AG490

Biological Activity

AG490 (Tyrphostin AG490) is a selective inhibitor of EGF receptor tyrosine kinase with an IC50 values of 2 and 13.5 μM for EGFR and ErbB2 respectively. AG490 is also a JAK3/STAT, JAK3/AP-1 and JAK3/MAPK signaling pathway inhibitor and also blocks JAK3 autophosphorylation. AG-490 significantly inhibits the constitutive activation of Stat3 in MOPC, MPC11, and S194 cells, leading to dramatic dose-dependent apoptosis. AG-490 (100 μM) inhibits Akt phosphorylation, inhibits the activation of nuclear factor-κB, and causes the activation of GSK-3β, leading to the reduction of c-Myc. AG-490 (50 μM) can induce apoptosis of imatinib-resistant BaF3 cells expressing T315I and E255K mutants of Bcr-Abl.

Product Citations

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  J Tradit Chin Med. 2019 April 15;39(2): 181-190.

  Efficacy of Zhonglun'a-decoction-containing serum on fibroblast-like synoviocyte apoptosis in rats with collagen-induced arthritis via inhibiting Janus kinase/signal transducer and activator of transcription signaling pathway.
  AG490 purchased from AbMole

• 

  Med Sci Monit. 2018 Sep 8;24:6255-6263.

  Inhibition of JAK2/STAT3 Signaling Pathway Suppresses Proliferation of Burkitt's Lymphoma Raji Cells via Cell Cycle Progression, Apoptosis, and Oxidative Stress by Modulating HSP70.
  AG490 purchased from AbMole

• 

  Nutr Res. 2018 Mar 18.

  Folic acid supplementation repressed hypoxia-induced inflammatory response via ROS and JAK2/STAT3 pathway in human pro-myelomonocytic cells
  AG490 purchased from AbMole

• 

  Int J Mol Med. 2017 Feb;39(2):307-316.

  Matrine induces the apoptosis of fibroblast-like synoviocytes derived from rats with collagen-induced arthritis by suppressing the activation of the JAK/STAT signaling pathway
  AG490 purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Med Sci Monit (2018). Figure 5. AG490 (Abmole Bioscience)
	Method	DCFH-DA assays
	Cell Lines	Raji cells
	Concentrations	5 μM
	Incubation Time	24 h, 36 h, 48 h, and 72 h
	Results	A result that was consistent with the effect of the JAK2/STAT3 signaling pathway inhibitor AG490 (P>0.05).

	Source	Med Sci Monit (2018). Figure 4. AG490 (Abmole Bioscience)
	Method	Annexin V-FITC/PI staining
	Cell Lines	Raji cells
	Concentrations	5 μM
	Incubation Time	24 h, 36 h, 48 h, and 72 h
	Results	According to the results of Annexin V-FITC/PI staining (Figure 4A), the HSP70 siRNA group (55.2±3.1%) and AG490 group (60.8±4.2%) showed a remarkably higher cell apoptosis rate than the Blank group (14.5±1.3%) and NC siRNA group.

	Source	Med Sci Monit (2018). Figure 3. AG490 (Abmole Bioscience)
	Method	MTT assay
	Cell Lines	Raji cells
	Concentrations	5 μM
	Incubation Time	24 h, 36 h, 48 h, and 72 h
	Results	As displayed by the MTT assay (Figure 3A), both HSP70 siRNA and AG490 could effectively reduce the proliferation of Raji cells (24 h, 36 h, 48 h, and 72 h) (all P<0.05), and no significant difference was found between the Blank group and NC siRNA group regarding Raji cell proliferation at different time points.

	Source	Med Sci Monit (2018). Figure 2. AG490 (Abmole Bioscience)
	Method	Western blotting
	Cell Lines	Raji cells
	Concentrations	5 μM
	Incubation Time	24 h
	Results	As shown by the results of Western blotting (Figure 2), AG490 down-regulated the expression of p-JAK2 and p-STAT3.

