TGFβRII; p38 MAPK (IC50 = 20 nM)

TA-02 (5 μM) inhibits the phosphorylation of MAPKAPK2 and HSP27, proteins that are phosphorylated by the p38α MAPK during cardiogenesis. Contrary to what would be predicted by a mechanism dependent on p38α MAPK inhibition, TA-02 at a concentration of 5 μM induces cardiogenesis while also increasing ATF-2 phosphorylation and MEF2C õexpression[1].
TA-02 induces T/Brachyury, whereas SB203580 addition increased MESP1 and T/Brachyury transcripts[1].
TA-02 significantly raises NKX2-5 expression when applied between days 0 and 8[1].
TA-02 is discovered to inhibit numerous targets with similar potency to p38α MAPK, including p38α, p38β, JNK3, JNK2, CIT, CK1ε, DMPK2, JNK1, DDR1 CK1δ, MEK5, and ERBB2[1].
Nuclear TCF/LEF-1 driven transcription of the DKK-1-like luciferase is inhibited by TA-02 and SB203580[1].
In vitro, TA-02 (5 nM–5 M) increases BDNF's anti-inflammation effects while inhibiting p38[2].

In vitro model and transfection [2]
The nerve cell line AGE1.HN was cultured in Dulbecco's modified Eagle's medium medium supplemented with 10% fetal bovine serum, 100 U/ml penicillin and 100 µg/ml streptomycin, and maintained at 37°C in a humidified atmosphere with 5% CO2. BDNF-pcDNA3.1 (forward, 5′-AGAAAAGCCAAFFAGTGAA-3′ and reverse, 5′-AAAAGGGGAAGATAGTGGATTTATGTT-3′) and negative-pcDNA3.1 negative mimic plasmids (forward, 5′-CCCCCCCCCCCCCCCCCC-3′ and reverse, 5′-CCCCCCCCCCCCCCCCCC-3′) were constructed by Sangon Biotech Co., Ltd.,. Cells were transfected with 100 ng BDNF plasmid or negative mimics using Lipofectamine 2000, according to the manufacturer's protocol. Following 48 h transfection, cells were induced with 100 ng/ml lipopolysaccharide for 4 h at 37°C. Cells were treated with 10 µM ANA-12, a TrkB inhibitor, or 5 nM TA-02, a p-38 inhibitor for 44 h at 37°C, and cells were inducted with 100 ng/ml LPS for 4 h at 37°C, respectively and then were induced by 100 ng/ml LPS for 4 h at 37°C. Negative group, cell was transfected with negative mimics.

[1]. Cardiomyocyte differentiation of pluripotent stem cells with SB203580 analogues correlates with Wnt pathway CK1 inhibition independent of p38 MAPK signaling. J Mol Cell Cardiol. 2015 Mar;80:56-70.

[[2]. The activation of BDNF reduced inflammation in a spinal cord injury model by TrkB/p38 MAPK signaling. Exp Ther Med. 2019 Mar;17(3):1688-1696.

The aim of the present study was to investigate the pro-inflammation effects of brain-derived neurotrophic factor (BDNF) signaling in promoting inflammation following spinal cord injury (SCI) in rats. Reverse transcription-quantitative polymerase chain reaction was used to detect the expression of BDNF in SCI rats. The Basso, Beattie and Bresnahan (BBB) test was used and the water content of spinal cord were assessed to determine the effects of BDNF on SCI. BDNF expression was increased in SCI rats. In an in vitro model, overexpression of BDNF induced the protein expression of tyrosine kinase receptor B (TrkB) and suppressed that of phosphorylated (p-)p38, and reduced inflammation, as indicated by tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-18, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 levels. Conversely, the TrkB inhibitor ANA-12 suppressed the protein expression of TrkB and induced that of p-p38, and promoted inflammation (as indicated by TNF-α, IL-1β, IL-6, IL-18, iNOS and COX-2 levels) in an in vitro model of SCI by BDNF overexpression. In addition, the p38 inhibitor TA-0, suppressed p38 protein expression and reduced inflammation in an in vitro model of SCI by BDNF overexpression. Together, these data suggest that the pro-inflammation effects of BDNF/TrkB promoted inflammation in SCI through p38 signaling in rats.[2]

