#TITLE_ALTERNATIVE#

• Main Author: RIYANTI NIM: 30713007, SORAYA
• Format: Dissertations
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/31038
• Institution: Institut Teknologi Bandung

The term diabetes mellitus describes a metabolic disorder of multiple aetiology <br /> <br /> <br /> <br /> characterized by chronic hyperglycaemia with disturbances of carbohydrate, fat and <br /> <br /> <br /> <br /> protein metabolism resulting from defects in insulin secretion, insulin action, or <br /> <br /> <br /> <br /> both.Almost all over the world people with diabetes mellitus always increase every <br /> <br /> <br /> <br /> year. Oral antidiabetic drugs that can inhibit the activity of the DPP-4 enzyme has <br /> <br /> <br /> <br /> been developed such as sitagliptin, vildagliptin, and saxagliptin. The mechanism of <br /> <br /> <br /> <br /> action of the DPP-4 inhibitor group is to increase the levels and action of Glucagon <br /> <br /> <br /> <br /> Like Peptide-1 (GLP-1) and Glucose-dependent Insulinotropic Polypeptide (GIP), <br /> <br /> <br /> <br /> increase insulin secretion according to blood glucose levels, and suppress glucagon <br /> <br /> <br /> <br /> secretion from pancreatic alpha cells. GLP-1 serves to stimulate insulin release and <br /> <br /> <br /> <br /> inhibit glucagon release so that blood sugar levels are maintained. <br /> <br /> <br /> <br /> Knowledge about the use of plants as traditional medicine had been known for <br /> <br /> <br /> <br /> generations. The use of plants in handling diabetes had long been practiced by people <br /> <br /> <br /> <br /> throughout the world. The purpose of this study was to observe and verify 21 types of <br /> <br /> <br /> <br /> plants which were commonly used in traditional medicine as blood glucose levels and <br /> <br /> <br /> <br /> also to find if there were any potential DPP-4 inhibitors in vitro and determined wich <br /> <br /> <br /> <br /> plants had the most active DPP-4 inhibitor activity and active compounds that was <br /> <br /> <br /> <br /> responsible for DPP-4 inhibitor activity of selected plant. <br /> <br /> <br /> <br /> This study began by collecting some plants that have been traditionally used to reduce <br /> <br /> <br /> <br /> blood glucose levels by the community including pacing leaves, mahogany seeds, bitter <br /> <br /> <br /> <br /> herbs, ciplukan/cecendet leaves, bidara upas tubers, bitter melon seeds, mimba leaves, <br /> <br /> <br /> <br /> angsana leaves, avocado seeds, jackfruit leaves, kencana ungu leaves, mala leaves, <br /> <br /> <br /> <br /> brotowali stems, bungur leaves, fenugreek seeds, yakon leaves, bodhi leaves, paitan <br /> <br /> <br /> <br /> leaves, pomegranate rind, patikan kebo herbs, and okra fruit. Extraction process <br /> <br /> <br /> <br /> carried out by reflux method with 96% ethanol, and testing of DPP-4 inhibitor activity <br /> <br /> <br /> <br /> was carried out in vitro with sitagliptin as a standard DPP-4 inhibitor. Plant that <br /> <br /> <br /> <br /> showed the highest inhibition percentage were selected and further research. Isolation <br /> <br /> <br /> <br /> of active compounds was guided by testing the DPP-4 inhibitor activity in vitro and the <br /> <br /> <br /> <br /> isolation process was performed using various chromatographic and other separation <br /> <br /> <br /> <br /> methods. Isolates were characterized using mass spectroscopy (MS), Nuclear <br /> <br /> <br /> <br /> Magnetic Resonance ( <br /> <br /> <br /> <br /> 1 <br /> <br /> <br /> <br /> H-NMR), ( <br /> <br /> <br /> <br /> 13 <br /> <br /> <br /> <br /> C-NMR), NMR-Heteronuclear Single Quantum <br /> <br /> <br /> <br /> Correlation (HSQC) and NMR-Heteronuclear Multiple Bond Correlation (HMBC). <br /> <br /> <br /> <br /> The isolates determined the activity of DPP-4 inhibitors and also as alpha-glucosidase <br /> <br /> <br /> <br /> inhibitors. <br /> <br /> <br /> <br /> There are six of plants that provide more than 50% DPP-4 inhibitory activity at 2.5 <br /> <br /> <br /> <br /> µg/mL, wich were yakon (Smallanthus sonchifolius (Poepp.) H.Rob.) leaves 52.84 ± <br /> <br /> <br /> <br /> 2.00 %; fenugreek (Trigonella foenum-graecum L) seeds 71.29 ± 0.33 %; bodhi (Ficus <br /> <br /> <br /> <br /> religiosa L.) leaves 68.98 ± 1.96 %, pomegranate (Punica granatum L.) rind 58.79 ± <br /> <br /> <br /> <br /> 2.29 %; brotowali (Tinospora crispa Miers ex Hoff.f) stem 65.86 ± 1.02%; and bungur <br /> <br /> <br /> <br /> (Lagerstroemia loudonii Teijsm. & Binn.) leaves 60.22 ± 2.01 %. Based on the DPP-4 <br /> <br /> <br /> <br /> inhibitory activity, easiness of obtaining plants and no previous studies related to DPP4 activity, bungur (Lagerstroemia loudonii Teijsm. & Binn.) was chosen as the plant <br /> <br /> <br /> <br /> to be further researched. <br /> <br /> <br /> <br /> Selected plant extracts (Lagerstroemia loudonii Teijsm. & Binn.) were fractionated <br /> <br /> <br /> <br /> using liquid-liquid extraction with water solvent, n-hexane and ethyl acetate. Testing <br /> <br /> <br /> <br /> of DPP-4 inhibitor activity was carried out on n-hexane, ethyl acetate and water <br /> <br /> <br /> <br /> fractions. The results of DPP-4 inhibitory activity on n-hexane fraction at 0.5; 1; 2.5; <br /> <br /> <br /> <br /> and 5 µg/mL showed that the percent inhibition were 13.37 ± 1.88 %; 18.85 ± 1.33 %; <br /> <br /> <br /> <br /> 12.21 ± 1.40 %; and 8.22 ± 1.31 % respectively. Ethyl acetate fraction at 0.5; 1; 2.5; <br /> <br /> <br /> <br /> and 5 µg/mL showed that the percent inhibition were 15.21 ± 0.72 %; 13.20 ± 1.64 %; <br /> <br /> <br /> <br /> 11.38 ± 1.20 %; and 13.20 ± 1.64 % respectively. Water fraction at 0.5 µg/mL did not <br /> <br /> <br /> <br /> show inhibitory activity, but at 1; 2.5; and 5 µg/mL demonstrated inhibition of 4.27 ± <br /> <br /> <br /> <br /> 1.56 %; 3.89 ± 0.26 %; and 6.50 ± 1.72 % respectively. Isolation of the DPP-4 inhibitor <br /> <br /> <br /> <br /> active compound from n-hexane fraction using vacuum liquid chromatography with <br /> <br /> <br /> <br /> gradient elution system using n-hexane-ethyl acetate-methanol at several compositions <br /> <br /> <br /> <br /> with increasing polarity as mobile phase and purification of isolates using column <br /> <br /> <br /> <br /> chromatography with an isocratic elution system using chloroform-ethyl acetate (9.5: <br /> <br /> <br /> <br /> 0.5). From the results of subfraction purification obtained LL isolates which have a <br /> <br /> <br /> <br /> melting point at 139.5-141.1 <br /> <br /> <br /> <br /> o <br /> <br /> <br /> <br /> C. <br /> <br /> <br /> <br /> Characterization results of LL isolates using spectroscopic methods <br /> <br /> <br /> <br /> 1 <br /> <br /> <br /> <br /> H-NMR, <br /> <br /> <br /> <br /> 13 <br /> <br /> <br /> <br /> C- <br /> <br /> <br /> <br /> NMR, NMR-Heteronuclear Single Quantum Correlation (HSQC) and NMR- <br /> <br /> <br /> <br /> Heteronuclear Multiple Bond Correlation (HMBC) note that LL compounds have 6 <br /> <br /> <br /> <br /> methyl groups, 1 double bond, 1 hydroxyl group, 11 methylene groups, 9 methane <br /> <br /> <br /> <br /> groups and 3 quaternary carbon. Therefore it was suggested that isolates leads to betasitosterol compounds. Confirmation of <br /> <br /> <br /> <br /> 1 <br /> <br /> <br /> <br /> H-NMR spectrum and <br /> <br /> <br /> <br /> 13 <br /> <br /> <br /> <br /> C-NMR LL isolates <br /> <br /> <br /> <br /> with beta-sitosterol isolates from Etlingera spaerocephala Var. grandiflora showed <br /> <br /> <br /> <br /> similar chemical shifts. The mass spectrum of LL isolates showed the presence of a <br /> <br /> <br /> <br /> molecular ion [M <br /> <br /> <br /> <br /> + <br /> <br /> <br /> <br /> -H] at 413.37. Therefore, it was concluded that the isolates obtained <br /> <br /> <br /> <br /> from bungur leaves were beta-sitosterol. Based on the results of this study, it can be <br /> <br /> <br /> <br /> concluded that of the 21 plants used traditionally to reduce blood glucose, there are <br /> <br /> <br /> <br /> six of plants that showed inhibitory activity greater than 50 %. Bungur leaf extract, its <br /> <br /> <br /> <br /> fraction and isolates had inhibitory activity against DPP-4 in vitro. Beta-sitosterol <br /> <br /> <br /> <br /> isolates have weak activity as DPP-4 inhibitors with IC50 of 224.66 µg/mL as compared <br /> <br /> <br /> <br /> to the standard inhibitor DPP-4 (sitagliptin) IC50 of 1.10 µg/mL. Beta-sitosterol <br /> <br /> <br /> <br /> isolates havd strong activity as alpha-glucosidase inhibitors with IC50 values of 60.14 <br /> <br /> <br /> <br /> µg/mL, whereas acarbose as a standard of alpha-glucosidase inhibitors had an IC50 <br /> <br /> <br /> <br /> value of 20.53 µg/mL. <br />