Derivative compound of benzimidazole-2-thioacetamide as well as preparation method and application of derivative compound
By integrating a thioacetamide group into the benzimidazole structure, the derivatives address the limitations of existing drugs by enhancing bioavailability and targeting multiple cancer pathways, exhibiting potent antitumor activity.
Patent Information
- Application Number
- CN202510255424.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-05
- Publication Date
- 2025-07-15
AI Technical Summary
The existing benzimidazole compounds have low bioavailability and poor metabolic stability in the anti-tumor field, and lack dual regulation capabilities for key oncogenic pathways such as PI3K/AKT/mTOR.
A series of benzimidazole-2-thioacetamide derivative compounds were synthesized by introducing functional thioacetamide functional groups into the 2nd position of the benzimidazole backbone, and a new compound was constructed through reasonable molecular design.
The compounds show good inhibitory effects on human tumor cell lines, especially on human chronic myeloid leukemia cell K562, providing a scientific basis for the research and development of anti-tumor drugs.
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Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of medicinal chemistry, and specifically relates to a derivative compound of benzimidazole-2-thioacetamide, a preparation method thereof, and an application thereof. Background Art
[0002] As a major disease threatening human health, the occurrence and development of malignant tumors are closely related to the continuous activation of abnormal signaling pathways (Oncogene 24.50(2005):7455-7464). Although targeted drugs represented by kinase inhibitors have significantly improved the clinical efficacy, acquired drug resistance and off-target toxicity still restrict their application prospects. Statistics show that more than half of cancer patients ultimately fail in treatment due to drug resistance problems (Mol Cancer17(2018):1-12). This current situation urgently requires the development of small molecule anti-tumor candidate drugs with novel mechanisms of action.
[0003] As an important drug skeleton, 2-substituted derivatives of benzimidazole compounds exhibit unique advantages in the anti-tumor field. Research shows that benzimidazole can serve as an excellent ligand unit to form hydrogen bonds, π-π conjugation, and hydrophobic interactions with target proteins or receptors (Crystengcomm 26.38(2024):5380-5392). Especially the introduction of sulfur element can not only enhance the hydrophobic binding of the molecule to the receptor, but also regulate enzyme activity through the participation of sulfur atoms in metal chelation (Cell Biochem Biophys 55.1(2009):1-23). However, existing benzimidazole compounds still face problems such as low bioavailability and poor metabolic stability, and their dual regulation ability for key oncogenic pathways such as PI3K / AKT / mTOR needs to be improved (Eur J Med Chem 271(2024):116425).
[0004] In recent years, the structure hybridization strategy has provided new ideas for optimizing lead compounds. The thioacetamide unit, due to its unique electronic effect and conformational flexibility, can serve as an ideal pharmacophore linker. Research has confirmed that it can maintain the rigid conformation of the molecule through an amide bond, thereby balancing the activity and druggability of the drug (J.Med.Chem 53.6(2010):2601-2611). These characteristics make it an ideal structural unit for constructing tumor-selective drugs.
[0005] Based on the above research background, in this study, the thioacetamide functional group was innovatively introduced into the 2-position of the benzimidazole skeleton, and a series of novel derivatives were constructed through reasonable molecular design, which not only provides new ideas for overcoming the limitations of existing benzimidazole drugs, but also lays a chemical foundation for the development of multi-target anti-tumor drugs. Summary of the Invention
[0006] The technical problem to be solved by the present invention is to provide a class of derivative compounds of benzimidazole-2-thioacetamide.
[0007] Another object of the present invention is to provide a compound containing the above compound or its isomers in the form of any mixture or its medicinal salt, as well as pharmaceutical auxiliaries or tumor preparations.
[0008] Another object of the present invention is to provide a method for preparing the above compound and its preparation method.
[0009] Another object of the present invention is to provide the anti-tumor use of the above compound or the composition.
[0010] The technical solution of the present invention is: a derivative compound of benzimidazole-2-thioacetamide, having a structure shown in the general formula (I):
[0011]
[0012] Wherein, R is an optionally substituted or unsubstituted phenyl or pyridyl group.
[0013] R is selected from a substituted or unsubstituted phenyl group, or a substituted or unsubstituted pyridyl group.
[0014] The substituents of the said substitution are halogen, methyl or methoxy group.
[0015] R is selected from 4-bromophenyl, 4-chlorophenyl, 3-chlorophenyl, 4-fluorophenyl, 3-fluorophenyl, 2-fluorophenyl, 2-chlorophenyl, 4-methylphenyl, 3-methylphenyl, 2-methylphenyl, 3-pyridyl, 2-pyridyl, 4-pyridyl or 3,4,5-trimethoxyphenyl.
