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.

This study aimed to investigate the role of the brain-derived neurotrophic factor (BDNF) signaling pathway in promoting the inflammatory response following spinal cord injury (SCI) in rats. The expression level of BDNF in the spinal cord tissue of SCI rats was detected using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The effects of BDNF on SCI were assessed using the Basso, Beattie, and Bresnahan (BBB) test and spinal cord water content measurement. The results showed that BDNF expression levels were elevated in the spinal cord tissue of SCI rats. In the in vitro model, BDNF overexpression induced tyrosine kinase receptor B (TrkB) protein expression, inhibited phosphorylated (p-)p38 protein expression, and reduced the inflammatory response, as indicated by the levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-18, inducible nitric oxide synthase (iNOS), and cyclooxygenase (COX)-2. Conversely, the TrkB inhibitor ANA-12 inhibited TrkB protein expression, induced p-p38 protein expression, and promoted an inflammatory response in an in vitro model of BDNF-induced spinal cord injury (SCI) (as indicated by TNF-α, IL-1β, IL-6, IL-18, iNOS, and COX-2 levels). Furthermore, the p38 inhibitor TA-0 inhibited p38 protein expression and reduced an inflammatory response in an in vitro model of BDNF-induced SCI. These data collectively suggest that the pro-inflammatory effects of BDNF/TrkB promote an inflammatory response in rat spinal cord injury (SCI) through the p38 signaling pathway. [2]

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Background: NRAS mutations activate the MAPK signaling pathway and are the oncogenic driver gene alterations in approximately 20% of melanoma cases in the United States. NRAS-mutant melanomas specifically rely on CRAF rather than BRAF to activate the downstream MEK/ERK signaling pathway. In BRAF-mutant melanoma, approved RAF-targeted therapies are typically used in combination with MEK inhibitors to exert clinical efficacy by inhibiting two targets in the oncogenic MAPK signaling pathway. Emerging data from pan-RAF inhibitors in early clinical development suggest efficacy regardless of combination with MEK inhibitors, but there are currently no approved targeted therapies for patients with NRAS-mutant melanoma. KIN-2787 is a novel oral pan-RAF inhibitor designed to be effective against RAF-dependent cancers, independent of RAF subtypes. Methods: At Reaction Biology, a kinase assay was performed on 688 kinases (including wild-type, atypical, and mutant kinases) using radioenzyme analysis. Cell viability was assessed by inhibiting downstream MAPK pathway signaling and inhibiting cell growth in human tumor cell lines. Combined cell growth inhibition studies were conducted using KIN-2787 and binimetinib in a 9×5 dose matrix. Incucyte imaging was used to assess prolonged cell growth inhibition. We evaluated the in vivo efficacy of KIN-2787 and its combination therapy in an NRAS-mutant xenograft model. Results: Kinomics analysis of KIN-2787 showed extremely high selectivity; at a concentration of 1 μM KIN-2787, only 2 out of 669 non-RAF family kinases were inhibited by more than 75%, and its selectivity windows for DDR1 and p38b were approximately 10-fold and 70-fold, respectively, relative to RAF kinases. We previously reported that KIN-2787 is active against BRAF, NRAS, and KRAS-mutant tumor cell lines, especially sensitive in BRAF models driven by class II and class III dimers. Here, we evaluated the potential of KIN-2787 in combination with binimetinib for the treatment of NRAS-mutant, BRAF wild-type melanoma. The results showed a synergistic effect between melanoma cell lines carrying NRAS hotspot mutations and binimetinib. In NRAS-mutant melanoma xenograft models, daily KIN-2787 combined with binimetinib significantly inhibited tumor growth compared to monotherapy, with inhibition of MAPK pathway biomarkers. Conclusion: KIN-2787 is a highly selective and potent next-generation pan-RAF inhibitor with activity against both BRAF and RAS-mutant human tumor cell models. Preclinical in vitro and in vivo studies of KIN-2787 combined with binimetinib have shown significant efficacy in NRAS-mutant melanoma models. Combined with its unique selectivity, these data support the use of KIN-2787 for combination therapy in this patient population. A phase I/Ib dose-escalation and expansion clinical trial evaluating the safety and efficacy of KIN-2787 is underway (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.)