Journal of Pharmaceutical Research International

32(46): 80-86, 2020; Article no.JPRI.65000 ISSN: 2456-9119 (Past name: British Journal of Pharmaceutical Research, Past ISSN: 2231-2919, NLM ID: 101631759)

Potential Hexokinase II Inhibition by Benzimidazole : , and

Hillary Le1 and Sung-Kun Kim1*

1Department of Natural Sciences, Northeastern State University, Broken Arrow, OK 74014, USA.

Authors’ contributions

This work was carried out in collaboration between both authors. Author HL wrote the first draft and managed the literature searches. Author SKK wrote and managed the manuscript. Both authors read and approved the final manuscript.

Article Information

DOI: 10.9734/JPRI/2020/v32i4631105 Editor(s): (1) Dr. Ana Cláudia Coelho, University of Trás-os-Montes and Alto Douro, Portugal. Reviewers: (1) Bharat B. Kashid, India. (2) Ahmed M. M. El-Saghier, Sohag University, Egypt. (3) Anju Khullar, General Shivdev Singh Diwan Gurbachan Singh Khalsa College, India. Complete Peer review History: http://www.sdiarticle4.com/review-history/65000

Received 12 November 2020 Review Article Accepted 19 January 2021 Published 05 February 2021

ABSTRACT

The nitrogen heterocycle benzimidazoles have been known to be drugs. Beyond that role, the benzimidazoles exhibit other pharmacological potential as anti-inflammatory, antiulcer, anti-hypertensive and anticancer agents. A growing body of evidence supports the anticancer efficiency via treatment with benzimidazoles. The target proteins for pharmacological effect appear to be cell microtubules. However, it has been reported that the benzimidazoles could inhibit hexokinase II, which is a critical factor in the glycolysis pathway in humans. Here, we discuss on the most common benzimidazoles such as albendazole, mebendazole and fenbendazole and focus on the potential anticancer activities of the target enzyme hexokinase II. This review would give better insight in development of target-specific benzimidazole derivatives as potential anticancer therapeutics.

Keywords: Benzimidazole; albendazole; mebendazole; fenbendazole; hexokinase.

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*Corresponding author: E-mail: [email protected];

Le and Kim; JPRI, 32(46): 80-86, 2020; Article no.JPRI.65000

1. INTRODUCTION hexokinase II, a key glycolytic enzyme in cancer cell metabolisms [14]. A nitrogen heterocycle benzimidazole was first synthesized by Hoebrecker [1], and after 80 The benzimidazoles have revolutionized the drug years the therapeutic activity as an anthelmintic discovery process by showing their diverse range drug was recognized [2]. Today, benzimidazoles of biological activities. The power of potential of are the most important anthelmintics serving as benzimidazoles has drawn interest in developing effective drugs for intestinal and tissue-dwelling potent benzimidazole derivatives containing parasites [3]. Benzimidazoles are bicyclic broad spectrum pharmacological activities. In this aromatic compounds with an ring review, we focus on the antitumor therapeutic fused with [4]. Although benzimidazoles effects of the following benzimidazole are primarily utilized as anthelmintic drugs, they anthelmintics: albendazole, mebendazole, and have also presented strong therapeutic fenbendazole by selectively surveying literatures application against pathogenic bacteria and fungi from 1984 to 2019 (Fig 1). In addition, we will [5]. Benzimidazole derivatives are also review the target enzyme hexokinase II for associated with a variety of types of anticancer effect by the above mentioned benzimidazoles. pharmacological activities such anti-inflammatory [6], antiulcer [7], anti-hypertensive [8] and even 2. ALBENDAZOLE possibly anticancer [9]. Albendazole is notably low in toxicity and The most popular benzimidazole anthelmintics considerably safer for treatment due to its such as albendazole, mebendazole, and benzimidazole carbamates. In 1975, albendazole fenbendazole have been used throughout the was invented by Robert J. Gyurik and Vassilios world since their introduction in the 1960s, J. Theodorides for treatment of parasites in showing efficacy against parasites in livestock sheep [15]. This drug became registered for use due to their high activity and low toxicity [10-11]. in humans in 1982 [16]. Albendazole (5- Researchers revealed some benzimidazoles (propylthio-1H-benzimidazol-2-yl) carbamic acid appear to lead to inhibition of microtubule methyl ester is insoluble in water and most polymerization as antimitotic drugs and result in organic solvents. It is poorly absorbed in the GI critical transition of the cell cycle leading to cell tract, and after absorption, albendazole is rapidly death [12-13]. Additionally, it has been reported converted by the liver to its primary metabolite, that a benzimidazole derivative may inhibit albendazole sulfoxide, which is

