A novel anticancer compounds containing a benzimidazole core for use in the treatment of different cancer derivatives
Novel benzimidazole compounds with specific substituents address the issue of chemotherapeutic side effects by providing high antiproliferative activity against breast, liver, and lung cancers, outperforming cisplatin in efficacy while minimizing harm to healthy cells.
Patent Information
- Authority / Receiving Office
- WO · WO
- Patent Type
- Applications
- Current Assignee / Owner
- SÜLEYMAN DEMİREL ÜNİVERSİTESİ İDARİ VE MALİ İŞLER DAİRE BAŞKANLIĞI GENEL SEKRETERLİK
- Filing Date
- 2025-12-05
- Publication Date
- 2026-06-11
AI Technical Summary
Current chemotherapeutic drugs for cancer treatment are effective but cause significant side effects on normal cells, necessitating the development of new antitumor drugs with high efficacy and reduced toxicity.
Synthesis of novel benzimidazole core compounds with specific substituents that exhibit high antiproliferative activity against breast, liver, and lung cancer cell lines, demonstrating cytotoxic effects on MCF-7, HepG2, DLD-1, and A549 cell lines, and are more effective than cisplatin.
The synthesized benzimidazole compounds show IC50 values two to twelve times lower than cisplatin, indicating significant cancer cell growth inhibition with minimal toxicity to healthy cells.
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Abstract
Description
[0001] DESCRIPTION
[0002] A NOVEL ANTICANCER COMPOUNDS CONTAINING A BENZIMIDAZOLE CORE FOR USE IN THE TREATMENT OF DIFFERENT CANCER DERIVATIVES
[0003] Technical Field
[0004] The present disclosure relates to novel anticancer compounds containing a benzimidazole core, which exhibit antiproliferative activity in breast, liver, colon and lung cell lines.
[0005] State of the Art
[0006] According to data provided by organisations such as the World Health Organization (WHO) and the International Agency for Research on Cancer (IARC), cancer continues to be a global health problem. Cancer incidence rates vary depending on region, gender and cancer-specific types. In 2020, approximately 19.3 million new cancer cases were diagnosed worldwide, and approximately 10 million people died from cancer in the same year. Due to the very high mortality rates, there is a need to discover new drug candidates.
[0007] Currently, there are hundreds of approved chemotherapeutic drugs worldwide. These drugs cover a wide range of treatment options, including conventional chemotherapy agents, targeted therapies, immunotherapies and hormone therapies. This diversity in chemotherapeutic drugs provides multiple pathways for the treatment of various cancer types, enabling more personalised and effective treatment strategies. However, chemotherapeutic drugs not only kill proliferating cancer cells but may also damage normal cells. For this reason, there is a need for new antitumour drug therapies that maintain or improve the efficacy of chemotherapy while reducing the severity of side effects.
[0008] Despite the use of hundreds of cancer drugs in the prior art, millions of people die every year due to cancer. Therefore, scientists continue their studies to discover new cancer drug candidates.
[0009] As a result, due to the above-mentioned disadvantages and deficiencies, a need has arisen to introduce an innovation in the relevant technical field. The Aim of the Invention
[0010] The present invention relates to novel anticancer compounds containing a benzimidazole core for use in the treatment of different cancer derivatives, which meet the above-mentioned requirements, eliminate all disadvantages, and provide additional advantages.
[0011] The aim of the invention is to present novel anticancer compounds containing a benzimidazole core that exhibit antiproliferative activity in breast, liver, colon and lung cell lines.
[0012] The aim of the invention is to develop new drug candidates with high efficacy against breast, liver, lung and colon cancer.
[0013] The aim of the invention is to present compounds that exhibit cytotoxic effects on MCF- 7, HepG2, DLD-1 , A549 and HEK-293T cell lines and have high antiproliferative activity, for use as chemotherapeutic drugs in cancer treatment.
[0014] In order to achieve the above-described aims, the invention is characterised by being an anticancer compound containing a benzimidazole core, having any of the following formulas, and exhibiting antiproliferative activity in breast, liver, colon and lung cell lines.
[0015] 3a 3 b
[0016] J
[0017] In order to achieve the aims of the invention, 1 -(4-methylbenzyl)-3-(2,3,4,5,6- pentamethylbenzyl)benzimidazolium chloride is synthesised by the process steps of:
[0018] • dissolving benzimidazole and potassium hydroxide (KOH) as a base in ethyl alcohol (10 mL), stirring the mixture at room temperature for 30 minutes, then adding 4-methylbenzyl chloride to the medium and refluxing for 6 hours, • cooling the reaction mixture to room temperature and filtering off the formed potassium chloride (KCI),
[0019] • purifying the obtained 1 -(4-methylbenzyl)benzimidazole by crystallisation in ethyl alcohol,
[0020] • transferring the synthesised 1-(4-methylbenzyl)benzimidazole (0.369 g) into a clean Schlenk tube and adding 5 mL of N,N-dimethylformamide (DMF),
[0021] • adding 2,3,4,5,6-pentamethylbenzyl chloride (0.326 g) to the reaction mixture and continuing the reaction by heating at 80 'C for 48 hours,
[0022] • removing the DMF from the medium under vacuum after completion of the reaction, and
[0023] • purifying the synthesised product by washing several times first with diethyl ether and then with hot benzene.
