cooperate with ABE to induce senescence of GC cells and increase anti-GC activity. Multi-target drugs can avoid the defects of combination therapy. In the present invention, the active inhibitory fragment of the BRD4 inhibitor JQ1 and the CDK4 / 6 inhibitor ribociclib were rationally spliced to obtain the JQ1-ribociclib hybrid, which has high gastric cancer inhibitory activity, providing a new approach for the treatment of gastric cancer. There is no relevant report in this regard. Summary of the Invention

[0006] The technical problem solved by the present invention is to provide a JQ1-ribociclib hybrid and its synthesis method. Another object of the present invention is to provide the application of the JQ1-ribociclib hybrid in the preparation of anti-gastric cancer drugs.

[0007] To solve the above technical problem, the present invention adopts the following technical solution. A JQ1-ribociclib hybrid, characterized in that the structural formula of the JQ1-ribociclib hybrid is shown in Formula I:

[0008]

[0009] The present invention also provides a synthesis method of the above JQ1-ribociclib hybrid, characterized in that the specific steps are as follows:

[0010] Step S1: Dissolve ribociclib, N-Boc-bromoethylamine and DIPEA (N,N-diisopropylethylamine) in DMF (N,N-dimethylformamide), react at 80-90 °C for 8-16 h. After the reaction is completed, add water, extract with dichloromethane, wash the organic phase successively with saturated sodium bicarbonate and saturated sodium chloride, dry over anhydrous sodium sulfate, filter, concentrate and purify to obtain Compound II;

[0011] Step S2: Dissolve Compound II obtained in Step S1 in an ethyl acetate solution of HCl, react at room temperature for 8-16 h. After the reaction is completed, evaporate the solvent to obtain Compound III;

[0012] Step S3: Dissolve Compound III obtained in Step S2 and JQ1-COOH in DMF, add HATU (polypeptide condensing agent, 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate) and DIPFA, react at room temperature for 8-16 h. After the reaction is completed, add water, extract with dichloromethane, wash the organic phase successively with saturated sodium bicarbonate and saturated sodium chloride, dry over anhydrous sodium sulfate, filter, concentrate and purify to obtain Compound I;

[0013] The corresponding reaction equations during the synthesis process are:

[0014]

[0015] The application of the JQ1-ribociclib hybrid described in the present invention in the preparation of anti-gastric cancer drugs.

[0016] The present invention has the following advantages and beneficial effects: The JQ1-ribociclib hybrid provided by the present invention has a strong inhibitory effect on the tested gastric cancer cells HGC-27 and MKN-45, and its IC 50 are 5.71 μM and 12.27 μM respectively, and can be further developed as anti-gastric cancer drugs. Specific Embodiments

[0017] The above content of the present invention will be further described in detail through the following examples. However, it should not be understood that the scope of the above subject matter of the present invention is limited to the following examples. All technologies implemented based on the above content of the present invention belong to the scope of the present invention.

[0018] Example 1

[0019] Synthesis of JQ1-Ribociclib Hybrid

[0020] Ribociclib (0.5 mmol, 217 mg), N-Boc-bromoethylamine (0.6 mmol, 134.4 mg) and DIPEA (1.5 mmol, 193.5 mg) were dissolved in 5 mL of anhydrous DMF and reacted at 85 °C for 12 h. After the reaction was complete, 20 mL of water was added, and the mixture was extracted 3 times with 60 mL of dichloromethane. The organic phase was washed successively with saturated sodium bicarbonate and brine, dried over anhydrous sodium sulfate, filtered, concentrated, and then separated by column chromatography to obtain 130 mg of pale yellow solid compound II, melting point: 100–103 °C, yield 37.5%. 1 HNMR (DMSO-d6, 400 MHz) δ (ppm): 9.47 (s, 1H), 8.79 (s, 1H), 8.16 (d, 1H, J = 8.8 Hz), 8.02 (s, 1H), 7.43 (d, 1H, J = 8.0 Hz), 6.71 (s, 1H), 6.61 (s, 1H), 4.78–4.70 (m, 1H), 3.11–3.07 (m, 12H), 2.55–2.39 (m, 8H), 1.98 (s, 4H), 1.64–1.63 (m, 2H), 1.22 (s, 9H); HRMS-ESI (m / z): calcd for C 30 H 44 N9O3 [M+H] + 578.3562, found: 578.3560.