[0016] The said derivative compound is selected from the following compounds:
[0017]
[0018] The preparation method of a derivative compound of benzimidazole-2-thioacetamide includes the following steps:
[0019]
[0020] A class of compositions containing the said compound or its isomers in the form of any mixture or its medicinal salt, as well as pharmaceutical auxiliaries or tumor preparations.
[0021] The application of the said compound and the said composition in the preparation of anti-tumor cell line drugs.
[0022] The tumor cell lines are human lung cancer cell line A549 (lung adenocarcinoma), human chronic myelogenous leukemia cell line K562 (chronic myelogenous leukemia), human prostate cancer cell line PC-3 (prostate cancer), and human bladder cancer cell line 5637 (bladder carcinoma).
[0023] Advantages of the present invention: Based on the benzimidazole structure, a series of compounds containing thioacetamide structural fragments are synthesized. These compounds have good inhibitory effects on human tumor cell lines, and have good inhibitory effects on human chronic myelogenous leukemia cell K562, providing an important scientific basis for the research and development of anti-tumor drugs.
[0024] Specifically, the MTT method was used to test the in vitro anti-proliferative activities of all target molecules against four human tumor cells, A549, PC-3, K562, and 5637. After treatment with a fixed concentration of 10 μM for 48 hours, some target compounds showed certain inhibitory effects on tumor cells (see Table 2). Among them, compound E12 showed the best inhibition rate against K562 cells, which was 62.71%. Further dose-dependent testing of E12 was carried out using two-fold serial dilution (1.25 - 20 μM), and the IC50 value of E12 against K562 cells was measured to be 11.07 μM (see Table 3). Detailed implementation mode
[0025] The present invention will be further described below through examples. It should be understood that the methods described in the embodiments of the present invention are only used to illustrate the present invention, rather than to limit the present invention. Any simple improvement of the preparation method of the present invention under the premise of the concept of the present invention belongs to the scope of the present invention. All raw materials and solvents used in the examples are commercially available products.
[0026] A preparation method of a benzimidazole-2-thioacetamide derivative compound, comprising the following steps:
[0027]
[0028] Example 1
[0029] (1) Preparation of intermediate C: N-(4-acetylphenyl)-2-chloroacetamide.
[0030] In a 250 ml round-bottom flask, 4-aminoacetophenone A (0.01 mol), anhydrous potassium carbonate (0.01 mol) and chloroacetyl chloride B (0.01 mol) were dissolved in N,N-dimethylformamide (DMF), and the mixture was stirred at room temperature for 24 hours. After the reaction, the reaction solution was poured into ice water for quenching. The resulting precipitate was collected by filtration, washed thoroughly and dried under vacuum to obtain intermediate C (N-(4-acetylphenyl)-2-chloroacetamide).
[0031] (2) Preparation of intermediate D: 2-(1H-benzimidazol-2-ylthio)-N-(4-acetylphenyl)acetamide.
[0032] In a 100 ml round-bottom flask, intermediate C (0.01 mol), 2-mercaptobenzimidazole (0.01 mol) and anhydrous potassium carbonate (0.02 mol) were dissolved in anhydrous acetone (20 mL), and the mixture was stirred at 25 °C for 6 hours. After the reaction, the resulting solid was collected by filtration, dried in air and recrystallized from ethanol to obtain intermediate D.
[0033] (3) Preparation of target compound E1: (E)-2-((1H-benzo[d]imidazol-2-yl)thio)-N-(4-(1-(2-(4-bromobenzoyl)hydrazonomethyl)ethyl)phenyl)acetamide.
[0034] In a 100 ml round-bottom flask, intermediate D (0.001 mol) and p-bromobenzoylhydrazine (0.001 mol) were dissolved in ethanol (20 mL), and the mixture was stirred at room temperature for 8 hours. The crude product formed by the reaction was collected by filtration, washed and recrystallized from ethanol to obtain the target derivative E1.
[0035] Other target compounds were synthesized with the corresponding raw materials or substituents by referring to step (3).
[0036] The physicochemical properties, 1H NMR and 13C NMR data of some synthesized derivatives containing a benzimidazole-2-thioacetamide are shown in Table 1
[0037] Table 1 Physicochemical properties and NMR spectroscopic data of some compounds of the present invention
[0038]
[0039]
[0040]
[0041]
[0042] Pharmacological Example 1:
[0043] MTT anti-tumor cell activity test
[0044] The inhibitory activities of the compound against human lung cancer cell A549, human prostate cancer cell PC-3, human chronic myeloid leukemia cell K562, and human bladder cancer cell 5637 were determined by the MTT method. The MTT colorimetric method is one of the most common standard methods for detecting cell proliferation and toxicity experiments in vitro.
