Project 4: Biophysics of Molecular Recognition and Transportation for Innovative Medicine
According to the population age structure diagram,
it can be predicted that Thailand will become an aged society for the next 20 years.
The proportion of the population with the ageover 65 years old will be increased from 10.4% in 2015 to 22.9% in 2035.
The non-infectious diseases such as heart disease, high/low blood pressure, diabetes, and cancer,
are the major cause of death for Thai population. Cancer has become a great impact in global health concern.
In recent years,a number of deathscaused by cancer,and patients have been increasing every year meanwhilethere are some limitations of cancer therapies.
The side effectsand resistance of anti-cancer drugs have been reported.
To develop the new technology and innovation for cancer therapy as well as new potent anti-cancer drugs,
the understanding at molecular level of the inhibition mechanism of cancer cells is needed. Consequently,
the new developments can increase the competitiveness of medicinal manufacturing industries.
Thailand 4.0 policy aims to drive Thailand by creating newinnovation and state-of-the-art technologies, especially for industrial uses.
The development of medicine technologies is involved in Thailand 4.0 policy. Therefore,
this present project contributes to construct the strong industrial basein the future.
Cancer is caused by abnormal cell growth via several mechanisms e.g. mutation of some proteins associated with cell division and uncontrolled cell apoptosis.
In addition, the attraction of a free radical leads to DNA damage, amino acid fragmentation, loss of protein-protein cross-linkages and lipid peroxidation .
The direct products of lipid peroxidation products such as malondialdehyde (MPA) act as carcinogenic .
Moreover, the lipid peroxidationcan inducebiologicalmembrane deformation,consequently allowing free radicals transport into the cell [2, 3].
Breast cancer is the most common in women worldwide. In Thailand, data from hospital based cancer registry,
the breast cancer is the most foundin Thai women. There are 926 patients in 2013 increased by 17% in comparison to the numbers of patients in 2009 (source: National cancer institutein 2013).
In general, breast cancer can be treated by surgery, radiation, chemotherapy etc. The suitable selection of cancer therapy technique depends on many factors,
for example level of tumour-associated antigens. The HER/2neuand Estrogen/Progesterone antigenslevel are high in cancer cells.
The anticancer drug such as trastuzumabor Herceptin® are commercially used.However, the drug resistance has been reported 
resulting in 90% failure in breast cancer therapy , especially for the patients who has no expression of
HER2/neu and Estrogen/Progesterone receptor.
Figure 4.1 Illustrating the DNA binding site of FOXM1 protein (PDB ID: 3G73)
One of the most used drugs using in breast cancer chemotherapy is Anthracyclineantibioticsclass i.e. Doxorubicin and Epirubicin.
The function of these drugs is DNA double strand break causing cell death [7, 8].
Normally, cell will stimulate DNA repairing when DNA is damaged via Cyclin B, P53protein.
The FOXM1 protein is essential in DNA repair process.For cancer cell, the FOXM1 protein is expressed and induced by chemotherapy .
The 3D structure of FOXM1 is depicted in Figure 4.1
The previous studies showed that the FOXM1 is highly expressed in several types of cancer cells,
respected to normal cells [10, 11]. Interestingly, the level of FOXM1 is extremely high for the chemo-resistant cancer cells [12, 13].
Afterwards, there are evidences indicating that the FOXM1 protein is importance in DNA repairing of chemo-resistant breast cancer cells
Therefore, the design of drugs against FOXM1 protein is interesting, especially for the breast cancerpatients who are during chemotherapy and have HER2/neu and Estrogen/Progesterone negative.
The antibiotic thiazole compound such as Siomycin A and Thiostrepton are extracted from Streptomyces sp..Bhat et al.
reported that the thiazole could inhibit FOXM1 protein resulting in apoptosis-stimulating .
Moreover, the thiazole could also inhibit the binding of FOXM1 and DNA affecting non-synthesis of essential proteins for DNA repair process [16, 17].
This result suggested that the thiazole acts as a competitive inhibitor to FOXM1.
The thiozole was bound to the FOXM1 protein at DNA binding domain; Forkhead/Winged-helix domain .
The thiozole is a potent FOXM1 inhibitor [19, 20], however the synthesis of thiazole compounds is difficult .
