mTORC2 (IC50 = 0.36 μM)

Good anti-GBM capabilities are exhibited by JR-AB2-011 (1 μM; 24 hours), which inhibits mTORC2 signaling and Rictor's interaction with mTOR [1]. Compared to CID613034, JR-AB2-011 (0.5–2 μM; 48 hours) exhibits less toxicity to normal neurons at concentrations up to 10 mM with no discernible cytotoxic effects [1].

Mouse models receiving JR-AB2-011 (4 mg/kg; i.p. daily for 10 days; 20 mg/kg; i.p. daily for 10 days) in each pilot protocol exhibited substantial effects on tumor development rates. Significant results (74% inhibition at the end of the 4 mg/kg/d treatment period; tumor growth delay of 10.0 days; 80% inhibition at the end of the 20 mg/kg/d drug period; tumor growth delay of 12.0 days) [1].

Examination of mTORC2 activity through in vitro kinase assay in rat primary microglia stimulated with rCCL17 or rCCL17 combined with AZD2098. To elucidate the underlying mechanism, the in vitro kinase assay was performed in primary microglia[2].

Apoptosis analysis [1]
Cell Types: U87 GBM cells; LN229 GBM cells
Tested Concentrations: 1 μM
Incubation Duration: 24 hrs (hours)
Experimental Results: Has good anti-GBM properties and blocks mTORC2 signaling and the association of Rictor with mTOR.

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Cytotoxicity assay [1]
Cell Types: normal mature human neurons
Tested Concentrations: 0.5, 1, 2 μM
Incubation Duration: 48 hrs (hours)
Experimental Results: Minimal toxicity to normal neurons, no obvious cytotoxic effect at concentrations up to 10 mM.

Animal/Disease Models: Female CB-17-scid (Taconic) mouse LN229 cells [1]
Doses: 4 mg/kg; 20 mg/kg
Route of Administration: daily intraperitoneal (ip) injection; 10-day
Experimental Results: compared with mice receiving vehicle alone Either dosage regimen demonstrated significant inhibition of tumor growth rate compared to mice.

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SAH rat models were assigned to receive recombinant CCL17 (rCCL17) or phosphate buffer saline (PBS). AZD2098 and JR-AB2-011 were applied to investigate the C-C motif chemokine receptor 4 (CCR4)/mammalian target of rapamycin complex 2 (mTORC2) axis in CCL17-mediated neuroprotection. To elucidate the underlying mechanism, the in vitro kinase assay was performed in primary microglia. Microglial-specific Rictor knockdown was administered via intracerebroventricular injection of adenovirus-associated virus. Brain water content, short-term neurobehavioural evaluation, western blot analysis, quantitative RT-PCR and histological staining were performed[2].

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In vivo Xenograft Experiments[3]
Institutional guidelines for animal welfare and experimental conduct were followed for all animal experiments, which were approved by the Institutional Animal Care and Use Committee and the Regional Administrative Authority under protocol G17/151. All animals received food and water ad libitum. Six weeks old male C57BL/6N mice (n = 20) underwent splenic injection of 2.5 × 105 B16 melanoma cells containing firefly luciferase-expressing plasmid pCHMWS_Luciferase under isoflurane anesthesia and 200 mg/kg metamizole pain treatment. Treatment with 20 mg/kg JR-AB2-011 intraperitoneal (i.p.) or solvent control for 13 days was started one day after tumor inoculation (n = 10). Thirteen days after intrasplenic injection, mice were terminated.

[1]. Benavides-Serrato A, et al. Correction: Specific blockade of Rictor-mTOR association inhibits mTORC2 activity and is cytotoxicin glioblastoma. PLoS One. 2019 Feb 6;14(2):e0212160.
[2]. Zhang A, et al. CCL17 exerts neuroprotection through activation of CCR4/mTORC2 axis in microglia after subarachnoid haemorrhage in rats. Stroke Vasc Neurol. 2022 Jul 26;8(1):4–16.
[3]. Guenzle J, et al. Pharmacological Inhibition of mTORC2 Reduces Migration and Metastasis in Melanoma. Int J Mol Sci. 2020 Dec 22;22(1):30.
[4]. Wu M, et al Dioscin ameliorates murine ulcerative colitis by regulating macrophage polarization. Pharmacol Res. 2021 Oct ; 172:105796.

