Small Molecule Inhibitor Therapeutic Target for Alzheimer's Disease.

Project SummaryThe objective of our proposed research is to develop a water soluble molecule that can reduce the levels of amyloid betaAand Drpand prevent abnormal interactions between Aand Drpin Alzheimers diseaseADaffected neuronsOur previous AD studies foundincreased production and accumulation of Aand increased expression of Drpandan abnormal interaction between Aand Drpin AD neurons that was associated with synaptic dysfunctionmitochondrialmtdamageand dysfunction of neurons affected by ADA therapeutic strategy for AD may involve treating AD affected neurons with molecules designed to reduce Aand Drplevels and to inhibit the interaction of Aand DrpVery few molecules have been developed to prevent ADbut those that are the most promising are insoluble in waterrendering them problematic treatments due to water insolubilityThereforein this project we are proposing to develop a water soluble molecule capable of reducing Aand Drplevels in AD neurons and of inhibiting the interaction between Aand DrpTo achieve this objectivewe have developed technology and have designed and producedmolecular crystal structuresincluding DDQdiethyldihydroxyphenethylaminoquinolinyl methylphosphonateWe selected DDQ for the proposed research to reduce Aand Drpinteraction at specific interacting sites in the ADrpcomplex and exhibits the best docking capabilities of allstructures and received the best docking scoreDDQ also readily bound to Drpindependently and while Drpinteracted with AWe synthesized DDQ by following retro synthesis analysis and analyzed its structure spectrallyWe tested DDQ in AD affected neuronsusing different methodologiesand measured the production of Aand expression of Drpafter DDQ docked with DrpOur preliminary studieswhich includedhuman cells incubated with Aand treated with DDQ andfurther transfected human cells with mutant APP cDNA and treated with DDQand characterized Aand DDQ treated cells and mutant APP transfected and DDQ treated cells for mRNAusing qRT PCRADrpinteractionsusing Co IPwestern blot analysisimmunocytochemistryand ultrastructural changesusing electron microscopyThese studies revealed that DDQ reduced Aand Drplevels as well as ADrpinteractions and protect AD affected cells from synaptic and mt toxicitiesThe current application seeksto determine the blood brain barrier crossing properties and pharmacokinetics of DDQ in wildtype miceto measure cognitive behavior in the DDQtreated anduntreated APP transgenic miceTglineandto determine the protective effects of DDQ in terms of reducing the interaction of Aand DrpThe outcome of our application will determine the drug delivery characteristics of DDQ and whether it is capable of targeting and binding to AD neurons and protecting them from Aand Drpinduced toxicitiesThe proposed research will also provide new information about DDQ for future AD clinical trials Project NarrativeThe elevated levels of Aand increased expressions of mitochondrial fission protein Drpand abnormal interactions between Aand Drphave been found to induce synaptic dysfunction and mitochondrial oxidative damagecausing neuronal damage in Alzheimers diseaseADneuronsThe primary objective of the proposed research is to develop the water soluble drug molecule DDQwhichbased on our preliminary studiesappears to be capable of reducing the Aand Drplevelsand of inhibiting interactions between Aand DrpIn our proposed experimentswe will determine whether DDQ enhances neuronal function and survival in AD affected neuronsand through pharmacokinetic and and animal studies of ADwe will characterize drug properties of DDQincluding whether DDQ can cross the blood brain barrierwhich will indicate whether DDQ is a promising drug molecule that may slow AD progression