Mirdametinib (PD0325901)

Licensed by Pfizer Catalog No.S1036

For research use only.

Mirdametinib (PD0325901) is a selective and non ATP-competitive MEK inhibitor with IC50 of 0.33 nM in cell-free assays, roughly 500-fold more potent than CI-1040 on phosphorylation of ERK1 and ERK2. Phase 2.

Mirdametinib (PD0325901) Chemical Structure

CAS No. 391210-10-9

Selleck's Mirdametinib (PD0325901) has been cited by 669 publications

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Biological Activity

Description Mirdametinib (PD0325901) is a selective and non ATP-competitive MEK inhibitor with IC50 of 0.33 nM in cell-free assays, roughly 500-fold more potent than CI-1040 on phosphorylation of ERK1 and ERK2. Phase 2.
Targets
MEK [1]
(Cell-free assay)
0.33 nM
In vitro

PD0325901 shows higher permeability than CI-1040, another MEK inhibitor. PD0325901 should be able to achieve higher systemic exposures than CI-1040. [1] PD0325901 is exquisitely specific and highly potent against purified MEK, revealing a Kiapp of 1 nM against activated MEK1 and MEK2. [2] PD0325901 is roughly 500-fold more potent than CI-1040 with respect to its cellular effects on phosphorylation of ERK1 and ERK2, displaying subnanomolar activity. [2] PD0325901 prevents the growth of melanoma cell lines. PD0325901 inhibits the growth of TPC-1 cells and K2 cells with GI50 of 11 nM and 6.3 nM, respectively. [3] PD0325901 significantly prevents the the growth of PTC cells harboring a BRAF mutation at very low concentration (10 nM) and only moderately increases the growth of the PTC cells carrying the RET/PTC1 rearrangement at the same concentration. PD0325901 effectively inhibits the phosphorylation of ERK1/2 in multiple PTC cell lines. [3]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
A549 NIP0R3pKdmirYnn0bY9vKG:oIF3FT|EhcW5iaIXtZY4> NVXhT|l2OTBidV2= MoS5NUBpeg>? M1LTb2lvcGmkaYTpc44hd2ZiTVXLNUBqdiCqdX3hckBCPTR7IHPlcIx{KGG|c3Xzd4VlKGG|IILl[JVkfGmxbjDpckBxTXKtMT:yJIxmfmWuIHH0JFExKHWPIHHmeIVzKDFiaIKgZpkhX2W|dHXyckBjdG:2dHnu[{BidmGueYPpdy=> MmftQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjV5Nk[2N|MoRjJ3N{[2OlM{RC:jPh?=
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A549 M17a[GlvcGmkaYTpc44hd2ZiTVXrNU8zKGmwIHj1cYFv NWjG[IdOPzVibXnudy=> M3LIOmlvcGmkaYTpc44hd2ZiTVXrNU8zKGmwIHj1cYFvKEF3NEmgZ4VtdHNiYYPz[ZN{\WRiYYOg[IVkemWjc3WgbY4hTVKNIIDoc5NxcG:{eXzheIlwdiCjZoTldkA4PSCvaX7zJIJ6KFenc4Tldo4h[myxdITpcoch[W6jbInzbZM> NFjrN209[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{M{O5PFQ2Oyd-MkOzPVg1PTN:L3G+
CCD-1070sk NVLpd|FqUW6qaXLpeIlwdiCxZjDNSYsyNzJiaX6gbJVu[W5? MnjGO|UhdWmwcx?= NXnOO4toUW6qaXLpeIlwdiCxZjDNSYsyNzJiaX6gbJVu[W5iQ1PEMVExPzC|azDj[YxteyCjc4Pld5Nm\CCjczDk[YNz\WG|ZTDpckBGWkticHjvd5Bpd3K7bHH0bY9vKGGodHXyJFc2KG2rboOgZpkhX2W|dHXyckBjdG:2dHnu[{BidmGueYPpdy=> MlrhQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjN|OUi0OVMoRjJ|M{m4OFU{RC:jPh?=
A375 MkXHTY5pcWKrdHnvckBw\iCPRXuxM|IhcW5iaIXtZY4> Ml;KO|UhdWmwcx?= MVvJcohq[mm2aX;uJI9nKE2Ha{GvNkBqdiCqdX3hckBCOzd3IHPlcIx{KGG|c3Xzd4VlKGG|IHTlZ5Jm[XOnIHnuJGVTUyCyaH;zdIhwenmuYYTpc44h[W[2ZYKgO|UhdWmwczDifUBY\XO2ZYLuJIJtd3S2aX7nJIFv[Wy7c3nz M4DzeVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ|M{m4OFU{Lz5{M{O5PFQ2OzxxYU6=
MCF7 NV;obJY4UW6qaXLpeIlwdiCxZjDNSYsyNzJiaX6gbJVu[W5? Ml3NO|UhdWmwcx?= NFfScoZKdmirYnn0bY9vKG:oIF3Fb|EwOiCrbjDoeY1idiCPQ1[3JINmdGy|IHHzd4V{e2WmIHHzJIRm[3KnYYPlJIlvKEWUSzDwbI9{eGixconsZZRqd25iYX\0[ZIhPzVibXnud{BjgSCZZYP0[ZJvKGKub4T0bY5oKGGwYXz5d4l{ MXe8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzN7OES1N{c,OjN|OUi0OVM9N2F-
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Assay
Methods Test Index PMID
Western blot p-ERK / p-AKT / Cyclin D1 / p27 ; p-p70S6K(T389) / p4EBP1(S65) / pS6(S235) ; cleaved PARP ; DR5 22573716 27699948 25867065
Immunofluorescence Fibronectin / Actin / pFAK ; ZO-1 28187762 25394671
In vivo The improved potency of PD0325901 relative to CI-1040 is evident. A single oral dose of PD0325901 (25 mg/kg) inhibits phosphorylation of ERK by more than 50% at 24 hours post-dosing. In contrast, CI-1040 at a much higher dose (150 mg/kg) only inhibit pERK levels for roughly 8 hours, returning to control levels by 24 hours after treatment. [2] Therefore, the dose required to produce a 70% incidence of complete tumor responses (C26 model) is 25 mg/kg/day versus 900 mg/kg/day for PD0325901 and CI-1040, respectively. Anticancer activity of PD 0325901 has been demonstrated for a broad spectrum of human tumor xenografts. [2] After 1 week of oral administration of PD0325901 (20–25 mg/kg/day) in mice, no tumor growth is detected in mice inoculated with PTC cells bearing a BRAF mutation. [3] For PTC with the RET/PTC1 rearrangement, the average tumor volume of the orthotopic tumor is decreased by 58% as compared with controls. In conclusion, PTC cells carrying a BRAF mutation are more sensitive to PD0325901 than are PTC cells carrying the RET/PTC1 rearrangement. [3]

