Discovery And Ootimizion Of Novel Diary Pyridine/furan Ketones As Metastasis Inhibitors And PROTAC-Akt Molecules For Akt Degradation

Malignant tumor is a major life-threatening and health-threatening disease worldwide.And epidemiological investigation showed that the tumor invasion and metastasis are the key factors influencing prognosis and recurrence,as the leading cause of death in most cancer patients.To date,for patients with tumor and tumor metastasis,there has been a variety of treatment or intervention strategies such as targeted therapy for specific genes or signaling proteins or tumor cells,or targeted degradation of proteins by proteolysis-targeting chimeras(PROTACAs),and a large number of small moleculars listed or under clinical trials.However,due to the complexity of the regulatory pathways and the challenge of tumor resistabce,research and development of potent moleculars with novel mechanisms against cancer and metastasis is urgently neededPhenotypic screening method of antimetastasis and protein degradation technology based on the PROTACs were applied to discovery of potent moleculars against metastasis and exploration of Akt degradation molecules,respectively.The specific researchs are as followsNovel diary pyridine/furan ketones as metastasis inhibitors:A variety of cytokines and signal transduction pathways are involved in the process of tumor metastasis.Many inhibitors against angiogenesis or MEK/ERK are used in patients with cancer metastasis,as a tumor suppression drugs rather than targeted inhibitors of cancer metastasis.And inhibitors targeting for metastasis have been in the preclinical stage Therefore,based on the results of the scratch test,diverse compounds selected in-house was tested about migration of tumor cells for the discovery of novel structures with unrevealed mechanism as metastasis inhibitors.1)Based on the inhibition of cell migration of H1299,intial screening results of 26 diverse compounds have shown that two compounds(H1217 and H1252;inhibition ratio of 5?M:60%and 50%),to some extent,can inhibit cell migration of H1299.2)This promoted us to optimize this kind of structure,through the application of scaffold hopping and the bioisosterism.23 compounds or four series consisting of furan-2(5H)-one,1H-pyrrole-2,5-dione,1,5-dihydro-2H-pyrrol-2-one,and pyridin-2-amine were synthesized and carried out the activity of antimetastasis(cell line:KHOS cell).Among them,two compounds(1-8,1-27)have a potent activity against tumor cell migration,and the corresponding structure-activity relationship has been obtained.3)Compound I-27 can significantly inhibit the occurrence of pulmonary metastases in murine 4T1 breast cancer of mice,and the inhibition rate reaches 61%(100mg/kg,PO).4)WSB-1 plays a key role in tumor metastasis1,2.Also,I-27 can significantly inhibit the expression of WSB-1 and has a inhibition rate of 72%in wsb-1 stable expression cell strain H1299,thereby affecting the ubiquitin degradation of RhoGDI2 mediated by WSB-1.5)To further research the protein targeted by compound I-27 and explore its molecular mechanism about migration of tumor cells,a linker containing alkynl group and photoaffinity group was introduced compound I-27 in the amino position which based on the corresponding SAR.And the resulting probe P1 could inhibit metastasis.Other important chemical biology research is still ongoing.In short,by a integrated strategy consisting of phenotype screening method for antimetastasis,reasonable structure optimization strategy,activity test on antimetastasis in vivo and in vitro,the mechanism research of compound I-27 and the design and synthesis of the probe P1 for proteins targeted by I-27,we got a series of novel structure with unrevealed mechanism as metastasis inhibitors.PROTAC-Akt molecules for Akt degradation:The occurrence of tumor is closely related to expression of its specific proteins,degradation of which has become a new strategy of tumor treatment recent year.Proteolysis targeting chimeric molecules PROTAC,are small molecules that eliminate the target proteins by the ubiquitin proteasome system.Specificity and paintability of PROTACs contribute to its potential application.In this paper,Compound H1150,our previous inhibitor of Akt which has a potent ability,was a starting material as the ligand of Akt.Different linkers were applied to combination of compound H1150 and Thalidomide structure in a rational way based on the corresponding SAR for the target moleculars P3 and P4.The westernblot results showed that both P3 and P4 could degrade Akt protein(P3 DC95<504nM;P4 DC95 ?482nM),and also had a potent inhibition of MM1S cell proliferation(P3 IC50=0.998?M,P4 IC50=0.569?M).In a word,PROTACs based on small molecular was firstly adopted to realize the Akt degredation of the PI3K/Akt/mTOR pathway,and the functional moleculars had a antitumor activity.Also,the optimization work for the degradation activity and the inhibition activity is still in progress.