[0024] In order to achieve the aims of the invention, 1 -(3-methylbenzyl)-3-(naphthalen-2- ylmethyl)benzimidazolium chloride is synthesised by the following process steps:
[0025] • dissolving benzimidazole and potassium hydroxide (KOH) as a base in ethyl alcohol,
[0026] • stirring the mixture at room temperature for 30 minutes, then adding 3-methylbenzyl chloride to the reaction medium and refluxing for 6 hours,
[0027] • cooling the reaction mixture to room temperature and filtering off potassium chloride (KCI),
[0028] • purifying the obtained 1 -(3-methylbenzyl)benzimidazole by crystallisation in ethyl alcohol,
[0029] • transferring the synthesised 1 -(3-methylbenzyl)benzimidazole (0.2 g) into a clean Schlenk tube and adding 4 mL of DMF,
[0030] • adding 2-bromomethyl naphthalene (0.199 g) to the reaction mixture and continuing the reaction by heating at 80 'C for 48 hours,
[0031] • removing the DMF from the medium under vacuum after completion of the reaction and purifying the synthesised product by crystallisation in ethyl alcohol.
[0032] The structural and characteristic features of the invention and all advantages thereof will be more clearly understood by means of the detailed description given below, and therefore the evaluation should be made by taking this detailed description into consideration. Detailed Description of the Invention
[0033] In this detailed description, novel anticancer compounds containing a benzimidazole core for use in the treatment of different cancer derivatives are described solely for better understanding of the subject matter and without creating any limiting effect.
[0034] The present disclosure relates to novel anticancer compounds containing a benzimidazole core, which exhibit antiproliferative activity in breast, liver, colon and lung cell lines. The feature of the invention is that the compounds exhibit cytotoxic effects on MCF-7, HepG2, DLD-1 , A549 and HEK-293T cell lines and have high antiproliferative activity.
[0035] Within the scope of the invention, two benzimidazolium salts with high antiproliferative activity were synthesised. The structures of the obtained compounds were characterised by NMR and IR, and both compounds were tested in vitro against four different cancer cell lines for different incubation periods. According to the obtained results, both designed compounds exhibited low IC50 values and significantly inhibited the growth of cancer cells. On the other hand, it was determined that the synthesised compounds were approximately two times more effective than cisplatin, which was used as a positive control drug, against breast cancer cells, approximately twelve times more effective against liver cancer cell lines, and approximately three times more effective against lung cancer cell lines.
[0036] The substituents used in the compounds synthesised within the scope of the invention render the prepared compounds novel and are decisive for their activity. Within the scope of this study, two new benzimidazolium salts were synthesised by changing the substituents.
[0037] Synthesis of 1-(4-methylbenzyl)-3-(2,3,4,5,6- oentamethylbenzvDbenzimidazolium chloride (3a):
[0038] Benzimidazole (1 ) and potassium hydroxide (KOH) as a base are dissolved in ethyl alcohol (10 mL). After the mixture is stirred at room temperature for 30 minutes, 4- methylbenzyl chloride is added to the medium and refluxed for 6 hours. Subsequently, the reaction mixture is cooled to room temperature and the formed potassium chloride (KCI) is filtered off. The obtained 1 -(4-methylbenzyl)benzimidazole is purified by crystallisation in ethyl alcohol. The synthesised 1 -(4-methylbenzyl)benzimidazole (0.369 g) compound (2a) is transferred into a clean Schlenk tube, and 5 mL of N,N- dimethylformamide (DMF) is added thereto. Subsequently, 2, 3, 4,5,6- pentamethylbenzyl chloride (0.326 g) is added to the obtained reaction mixture, and the reaction is continued by heating at 80 D for 48 hours. After completion of the reaction, the DMF in the medium is removed under vacuum. The synthesised product (3a) is purified by washing several times first with diethyl ether and then with hot benzene.
[0039] MA: 419.01 g / mol; M.P.: 185-187SC, Colour: White.
[0040] IR: 1550 (C=N); 3472, 3393, 3100, 3022, 2951 and 2924 cm1(C-H).