Fig. 1. Benzimidazoles. 2D structures of molecules: Albendazole (a), Mebendazole (b) and Fenbendazole (c)

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known to be responsible for therapeutic activity benzimidazole carbamate to be used on humans of albendazole. Albendazole inhibits [22]. Mebendazole halts glucose uptake and can polymerization of parasite tubulin into also be prescribed to treat parasitic pinworm microtubules with a higher affinity to the parasite infections including threadworm, tapeworms, tubulin than host tubulin, effectively and roundworms, and other nematode infections in specifically targeting the parasite. This property humans and livestock [23]. Mebendazole has of albendazole results in prevention of the typically been used in low dosages for local gut parasite from maintaining energy production, helminthic infections, but requires increasingly while also promoting parasite destruction [17]. high amounts in months to years for chronic echinococcus infections. Current research Albendazole can treat neurocysticercosis, which indicates that mebendazole has potential is an infection by the pork tapeworm in the antitumor properties [24]. This drug shows early muscles, brain, and eyes. This condition has the clinical efficacy in glioblastoma and potential to cause seizures, brain swelling, and medulloblastoma. In experiments performed by vision problems [18]. This drug is commonly Bai et al. (2015), researchers explored three coupled with surgery to target cystic hydatid different mebendazole polymorphs (A, B, and C) disease, an infection caused by dog tapeworm in that can penetrate the brain [25]. Results the liver, lung, and lining of the abdomen. observed by polymorph C had the highest Human hydatid cyst (cystic echinococcosis) is a efficacy by therapeutically reaching the mouse zoonotic disease that is recognized as a major brain tissue and tumor with fewer side effects public health issue on a global scale. Human when coupled with elacridar than cases observed hydatid cysts is the result of ingestion of with other therapeutics [25]. Customized Echinococcus granulosus eggs, and as the mebendazole formulated tablets were observed worms enter the larval stage, hydatid cysts form for the polymorph content by IR spectroscopy in the liver, lungs, and other organs in the human and for efficacy and tolerability. Polymorph A body. This life-threatening disease is typically presented lower levels in the plasma and brain, treated with hydatid cyst open surgery, but there and B and C showed greater toxicity and is a high-risk recurrence. Albendazole has been increased survival. Elacridar is a drug that acts used alone or coupled with surgery to treat the as a bioenhancer that inhibits multiple drug condition of hydatid cysts. Efficacy is still not resistant tumors. Therefore, mebendazole, which well documented, and hydatid cyst treatment with is enhanced by elacridar, has high efficacy and is albendazole is still undergoing further research in a better choice of brain cancer therapy [25]. case studies and clinical trials in humans and animals [19]. According to Fattahi Masoom et al. 4. FENBENDAZOLE (2017), albendazole may have benefits for patients with inoperable cystic echinococcosis Fenbendazole (methyl N-(6-phenylsulfanyl-1H- with multiple cysts [20]. There were 164 patients benzimidazol-2-yl) carbamate) is a broad- with echinococcosis referred to Ghaem hospital spectrum benzimidazole anthelmintic that between 2001 and 2013 who were diagnosed contains anti-proliferative activity and is typically with alveolar echinococcosis. Patients who used in veterinary medicine to target parasitic underwent higher phases had notably increased pinworms. Fenbendazole is also known to have chances of better response. Albendazole has low degree of toxicity and high degree of safety shown promising results in pharmacological in animals [26-30]. Fenbendazole is used to application of treating human hydatid cysts in target gastrointestinal parasites including: neurocysticercosis. giardia, roundworms, hookworms, whipworms, the tapeworm genus Taenia - excluding 3. MEBENDAZOLE dipylidium caninum, which is a common dog tapeworm - , pinworms, aelurostrongylus, Mebendazole (2-benzimidazolecarbamic acid paragonimiasis, strongyles, and strongyloides. methyl ester), another class of benzimidazoles Fenbendazoles can be administered to that came into use in 1971, was developed by experimental animals such as sheep, cattle, Janssen Pharmaceutica in Belgium [21]. horses, dogs, cats, and rabbits [31]. This broad- Mabendazole is on the World Health spectrum anthelmintic disrupts microtubules and Organization's List of Essential Medicines and is proteasomes, reduces glucose uptake from available as a generic medication. Compared to glucose transporter isoform 4 (GLUT4), albendazole, which was moderately toxic to expresses glycolytic enzymes such as humans, Mebendazole was the first hexokinase II, starving cancer cells [14]. When

