SPHINX31

SKU: orb1302007

Description

SPHINX31 is a highly potent and selective SRPK1 inhibitor with an IC50 of 5.9 nM. This compound is a valuable chemical probe for studying SRPK1 function in angiogenesis and splicing regulation in both cellular and animal models of cancer and ocular disease.

Research Area

Cardiovascular Research, Cell Biology, Metabolism Research, Signal Transduction

Key Properties

• CAS Number: 1818389-84-2
• MW: 507.51
• Purity: >99.99% (May vary between batches)
• Formula: C₂₇H₂₄F₃N₅O₂
• SMILES: FC(F)(F)c1ccc(N2CCN(Cc3ccccn3)CC2)c(NC(=O)c2ccc(o2)-c2ccncc2)c1
• Target: VEGFR,Serine/threonin kinase
• Solubility: Ethanol:10 mg/mL (19.7 mM);H2O:Insoluble;DMSO:18.85 mg/mL (37.14 mM);10% DMSO+40% PEG300+5% Tween 80+45% Saline:1 mg/mL (1.97 mM)

Bioactivity

• Target IC50:
  SRPK1:5.9 nM (cell free)
• In Vivo:
  In vivo, SPHINX31 (2 μg per eye) inhibits blood vessel growth and macrophage infiltration in the eyes of a mouse model of choroidal neovascularization . Injection of 0.8 mg/kg SPHINX31 (i.p.) into DBA2J mice resulted in a concentration of 0.225 ± 0.036 μM in plasma after 24 h. In xenotransplanted RAIL mice with MOLM-13, THP-1 cells or first passage patient-derived AMLs, SPHINX31 (0.8 or 2.0 mg/kg) led to a significant reduction in leukemic cell growth and a dose-dependent prolongation of survival of mice given MOLM-13, THP-1, and patient-derived MLL-X AMLs .
• In Vitro:
  SPHINX31 inhibits phosphorylation of serine/arginine-rich splicing factor 1 (SRSF1), an SRPK1 substrate, in PC3 cells (EC50: 360 nM) and increases expression of the anti-angiogenic VEGF-A165b splice variant in retinal pigment epithelial (RPE) cells . SPHINX31 inhibited the growth of MLL-mutant AML cell lines with an IC50 >1 order of magnitude lower than for other AML lines. There was no impact of SPHINX31 on the clonogenic potential of normal mouse hematopoietic stem-progenitor cells (HSPCs). 1.5, 3, and 6 μM SPHINX31 did not affect the colony-forming ability of normal human cord blood CD34+ cells .
• Cell Research:
  Cells were transduced with gRNA vectors or treated with SPHINX31 and stained at the indicated time points with anti-mouse CD11b PE/Cy5 and anti-human CD11b PE or anti-human CD13 FITC. Data were analyzed by using LSRFortessa and FlowJo. Apoptosis levels were measured in human and/or mouse AML cells transduced with dual gRNA vectors (against SRPK1 and 3' BCL2 enhancer) and/or treated with 1 or 3 μM SPHINX31 at indicated time points, by using Annexin V. Data were analyzed by using LSRFortessa instruments. Cell cycle stages were measured in human and/or mouse AML cells transduced with dual gRNA vectors against SRPK1 and/or treated with 1 or 3 μM SPHINX31 at indicated time points, using Propidium Iodide. Data were analyzed using LSRFortessa instruments .
• Animal Research:
  For in vivo experiments, 6–10-week-old female Rosa26Cas9/+ mice were treated triweekly for two weeks with either vehicle or 2 mg/kg SPHINX31. Four weeks post-treatment, bone marrow cells from these mice were freshly extracted (as mentioned above) and blocked with anti-mouse CD16/32 and 10% mouse serum. For the identification of LK/LSK, LT-HSC, myeloid and B-cell subpopulations, staining was performed using CD4 PE/Cy5, CD5 PE/Cy5, CD8a PE/Cy5, CD11b PE/Cy5, B220 PE/Cy5, TER-119 PE/Cy5, GR-1 PE/Cy5, SCA-1 Pacific Blue, CD150 PE/Cy7, CD34 FITC and CD117 APC-eFluor780. In each of the multi-colour flow cytometry experiments, we included the fluorescence minus one (FMO) controls. FMO controls provide a measure of spillover in a given channel. This allows for correct gating and selects only the stained cells in the experimental sample. Flow cytometry analysis was performed using a LSRFortessa instrument and resulting data were subsequently analyzed using FlowJo .

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

Inhibitor, eye disease, HuCCA-1, neovascular, inhibit, Norway Brown rats, diabetics, angiogenic, SPHINX 31, SPHINX31, SPHINX-31, SRPK, SRPK1, retinal permeability, Serine kinase, threoninkinase, threonin kinase, Serinekinase, Serine-arginine protein kinases, VEGFR, Vascular endothelial growth factor receptor

Compound Identifiers

PubChem CID

91972002