Cell Imagine Phenotype Profiles Of Drugs And Candidate Drugs

The insufficient safety and efficacy are the main reasons for the failure incandidate drugs clinical trials recent years. Therefore, to fully understand themechanisms of candidate drugs and to enhance their quality become the key tosuccess in the pre-clinical research. Multi-phenotype analysis to small molecules isthe most effective way to probe their mechanisms and explain the therapeutic effectsof compounds and their biological mechanisms at the systematic level. Cells, as thesmallest unit of life system, are also the fundamental and functional unit of drugresponse. The multi-phenotype analysis of cells level can quantitatively anddynamically determine the reaction, targets and signaling pathways of drugs.Compared with the technique of genomics, proteomics, metabonomics andphosphorylation proteomics, assays based on cell phenotype profile could providericher and more direct information about drug effect and is a revolutionized techniqueto optimize candidate effect in recent years’ new drugs development. The majorproblem at present is how to use the phenotype information based on the cell level toincrease the drug’s effect and decrease its cyto-toxicity. The research purpose of thisproject is to fully understand the pharmacological features of the drug and candidatedrugs so as to provide experimental data for the development and repositioning ofdrugs or candidate drugs through cell phenotype analysis; to explore the researchways to establish drug cell phenotype profile so as to provide more valuableinformation for drugs’ prediction research through the analysis to the relationshipbetween scaled tested drugs’ structure or targets and cell’s drug effect phenotype andcyto-toxicity phenotype.This project adopts the high content analysis/screening technique, choosesspecial expressive drugs targets and the combination between30analytical cell line ofcertain signal access and2groups of multi-phenotype cyto-toxicity profiles, examines the influence of approximately100already-marketed drugs and candidate drugscovering various targets to40kinds of cell targets/signal access phenotype （coveringtypical targets and major signal pathways） and21kinds of cell cyto-toxicityphenotype （covering major cell cyto-toxicity mechanism and cell morphologicalchange）. According to the EC⁵⁰/IC⁵⁰and EC²⁵/IC²⁵of agonism or antagonism cellphenotype of tested drugs, the project analyzes the off-target effects, hidden effectsand side effects; analyzes the relationship between the tested drugs’ targets orstructure and cell targets/signal pathway phenotype or cell cyto-toxicity phenotypethrough the pharmacological network of social analysis, AP-cluster analysis and thepharmacophore analysis, etc..The research finds:（1）45%of tested drugs have more than1exact outside-targeteffect, the proportion of which is close in candidate drugs and already-marketed drugs.The onset concentration of The overwhelming majority of outside-target effect ismuch higher than that of the inside-target effect;（2） in12candidate drugs, onlyLL-SGC-001in high concentration can stimulate oxidative stress, other candidatedrugs did not have pharmacological non-targets effect and cyto-toxicity effect;（3） theanti-tumor candidate drugs PAC-1and YCL-0426targeted at pre-caspase3have thefunction to induce Hypoxia injury and DNA DSB injury; sunitinib, the multi-targetsRTKs inhibitor, has the function to extensively inhibit signaling pathways related withcell proliferation and potential inflammatory reaction;（4） HMG-CoA Reductaseinhibitor lovastatin,-glucosidase inhibitor acarbose, L-Ca²⁺channel blockerlercanidipine and amlodipine and PPAR agonist fenofibrate have pharmacologicalnon-target effect;（5） spirolactone, lercanidipine, isoproterenol, histamine anddexamethasone has clear function to activate AR （androgen receptor）;（6） S1PR1andCBR1are the receptors having most off-target effect, the off-target elements mostlyinclude M-R, L-Ca²⁺channel and hepatitis B virus ligand;（7） PPAR agonist affectsCBR1、S1PR1、IGFR/AKT and Insulin/GLUT4phenotypes, but it did not affect GR,ERα, PR and AR cell phenotype which also belongs to nucleus receptors. PPARδ’sagonist may have the activity to activate transcriptional factor NF-kB （P65subunit）;（8） the relevance between the tested drugs targets and cell targets/signalpathways phenotype is comparatively high; the relevance between the structure of thetested drugs and the cell cyto-toxicity phenotype is comparatively high;（9） Theresults from compounds with similar phenotype profile showed that: M-R, L-Ca²⁺channels, hepatitis B virus nucleocapsid would share the similar ligand binding sites with CBR1; PPAR and steroid hormone receptor （GR, ER, PR and AR） did notshare the similar ligand binding sites, PPAR agonists might affect CBR1, S1PR1,IGFR/AKT and Insulin/GLUT4phenotype; the crosstalk occurred in different classesof steroid hormone receptors or different steroid hormones; PPAR agonists mayactivate P65（NF-kB）;（10） cluster analysis can better show the relevance between cellcyto-toxicity phenotype and the structure of the tested drugs, pharmacologicalnetwork of social analysis may better show the relevance between cell cyto-toxicitymechanism and drugs target type.To sum up, this research explores and establishes the research method aboutdrugs cell phenotype profile and provide experimental data for the optimization ofcandidate drugs and repositioning of the old drugs.