JPH203 (KYT-0353)
(Synonyms: KYT-0353) 目录号 : GC32690
JPH203 (KYT-0353) 是一种选择性 L 型氨基酸转运蛋白 1 抑制剂，显着抑制 HT-29 YD-38 和白血病细胞的亮氨酸摄取和细胞生长，IC50 值分别为 0.06 μM 和 4.1 μM。
Cas No.:1037592-40-7
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
Saos2 cells |
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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. |
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Reaction Conditions |
Treatment with 100 µM JPH203 (KYT-0353) for 72 hours |
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Applications |
JPH203 (KYT-0353) inhibited colony formation in Saos2 cells. |
| Animal experiment [2]: | |
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Animal models |
Six-week-old male athymic BALB/c nude mice |
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Preparation Method |
Mice in treatment group were injected with JPH203 (KYT-0353) intravenously every day for 20 days. |
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Dosage form |
6.3 mg/kg; 12.5 mg/kg; 25 mg/kg JPH203 (KYT-0353) for 20 days |
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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 . |
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References: [1]. 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. [2]. 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. |
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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.
JPH203 (KYT-0353) 是一种选择性 L 型氨基酸转运蛋白 1 抑制剂，显着抑制 HT-29 YD-38 和白血病细胞的亮氨酸摄取和细胞生长，IC50 值分别为 0.06 μM 和 4.1 μM [1,5]。
JPH203 (KYT-0353) 可上调 Bad、Bax、Bak 和 caspase-9 等促凋亡因子的激活形式，下调 Bcl-2、Bcl-xl、激活线粒体依赖性凋亡信号通路 JPH203 (KYT-0353)，可区分 LAT1 和 LAT2 的相对丰度，对 LAT1[2] 具有高选择性。 JPH203 (KYT-0353) 在小鼠、大鼠、狗、猴子和人类的肝微粒体中代谢稳定[3]。 JPH203 (KYT-0353) 诱导 G2/M 和 G0/G1 细胞周期停滞，并减少 S 期，伴随着细胞周期进程中蛋白质表达的改变:细胞周期蛋白 D1、CDK4 和 CDK6[4 ]。白血病研究报道 JPH203 (KYT-0353) 诱导 ?? 型细胞死亡、自噬，并在处理后 48 h 发生 caspase-3 依赖性细胞凋亡[6]。JPH203 (KYT-0353) 抑制 LAT1 0353) 通过下调 mTOR 增强细胞衰老，使癌细胞对辐射敏感[7]。
体内模型，使用静脉内给药的 JPH203 (KYT-0353)（12.5 和 25.0 mg/kg），对移植到裸鼠的 HT-29 肿瘤的生长有统计学意义的显着抑制，体重仅略有下降[4].
| Cas No. | 1037592-40-7 | SDF | |
| 别名 | KYT-0353 | ||
| Canonical SMILES | N[C@@H](CC1=CC(Cl)=C(C(Cl)=C1)OCC2=C(OC(C3=CC=CC=C3)=N4)C4=CC(N)=C2)C(O)=O | ||
| 分子式 | C23H19Cl2N3O4 | 分子量 | 472.32 |
| 溶解度 | 0.1N hydrochloric acid : 7.3 mg/mL (15.46 mM);Methanol : 6.4 mg/mL (13.55 mM);DMSO : < 1 mg/mL (insoluble or slightly soluble) | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.1172 mL | 10.586 mL | 21.1721 mL |
| 5 mM | 0.4234 mL | 2.1172 mL | 4.2344 mL |
| 10 mM | 0.2117 mL | 1.0586 mL | 2.1172 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
L-type amino acid transporter 1 inhibitors inhibit tumor cell growth
Cancer Sci 2010 Jan;101(1):173-9.PMID:19900191DOI:10.1111/j.1349-7006.2009.01386.x.