Protocol (from reference)

Kinase Assay:

[1]

  • In vitro cascade assay:

    Incorporation of 32P into myelin basic protein (MBP) is assayed in the presence of a glutathione S-transferase fusion protein containing p44MAP kinase (GST-MAPK) and a glutathione S-transferase protein containing p45MEK (GST-MEK). The assay solution contained 20 mM HEPES, pH 7.4, 10 mM MgCl2, 1 mM MnCl2, 1 mM EGTA, 50 mM [gamma-32P]ATP, 10 mg GST-MEK, 0.5 mg GST-MAPK and 40 mg MBP in a final volume of 100 mL. Reactions are stopped after 20 minutes by addition of trichloroacetic acid and filtered through a GF/C filter mat. 32P retained on the filter mat is determined using a 1205 Betaplate. PD0325901 is assessed at various dose ranges in order to determine dose response curves.

Cell Research:

[3]

  • Cell lines: PTC cells
  • Concentrations: 0.1 nM- 1 μM
  • Incubation Time: 48 hours
  • Method:

    PTC cells (1 × 104) are seeded in 24-well plates with 1 mL of medium for 4 days in a 37 °C incubator. MEK inhibitor PD0325901 at varying concentrations is added to the cells in triplicate on day 0. MTT dissolved in 0.8% NaCl solution at 5 mg/mL is added to each well (0.2 mL) on day 2 to test GI50 or every day for cell growth curves. The cells are incubated at 37 °C for 3 hours with MTT. The liquid is then aspirated from the wells and discarded. Stained cells are dissolved in 0.5 mL of DMSO and their absorption at 570 nm is measured using a Synergy HT multidetection microplate reader. For GI50, cell growth is calculated as 100 × (T − T0)/(C − T0), where T is the optical density of the wells treated with inhibitors after a 48-hour period, T0 is the optical density at time zero, and C is the control optical density with DMSO only.

Animal Research:

[3]

  • Animal Models: Ncr-nu/nu mice bearing PTC cells
  • Dosages: 20-25 mg/kg
  • Administration: Oral gavage

Solubility (25°C)

In vitro

In vivo

Add solvents to the product individually and in order
(Data is from Selleck tests instead of citations):
5%DMSO+40%PEG300+5%Tween80+50%ddH2O
For best results, use promptly after mixing.

5mg/mL

Chemical Information

Molecular Weight 482.19
Formula

C16H14F3IN2O4

CAS No. 391210-10-9
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles C1=CC(=C(C=C1I)F)NC2=C(C=CC(=C2F)F)C(=O)NOCC(CO)O

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

mg/kg g μL

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Molarity Calculator

Mass Concentration Volume Molecular Weight

Clinical Trial Information

NCT Number Recruitment Interventions Conditions Sponsor/Collaborators Start Date Phases
NCT02510001 Completed Drug: PF-02341066|Drug: PD-0325901|Drug: Binimetinib Solid Tumor|Colorectal Cancer University of Oxford|Queen''s University Belfast|Oxford University Hospitals NHS Trust|Velindre NHS Trust|University Hospital Antwerp|Hospital Vall d''Hebron|Saint Antoine University Hospital|European Georges Pompidou Hospital|Pfizer|University of Turin Italy|Belfast Health and Social Care Trust|Beaumont Hospital|European Commission|Array BioPharma|University of Paris 5 - Rene Descartes November 2014 Phase 1
NCT02096471 Completed Drug: PD-0325901 Neurofibromatosis Type 1 and Growing or Symptomatic Inoperable PN University of Alabama at Birmingham June 2014 Phase 2
NCT02022982 Active not recruiting Drug: Palbociclib|Drug: PD-0325901 KRAS Mutant Non-Small Cell Lung Cancer|Solid Tumors Dana-Farber Cancer Institute January 2014 Phase 1|Phase 2
NCT01347866 Terminated Drug: PF-05212384|Drug: PD-0325901|Drug: irinotecan Advanced Cancer Pfizer October 2011 Phase 1
NCT00174369 Terminated Drug: PD-0325901 Carcinoma Non-Small-Cell Lung Pfizer November 2005 Phase 2

(data from https://clinicaltrials.gov, updated on 2022-08-01)

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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Frequently Asked Questions

Question 1:
Whether the inhibitor PD0325901 interacts with other targets other than MEK?

Answer:
PD0325901 has very high selectivity to MEK. In addition, it can also inhibits VEGF activity according to the reference: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2713590/

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