MF63

Biological Activity

MF63 potently inhibited the human mPGES-1 enzyme (IC50=1.3 nM), with a high degree (>1000-fold) of selectivity over other prostanoid synthases. Microsomal prostaglandin E synthase-1 (mPGES-1) is a terminal prostaglandin E (2) (PGE (2)) synthase in the cyclooxygenase pathway. Inhibitors of mPGES-1 may block PGE (2) production and relieve inflammatory symptoms. In rodent species, MF63 strongly inhibited guinea pig mPGES-1 (IC50=0.9 nM) but not the mouse or rat enzyme. In addition, MF63 did not cause NSAID-like gastrointestinal toxic effects, such as mucosal erosions or leakage in the KI mice or nonhuman primates, although it markedly inhibited PGE2 synthesis in the KI mouse stomach.

Product Citations

• 

  Oncotarget. 2016 Jul 12;7(28):44350-44364.

  Linking microsomal prostaglandin E Synthase-1/PGE-2 pathway with miR-15a and -186 expression: Novel mechanism of VEGF modulation in prostate cancer
  MF63 purchased from AbMole

• 

  Am J Physiol Gastrointest Liver Physiol. 2016 Jun 1;310(11):G1071-80.

  Microsomal prostaglandin E synthase-1 protects against Fas-induced liver injury.
  MF63 purchased from AbMole

• 

  Endocr Relat Cancer. 2015 Aug;665-78.

  mPGES-1 in prostate cancer controls stemness and amplifies epidermal growth factor receptor-driven oncogenicity.
  MF63 purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Oncotarget (2016). Figure 4. MF63 (Abmole Bioscience, Houston, TX, USA)
	Method
	Cell Lines	mPGES-1+/+ cells
	Concentrations	10 μM
	Incubation Time	18 h
	Results	Moreover, MF63 increased levels of both miRs in mPGES-1+/+ cells (Figure 4C), and miR-15a and miR-186 were up-regulated with respect to mPGES-1+/+ tumors in DU145 mPGES-1-/- xenografts in vivo (Figure 4D).

	Source	Oncotarget (2016). Figure 3. MF63 (Abmole Bioscience, Houston, TX, USA)
	Method
	Cell Lines	mPGES-1+/+ cells
	Concentrations	10 μM
	Incubation Time
	Results	Consistently,MF63 (10 μM) significantly increased Dicer expression in mPGES-1+/+ cells (Figure 3B), whereas exogenous PGE-2(1 μM, 2–24 h) decreased in mPGES-1-/- cells (Figure 3C).

	Source	Oncotarget (2016). Figure 1. MF63 (Abmole Bioscience, Houston, TX, USA)
	Method
	Cell Lines	mPGES-1+/+ cells
	Concentrations	10 μM
	Incubation Time
	Results	MF63 (10 μM), a reversible mPGES-1 inhibitor, significantly decreased VEGF expression in mPGES-1+/+ cells with maximum effect at 8–18 h (Figure 1D), while exogenous PGE-2 (1 μM) increased VEGF release and expression in mPGES-1-/- cells time-dependently (Figure 1B, 1E).

	Source	Am J Physiol Gastrointest Liver Physiol (2016) . Figure 4.mPGES-1 inhibitor MF63 (Abmole bioscience, Houston, TX)
	Method	Western blotting analysis
	Cell Lines
	Concentrations	1µM
	Incubation Time	6 h
	Results	The above effects were attenuated by pretreatment with the mPGES-1 inhibitor, MF63, in Jo2-treated mPGES-1 Tg mice (Fig. 4A). These findings indicate an important role of EGFR/Akt and associated apoptosis-regulatory molecules in mPGES-1-mediated protection against liver injury.

	Source	Am J Physiol Gastrointest Liver Physiol (2016) . Figure 1.mPGES-1 inhibitor MF63 (Abmole bioscience, Houston, TX)
	Method	mPGES-1 Tg mice oral gavage (twice every 12 h)
	Cell Lines
	Concentrations	50µg/g body weight
	Incubation Time	5 h
	Results	Two doses of MF63 were administrated via oral gavage (every 12h) before Jo2 injection and the mice were sacrificed 5 hours after Jo2 injection (outlined in Fig. 3A). We observed that MF63 pretreatment partially reversed the resistance of mPGES-1 Tg mice to Jo2-induced liver injury (Fig. 3B-F).These findings further support the role of mPGES-1 in modulation of Fas-induced hepatocyte apoptosis and liver injury.

	Source	Endocrinology (2015) . Figure 4. MF63 was from AbMole.
	Method	EIA assay and western blot analysis
	Cell Lines	DU145, PC-3
	Concentrations	10 µ M
	Incubation Time	48 h
	Results	MF63 (10 µmol l-1, 24-48 h) significantly inhibited PGE2 production and reversed the mesenchymal phenotype in DU145 and PC-3 cells, promoting E-cadherin and inhibiting vimentin expression (Fig. 4A, B and C). MF63 also upregulated α6-integrin and significantly reduced cell clonogenicity (Fig.4D, and E).

Protocol (for reference only)

Cell Experiment
Cell lines	A549 cells
Preparation method	A549 cells were seeded in 96-well dishes at a density of 25,000 cells/well and incubated for 20 h in RPMI 1640 medium supplemented with 2% fetal bovine serum (FBS). The cells were then incubated with 10 ng/ml IL-1 for 24 h in the presence of a vehicle or inhibitor from a 100-fold concentrated DMSO stock solution in a medium containing 2 or 50% FBS. Concentrations of PGE2 and PGF2 in the cell-free medium were measured by EIA (Assay Designs Inc.). IC50 values were derived from eight-point titrations.
Concentrations	0.01~100 µ M
Incubation time	24 h

Animal Experiment
Animal models	mPGES-1 Knock-In Mice
Formulation	a suspension in 0.5% methylcellulose
Dosages	100 mg/kg
Administration	oral gavage

Chemical Information

Molecular Weight	378.81
Formula	C23H11ClN4
CAS Number	892549-43-8
Solubility (25°C)	DMSO 42 mg/mL
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month

Conversion of different model animals based on BSA (PMID: 27057123)

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.

References

[1] Xu et al. J Pharmacol Exp Ther. MF63 [2-(6-chloro-1H-phenanthro[9,10-d]imidazol-2-yl)-isophthalonitrile], a selective microsomal prostaglandin E synthase-1 inhibitor, relieves pyresis and pain in preclinical models of inflammation.