MSC DRUG DESIGN AND BIOMEDICAL SCIENCE/ MSc BIOMEDICAL SCIENCE/ MSc PHARMACEUTICAL SCIENCE/ MSc PHARMACEUTICAL SCIENCE
BMS11105 DRUG DESIGN AND CHEMOTHERAPY 2020
LABORATORY PRACTICAL WRITE-UP TEMPLATE
| 1. Background: G-quadruplex is units of the noncanonical structure attached to the telomeric deoxyribonucleic acid of every human. They function to increase the structure of the deoxyribonucleic (Hu et al., 2017). Moreover, G-quadruplex is useful in biological synthesis individually as building blocks or as elements used to perform various functions (Kolesnikova and Curtis, 2019). The synthesis of G-quadruplex with higher-order is also crucial for the research and manufacture of cancer drugs with limited side effects to patients. Else ways, telomerase is a repetitive ribonucleic protein at the three ends of every telomere. More specifically, a telomere is a point repetition on every chromosome of a eukaryote (Telomeres and telomerase 2020). Derivatives of anthraquinones have proved useful for researchers due to their ability to treat cancerous tumours, inflammations, malaria, viral and bacterial diseases (Nor et al., 2013). Furthermore, researchers realized that substituting amino with anthraquinones can cure cancer in human and other mammals. Additionally, there are several derivatives of aminoanthraquinone that can readily combine with human nucleotide, and they include reactive blue 2, and the acid blue 129. The most researched and used protein derivative is reactive blue two since it is hydrophobic and react efficiently with the electron of the aromatic systems. Furthermore, the protein derivative exhibits hydrophobic interaction between hydrogen and nitrogen hence can block specific purinergic receptors. On the other hand, chemotherapy is the processes which entail the use of drugs to treat cancer in human beings. The use of chemotherapy as a method for managing cancer always aims at curing palliating or controlling the cancerous cells (How is chemotherapy used to treat cancer 2019). More often, medical doctors use the term curative intent whenever they subscribe a medicine with higher chances of curing cancer since it might not cure cancer of a patient completely. Control of cancer involves stopping the spread of cancer cells to other parts of a patient body or reducing the size of the tumour. Palliating cancer involves relieving pain from, and it is useful during the advanced stages of the disease. The experiment aimed to synthesize aminoanthraquinones to be used as a stabilizer for G-quadruplex.
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| 2. References Arvindekar, A.U., Pereira, G.R. and Laddha, K.S., 2015. Assessment of conventional and novel extraction techniques on extraction efficiency of five anthraquinones from Rheum emodi. Journal of food science and technology, 52(10), pp.6574-6582. Hu, M.H., Chen, S.B., Wang, B., Ou, T.M., Gu, L.Q., Tan, J.H. and Huang, Z.S., 2017. Specific targeting of telomeric multimeric G-quadruplexes by a new triaryl-substituted imidazole. Nucleic acids research, 45(4), pp.1606-1618. Kolesnikova, S. and Curtis, E.A., 2019. Structure and Function of Multimeric G-quadruplexes. Molecules, 24(17), p.3074. Mohanlall, V., Steenkamp, P. and Odhav, B., 2011. Isolation and characterization of anthraquinone derivatives from Ceratotheca triloba (Bernh.) Hook. f. Journal of Medicinal Plants Research, 5(14), pp.3132-3141. Nor, S.M.M., Sukari, M.A.H.M., Azziz, S.S.S.A., Fah, W.C., Alimon, H. and Juhan, S.F., 2013. Synthesis of new cytotoxic aminoanthraquinone derivatives via nucleophilic substitution reactions. Molecules, 18(7), pp.8046-8062. |
| 3. Experimental: Asimina triloba, Belonites bispinosa and Eremostachys vicaryi Benth was collected from Pretoria forests. The plants were sorted, and only healthy roots, stems, leaves and flowers were sun-dried on the roof of the laboratory for eight days before being grounded to powdered form using a blender. Homogenization of the powdered plant was done using a mixture of ethanol and water at a ratio of 4 to 1 for five minutes (Mohanlall et al., 2011). After separating filtrate from the residue, the filtrate was concentrated to a volume of 0.1 then mixed with two molar sulfuric acids. The extraction of the filtrate was conducted using three molar chloroform, and it resulted in immiscible layers. Ammonium hydroxide was added to the aqueous layer until a pH of ten was reached. A mixture of chloroform and methanol at a ratio of 3 to 1 was used for further extraction to obtain an alkaloid. Merck thin layer of chromatography paper was used, and streaking was prevented by adding 1 per cent ammonium chloride. The chromatography was conducted quickly to curb oxidation, and the extract on the chromatograph was viewed against ultraviolet light. Proper formation of the colours was initiated by spraying the chromatography paper with p-anisaldehyde. Add water, and 93 per cent concentrated sulfuric acid to 5 nitro anthraquinone in the ratio of 40:3. The solution was boiled while at high temperature, a mixture of sodium chloride, sodium chlorate and water was added for 15 hours. The resulting product chloroanthraquinone was washed, and it resembles the ideal product. |
| 4. Results and discussion: (i) T.L.C.: (i)The results from the chromatograph were carefully analyzed, and it was realized that anthraquinone varied in quantity depending on parts of the plant investigated (Arvindekar, 2015). For instance, only six was present in other parts of the plant with 13 present in the roots of the plants. The starting Rf values for anthraquinone from the roots, stem, flower and seed pod was 0.24, 0.35 and 0.40, as shown in figure 1.