	Source	Nutr Res (2018). Figure 4. AG490 (AbMole BioScience, Houston, TX, USA)
	Method	Cell viability assay
	Cell Lines	THP-1 cells
	Concentrations	10 μM
	Incubation Time	24 h
	Results	p-STAT3 protein levels were repressed by folic acid and AG490, and the trend for repression was consistent with p-JAK2.

	Source	China Tropical Medicine (2017). Figure 5~6. AG490 (Abmole Bioscience, Shanghai, China)
	Method
	Cell Lines	THP-1 cell
	Concentrations	10 μmol/L
	Incubation Time	30 min
	Results	The expression of NF-κB p65 protein in p-JAK2, p-STAT3 and nucleus was significantly lower than that in hypoxia group (P <0.05), while the expression of JAK2 and STAT3 The protein expression did not affect.

	Source	China Tropical Medicine (2017). Figure 1~4. AG490 (Abmole Bioscience, Shanghai, China)
	Method
	Cell Lines	THP-1 cell
	Concentrations	10 μmol/L
	Incubation Time	30 min
	Results	The secretion and mRNA expression of IL-1β and TNF-α were significantly lower than those of hypoxia group (P <0.05), and the difference was statistically significant (P <0.05)

	Source	Int J Mol Med (2017) . Figure 6. AG490 (Abmole Bioscience, Houston, TX, USA)
	Method	qRT-PCR
	Cell Lines	Fibroblast-like synoviocytes (FLS)
	Concentrations	25 μM
	Incubation Time	24 h
	Results	Treatment with matrine, AG490, or matrine+AG490 markedly increased the mRNA expression of Bax and caspase-3, and decreased expression of Bcl-2 (P<0.01 vs. CIA alone).

	Source	Int J Mol Med (2017) . Figure 5. AG490 (Abmole Bioscience, Houston, TX, USA)
	Method	Western blot
	Cell Lines	Fibroblast-like synoviocytes (FLS)
	Concentrations	25 μM
	Incubation Time	24 h
	Results	Protein expression levels of p-JAK2, p-STAT1 and p-STAT3 in FLS isolated after 24 h of drug treatment were only faintly detectable in the control group, and markedly higher in the CIA group (P<0.01 vs. control; Fig. 5).

	Source	Int J Mol Med (2017) . Figure 4. AG490 (Abmole Bioscience, Houston, TX, USA)
	Method	propidium iodide (PI) staining and flow cytometry
	Cell Lines	Fibroblast-like synoviocytes (FLS)
	Concentrations	25 μM
	Incubation Time	24 h
	Results	All drug treatments resulted in signifcantly higher apoptosis rates than that observed for the CIA group (P<0.01; matrine, 20.10±0.77%; AG490, 20.42±0.91%; matrine+AG490, 25.11±1.00%).

	Source	Int J Mol Med (2017) . Figure 3. AG490 (Abmole Bioscience, Houston, TX, USA)
	Method	propidium iodide (PI) staining and flow cytometry
	Cell Lines	Fibroblast-like synoviocytes (FLS)
	Concentrations	25 μM
	Incubation Time	24 h
	Results	Compared with the CIA group, the percentages of FLS in the S phase were significantly decreased in cells treated with matrine, the specifc JAK2 inhibitor AG490 (inhibits phosphorylation of STAT1/3 in RA-FLS), or both for 24 h (P<0.01; Fig. 3).

Protocol

Cell Experiment
Cell lines	ALL cells
Preparation method	cell proliferation assay as described previously
Concentrations	0~50μM
Incubation time	16 h

Animal Experiment
Animal models	ALL cells tumour xenograft SCID mice
Formulation	DMSO
Dosages	0.85mg/kg +0.5mg/kg daily
Administration	i.p.

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

Chemical Information

References

[1] Fernandez-Sanchez et al. Curr Med Chem. AG490 Promotes HIF-1α Accumulation by Inhibiting Its Hydroxylation.

[2] Davoodi-Semiromi et al. J Clin Immunol. Tyrphostin AG490 Agent Modestly but Significantly Prevents Onset of Type 1 in NOD Mouse; Implication of Immunologic and Metabolic Effects of a Jak-Stat Pathway Inhibitor.