[0041] 1H NMR (400.13 MHz, CDCI3, 298 K), 5: 2.01 , 2.23, 2.26 and 2.28 [s, 18 H, NCH2C6H4(CH3)-4, NCH2C6(CH3)5-2,3,4,5,6]; 5.85 and 5.89 [s, 4 H, NCH2C6H4(CH3)- 4, NCH2C6(CH3)5-2,3,4,5,6]; 7.12-7.52 (m, 8 H, Ar-H); 11.31 (s, 1 H, NCHN).
[0042] 13C NMR (100.13 MHz, CDCI3, 298 K), 5: 16.98, 17.14, 17.32, 21 .15 [NCH2C6H4(CH3)-
[0043] 4, NCH2C6(CH3)5-2,3,4,5,6]; 44.33, 51.37 [NCH2C6H4(CH3)-4, NCH2C6(CH3)5- 2, 3, 4, 5, 6]; 113.62, 113.75, 124.91 , 126.82, 126.92, 128.11 , 129.90, 130.08, 131.53, 131.62, 133.57, 133.93, 137.28 and 138.98, (Ar-C); 143.56 (NCHN).
[0044] Synthesis of 1 -(3-methylbenzyl)-3-(naohthalen-2-ylmethyl)benzimidazolium chloride (3b):
[0045] Benzimidazole (1 ) and KOH are dissolved in ethyl alcohol. After the mixture is stirred at room temperature for 30 minutes, 3-methylbenzyl chloride is added to the reaction medium and refluxed for 6 hours. Subsequently, the reaction mixture is cooled to room temperature and the formed KCI is filtered off. The obtained 1 -(3- methylbenzyl)benzimidazole (2b) is purified by crystallisation in ethyl alcohol. The synthesised 1 -(3-methylbenzyl)benzimidazole (0.2 g) compound (2b) is transferred into a clean Schlenk tube, and 4 mL of DMF is added thereto. Subsequently, 2- bromomethyl naphthalene (0.199 g) is added to the obtained reaction mixture, and the reaction is continued by heating at 80 'C for 48 ho urs. After completion of the reaction, the DMF in the medium is removed under vacuum. The synthesised product is purified by crystallisation in ethyl alcohol.
[0046] 1 3b 3
[0047] MA: 419.01 g / mol; M.P.: 185-187SC, Colour: White.
[0048] IR: 1550 (C=N); 3472, 3393, 3100, 3022, 2951 and 2924 cm1(C-H).
[0049] 1H NMR (400.13 MHz, CDCI3, 298 K), 5: 2.01 , 2.23, 2.26 and 2.28 [s, 18 H, NCH2C6H4(CH3)-4, NCH2C6(CH3)5-2,3,4,5,6]; 5.85 and 5.89 [s, 4 H, NCH2C6H4(CH3)- 4, NCW2C6(CH3)5-2,3,4,5,6]; 7.12-7.52 (m, 8 H, Ar-H); 11.31 (s, 1 H, NCHN).
[0050] 13C NMR (100.13 MHz, CDCI3, 298 K), 5: 16.98, 17.14, 17.32, 21 .15 [NCH2C6H4(CH3)- 4, NCH2C6(CH3)5-2,3,4,5,6]; 44.33, 51.37 [NCH2C6H4(CH3)-4, NCH2C6(CH3)5- 2, 3, 4, 5, 6]; 113.62, 113.75, 124.91 , 126.82, 126.92, 128.11 , 129.90, 130.08, 131.53, 131.62, 133.57, 133.93, 137.28 and 138.98, (Ar-C); 143.56 (NCHN).
[0051] Cytotoxic activity studies of the synthesised compounds (3a, 3b): The human epithelial breast adenocarcinoma cell line (MCF-7) (ATCC® HTB-22™), human liver hepatocellular carcinoma cell line (HepG2) (ATCC® HB-8065™), human epithelial colon adenocarcinoma cell line (DLD-1 ) (ATCC® CCL-221 ™), and human epithelial lung cancer cell line (A549) (ATCC® CCL-185™) were purchased from the American Type Culture Collection. HEK-293T was used as a healthy human cell line. HepG2, DLD-1 , A549 and HEK-293T cells were cultured in DMEM medium containing 10% fetal bovine serum (FBS) and 1 % GlutaMAX. RPMI was used instead of DMEM for the MCF-7 cell line. After reaching 90% cell confluence, the culture medium was removed and the cells were washed twice with PBS. Three millilitres of trypsin-EDTA were added to the flask and the cells were incubated for 5 minutes. The detached cells were centrifuged at 1450 rpm for 5 minutes and the supernatant was discarded. Five millilitres of fresh culture medium were added to the pellet, and cell counting was performed using trypan blue staining. HepG2, DLD-1 , MCF-7, A549 and HEK-293T cells were seeded into sterile 96-well plates at a density of 5 x 103cells per well. After 24 hours, the cells (HepG2, DLD-1 , HEK-293T) were exposed to the synthesised compounds at different concentrations (200, 100, 50, 25 pM) for 72 hours. In the MCF- 7 cell line, the compounds were tested at concentrations of 200, 100, 50, 25 and 12.5 pM. In the A549 cell line, the compounds were tested at concentrations of 200, 100, 50, 25, 12.5 and 6.25 pM. Cisplatin, an anticancer agent, was used as a positive control. After completion of this period, MTT stock solution (50 pL, 5 mg / mL) was added to each well and the plates were incubated for 2 hours. Subsequently, the dye in each well was carefully removed and replaced with 200 pL of DMSO. After the 96-well plates were mixed on a shaker for half an hour, absorbance values were measured at 590 nm using a Promega plate reader. IC50 values were calculated using GraphPad Prism software 5. The obtained results are presented in Table 1 .