Most tumor cell membranes overexpress L-type amino acid transporter 1, while normal cell membranes contain l-type amino acid transporter 2; both are Na(+)-independent amino acid transporters. Therefore, compounds that selectively inhibit L-type amino acid transporter 1 offer researchers with a novel cancer molecular target. Synthetic chemistry efforts and in vitro screening have produced a variety of novel compounds possessing high in vitrol-type amino acid transporter 1 selectivity; KYT-0353 was one such compound. The present studies illustrate that KYT-0353 inhibited (14)C-leucine uptake and cell growth in human colon cancer-derived HT-29 cells; IC(50)s were 0.06 microm and 4.1 microm, respectively. KYT-0353 also inhibited (14)C-leucine uptake in mouse renal proximal tubule cells expressing l-type amino acid transporter 1, and inhibited cell growth; IC(50)s were 0.14 microm and 16.4 microm, respectively. Compared to control animals, intravenously administered KYT-0353 (12.5 mg/kg and 25.0 mg/kg) showed statistically significant growth inhibition against HT-29 tumors transplanted to nude mice with maximal inhibition ratios of 65.9% and 77.2%, respectively. Body weight increase with time--a safety indicator--was slightly depressed at 12.5 mg/kg and 25.0 mg/kg with maximal ratios of 3.7% (day 2) and 6.3% (day 11), respectively. Thus, KYT-0353 showed significant growth inhibitory effects on HT-29 cells both in vitro and in vivo, whereas it only caused a slight body weight depression. Therefore, KYT-0353 appears to have potential as a novel anti-tumor agent, presumably via selective in vivol-type amino acid transporter 1 inhibition.
Investigation of the role of transporters on the hepatic elimination of an LAT1 selective inhibitor JPH203
J Pharm Sci 2013 Sep;102(9):3228-38.PMID:23712732DOI:10.1002/jps.23601.
JPH203 has been developed as an anticancer drug that inhibits L-type amino acid transporter 1-mediated essential amino acid uptake into tumor cells. This study sought to elucidate which drug transporters may be involved in JPH203 hepatic elimination, and to estimate human hepatic clearance. In Sprague-Dawley rats, JPH203 total body clearance approached blood flow rate. JPH203 biotransformation via phase II metabolism produces N-acetyl-JPH203 (NAc-JPH203). NAc-JPH203 accumulates in the bile, and NAc-JPH203 canalicular efflux was significantly decreased in Mrp2-deficient mutant rats (Eisai hyperbilirubinemic rats). JPH203 and NAc-JPH203 are organic anion transporters [organic anion transporting polypeptide (OATP)1B1, OATP1B3, OATP2B1, and OAT3] substrates. In human cryopreserved hepatocytes, JPH203 uptake was saturable and inhibited by rifampicin, a prototypical OATP inhibitor. JPH203 metabolic clearance was larger than influx clearance and eventually passive clearance; JPH203 uptake appears to be the rate-determining process in overall hepatic elimination. Furthermore, unlike rats, the human hepatic clearance was predicted to be intrinsic clearance rate limited. These results suggest that the hepatic uptake transporters are determinant factors to determine JPH203 systemic exposure.
JPH203, a newly developed anti-cancer drug, shows a preincubation inhibitory effect on L-type amino acid transporter 1 function
J Pharmacol Sci 2020 Sep;144(1):16-22.PMID:32653341DOI:10.1016/j.jphs.2020.06.006.
JPH203 is a novel anti-cancer drug targeting L-type amino acid transporter 1 (LAT1), which plays a primary role in the uptake of essential amino acids in tumor cells. Although a co-incubation inhibitory effect of JPH203 has been shown in a conventional uptake assay, its preincubation inhibitory effects have remained undetermined. Therefore, we aimed to characterize the preincubation inhibitory effects of JPH203 on LAT1 function using leucine uptake assays in LAT1-positive human colon cancer HT-29 cells. Preincubation of the cells with JPH203 (0.3 μM for 120 min) decreased the activity level to 30% of that in dimethylsulfoxide-treated cells. Similarly, in time-dependency analysis, preincubation of HT-29 cells with 10 μM JPH203 for 30, 60, and 120 min decreased the leucine uptake activity (42%, 32%, and 28% of that in control cells, respectively). Furthermore, the IC50 value of the combination of preincubation and co-incubation effects was lower than that of co-incubation inhibition alone (34.2 ± 3.6 nM vs. 99.2 ± 11.0 nM). In conclusion, we revealed that JPH203 has the capability to inhibit LAT1 function through preincubation effects. Moreover, preincubation synergistically enhances the co-incubation inhibitory effects. These findings provide a novel insight into the anti-cancer effects of JPH203 in cancer therapy.
