c-Met (Ki = 355 nM)

ARQ-197 treatment results in decreased tumor growth in all three xenograft models: 66% in the HT29 model, 45% in the MKN-45 model, and 79% in the MDA-MB-231 model. Following oral administration of ARQ-197 at 200 mg/kg, no appreciable changes in body weight are seen in these xenograft studies. A significant decrease in c-Met autophosphorylation occurs 24 hours following an oral dosage of 200 mg/kg of ARQ-197, indicating that the compound has a pharmacodynamically strong inhibitory effect on c-Met phosphorylation in human colon xenograft tumors (HT29). The application of the same dosage in mice results in tumor xenografts being exposed to sustained plasma levels of ARQ-197, which is in line with the pharmacodynamic inhibition of c-Met phosphorylation and the inhibition of the growth of cancer cell lines harboring c-Met. Ten hours following dosage, plasma levels of ARQ-197 are found to be 1.3 μM, which is more than three times higher than the drug's biochemical inhibitory constant for c-Met. Consequently, in vivo suppression of the target by ARQ-197 is achieved in the xenografted human tumor tissue. Finally, it can be said that ARQ-197 prevents c-Met-dependent xenografted human tumors from growing.[1]

For thirty minutes at room temperature, recombinant c-Met protein (100 ng) is preincubated with increasing concentrations of ARQ-197. After preincubation, the reaction mixture is mixed with different concentrations of ATP containing 5 μCi of [γ-32P]ATP and 100 μM of poly-Glu-Tyr substrate. After five minutes of room temperature incubation, the reaction is halted by adding five microliters of SDS-polyacrylamide gel, which reduces the sample buffer. After that, the samples are put onto a 7.5% acrylamide gel, and SDS-PAGE is carried out. The final method used to visualize the phosphorylated poly-Glu-Tyr substrates is autoradiography. Densitometry is used to measure c-Met activity.

In black 96-well plates, 5 × 10³ cells are seeded per well and left overnight in a medium containing 10% FBS for HT29, MKN-45, and MDA-MB-231 cells. Next day, cells are exposed to progressively higher concentrations of ARQ-197 (0.03-10 μM) at 37 °C for 24, 32, and 48 hours. Following treatment with ARQ-197, the drug-containing medium is removed, and the cells are incubated in a labeling solution (10 mM HEPES, 140 mM NaCl, and 6 mM CaCl₂) containing 1 μg/mL propidium iodide (red channel), 500-times diluted Annexin V-FITC (green channel), and 2 μg/mL Hoescht 33342 (blue channel). This process continues for at least 10 minutes. Acquisition and analysis of high-content images are done. Four pictures are supposed to be taken per well by the program. For the 4,6-diamidino-2-phenylindole, FITC, and rhodamine channels, the exposure times are set at 16.7 ms/10% gain, 500 ms/35% gain, and 300 ms/30% gain, respectively. After processing the images, the quantity of positive cells in each channel and condition is calculated. Furthermore, the identical protocols are carried out when HT29 cells are treated for 32 hours with varying doses of ARQ-197 in the presence or absence of 25, 50, and 100 μM ZvAD-FMK (irreversible general caspase inhibitor). Each experiment is carried out three times. The impact of ARQ-197 when glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and c-Met are knocked down using siRNA is examined in order to ascertain whether the apoptotic effect is caused by c-Met inhibition. SiRNAs that are nontargeted, gapgh-targeted, or met-targeted are transfected into HT29, MKN-45, and MDA-MB-231 cells. Following three days, particular antibodies are used to measure the expression levels of β-actin, GAPDH, and c-Met. After transfecting HT29, MKN-45, and MDA-MB-231 cells with a met-targeted siRNA for two days, the cells are cultured either in the absence or with increasing concentrations of ZvAD-FMK for an additional day to ascertain whether the effect is caspase dependent. Also transfected in parallel as controls are a nontargeted siRNA and a gapgh-targeted siRNA (siRNA GAPDH). Annexin V-FITC and propidium iodide are then used to stain the cells, allowing for the calculation of the percentage of apoptotic cells.

[1]. Mol Cancer Ther . 2010 Jun;9(6):1544-53.

[2]. Nat Rev Drug Discov . 2008 Jun;7(6):504-16.

LSM-1131 is a member of indoles.
Tivantinib has been investigated in Solid Tumors.
Tivantinib is an orally bioavailable small molecule inhibitor of c-Met with potential antineoplastic activity. c-Met inhibitor ARQ 197 binds to the c-Met protein and disrupts c-Met signal transduction pathways, which may induce cell death in tumor cells overexpressing c-Met protein or expressing constitutively activated c-Met protein. c-Met protein, the product of the proto-oncogene c-Met, is a receptor tyrosine kinase also known as hepatocyte growth factor receptor (HGFR); this protein is overexpressed or mutated in many tumor cell types and plays key roles in tumor cell proliferation, survival, invasion, and metastasis, and tumor angiogenesis.

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Drug Indication
Treatment of hepatoblastoma

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Mechanism of Action
Tivantinib mediates its effects by inhibiting the activity of c-Met, a receptor tyrosine kinase that plays multiple key roles in human cancer, including cancer cell growth, survival, angiogenesis, invasion and metastasis. C-Met is abnormally activated in most cancers and is believed to control multiple signal transduction pathways involved in tumor growth and metastasis.

C23H19N3O2

369.42

369.147

C, 74.78; H, 5.18; N, 11.37; O, 8.66

905854-02-6

(3S,4S)-Tivantinib;905854-03-7;(Rac)-Tivantinib;1239986-50-5;(rel)-Tivantinib;905853-99-8

11494412

white solid powder

1.5±0.1 g/cm3

716.0±60.0 °C at 760 mmHg

386.8±32.9 °C

0.0±2.3 mmHg at 25°C

1.797

3.26

2

2

2

28

666

2

O=C1NC(=O)[C@@H](C2=CNC3C=CC=CC2=3)[C@@H]1C1=CN2CCCC3C2=C1C=CC=3

UCEQXRCJXIVODC-PMACEKPBSA-N

InChI=1S/C23H19N3O2/c27-22-19(16-11-24-18-9-2-1-7-14(16)18)20(23(28)25-22)17-12-26-10-4-6-13-5-3-8-15(17)21(13)26/h1-3,5,7-9,11-12,19-20,24H,4,6,10H2,(H,25,27,28)/t19-,20-/m0/s1

(3R,4R)-3-(1-azatricyclo[6.3.1.04,12]dodeca-2,4,6,8(12)-tetraen-3-yl)-4-(1H-indol-3-yl)pyrrolidine-2,5-dione

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.77 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 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 25.0 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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 (Please use freshly prepared in vivo formulations for optimal results.)