| CAS |
1004990-28-6 |
| English Name |
PF-AKT400 |
| Synonyms |
AKTproteinkinaseinhibitor2-Acetamido-5-Nitrothiazole;N-(5-Nitrothiazol-2-yl)acetamide |
| Molecular Formula |
C20H22F2N6O |
| Molecular Weight |
400.43 |
| Solubility |
Soluble in DMSO ≥5mg/mL |
| Purity |
≥98% |
| Appearance |
Solid |
| Storage |
Powder:2-8℃，2 years;Insolvent(Mother Liquid):-20℃，6 months;-80℃，1 year |
| SMILES |
CCC1=CNC2=C1C(=NC=N2)N3CC[C@@](C3)(CNC(=O)C4=C(C=C(C=C4)F)F)N |
| InChIKey |
MOZRQQTUYAYCQT-FQEVSTJZSA-N |
| InChI |
InChI=1S/C20H22F2N6O/c1-2-12-8-24-17-16(12)18(27-11-26-17)28-6-5-20(23，10-28)9-25-19(29)14-4-3-13(21)7-15(14)22/h3-4，7-8，11H，2，5-6，9-10，23H2，1H3，(H，25，29)(H，24，26，27)/t20-/m0/s1 |
| PubChem CID |
25061501 |
| Target Point |
Akt |
| Passage |
PI3K/Akt/mTOR |
| Background |
PF-AKT400 is a potent, ATP-competitive and selective Akt inhibitor. |
| Biological Activity |
PF-AKT400 is a broadly selective， potent， ATP-competitive Akt inhibitor， displays 900-fold greater selectivity for PKBα (IC50=0.5 nM) than PKA (IC50=450 nM).[1-2] |
| IC50 |
PKBα:0.5nM;PKA:450nM[1] |
| In Vitro |
PF-AKT400(Compound 42)provides significantly enhanced selectivity for Akt relative to earlier leads such as spiroindoline 2. Free IC50 and EC50 values are estimated for phospho-S6 reduction(110 nM)and Akt hyperphosphorylation(216 nM)，respectively. These values corresponded well to the cellular IC50 for PF-AKT400 in U87 cells measuring p-GSK-3α(310 nM)[2]. |
| In Vivo |
PF-AKT400 is subsequently evaluated for modulation of Akt in tumors and in multiple in vivo mouse models of antitumor efficacy. It is active in a PC3 prostate carcinoma xenograft experiment，with 75% TGI observed at 100 mg/kg b.i.d. dosing for 10 days. In a colorectal carcinoma(Colo205)xenograft study，PF-AKT400 produces 60% TGI at 150 mg/kg b.i.d. after 10 days. Most intriguingly，in combination with Rapamycin(10 mg/kg，ip)，75 mg/kg b.i.d.(10 days)of PF-AKT400 results in 98% TGI in an additional PC3 prostate carcinoma xenograft study compared to 56% TGI and 66% TGI with PF-AKT400 and Rapamycin as single agents. To define the in vivo potency of PF-AKT400(Compound 42)in the PC3 xenograft model，oral administration of 25，75，and 100 mg/kg PF-AKT400 is performed with blood and tumor sampling over time. Immunoblot analysis of detergent-soluble extracts derived from PC3 tumors shows a significant reduction of S6 phosphorylation，and hyperphosphorylation of Akt upon treatment at doses that produced significant tumor growth inhibition. Plasma drug concentrations peak rapidly after oral administration of doses between 25-100 mg/kg(Tmax=0.5 h). Peak PD responses of phospho-S6 and phospho-Akt are observed at approximately 2-4h and 1h post-administration of PF-AKT400，respectively. The time-course of PD marker response is well described by a PK/PD model at doses that ranged from no efficacy(25 mg/kg)to maximal efficacy(100 mg/kg)[2]. |
| Animal Experiment |
Studies to describe the PK/PD relationship for PF-AKT400 are performed in male SCID/Beige mice bearing subcutaneous PC3 prostate carcinoma xenografts. Once tumors reach about ~300mm3 in size，PF-AKT400 is formulated in 0.5% methylcellulose vehicle and administered orally to 3 mice per dose group. Plasma and tumors are harvested over time，tumor lysates prepared，and the levels of phospho S6 reduction and phospho Akt induction are evaluated by immunoblot.[1] |
| Kinase Experiment |
A fluorescence polarization IMAP type assay is used. An amount of 15 μL of diluted PF-AKT400(Compound 42)in DMSO is mixed with 60 μL of reaction buffer(10 mM Tris-HCl，pH 7.5，10 mM MgCl2，0.1 mM EGTA，0.01% Triton-X100，1 mM DTT). Then 5 μL of the compound/buffer mixture，10 μL of a solution containing 4 μM ATP and 40 nM fluorescent-labeled Crosstide(Tamara-labeled GRPRTSSFAEG peptide)，and 5 μL of Akt1 protein(lacking the pleckstrin homology(PH)domain，containing an Asp at position 473，and prephosphorylated at Thr 308)in reaction buffer are combined. After a 90 min incubation，IMAP beads are added and plates are read(lamp filter，544 nm; emission filter，615 nm). The same procedure can be applied to full length Akt1 to provide similar results. All IC50 values are the geometric mean of at least n=2 determinations[2]. |
| Data Literature Source |
[1]. Chen SF，et al. Binding selectivity studies of PKBα using molecular dynamics simulation and free energy calculations. J Mol Model. 2013 Nov;19(11):5097-5112.
[2]. Freeman-Cook KD，et al. Design of selective，ATP-competitive inhibitors of Akt. J Med Chem. 2010 Jun 24;53(12):4615-4622. |
| Unit |
Bottle |
| Specification |
1mg |
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