[0052] Table 1 : Effects of the compounds (3a, 3b) synthesised within the scope of the invention on human cell lines
[0053] N.T.*: Not tested As can be understood from Table 1 , the IC50 value of compound 3a for 72 hours was determined to be 5.27, 2.93, 4.68 and 2.04 pM for MCF-7, HepG2, DLD-1 and A549, respectively. The IC50 value of compound 3b was found to be 4.57, 2.96, 2.84 and 4.74 pM for MCF-7, HepG2, DLD-1 and A549, respectively. In addition, the toxic effects against the healthy human cell line (HEK-293T) were also investigated.
Claims
CLAIMS1. An anticancer compound characterized it that; containing a benzimidazole core, having any of the following formulas, and exhibiting antiproliferative activity in breast, liver, colon and lung cell lines.J2. A method for synthesising 1 -(4-methylbenzyl)-3-(2,3,4,5,6 pentamethylbenzyl)benzimidazolium chloride (3a) mentioned in claim 1 characterized by comprising the process steps of:• dissolving benzimidazole and potassium hydroxide (KOH) as a base in ethyl alcohol (10 mL), stirring the mixture at room temperature for 30 minutes, then adding 4-methylbenzyl chloride to the medium and refluxing for 6 hours,• cooling the reaction mixture to room temperature and filtering off the formed potassium chloride (KCI),• purifying the obtained 1 -(4-methylbenzyl)benzimidazole by crystallisation in ethyl alcohol,• transferring the synthesised 1 -(4-methylbenzyl)benzimidazole (0.369 g) compound into a clean Schlenk tube and adding 5 mL of N,N- dimethylformamide (DMF),• adding 2,3,4,5,6-pentamethylbenzyl chloride (0.326 g) to the obtained reaction mixture and continuing the reaction by heating at 80 'C for 48 hours,• removing the DMF from the medium under vacuum after completion of the reaction, and• purifying the synthesised product (3a) by washing several times first with diethyl ether and then with hot benzene.
3. A method for synthesising 1 -(3-methylbenzyl)-3-(naphthalen-2- ylmethyl)benzimidazolium chloride (3b) mentioned in claim 1 characterized by comprising the process steps of:• dissolving benzimidazole and potassium hydroxide (KOH) as a base in ethyl alcohol,• stirring the mixture at room temperature for 30 minutes, then adding 3- methylbenzyl chloride to the reaction medium and refluxing for 6 hours,• cooling the reaction mixture to room temperature and filtering off potassium chloride (KCI),• purifying the obtained 1 -(3-methylbenzyl)benzimidazole by crystallisation in ethyl alcohol,• transferring the synthesised 1 -(3-methylbenzyl)benzimidazole (0.2 g) into a clean Schlenk tube and adding 4 mL of DMF,• adding 2-bromomethyl naphthalene (0.199 g) to the obtained reaction mixture and continuing the reaction by heating at 80 C for 48 hours, and• removing the DMF from the medium under vacuum after completion of the reaction and purifying the synthesised product (3b) by crystallisation in ethyl alcohol.
4. The method according to claim 2 characterized in that; the IC50 value of 1-(4- methylbenzyl)-3-(2,3,4,5,6-pentamethylbenzyl)benzimidazolium chloride (3a) synthesised by the method is 5.27, 2.93, 4.68 and 2.04 pM for MCF-7, HepG2, DLD-1 and A549, respectively.
5. The method according to claim 3 characterized in that; the IC50 value of 1-(3- methylbenzyl)-3-(naphthalen-2-ylmethyl)benzimidazolium chloride (3b) synthesised by the method is 4.57, 2.96, 2.84 and 4.74 pM for MCF-7, HepG2, DLD-1 and A549, respectively.