title: The preclinical evaluation of the dual mTORC1/2 inhibitor INK-128 as a potential anti-colorectal cancer agent.
abstract: The colorectal cancer is the leading contributor of cancer-related mortality. Mammalian target of rapamycin (mTOR), existing in 2 complexes (mTORC1/2), is frequently dysregulated and constitutively activated in colorectal cancers. It represents an important drug target. Here we found that INK-128, the novel ATP-competitive kinase inhibitor of mTOR, blocked both mTORC1 and mTORC2 activation in colorectal cancer cells (both primary and transformed cells). The immunoprecipitation results showed that the assembly of mTORC1 (mTOR-Raptor association) and mTORC2 (mTOR-Rictor-Sin1 association) was also disrupted by INK-128. INK-128 inhibited colorectal cancer cell growth and survival, and induced both apoptotic and non-apoptotic cancer cell death. Further, INK-128 showed no effect on Erk/MAPK activation, while MEK/Erk inhibition by MEK-162 enhanced INK-128-induced cytotoxicity in colorectal cancer cells. Meanwhile, INK-128 downregulated Fascin1 (FSCN1)/E-Cadherin expressions and inhibited HT-29 cell in vitro migration. In vivo, daily INK-128 oral administration inhibited HT-29 xenograft growth in mice, which was further enhanced by MEK-162 administration. Finally, we found that INK-128 sensitized 5-fluorouracil-(5-FU)-mediated anti-HT-29 activity in vivo and in vitro. Thus, our preclinical studies strongly suggest that INK-128 might be investigated for colorectal cancer treatment in clinical trials.
authors.affiliation: a Department of Gastroenterology; Xuzhou Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Chinese Medicine ; Xuzhou , China.
authors.forename: Chen
authors.initials: C
authors.lastname: Li
authors.forename: Jian-Feng
authors.initials: JF
authors.lastname: Cui
authors.forename: Min-Bin
authors.initials: MB
authors.lastname: Chen
authors.forename: Chao-Ying
authors.initials: CY
authors.lastname: Liu
authors.forename: Feng
authors.initials: F
authors.lastname: Liu
authors.forename: Qian-De
authors.initials: QD
authors.lastname: Zhang
authors.forename: Jian
authors.initials: J
authors.lastname: Zou
authors.forename: Pei-Hua
authors.initials: PH
authors.lastname: Lu
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001583:Benzoxazoles / Q000008:administration & dosage / Q000494:pharmacology*; D015820:Cadherins / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D015179:Colorectal Neoplasms / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical; D004357:Drug Synergism; D004789:Enzyme Activation / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors; D005472:Fluorouracil / Q000494:pharmacology; D019073:HT29 Cells; D006801:Humans; D008297:Male; D000076222:Mechanistic Target of Rapamycin Complex 1; D000076225:Mechanistic Target of Rapamycin Complex 2; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D046912:Multiprotein Complexes / Q000037:antagonists & inhibitors*; D047428:Protein Kinase Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D011743:Pyrimidines / Q000008:administration & dosage / Q000494:pharmacology*; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors*; D023041:Xenograft Model Antitumor Assays; D000225:Adenine / Q000031:analogs & derivatives
chemical_list: D000970:Antineoplastic Agents; D001583:Benzoxazoles; D015820:Cadherins; D002352:Carrier Proteins; D048049:Extracellular Signal-Regulated MAP Kinases; D005472:Fluorouracil; D000076222:Mechanistic Target of Rapamycin Complex 1; D000076225:Mechanistic Target of Rapamycin Complex 2; D008840:Microfilament Proteins; D046912:Multiprotein Complexes; D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; D058570:TOR Serine-Threonine Kinases; C502658:FSCN1 protein, human; C572449:sapanisertib; D000225:Adenine
keywords: (S6K), p70S6K1; (mTOR), mammalian target of rapamycin; (mTORC1), mTOR complex 1; (mTORC2), mTOR complex 2; Co-IP, co-immunoprecipitation; ECL, enhanced chemiluminescence; FSCN10, Fascin1; HRP, horseradish peroxidase; INK-128; PI, propidium iodide; SD, standard deviation (SD); cell growth and migration; colorectal cancer; mTOR