[0021] The above compound II was dissolved in 5 mL of an ethyl acetate solution of 4.0 M HCl and reacted at room temperature for 8 h. After the reaction was completed, the solvent was evaporated to dryness to obtain 100 mg of yellow solid compound III, yield 93%. The product was used directly for the next step without further purification.

[0022] Dissolve the above compound III (0.21 mmol, 100 mg) and JQ1-COOH (0.21 mmol, 84 mg) in 5 mL of DMF, add HATU (0.315 mmol, 120 mg) and DIPEA (1.26 mmol, 163 mg), and react at room temperature for 12 h. After the reaction is completed, add 20 mL of water and extract 3 times with 60 mL of dichloromethane. The organic phase is washed successively with saturated sodium bicarbonate and brine, dried over anhydrous sodium sulfate, filtered, concentrated, and then separated by column chromatography to obtain 41 mg of a pale yellow solid compound I, namely JQ1-ribociclib hybrid, melting point: 190–194 °C, yield 22.7%. 1 1H NMR (DMSO-d6, 400 MHz) δ (ppm): 9.40 (s, 1H), 8.77 (s, 1H), 8.23–8.17 (m, 2H), 8.15 (s, 1H), 7.49–7.42 (m, 4H), 6.60 (s, 1H), 4.78–4.69 (m, 1H), 4.54–4.50 (m, 1H), 3.31–3.06 (m, 16H), 2.59–2.40 (m, 13H), 1.98 (s, 4H), 1.71–1.56 (m, 5H); HRMS-ESI (m / z): calcd for C 44 1 51 6 13 4 + 4

[0023] Example 2

[0024] Anti-proliferative activity assay of JQ1-ribociclib hybrid

[0025] After routine trypsin digestion of HGC-27 and MKN-45 cells in the logarithmic growth phase, they were evenly inoculated into a 96-well culture plate and cultured overnight in a cell culture incubator at 37 °C and 5% CO2 by volume. Different concentrations of compound I diluted in multiples were added, and after culturing in a 37 °C, CO2 incubator for 72 hours, the supernatant was discarded, MTT was added and the cells were cultured for another 4 hours. After dissolution with DMSO, the absorbance A value of each well was measured at a wavelength of 570 nm using a full-wavelength multifunctional reader. The results showed that the JQ1-ribociclib hybrid had a strong inhibitory effect on the tested gastric cancer cells HGC-27 and MKN-45, and its IC 50 values were 5.71 μM and 12.27 μM, respectively.

[0026] The above has shown and described the basic principles, main features and advantages of the present invention. Without departing from the spirit and scope of the present invention, there are various changes and improvements to the present invention, and these changes and improvements all fall within the scope of the present invention claimed.

Claims

1. A JQ1-ribociclib heterocomplex, characterized in that The structural formula of the JQ1-ribociclib hybrid is shown in Formula I:

2. A method for synthesizing the JQ1-Riboxib hybrid according to claim 1, characterized in that The specific steps are: Step S1: dissolving Riboxib, N-Boc-bromoethylamine and DIPEA in DMF, reacting at 80-90° C. for 8-16 h. After the reaction is completed, adding water, extracting with dichloromethane, washing the organic phase with saturated sodium bicarbonate and saturated sodium chloride in sequence, drying with anhydrous sodium sulfate, filtering, concentrating and purifying to obtain compound II; Step S2: dissolving the compound II obtained in step S1 in an ethyl acetate solution of HCl, reacting at room temperature for 8 to 16 hours, and evaporating the solvent after the reaction to obtain compound III; Step S3: dissolving the compound III and JQ1-COOH obtained in step S2 in DMF, adding HATU and DIPFA, and reacting at room temperature for 8 to 16 hours. After the reaction is completed, adding water, extracting with dichloromethane, washing the organic phase with saturated sodium bicarbonate and saturated sodium chloride in turn, drying with anhydrous sodium sulfate, filtering, concentrating, and purifying to obtain compound I; The corresponding reaction equation in the synthesis process is:

3. Use of the JQ1-ribociclib hybrid according to claim 1 in the preparation of anti-gastric cancer drugs.

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