Figure 1. Chromatograph of anthraquinone extract from different plant parts The impurities present during the extraction process of the plants material included the insoluble solid parts which could reduce the rate of diffusion of the extracts. Non-polar solutions slow down the rate of diffusion of extracts on the chromatography paper while polar solutions diffuse faster. (ii) Percentage Yield Calculation (ii) C14H7ClO2 + C4H11NO2 à C18H17O4N + HCl (l) (l) (s) (aq) 242.65g/mol 105.14 g/mol 223.23 g/mol 36.47 g/mol Theoretically, 1 mole of 1 – chloroanthraquinone reacts with 1 mole of 2-(2-aminoethoxy)ethanol to produce 1 mole of red solid 4-(4 Butylbenzoyloxy)benzoic acid) Mole = given mass/molecular weight Given 0.25 grams of 1-chloroanthraquinone as starting material, the number of moles present is: 0.25/242.65 = 1.0302905E-3 The ratio of moles of the material to the product in the reaction was 1:1, therefore 1.0302905E-3 moles of 1 – chloroanthraquinone yields to 1.0302905E-3 moles of the product. The mass of the product is 1.0302905E-3 * 223.23 = 0.2300 grams (iii) Comment on the purity and % yield: The purity of the extract was low since most of the compound were found in inactive forms from plants part. Moreover, the yield was low because most of the product was sieved as residue. The best method to improve the purity and yield of the product is through chemical synthesis method. 5. Summary conclusion. The three recovered anthraquinone derivatives from the plant’s extract were similar to mitoxanthrone in structure. Mitoxanthrone is important for the chemotherapy of leukaemia, prostate and breast cancer. Hence, the derivatives also have the potential for use in the management of cancerous tumours. Moreover, they can be used to stabilize G-quadruplex effectively. |
| Question section (show all calculations in full) |
| Q 1 Msc-A 255.23g/mol Msc-B 288.23g/mol |
| (Q2) (a) The relative molecular weight of 1-chloroanthraquinone, 242.66 g/mol Given mass/molecular weight = Mole 0.25/242.66 = 1.030248084 E-3 moles
b) Mass = density * volume 4.38g = 1.46 * 3
c)4.38/104=0.04212moles d)1-chloroanthraquinone e)0.04212-1.030248084E-3=0.04109moles f) It was important to use excess amine to ensure complete oxidation of Anthraquinone |
| Q3. (a) The molar reaction ratio for C4H11NO2 and C2H6OS is 1:1 Moles of Msc-B = 0.0016/288.23 = 5.551122368E-6 Relative molecular mass of dimethylsulfoxide is 78.14 g/mol. Reacted mass of dimethylsulfoxide = 78.14 * 5.551122368E-6 = 4.337647018 E -4 g Density of dimethylsulfoxide = 1.1 g/cm3 Volume of dimethylsulfoxide = 4.337647018 E -4/1.1 = 0.394331547 mL |
| (b) Molarity = 400 E-6/1 E -3 = 0.4 mol/L (c) 0.0016 grams of Msc-B was weighed then dilute the solute with dimethylsulfoxide until one liter was obtained (Ryan, n.d).More specifically 0.394331547 milliliter of dimethylsulfoxide was added to make the solution. (Q4.a) The cationic structural feature of deoxyribonucleic acid is vital for the process of telomerase (Yang & Okamoto, 2010). Cancer cells can multiply through telomerase, and the cationic feature makes them vulnerable to chemotherapy drugs. (b) Compound that targets the G-quadruplex can limit the formation of the structure (Lin & Yang, 2017). For instance, potassium ion can induce the deoxyribonucleic acid there inhibiting G-quadruplex formation. (b (i) Screening by phytochemical is another technique that was used to test the purity of the sample. (c) Di-substituted anthraquinones have two carbon spacing that exists between its amine, which is a crucial structural portion for inhibition of telomerase activities (Huang et al., 2008). Furthermore, di-substituted anthraquinone provides an essential site for the binding of the remnant parts of a G-quadruplex. References for Q 4. Huang, H. S., Huang, K. F., Li, C. L., Huang, Y. Y., Chiang, Y. H., Huang, F. C., & Lin, J. J. (2008). Synthesis, human telomerase inhibition and anti-proliferative studies of a series of 2, 7-bis-substituted amido-anthraquinone derivatives. Bioorganic & medicinal chemistry, 16(14), 6976-6986. Lin, C., & Yang, D. (2017). Human telomeric G-quadruplex structures and G-quadruplex-interactive compounds. In Telomeres and Telomerase (pp. 171-196). Humana Press, New York, NY. Yang, D., & Okamoto, K. (2010). Structural insights into G-quadruplexes: towards new anticancer drugs. Future medicinal chemistry, 2(4), 619-646.
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