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insulin initiates the glucose uptake in cells, post binding. The open structure of hexokinase is GLUT4 translocates from the intracellular shown in Fig 2a. Once glucose is bound, vesicles directly to the plasma membrane where hexokinase undergoes a conformation change as glucose is taken up. The linear movement of shown in Fig 2b. Due to the possibility of GLUT4 towards the plasma membrane takes substantial conformational change, some place along microtubules. When microtubules benzimidazoles may have an opportunity to bind are disrupted with fenbendazole, GLUT4 is the active site of hexokinases. For example, disrupted simultaneously, thus reducing insulin- Zeng et al. (2015) revealed the complex structure stimulated glucose absorption [32]. between human hexokinase II and a potential Fenbendazole can also reactivate the p53 gene inhibitor 2,6-disubstituted glucosamine, indicating that functions to suppress tumors and inhibit that the active site of hexokinase is potentially a some cancers [33]. Accumulation of ubiquitylated good binding target for finding inhibitors [35]. derivatives including p53 appears to result in apoptosis in the mitochondrial pathway. There are four isoforms of hexokinase: hexokinases I, II, III, and IV. Hexokinases I, II, Recent clinical studies explore future therapies and III are inhibited noncompetitively by glucose that can lead to anticancer effects. A research 6-phosphate and competitively by ATP. experiment demonstrated the inhibition of Hexokinase I catalyzes the first step in glycolysis. proteasome function and introduction of Hexokinase inhibits glycolysis by 2-deoxy-D- apoptosis in human lung cancer cells by glucose (2-DG), a glucose analog, which is the fenbendazole [34]. Fluorogenic substrates were rate-limiting step of glycolysis via hexokinase. 2- used to show that fenbendazole results in the DG assists in the development of anticancer decrease of proteasomal activity in the cells. strategies that include radio, chemosensitization, Non-small cell lung carcinoma (NSCLC) were and oxidative stress. Hexokinase phosphorylates treated with fenbendazole and accumulated this analog to 2-DG-phosphate (2-DG-P) and green fluorescent protein with an increased half- metabolism is disrupted by phosphoglucose life. Apoptosis regulatory proteins are typically isomerase [36]. This results in 2-DG-P degraded by ubiquitin-proteasome pathway and accumulation and ATP depletion in cells. accumulate within cells treated by fenbendazole. Furthermore, 2-DG-P increases the activity of NSCLC cells with fenbendazole lead to autophagy, enhances reactive oxygen species endoplasmic reticulum stress, reactive oxygen production, and activates AMP-activated protein species production, decreased mitochondrial kinase-9. membrane potential, and cytochrome c release [34]. Fenbendazole leads to cancer cell death, Hexokinase II takes part in glycolysis and in the which may lead to new anticancer agents with transports into and out of the mitochondrial high efficacy and specificity for tumor cells by intermembrane space through the association inhibition of hexokinase in glycolysis. with a voltage-dependent anion channels; thus, the depletion of hexokinase II may inhibit the 5. HEXOKINASE glycolytic pathway [37]. There are several compounds synthesized in order to inhibit Hexokinases play a key role in cellular glucose hexokinase II. The compound 2,6-disubstituted metabolism. Hexokinase is the first step in glucosamine was found by a high throughput glycolysis and uses an ATP molecule in the initial screen, and the compound showed a competitive process, influencing the direction of glucose flux inhibition with a micro-range inhibition equilibrium within the cell. This is the first step of all major constant (Ki = 2.9 μM) [35]; the complex pathways of glucose usage including glycolysis, structure is shown in Fig 2c. As another example, pentose phosphate pathway, and glycogenesis. 3-bromopyruvic acid showed its amazing ability Glycolysis and gluconeogenesis are critical for to inhibit hexokinase II and became an antitumor energy metabolism and storage. Hexokinases agent with high efficacy [38]. In addition, transfer a phosphate from ATP to glucose, which researchers synthesized a compound called as yields glucose-6-phosphate. This results in a lonidamine (LND), which serves as a hexokinase downstream effect of the concentration gradient inhibitor, antitumor agent, mitochondrial pyruvate and allows facilitation of cellular glucose influx. carrier, and plasma membrane monocarboxylate This reaction must be shielded from water in transporters inhibitor [39]. More specifically, order to keep ATP molecules from being cleaved lonidamine disrupts energy metabolism of cancer off by water. As a result, hexokinase performs an cells, specifically inhibiting aerobic glycolytic induced fit, which encloses glucose and ATP activity affecting the mitochondrial complex II,