Despite recent advances in therapy, liver metastasis from melanoma is still associated with poor prognosis. Although targeting the mTOR signaling pathway exerts potent anti-tumor activity, little is known about specific mTORC2 inhibition regarding liver metastasis. Using the novel mTORC2 specific inhibitor JR-AB2-011, we show significantly reduced migration and invasion capacity by impaired activation of MMP2 in melanoma cells. In addition, blockade of mTORC2 induces cell death by non-apoptotic pathways and reduces tumor cell proliferation rate dose-dependently. Furthermore, a significant reduction of liver metastasis was detected in a syngeneic murine metastasis model upon therapy with JR-AB2-011 as determined by in vivo imaging and necropsy. Hence, our study for the first time highlights the impact of the pharmacological blockade of mTORC2 as a potent novel anti-cancer approach for liver metastasis from melanoma.[3]

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Restoring immune balance by targeting macrophage polarization is a potentially valuable therapeutic strategy for ulcerative colitis (UC). Dioscin is a steroidal saponin with potent anti-inflammatory, immunoregulatory, and hypolipidemic effects. This study examined the protective effect of Dioscin on UC in mice and explored the underlying mechanisms. Mice were induced colitis by dextran sulfate sodium (DSS) and concurrently treated with Dioscin oral administration. RAW264.7 cells were skewed to M1 macrophage polarization by lipopolysaccharide (LPS) and interferon-γ (INF-γ) in vitro, and received Dioscin treatment. The results showed that Dioscin ameliorated colitis in mice, reduced macrophage M1 polarization, but markedly promoted M2 polarization in mice colon. Dioscin inhibited mammalian target rapamycin complex 1 (mTORC1)/hypoxia-inducible factor-1α (HIF-1α) signaling and restrained glycolysis in RAW264.7; however, it activated mammalian target rapamycin complex 2 (mTORC2)/peroxisome proliferator-activated receptor-γ (PPAR-γ) signal and facilitated fatty acid oxidation (FAO). The modulation of mTORs signaling may inhibit M1, but promote M2 polarization. Furthermore, the effect of Dioscin on M2 polarization was neutralized by the FAO inhibitor Etomoxir and the mTORC2 inhibitor JR-AB2-011. In parallel, the inhibitory effect of Dioscin on M1 polarization was mitigated by the mTORC1 agonist L-leucine. Both JR-AB2-011 and L-leucine blocked the therapeutic effect of Dioscin in mice with UC. Therefore, Dioscin ameliorated UC in mice, possibly by restraining M1, while skewing M2 polarization of macrophages. Regulation of mTORC1/HIF-1α and mTORC2/PPAR-γ signals is a possible mechanism by which Dioscin inhibited aerobic glycolysis and promoted FAO of macrophages. In summary, Dioscin protected mice against DSS-induced UC by regulating mTOR signaling, thereby adjusting macrophage metabolism and polarization.[4]

C17H14CL2FN3OS

398.281963825226

397.021

C, 51.27; H, 3.54; Cl, 17.80; F, 4.77; N, 10.55; O, 4.02; S, 8.05

2411853-34-2

2411853-34-2;329182-61-8 (wrong);

138319699

Typically exists as white to off-white solids at room temperature

5.2

1

4

2

25

516

0

TWTNZYABDOSOSR-UHFFFAOYSA-N

InChI=1S/C17H14Cl2FN3OS/c1-10-9-21-17(25-10)23(13-5-2-11(20)3-6-13)16(24)22-12-4-7-14(18)15(19)8-12/h2-8,10H,9H2,1H3,(H,22,24)

(Z)-N-(3,4-dichlorophenyl)-2-((4-fluorophenyl)imino)-5-methylthiazolidine-3-carboxamide

JRAB2011 JR AB2 011 JR-AB2-011

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~62.5 mg/mL (~156.92 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (5.22 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (5.22 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)