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Milciclib CDK inhibitor

Cat.No.S2751

Milciclib is a potent, ATP-competitive CDK inhibitor for CDK2 with IC50 of 45 nM. It is >3-fold more selective for CDK2 than CDK1, 2, 4, 5, and 7, it's also an inhibitor of TRKA with IC50 of 53nM. This compound induces cell death through autophagy. Phase 2.
Milciclib CDK inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 460.57

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Quality Control

Cell Culture, Treatment & Working Concentration

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
human A2780 cells Proliferation assay 72 h Antiproliferative activity against human A2780 cells after 72 hrs by cell Titer_Glo assay, IC50=0.2 μM
human A2780 cells Function assay Inhibition of CDK2 in human A2780 cells assessed as reduction of hyperphosphorylated form of retinoblastoma protein at 1 uM relative to control
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Chemical Information, Storage & Stability

Molecular Weight 460.57 Formula

C25H32N8O

 Storage (From the date of receipt)
CAS No. 802539-81-7 Download SDF Storage of Stock Solutions

Synonyms PHA-848125 Smiles CC1(CC2=CN=C(N=C2C3=C1C(=NN3C)C(=O)NC)NC4=CC=C(C=C4)N5CCN(CC5)C)C

Solubility

In vitro
Batch:

DMSO : 17 mg/mL (36.91 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molarity Calculator

Mass Concentration Volume Molecular Weight

In vivo
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Mechanism of Action

Targets/IC50/Ki
CDK2/CyclinA [1]
(Cell-free assay)
45 nM
TrkA [1]
(Cell-free assay)
53 nM
CDK7/CyclinH [1]
(Cell-free assay)
150 nM
CDK4/CyclinD1 [1]
(Cell-free assay)
160 nM
CDK5/p35 [1]
(Cell-free assay)
265 nM
CDK2/CyclinE [1]
(Cell-free assay)
363 nM
CDK1/CyclinB [1]
(Cell-free assay)
398 nM
In vitro
PHA-848125 inhibits, although with lower potency, the activities of cyclin H/CDK7, cyclin D1/CDK4, p35/CDK5, as well as cyclin E/CDK2 and cyclin B/CDK1 with IC50 values of 0.15, 0.16, 0.265, 0.363, 0.398 μM, respectively. [1] Thropomyosin receptor kinase A is also inhibited by this compound in the same nanomolar range as CDKs. In the most PHA-848125-sensitive cell line, this compound induces a concentration-dependent G(1) arrest.[2] This compound also impairs phosphorylation of the retinoblastoma protein at CDK2 and CDK4 specific sites, reduces retinoblastoma protein and cyclin A levels, and increases p21(Cip1), p27(Kip1) and p53 expression. This chemical is added to the cells 48 h after TMZ and cell growth is evaluated after 3 additional days of culture. [2] A drug combination of TMZ, BG and this compound induces an additive or synergistic effect on cell growth, depending on the cell line. [2] In the absence of BG, the combination is still more active than the single agents in cell lines moderately sensitive to TMZ, but comparable to this compound alone in the two TMZ-resistant cell lines. When TMZ plus BG are used in combination with this chemical against cultured normal melanocytes, neither synergistic nor additive antiproliferative effects are observed.[2]
Kinase Assay
Biochemical kinase inhibition assays
Inhibition of kinase activity by this compound is assessed using a strong anion exchanger (Dowex 1X8 resin)–based assay in robotized format run on 384-well plates. In this assay, specific peptides or protein substrates are transphosphorylated by their specific kinase in the presence of ATP traced with [γ-33P]ATP using optimal buffers and cofactors. The potency of this chemical toward CDKs and 38 additional kinases belonging to an in-house Kinase Selectivity Screening panel is evaluated, and the relevant IC50s are determined. For each enzyme, the absolute KM values for ATP and the specific substrate are calculated and each assay is then run at optimized ATP (2KM) and substrate (5KM) concentrations. This setting enables direct comparison of IC50 values of this compound across the panel for evaluation of its biochemical profile.
In vivo
In the preclinical xenograft A2780 human ovarian carcinoma model, this compound reveals good efficacy and is well tolerated upon repeated daily treatments. Treatment of K-Ras(G12D)LA2 mice with this chemical (40 mg/kg twice daily for 10 days) results in significant tumor growth inhibition at the end of the treatment and is accompanied by a reduction in the cell membrane turnover.[3] On the other hand, following oral administration, it shows significant antitumor activity in various human xenografts, carcinogen-induced tumors and in disseminated primary leukemia models; the plasma concentrations in rodents being in the same range as those found inhibiting cancer cell proliferation.[4]
References

Applications

Methods Biomarkers Images PMID
Western blot pRb / P-pRb / Cdc6 / Cyclin A / Cyclin B1 p-TrkA / TrkA / p-MAPK / MAPK / p-AKT p-IGF-1Rβ / p-EGFR / p-FGFR3 / p-c-Yes / p-Src / p-S6 S2751-WB1 23347136

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