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a b c

Fig. 2. The structural change in hexokinases. The open structure of yeast hexokinase PII without glucose (a), the closed structure of hexokinase from schistosoma mansoni in the presence of glucose (colored in blue) (b), and human hexokinase II structure with a 2,6- disubstituted glucosamine (c). The PDB ID: 1IG8 (a) and 1BDG (b) 5HG1 (c) and hence, LND might impair energy-requiring ETHICAL APPROVAL processes [39]. Fenbendazole also impairs enzymatic activity of hexokinase II, eventually It is not applicable. leading to the activation of apoptotic signals [14]. ACKNOWLEDGEMENTS 6. CONCLUSION The authors would like to thank Rachel Benzimidazoles that have historically been used Hoffmeister for reviewing this manuscript. in veterinary medicine may be beneficial in treating cancer in humans. Traditionally, they COMPETING INTERESTS have been prescribed as antimicrobial and agents, but clinical studies show Authors have declared that no competing these agents offer anticancer effects with low interests exist. toxicity and are highly effective by starving cells of glucose through apoptosis. By interrupting the REFERENCES energy metabolism of parasites by binding to tubulin, microtubule polymerization can be 1. Keri RS, Hiremathad S, Budagumpi S, disrupted causing nutrient uptake inhibition and Nagaraja BM. Comprehensive review in prevention of the major pathways of glucose by current developments of benzimidazole- hexokinases. Glycolysis is prevented by based medicinal chemistry. Chem Biol hexokinase and therefore inhibits glucose Drug Des. 2015;86:19-65. pathways. Benzimidazoles such as albendazole, 2. McFarland JW. The chemotherapy of mebendazole, and fenbendazole have been intestinal nematodes. Prog Drug Res. shown to inhibit the progression of difficult-to- 1972;16:157–193. treat cancer types. By inhibiting glucose from 3. Amari M, Fodili M, Nedjar-kolli B. being upregulated by the enzyme hexokinases Reactivity studies on 4-aminopyrones: and the binding site of the protein, the cells Access to benzimidazole and benzimid cannot complete the glycolysis pathway and die. azolone derivatives. J Heterocycl Chem. Thus, researchers across the world need to 2002;39:811. rd design new benzimidazole derivatives that could 4. Bansal RK. Herocyclic Chemistry. 3 ed. specifically target hexokinase II in an attempt to New Delhi, New Age International; 2002. treat cancerous cells with low toxicity and no 5. Mavrova AT, Vuchev D, Anichina K, adverse effects. Vassilev N. Synthesis, antitrichinnellosis and activity of some novel CONSENT thieno [2,3-d] pyrimidin-4(3H)-ones containing benzimidazole ring. Eur J Med It is not applicable. Chem. 2010;45:5856.

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