PMID- 36253285 OWN - NLM STAT- MEDLINE DCOM- 20230111 LR - 20230222 IS - 2095-4964 (Print) VI - 21 IP - 1 DP - 2023 Jan TI - In vitro and in vivo anticancer potential and molecular targets of the new colchicine analog IIIM-067. PG - 62-76 LID - S2095-4964(22)00108-X [pii] LID - 10.1016/j.joim.2022.09.006 [doi] AB - OBJECTIVE: The current study evaluated various new colchicine analogs for their anticancer activity and to study the primary mechanism of apoptosis and in vivo antitumor activity of the analogs with selective anticancer properties and minimal toxicity to normal cells. METHODS: Sulforhodamine B (SRB) assay was used to screen various colchicine analogs for their in vitro cytotoxicity. The effect of N-[(7S)-1,2,3-trimethoxy-9-oxo-10-(pyrrolidine-1-yl)5,6,7,9-tetrahydrobenzo[a] heptalene-7-yl] acetamide (IIIM-067) on clonogenicity, apoptotic induction, and invasiveness of A549 cells was determined using a clonogenic assay, scratch assay, and staining with 4',6-diamidino-2-phenylindole (DAPI) and annexin V/propidium iodide. Mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) levels were observed using fluorescence microscopy. Western blot analysis was used to quantify expression of proteins involved in apoptosis, cell cycle, and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling. Pharmacokinetic and in vivo efficacy studies against Ehrlich ascites carcinoma (EAC) and Ehrlich solid tumor models were conducted using Swiss albino mice. RESULTS: IIIM-067 showed potent cytotoxicity and better selectivity than all other colchicine analogs screened in this study. The selective activity of IIIM-067 toward A549 cells was higher among other cancer cell lines, with a selectivity index (SI) value of 2.28. IIIM-067 demonstrated concentration- and time-dependent cytotoxicity against A549 cells with half-maximal inhibitory concentration values of 0.207, 0.150 and 0.106 mumol/L at 24, 48 and 72 h, respectively. It also had reduced toxicity to normal cells (SI > 1) than the parent compound colchicine (SI = 1). IIIM-067 reduced the clonogenic ability of A549 cells in a dose-dependent manner. IIIM-067 enhanced ROS production from 24.6% at 0.05 mumol/L to 82.1% at 0.4 mumol/L and substantially decreased the MMP (100% in control to 5.6% at 0.4 mumol/L). The annexin V-FITC assay demonstrated 78% apoptosis at 0.4 mumol/L. IIIM-067 significantly (P < 0.5) induced the expression of various intrinsic apoptotic pathway proteins, and it differentially regulated the PI3K/AKT/mTOR signaling pathway. Furthermore, IIIM-067 exhibited remarkable in vivo anticancer activity against the murine EAC model, with tumor growth inhibition (TGI) of 67.0% at a dose of 6 mg/kg (i.p.) and a reduced mortality compared to colchicine. IIIM-067 also effectively inhibited the tumor growth in the murine solid tumor model with TGI rates of 48.10%, 55.68% and 44.00% at doses of 5 mg/kg (i.p.), 6 mg/kg (i.p.) and 7 mg/kg (p.o.), respectively. CONCLUSION: IIIM-067 exhibited significant anticancer activity with reduced toxicity both in vitro and in vivo and is a promising anticancer candidate. However, further studies are required in clinical settings to fully understand its potential. CI - Copyright (c) 2022 Shanghai Changhai Hospital. Published by Elsevier B.V. All rights reserved. FAU - Malik, Sumera AU - Malik S AD - Pharmacology Division, Council of Scientific and Industrial Research (CSIR)-Indian Institute of Integrative Medicine, Jammu 180001, Jammu & Kashmir, India; Academy of Scientific & Innovative Research (AcSIR), Ghaziabad 201002, Uttar