Design,Synthesis And Preliminary Bioacitivity Of5-Aryl-3-Substituted Mercapto-1,2,4-triazoles And Diarylamides

This dissertation is focused on the discovery of new glucokinase activators (GKAs) and mitochondrial uncouplers based on Glucokinase(GK) and Mitochondrial targets, and the dissertation is divided into the following two parts:Part1. Design, synthesis, characterization and bioactivities of new GKAsLGH00230is a hit compound for glucokinase obtained from a high throughput screening of56,000compounds by Prof Jia Li’s Group of the national Center for Drug Screening (Shanghai). Comparing with the normal GKAs’structures, this hit compound is of a novel skeleton. In this dissertation, the structure modification of LGH00230and the structure-activity relationships were preliminarily investigated.1. The general methods for synthesis of LGH00230and its derivatives were developed, and thirty three new compounds were designed and prepared by modification of the structure unit A of LGH00230. All the structures were characterized by1H NMR,13CNMR and HRMS (ESI) spectra. The activity tests show that the compound2-8and2-12, which linked a propargyl group and a benzyl group, respectively, at the mercapto group, are more potent than the hit compound LGH00230. In particular, compound2-8possesses an EC50value of2.36μM with a high activation fold of3.53(versus control level), while compound2-12has a higher potency (ECso,4.08μM) with an activation fold of3.28. When a cyanomethyl group linked to the mercapto group (2-9), the potency is also slightly higher than that of LGH00230. In addition, the structure-activity relationship (SAR) study showed that the maintaining of the sulfide chain is useful to the GK activation.2. Five compounds were designed, synthesized and characterized by modifying the structure unit B. The assay shows that the potency in GK activation are lost when the N-H bond is substituted by N-propargyl (2-32), N-methyl (2-34), or N-amino (2-35) except for the substitution of N-acetyl (2-33) which shows much less activity. These results indicate that the presence of the N-H bond in these compounds is very important. Insertion of a methylene group between Aryl and triazole group affords compound2-43which is invalid in GK activation, indicating the significance of the conjugated scaffold.3. Based on modifying structure unit C of the hit compound LGH00230, thirty eight compounds were designed, synthesized and characterized. While change length and type of the ortho ether chain, different GK potency has been obtained with almost equal activation fold except for compound2-49. The best compound is propyloxy substituted derivative2-8. Further, the potency in GK activation is lost while modify the ether chain with alkyl or thioether chain or replace the phenyl group with pyridyl group.4. Compounds2-8,2-13,2-47and compound GKA25AstraZeneca’s reported stimulated insulin release test, the results showed that compounds2-8,2-13and2-47INS-1E cells have the ability of insulin secretion, espercially the compounds2-8and2-13are stronger; Compounds2-8,2-13and2-47have nontoxicity to INS-1E cells.Part2. Design, synthesis, characterization and bioactivity of new mitochondrial uncouplersBased on the previous work of our group, we are mainly focused on the further modification of two active compounds, AUK2029and AUK1029, and preliminary exploration of the structure-activity relationship of this kind of compounds. 1. Nineteen new compounds were designed and synthesized by modifying the structure unit A, B and C of AUK2029and AUK1029. All the structures were characterized by1H NMR,13C NMR and HRMS (ESI) spectra.2. Mitochondrial membrane potential (MMP) tests on L6myotube were set as the preliminary evaluation method for uncoupling activity. Almost all the compounds significantly affect the activity of reducing MMP at the starting concentration2.5μM except for compound LBR1225, which reduced MMP moderately.(1) When region B is an amide, modification of structure unit A with5-methyl group and4-chloro group afford LBR1220and LBR1219, which show much potency on reducing MMP at2.5μM concentration while comparing with10μM of CCCP. LBR1224, with a hydroxyl group at5-position of region A, moderately decreases the MMP at low concentration and exhibits good concentration gradient manner.(2) When region B is an ester bond, hydroxyl substitution at ortho position of region A is not necessary since compounds LBR1216and LBR1217, which bearing a o-methoxy and o-nitro group respectively, also can significantly reduce the MMP of L6myotube at lower concentration. This conclusion differs from the previous results upon AUK1029derivatives.(3) Increasing the volume of the substituent at region A is not helpful in reducing the MMP significantly.(4) The capacity on decreasing MMP is significantly weakened while replacing the hydroxyl group by an amino group at region B.(5) The introduction of one more hydroxyl group at5-position of region C exhibits moderate reduction ability of MMP. 3. Preliminary tests on promoting the glucose consumption and lactate release of L6myotube show that5μM of LBR1216and LBR1217be of similar activity with10μM of Berberine. The glucose consumption of LBR1216and LBR1217is greater than Berberine at10μM concentration, and at the same concentration, the lactate release of LBR1216and LBR1217is almost the same as Berberine.4. Glucose consumption and glucose uptake analysis of LBR1216and LBR1217as well as AUK1029indicated the similar promotion ability of these three compounds. At the same concentration (10μM) the promotion ability of LBR1216and LBR1217on glucose consumption is stronger than Berberine.5.3H-labeled glucose absorption experiment indicated that the compound LBR1216at2.5μM concentration has more potency on promoting glucose uptake than5μM of Berberine, and LBR1217shows the similar ability on glucose uptake at lower concentration (0.525μM) with5μM of Berberine. LBR1217promotes glucose uptake significantly at5μM concentration, and the activity is stronger than that of Berberine.