Induction of CTH expression in response to amino acid starvation confers resistance to anti-LAT1 therapy in MDA-MB-231 cells
Sci Rep 2022 Jan 19;12(1):1021.PMID:35046465DOI:10.1038/s41598-022-04987-5.
L type amino acid transporter 1 (LAT1) is an attractive molecular target for cancer therapy because of its overexpression in many cancer cells. JPH203, a selective LAT1 inhibitor, causes amino acid deprivation and suppresses cancer cell proliferation. However, several cancer cells showed resistance to amino acid deprivation. In this study, we aimed to elucidate the molecular mechanism of different sensitivity between 2 breast cancer cells to anti-LAT1 therapy. MDA-MB-231 cells were more resistant to growth suppression effect of JPH203 than T-47D cells (IC50 was 200 ± 12.5 μM for MDA-MB-231, and 5 ± 1.1 μM for T-47D cells; p < 0.05). Transcriptome and biochemical analysis were done in these cells in the presence/absence of JPH203. JPH203 induced intracellular amino acid deprivation stress in both cells, but it upregulated cystathionine γ lyase (CTH), an enzyme for synthesis of antioxidants, only in MDA-MB-231 cells. Moreover, siRNA-mediated CTH knockdown induced oxidative stress in response to JPH203 leading to decreased cell viability in MDA-MB-231 cells. These results suggest that activation of anti-oxidation pathways in response to amino acid deprivation confers resistance to anti-LAT1 therapy.
Yeast Cell-Based Transport Assay for the Functional Characterization of Human 4F2hc-LAT1 and -LAT2, and LAT1 and LAT2 Substrates and Inhibitors
Front Mol Biosci 2021 May 28;8:676854.PMID:34124158DOI:10.3389/fmolb.2021.676854.
In mammalian cells, the L-type amino acid transporters (LATs) LAT1 (SLC7A5) and LAT2 (SLC7A8) form heterodimeric amino acid transporters (HATs) with the ancillary protein 4F2hc and are involved in the cellular uptake of specific amino acids. The HAT 4F2hc-LAT1 is found upregulated in various cancer cell types, while 4F2hc-LAT2 is a transporter for non-cancer cells. Preclinical studies have highlighted that 4F2hc-LAT1 plays an important role in tumor progression representing a valid anticancer target. Consequently, current research is focusing on the development of potent and specific human 4F2hc-LAT1 inhibitors. On the other hand, 4F2hc-LAT2 is emerging as target of other diseases, thus also gaining clinical interest. To determine affinity and specificity of substrates and inhibitors for 4F2hc-LAT1 or 4F2hc-LAT2, robust transport cell assays are indispensable. We have optimized and validated a transport assay using cells of the methylotrophic yeast Pichia pastoris stably overexpressing the human HATs 4F2hc-LAT1 or -LAT2, and the LATs LAT1 or LAT2 alone. The radioligand [3H]L-leucine was used as reporter and the substrates L-leucine, triiodothyronine (T3) and thyroxine (T4) as well as the inhibitors BCH and JPH203 (KYT-0353) for assay validation. Obtained half-maximal inhibitory concentrations also provided new insights, e.g., into the LAT specificity of the potent inhibitor JPH203 and on the potency of the thyroid hormones T3 and T4 to inhibit transport through human 4F2hc-LAT2. The LAT1 and LAT2 assays are of particular interest to determine possible implications and influences of 4F2hc in ligand binding and transport. In summary, the presented assays are valuable for characterization of ligands, e.g., towards 4F2hc-LAT1 specificity, and can also be applied for compound screening. Finally, our established approach and assay would also be applicable to other HATs and LATs of interest.