Cell lines

Saos2 cells

Preparation Method

Colony formation assays were performed by seeding 300 cells/well into 6-well plates for 24 h after growth, cells were treated with 100 M JPH203 (KYT-0353) for 72 h to remove the JPH203 (KYT-0353) treatment and cultured for 10 days with fresh medium.

Reaction Conditions

Treatment with 100 μM JPH203 (KYT-0353) for 72 hours

Applications

JPH203 (KYT-0353) inhibited colony formation in Saos2 cells.

Animal models

Six-week-old male athymic BALB/c nude mice

Preparation Method

Mice in treatment group were injected with JPH203 (KYT-0353) intravenously every day for 20 days.

Dosage form

6.3 mg/kg; 12.5 mg/kg; 25 mg/kg JPH203 (KYT-0353) for 20 days

Applications

JPH203 (KYT-0353) (KYT-0353) was administered intravenously daily for 20 days at three different doses starting at day 3 after the injection of cancer cells. On the days 18 and 21, JPH203 (KYT-0353) showed dose-dependent inhibition on tumor growth with significantly inhibited tumor growth in the groups of JPH203 (KYT-0353) at 12.5 mg/kg and 25 mg/kg .

JPH203 (KYT-0353), a selective L-type amino acid transporter 1 inhibitor, significantly inhibited leucine uptake and cell growth in HT-29 YD-38 and leukemia cells, with IC50 values of 0.06 μM and 4.1 μM, respectively^[1,5].

JPH203 (KYT-0353) can up-regulate the activated forms of pro-apoptotic factors such as Bad, Bax, Bak and caspase-9 and down-regulate anti-apoptotic factors such as Bcl-2,Bcl-xl, activation of mitochondria dependent apoptotic signaling pathway JPH203 (KYT-0353), can distinguish the relative abundance of LAT1 and LAT2, and it has high selectivity for LAT1^[2]. JPH203 (KYT-0353) is metabolically stable in liver microsomes of mice, rats, dogs, monkeys and humans^[3]. JPH203 (KYT-0353) induced both G2/M and G0/G1 cell cycle arrest, as well as reduced the S phase accompanied by altered expression of the proteins in cell cycle progression: cyclin D1, CDK4, and CDK6^[4]. Study on leukemia reported that JPH203 (KYT-0353) induced type ?? cell death, autophagy, followed by caspase-3-dependent apoptosis at 48 h after treatment^[6].LAT1 inhibition by JPH203 (KYT-0353) sensitizes cancer cells to radiation by enhancing cellular senescence via mTOR downregulation^[7].

In vivo model, using intravenously administered JPH203 (KYT-0353) (12.5 and 25.0 mg/kg), a statistically significant inhibition of growth of HT-29 tumors transplanted to nude mice with only a slight decrease in body weight^[4].

References:
[1]: Yun DW, Lee SA, et,al. JPH203, an L-type amino acid transporter 1-selective compound, induces apoptosis of YD-38 human oral cancer cells. J Pharmacol Sci. 2014;124(2):208-17. doi: 10.1254/jphs.13154fp. Epub 2014 Feb 4. PMID: 24492461.
[2]: Choi DW, Kim DK, et,al. JPH203, a selective L-type amino acid transporter 1 inhibitor, induces mitochondria-dependent apoptosis in Saos2 human osteosarcoma cells. Korean J Physiol Pharmacol. 2017 Nov;21(6):599-607. doi: 10.4196/kjpp.2017.21.6.599. Epub 2017 Oct 30. PMID: 29200902; PMCID: PMC5709476.
[3]: Wempe MF, Rice PJ, et,al. Metabolism and pharmacokinetic studies of JPH203, an L-amino acid transporter 1 (LAT1) selective compound. Drug Metab Pharmacokinet. 2012;27(1):155-61. doi: 10.2133/dmpk.dmpk-11-rg-091. Epub 2011 Sep 13. PMID: 21914964.
[4]: Yothaisong S, Dokduang H, et,al. Inhibition of l-type amino acid transporter 1 activity as a new therapeutic target for cholangiocarcinoma treatment. Tumour Biol. 2017 Mar;39(3):1010428317694545. doi: 10.1177/1010428317694545. PMID: 28347255.
[5]: Oda K, Hosoda N, et,al. L-type amino acid transporter 1 inhibitors inhibit tumor cell growth. Cancer Sci. 2010 Jan;101(1):173-9. doi: 10.1111/j.1349-7006.2009.01386.x. Epub 2009 Oct 8. PMID: 19900191.
[6]: Rosilio, C, Nebout, M,et al. L-type amino-acid transporter 1 (LAT1): a therapeutic target supporting growth and survival of T-cell lymphoblastic lymphoma/T-cell acute lymphoblastic leukemia. Leukemia 29, 1253–1266 (2015). https://doi.org/10.1038/leu.2014.338
[7]: Bo T, Kobayashi S, et,al. LAT1 inhibitor JPH203 sensitizes cancer cells to radiation by enhancing radiation-induced cellular senescence. Transl Oncol. 2021 Nov;14(11):101212. doi: 10.1016/j.tranon.2021.101212. Epub 2021 Aug 27. PMID: 34461558; PMCID: PMC8405945.