Pradesh, India. FAU - Mintoo, Mubashir J AU - Mintoo MJ AD - Pharmacology Division, Council of Scientific and Industrial Research (CSIR)-Indian Institute of Integrative Medicine, Jammu 180001, Jammu & Kashmir, India; Academy of Scientific & Innovative Research (AcSIR), Ghaziabad 201002, Uttar Pradesh, India. FAU - Reddy, Chilakala Nagarjuna AU - Reddy CN AD - Academy of Scientific & Innovative Research (AcSIR), Ghaziabad 201002, Uttar Pradesh, India; Medicinal Chemistry Division, CSIR-Indian Institute of Integrative Medicine, Jammu 180001, Jammu & Kashmir, India. FAU - Kumar, Rajesh AU - Kumar R AD - Pharmacology Division, Council of Scientific and Industrial Research (CSIR)-Indian Institute of Integrative Medicine, Jammu 180001, Jammu & Kashmir, India. FAU - Kotwal, Pankul AU - Kotwal P AD - Academy of Scientific & Innovative Research (AcSIR), Ghaziabad 201002, Uttar Pradesh, India; Pharmacokinetics-Pharmacodynamics (PK-PD), Toxicology Division, CSIR-Indian Institute of Integrative Medicine, Jammu 180001, Jammu & Kashmir, India. FAU - Bharate, Sandip B AU - Bharate SB AD - Academy of Scientific & Innovative Research (AcSIR), Ghaziabad 201002, Uttar Pradesh, India; Medicinal Chemistry Division, CSIR-Indian Institute of Integrative Medicine, Jammu 180001, Jammu & Kashmir, India. FAU - Nandi, Utpal AU - Nandi U AD - Academy of Scientific & Innovative Research (AcSIR), Ghaziabad 201002, Uttar Pradesh, India; Pharmacokinetics-Pharmacodynamics (PK-PD), Toxicology Division, CSIR-Indian Institute of Integrative Medicine, Jammu 180001, Jammu & Kashmir, India. FAU - Mondhe, Dilip M AU - Mondhe DM AD - Pharmacology Division, Council of Scientific and Industrial Research (CSIR)-Indian Institute of Integrative Medicine, Jammu 180001, Jammu & Kashmir, India; Academy of Scientific & Innovative Research (AcSIR), Ghaziabad 201002, Uttar Pradesh, India. Electronic address: mondhe33@gmail.com. FAU - Shukla, Sanket K AU - Shukla SK AD - Pharmacology Division, Council of Scientific and Industrial Research (CSIR)-Indian Institute of Integrative Medicine, Jammu 180001, Jammu & Kashmir, India; Academy of Scientific & Innovative Research (AcSIR), Ghaziabad 201002, Uttar Pradesh, India. Electronic address: sanket.shukla@iiim.res.in. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20220930 PL - Netherlands TA - J Integr Med JT - Journal of integrative medicine JID - 101603118 RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) RN - 0 (Antineoplastic Agents, Phytogenic) RN - EC 2.7.1.- (Phosphatidylinositol 3-Kinases) RN - 0 (Reactive Oxygen Species) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - SML2Y3J35T (Colchicine) SB - IM MH - Animals MH - Mice MH - *Proto-Oncogene Proteins c-akt/metabolism MH - *Antineoplastic Agents, Phytogenic/pharmacology MH - Phosphatidylinositol 3-Kinases/metabolism MH - Reactive Oxygen Species/metabolism MH - TOR Serine-Threonine Kinases/metabolism MH - Colchicine/pharmacology MH - Apoptosis MH - Cell Line, Tumor MH - Cell Proliferation MH - Mammals/metabolism OTO - NOTNLM OT - A549 cells OT - Antineoplastic agents OT - Apoptosis OT - Carcinoma OT - Colchicine OT - Ehrlich tumor OT - IIIM-067 EDAT- 2022/10/18 06:00 MHDA- 2023/01/12 06:00 CRDT- 2022/10/17 22:07 PHST- 2022/01/25 00:00 [received] PHST- 2022/06/14 00:00 [accepted] PHST- 2022/10/18 06:00 [pubmed] PHST- 2023/01/12 06:00 [medline] PHST- 2022/10/17 22:07 [entrez] AID - S2095-4964(22)00108-X [pii] AID - 10.1016/j.joim.2022.09.006 [doi] PST - ppublish SO - J Integr Med. 2023 Jan;21(1):62-76. doi: 10.1016/j.joim.2022.09.006. Epub 2022 Sep 30.