Cell lines

Human cell MDA PCa 2b, PC-3 lines.

Preparation method

The solubility of this compound in DMSO is<10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months..

Reacting condition

24h; 4 μM

Applications

A crucial step in the transduction of TGF-β 1 signals is the phosphorylation of receptor-activated Smad2 and Smad3. We thus assessed the phosphorylation of Smad2 in lysates of MDA PCa 2b cells, PC-3 cells, and PMOs treated with rhTGF- β1. We found that TGF-β1 induces phosphorylation of Smad2 in PC-3 cells and PMOs but not in MDA PCa 2b cells. Further, treatment with LY2109761 reverses the Smad2 phosphorylation induced by rhTGF-β1. In other words, LY2109761 inhibits TGF-β1–induced Smad2 activation in PC-3 cells and PMOs.

Animal models

Male SCID mice

Dosage form

200 mg/kg/day; oral taken

Application

After 3 weeks of treatment, X-ray analysis of the vehicle control group revealed two broken bones and loss of 30%–70% of the radiopaque areas in the tumor-bearing bones. MRI analysis showed a significantly smaller tumor volume in the treated group than in the controls (p =0.012). Micro-CT analysis of the tumor-bearing bones of the controls and treated mice demonstrated significantly lower BV (p=0.00043), BMC (p =0.000132), and BMD (p = 0.000085) in the control mice. Furthermore, BV, BMC, and BMD in the treated group were restored to values found in the normal (uninjected) femurs, which supports the efficacy of treatment. Finally, bone histomorphometric analysis demonstrated that LY2109761 inhibited PC-3-induced activation of osteoclasts.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

LY2109761 is a small-molecule inhibitor selectively targeting both TGF-β receptor type I and II ((TβRI/II)) with Ki of 38 nM and 300 nM, respectively [1]. It gave IC50 value of 69 nM in TβRI enzymatic assay [2]. Crystal structure showed the binding of LY2109761 to the ATP binding site of the TGF-β R1 kinase domain [2]. Weak inhibitory activities were reported for other kinases, including Lck, Sapk2α, MKK6, Fyn, and JNK3 (18–89% inhibition at 20 μM) [2].

LY2109761 has shown potential anti-tumor activity in preclinical studies. Deregulation of TGF-β signaling pathway is correlated with various malignant. LY2109761 suppressed proliferation, migration and invasion, and induced apoptosis of pancreatic cancer cells [1]. When combined with gemcitabine, it inhibited tumor growth and metastasis in a mouse model of metastatic pancreatic cancer [1]. It also inhibited the anti-apoptotic effects of TGF-beta1 in myelo-monocytic leukaemic cells [3]. Results in GBM cell lines and an orthotopic intracranial model indicated that LY2109761 increased radiosensitivity and resulted in prolonged survival in glioblastoma [4]. In addition, LY2109761 reduced radiation-induced pneumonitis and pulmonary fibrosis in a murine model [5].

References:
[1]Melisi D, Ishiyama S, Sclabas GM et al. LY2109761, a novel transforming growth factor beta receptor type I and type II dual inhibitor, as a therapeutic approach to suppressing pancreatic cancer metastasis. Mol Cancer Ther 2008; 7: 829-840.[2]Li HY, McMillen WT, Heap CR et al. Optimization of a dihydropyrrolopyrazole series of transforming growth factor-beta type I receptor kinase domain inhibitors: discovery of an orally bioavailable transforming growth factor-beta receptor type I inhibitor as antitumor agent. J Med Chem 2008; 51: 2302-2306.[3]Xu Y, Tabe Y, Jin L et al. TGF-beta receptor kinase inhibitor LY2109761 reverses the anti-apoptotic effects of TGF-beta1 in myelo-monocytic leukaemic cells co-cultured with stromal cells. Br J Haematol 2008; 142: 192-201.[4]Zhang M, Kleber S, Rohrich M et al. Blockade of TGF-beta signaling by the TGFbetaR-I kinase inhibitor LY2109761 enhances radiation response and prolongs survival in glioblastoma. Cancer Res 2011; 71: 7155-7167.[5]Flechsig P, Dadrich M, Bickelhaupt S et al. LY2109761 attenuates radiation-induced pulmonary murine fibrosis via reversal of TGF-beta and BMP-associated proinflammatory and proangiogenic signals. Clin Cancer Res 2012; 18: 3616-3627.