[0045] Implementation steps: The MTT colorimetric method was used to evaluate the anti-proliferative activities of the target compound against A549, 5637, PC-3, and K562, and the experiment followed the established standard operating procedure. Specifically, after the cells were pre-cultured in a 96-well plate for 24 hours, the test compound was added and incubated for 48 hours, with 5-fluorouracil and imatinib as clinical reference drugs. The initial screening concentration was 10 μM, and then the dose-dependent test of compound E12 was carried out using a two-fold serial dilution (1.25–20 μM). After the treatment, MTT solution (5 mg / mL, 10% v / v) was added to the cell culture medium and incubated for 4 hours. Subsequently, the supernatant was removed, and the formazan crystals were dissolved in DMSO (150 μL / well) by orbital shaking in the dark. The optical density (OD) was measured at 490 nm using a Tecan Infinite M200Pro microplate reader, and all experiments were repeated three times to ensure the reliability of the data. The cell growth inhibition rate = 1 - (OD value of the drug-treated group - OD value of the blank group) / (OD value of the negative group - OD value of the blank group) × 100%.
[0046] The examples of the present invention are used to illustrate the technical solutions of the present invention, but the content of the examples is not limited thereto. The experimental results of some target compounds are shown in the following table.
[0047] Table 2 In vitro anti-proliferative inhibition rates of the compounds of the present invention against four tumor cell lines
[0048]
[0049]
[0050] Data are expressed as mean ± standard deviation.
[0051] a Growth inhibitory effect at a fixed concentration (10 μM, 48 hours)
[0052] b Reference drug: 5-Fu: 5-fluorouracil
[0053] Table 3 In vitro inhibitory effect of compound E12 on chronic myeloid leukemia cell K562
[0054]
[0055] a IC50 (μM): The concentration required to inhibit cell proliferation by 50% relative to the solvent-treated control group after 48 hours of compound exposure.
[0056] b Reference drugs: 5-Fu: 5-fluorouracil; IMA: imatinib
[0057] The in vitro anti-proliferative activities of all target molecules against four human tumor cell lines, A549, PC-3, K562, and 5637, were tested using the MTT method. After treatment with a fixed concentration of 10 μM for 48 hours, some target compounds showed certain inhibitory effects on tumor cells (see Table 2). Among them, compound E12 showed the best inhibitory rate against K562 cells, which was 62.71%. Further dose-dependent testing of E12 was carried out using two-fold serial dilution (1.25–20 μM), and the IC50 value of E12 against K562 cells was determined to be 11.07 μM (see Table 3).
[0058] For parts or structures not specifically described in the present invention, existing technologies or existing products can be used, and no further elaboration will be provided here.
[0059] The above are only embodiments of the present invention and do not limit the patent scope of the present invention. Any equivalent structural or equivalent process transformation made using the content of the specification of the present invention, or directly or indirectly applied in other related technical fields, shall be equally included in the patent protection scope of the present invention.
Claims
1. A derivative compound of benzimidazole-2-thioacetamide, characterized in that: It has the structure shown in the general formula (I): Among them, R is an optionally substituted or unsubstituted phenyl or pyridyl group.
2. A derivative compound of benzimidazole-2-thioacetamide according to claim 1, characterized in that: R is selected from a substituted or unsubstituted phenyl group, or a substituted or unsubstituted pyridyl group.
3. A derivative compound of benzimidazole-2-thioacetamide according to claim 1 or 2, characterized in that: The substituents of the said substitution are halogen, methyl or methoxy.
4. A derivative compound of benzimidazole-2-thioacetamide according to claim 1, characterized in that: R is selected from 4-bromophenyl, 4-chlorophenyl, 3-chlorophenyl, 4-fluorophenyl, 3-fluorophenyl, 2-fluorophenyl, 2-chlorophenyl, 4-methylphenyl, 3-methylphenyl, 2-methylphenyl, 3-pyridyl, 2-pyridyl, 4-pyridyl or 3,4,5-trimethoxyphenyl.
5. A derivative compound of benzimidazole-2-thioacetamide according to claim 1, characterized in that: The said derivative compound is selected from the following compounds:
6. A method for preparing a derivative compound of benzimidazole-2-thioacetamide according to any one of claims 1-5, characterized in that: Comprising the following steps:
7. A class of compositions, characterized in that: Containing the compound according to any one of claims 1-5 or an isomer or any mixture thereof in the form thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutical adjuvant or an antitumor preparation.
8. Use of the compound according to any one of claims 1-5, or the composition according to claim 7 in the preparation of a drug for an anti-tumor cell line.
9. The application according to claim 8, wherein The said tumor cell lines are human lung cancer cell line A549 (lung adenocarcinoma), human chronic myeloid leukemia cell line K562 (chronic myelogenous leukemia), human prostate cancer cell line PC-3 (prostate cancer), human bladder cancer cell line 5637 (bladder carcinoma).