Prexasertib dihydrochloride

SKU: orb1302948

Description

Prexasertib dihydrochloride (LY2606368) is a potent ATP-competitive inhibitor of CHK1 (Ki: 0.9 nM), also showing activity against CHK2 and RSK in biochemical assays. This small molecule is widely used in cancer research to study DNA damage response pathways and has demonstrated antitumor efficacy in both cellular and animal models.

Research Area

Cell Biology, Signal Transduction

Key Properties

• CAS Number: 1234015-54-3
• MW: 438.31
• Purity: 99.82% (May vary between batches)
• Formula: C₁₈H₂₁Cl₂N₇O₂
• SMILES: Cl.Cl.COc1cccc(OCCCN)c1-c1cc(Nc2cnc(cn2)C#N)n[nH]1
• Target: Chk,Apoptosis,S6 Kinase
• Solubility: DMSO:4.39 mg/mL (10.02 mM)

Bioactivity

• Target IC50:
  RSK:9 nM|Chk1:0.9 nM|Chk2:8 nM
• In Vivo:
  In cancer xenografts, LY2606368 inhibits tumor growth by monotherapy and combined with other agents. In an orthotopic SKOV3 ovarian cancer model, LY2606368 inhibits the growth of primary tumors and markedly reduces the incidence of metastases and ascites accumulation. In an SW1990 orthotopic pancreatic cancer model, LY2606368 also causes a 92% inhibition of primary tumor growth and the elimination of metastases to the lymph node, spleen, and intestine.
• In Vitro:
  LY2606368 induces DNA damage and increases in pH2A.X. In cells, LY2606368 causes the rapid appearance of TUNEL and pH2AX-positive double-stranded DNA breaks in the S-phase cell population. In a functional assay, LY2606368 effectively abrogates the G2–M checkpoint activated by doxorubicin in p53-deficient HeLa cells (EC50: 9 nM). LY2606368 was broadly antiproliferative in the most sensitive cell lines (IC50s<50 nM) with a minority of cell lines showing considerable resistance (IC50s >1,000 nM). LY2606368 requires CDK2 and CDC25A to cause DNA damage.
• Cell Research:
  HeLa cells were plated onto T25 flasks and allowed to recover for 24 hours.LY2606368 was then added to give final concentrations of 33 or 100 nmol/L.In some experiments,20 μMol/L Z-VAD-FMK was included during the drug treatment.Cells were treated for 12 hours,and during the last 2 hours,colchicine was added to 1 μg/mL.Fixation of nuclei for metaphase spreads was done following the method of Bayani and Squire.Chromosome spreads were done.A 12-μL volume of cell suspension in 3:1 methanol/acetic acid fixative was dropped from a height of 3 cm onto dry glass slides or coverslips.The slides were then heated for 45 seconds on a 43°C metal block,before being removed to allow drying to complete at room temperature.Coverslips were mounted on slides with Vectashield Hard Set mounting medium with DAPI.Slides were examined with a Leica DMR fluorescence microscope and images were captured using a SPOT RT3 Slider camera.
• Animal Research:
  LY2606368 is formulated in a vehicle consisting of 20% Captisol.Female CD-1 nu-/nu- mice (26-28 g) are used for this study. Tumor growth is initiated by subcutaneous injection of 1×106 Calu-6 cells in a 1:1 mixture of serum-free growth medium and Matrigel in the rear flank of each subject animal. When tumor volumes reach approximately 150 mm3 in size, the animals are randomized by tumor size and body weight and placed into their respective treatment groups. The vehicle consisting of 20% Captisol pH4 or LY2606368 is administered by subcutaneous injection in a volume of 200 μL. Four, eight, 12, 24, and 48 hours after drug administration, blood for plasma drug exposure is extracted via cardiac puncture and assayed on a Sciex API 4000 LC/MS-MS system. The xenograft tissue is promptly removed and prepared. Lysates were analyzed by immunoblot analysis for protein phosphorylation levels.

Storage & Handling

• Storage: Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice/Shipping at ambient temperature.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

S296, S516, S6 Kinase, S6Kinase, RSK1, Ribosomal S6 Kinase (RSK), SIK, Prexasertib 2HCl, Prexasertib HCl, H2AX, MELK, LY2606368 (dihydrochloride), LY2606368 2HCl, Apoptosis, ARK5, BRSK2, double-stranded, CDC25A, Chk2, Chk1, Checkpoint Kinase (Chk)

Compound Identifiers

PubChem CID

46700755