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Background: NRAS mutations which activate MAPK signaling represent oncogenic driver alterations in approximately 20% melanoma cases in the US. NRAS mutant melanomas are uniquely dependent upon CRAF rather than BRAF for activation of downstream MEK/ERK signaling. In BRAF mutant melanoma, approved RAF-targeted therapies are commonly used in combination with a MEK inhibitor which provides clinical benefit by inhibition of two targets within the oncogenic MAPK signaling pathway. Emerging data with pan-RAF inhibitors in early clinical development suggests benefit with and without combined MEK inhibition, yet no approved targeted therapy exists for NRAS mutant melanoma patients. KIN-2787 is a novel, orally available pan-RAF inhibitor designed to be effective in RAF-dependent cancers, regardless of isoform. Methods: Kinome profiling was evaluated by radiometric enzyme assay at Reaction Biology across 688 kinases (including wild type, atypical, and mutant). Cellular activity was assessed by suppression of downstream MAPK pathway signaling and cell growth inhibition in human tumor cell lines. Combination cell growth inhibition studies were performed in 9x5 dose matrices with KIN-2787 and binimetinib, respectively. Extended cell growth inhibition effects were assessed by Incucyte imaging. In vivo KIN-2787 and combination efficacy was evaluated in NRAS mutant xenograft models. Results: Kinome profiling of KIN-2787 revealed exquisite selectivity with only 2 of 669 non-RAF family kinases inhibited > 75% at 1 M KIN-2787 and retained ̃10x and 70x selectivity window against those two kinases, DDR1 and p38b, respectively, relative to RAF kinases. We previously reported KIN-2787 activity across BRAF, NRAS, and KRAS mutant tumor cell lines with greatest sensitivity in Class II and III dimer-driven BRAF models. Here, we evaluated NRAS mutant, BRAF WT melanoma for combination potential with binimetinib. Melanoma tumor cell lines bearing NRAS hotspot mutations demonstrated synergistic benefit with KIN-2787 combined with binimetinib. Daily KIN-2787 plus binimetinib treatment in NRAS mutant melanoma xenograft models resulted in significant tumor growth inhibition benefit relative to either agent alone and was associated with added MAPK pathway biomarker suppression. Conclusions: KIN-2787 is a highly selective, potent, next-generation, pan-RAF inhibitor with activity across BRAF and RAS mutant human tumor cell models. Preclinical in vitro and in vivo studies using KIN-2787 in combination with binimetinib demonstrated significant combination benefit in NRAS mutant melanoma models. Taken together with its unique selectively, these data support use of KIN-2787 in combination therapy in this patient segment. A Phase 1/1b dose escalation and expansion clinical trial evaluating the safety and efficacy of KIN-2787 is ongoing (NCT04913285). [1]

C20H13F2N3

333.3341

333.107

C, 72.06; H, 3.93; F, 11.40; N, 12.61

1784751-19-4

TA-01;1784751-18-3

91691130

White to off-white solid powder

1.3±0.1 g/cm3

515.7±50.0 °C at 760 mmHg

265.7±30.1 °C

0.0±1.3 mmHg at 25°C

1.619

4.99

1

4

3

25

421

0

FC1=C([H])C([H])=C([H])C([H])=C1C1=NC(C2C([H])=C([H])C(=C([H])C=2[H])F)=C(C2C([H])=C([H])N=C([H])C=2[H])N1[H]

QIFJOFNVIVQRNJ-UHFFFAOYSA-N

InChI=1S/C20H13F2N3/c21-15-7-5-13(6-8-15)18-19(14-9-11-23-12-10-14)25-20(24-18)16-3-1-2-4-17(16)22/h1-12H,(H,24,25)

4-[2-(2-fluorophenyl)-4-(4-fluorophenyl)-1H-imidazol-5-yl]pyridine

TA 02; TA02; TA-02; 1784751-19-4; 4-(2-(2-Fluorophenyl)-4-(4-fluorophenyl)-1H-imidazol-5-yl)pyridine; 4-[2-(2-fluorophenyl)-4-(4-fluorophenyl)-1H-imidazol-5-yl]pyridine; MFCD29924749; SCHEMBL17002317; TA-02

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)

DMSO: 25~67 mg/mL (75.0~201.0 mM)
Ethanol: ~3 mg/mL (~9.0 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.50 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 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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Solubility in Formulation 2: ≥ 2.5 mg/mL (7.50 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 25.0 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.)