The stability and water solubility of thaizole are poor [22, 23]. Chen et al.
designed the new antibiotic agents aiming to exceed the limitation of thiazole .
Figure 4.2 Structure of beta cyclodextrin consisting of hydrophobic nanocavity and hydrophilic outer surface.
Cyclodextrins (CDs) are potent drugs carrier which can improve solubility and stability of drugsandconsequently enhance drug therapeutic efficiency [25-29]. CDs are natural product, synthesized mainly from potato flour by CyclodextrinGlycsyltransferase (CG Tase catalytic enzyme).
The structure of CDs consisting of 1,4-linkged glucopyranose polymer formed in truncated cone shape (see in Figure 4.2
) The cavity interior is hydrophobic and the outer surface is hydrophilic. By the CDs' structure,
the poor water soluble drugs prefer to bind inside the CDs' cavity. In drug delivery system (DDS) based-cyclodextrins,
there are two key factorsfor the development ofdeliver efficacy i.e. i) stability and solubility of drugs and ii) membrane permeability [30, 31].
Hence, the aim of this project is to investigate the binding mechanism of
thiazole antibiotic compounds to FOXM1 protein using both experimental and theoretical studies. Molecular dynamics
simulation can provide deep understanding in molecular recognition of protein−ligand complexes.
In addition, the transportation of thiazole as well as thaizole-cyclodextrin complexes across cell membrane will bestudied.
After screening thiazole compounds by theoretical studies, the cell testing is used for the candidatethiazole.
The obtained information from this project is expected to be benefit in new potent anticancer drug design.
The present study integrated physics aiming for development of biological and medicinal technologies. Moreover,
the quality graduates and young researchers are also the project's objective.
The project overview and research plan are presented in Figures 4.3
and Figures 4.4
Figure 4.3 Project overview.
Figure 4.4 Research plan for 3 years using both theoretical and experimental studies.
1. Dahiya, P., et al., Reactive oxygen species in periodontitis
. Journal of Indian Society of Periodontology, 2013. 17(4): p. 411-416.
2. Van der Paal, J., et al., Effect of lipid peroxidation on membrane permeability of cancer and normal cells subjected to oxidative stress
. Chemical Science,
2016. 7(1): p. 489-498.
3. Boonnoy, P., et al., Bilayer Deformation, Pores, and Micellation Induced by Oxidized Lipids
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4. Riedel, R.F., et al., A genomic approach to identify molecular pathways associated with chemotherapy resistance
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5. Longley, D. and P. Johnston, Molecular mechanisms of drug resistance
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6. Littler, D.R., et al., Structure of the FoxM1 DNA-recognition domain bound to a promoter sequence
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7. Capranico, G., et al., Role of DNA breakage in cytotoxicity of doxorubicin, 9-deoxydoxorubicin, and 4-demethyl-6-deoxydoxorubicin in murine leukemia P388 cells
. Cancer research, 1989. 49(8): p. 2022-2027.
8. Yang, F., C.J. Kemp, and S. Henikoff, Anthracyclines induce double-strand DNA breaks at active gene promoters
. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 2015. 773: p. 9-15.
9. Zona, S., et al., FOXM1: an emerging master regulator of DNA damage response and genotoxic agent resistance
. Biochimica et Biophysica Acta (BBA)-Gene Regulatory Mechanisms, 2014. 1839(11): p. 1316-1322.
10. Myatt, S.S. and E.W.-F. Lam, Targeting foxm1
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11. Halasi, M. and A.L. Gartel, Targeting FOXM1 in cancer
. Biochemical pharmacology, 2013. 85(5): p. 644-652.
12. Monteiro, L.J., et al., The Forkhead Box M1 protein regulates BRIP1 expression and DNA damage repair in epirubicin treatment
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13. Wang, Y., et al., FoxM1 expression is significantly associated with cisplatin-based chemotherapy resistance and poor prognosis in advanced non-small cell lung cancer patients.
Lung cancer, 2013. 79(2): p. 173-179.
14. Khongkow, P., et al., FOXM1 targets NBS1 to regulate DNA damage-induced senescence and epirubicin resistance
. Oncogene, 2014. 33(32): p. 4144-4155.
15. Bhat, U.G., M. Halasi, and A.L. Gartel, Thiazole antibiotics target FoxM1 and induce apoptosis in human cancer cells
. PloS one, 2009. 4(5): p. e5592.
16. Gartel, A.L., Thiazole Antibiotics Siomycin a and Thiostrepton Inhibit the Transcriptional Activity of FOXM1
. Frontiers in Oncology, 2013. 3: p. 150.
17. Halasi, M., et al., Thiazole antibiotics against breast cancer
. Cell Cycle, 2010. 9(6): p. 1214-1217.
18. Hegde, N.S., et al., The transcription factor FOXM1 is a cellular target of the natural product thiostrepton
. Nature chemistry, 2011. 3(9): p. 725-731.
19. Radhakrishnan, S.K., et al., Identification of a chemical inhibitor of the oncogenic transcription factor forkhead box M1
. Cancer research,
2006. 66(19): p. 9731-9735.
20. Chan, D.W., et al., Targeting GRB7/ERK/FOXM1 signaling pathway impairs aggressiveness of ovarian cancer cells
. PloS one, 2012. 7(12): p. e52578.
21. Nicolaou, K., et al., Constructing molecular complexity and diversity: total synthesis of natural products of biological and medicinal importance
. Chemical Society reviews, 2012. 41(15): p. 5185-5238.
22. Bodanszky, M., et al., Thiostrepton. Degradation products and structural features
. Journal of the American Chemical Society, 1964. 86(12): p. 2478-2490.
23. Zhang, F. and W.L. Kelly, In vivo production of thiopeptide variants
. Methods Enzymol, 2012. 516: p. 3-24.
24. Chen, Y., et al., In silico investigation of FOXM1 binding and novel inhibitors in epithelial ovarian cancer
. Bioorganic & medicinal chemistry,
2015. 23(15): p. 4576-4582.
25. Danciu, C., et al., Genistein in 1: 1 inclusion complexes with ramified cyclodextrins: theoretical, physicochemical and biological evaluation
. International journal of molecular sciences, 2014. 15(2): p. 1962-1982.
26. Patro, N.M., et al., Comparison and correlation of in vitro, in vivo and in silico evaluations of alpha, beta and gamma cyclodextrin complexes of curcumin
. Journal of Inclusion Phenomena and Macrocyclic Chemistry, 2014. 78(1-4): p. 471-483.
27. Zhang, Q.-F., et al., Aqueous solubility and stability enhancement of astilbin through complexation with cyclodextrins
. Journal of agricultural and food chemistry, 2012. 61(1): p. 151-156.
28. Dong, N., et al., Cucurbit [n] urils (n= 7, 8) binding of camptothecin and the effects on solubility and reactivity of the anticancer drug
. Supramolecular Chemistry, 2008. 20(7): p. 663-671.
29. Tommasini, S., et al., Improvement in solubility and dissolution rate of flavonoids by complexation with β-cyclodextrin
. Journal of pharmaceutical and biomedical analysis, 2004. 35(2): p. 379-387.
30. Khuntawee, W., et al., Molecular dynamics simulations of the interaction of beta cyclodextrin with a lipid bilayer
. Journal of chemical information and modeling, 2015. 55(9): p. 1894-1902.
31. Amidon, G.L., et al., A theoretical basis for a biopharmaceutic drug classification: the correlation of in vitro drug product dissolution and in vivo bioavailability
. Pharmaceutical research, 1995. 12(3): p. 413-420.
Associate Professor Dr. Jirasak Wong-ekkabut 1)
Dr. Saree Phongphanphanee 2)
, Dr. Thana Sutthibutpong 3)
, Assist. Prof. Dr. Prapasiri Pongprayoon4)
, Assoc. Prof. Dr. Wanwipa Vongsangnak5)
Dr. Mesayamas Kongsema 5)
, Dr. Wasinee Khuntawee 1)
1) Department of Physics, Faculty of Science, Kasetsart University, 2) Department of Meterials Science, Faculty of Science, Kasetsart University, 3) Department of Physics, Faculty of Science, King Mongkut's University of Technology Thonburi, 4) Department of Chemistry, Faculty of Science, Kasetsart University, 5) Department of Zoology, Faculty of Science, Kasetsart University