CH5132799

• Catalog No.: S2699
• CAS Number: 1007207-67-1
• M. F.: C15H19N7O3S
• M. W.: 377.42146
• Storage: -20°C

SYNONYMS

• IUPAC name: 5-[7-methanesulfonyl-2-(morpholin-4-yl)-5H,6H,7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrimidin-2-amine
• IUPAC Traditional name: 5-[7-methanesulfonyl-2-(morpholin-4-yl)-5H,6H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrimidin-2-amine

DATABASE IDS

• CAS Number: 1007207-67-1

PROPERTIES

• Target: PI3K
• Salt Data: Free Base
• Storage Condition: -20°C

DETAILS

Description (English)

Biological Activity

• Description: CH5132799 exhibits a strong inhibitory activity especially against PI3Kα with IC50 of 14 nM.
• Targets: PI3Kα; PI3Kβ; PI3Kδ; PI3Kγ
• IC50: 0.014 μM; 0.12 μM; 0.50 μM; 0.036 μM ^[1]
• In Vitro: CH5132799 selectively inhibits class I PI3Ks, PI3Kα (IC50 = 0.014 μM ), PI3Kβ (IC50 = 0.12 μM ), PI3Kδ (IC50 = 0.50 μM ), PI3Kγ (IC50 = 0.036 μM ), but shows less inhibition of class II PI3Ks, class III PI3k and mTOR and also no inhibitory activity (IC50 > 10 μM) against 26 protein kinases. CH5132799 exhibits more inhibitory activities against PI3Kα with oncogenic mutations E542K (IC50 = 6.7 nM), E545K (IC50 = 6.7 nM) and H1047R (IC50 = 5.6 nM) than wild-type PI3Kα. CH5132799 treated breast cnacer KPL-4 cells, which harbor the PIK3CA mutation, phosphorylation of Akt and its direct substrates, PRAS40 and FoxO1/3a and phosphorylation of downstream factors, including S6K, S6 and 4E-BP1, are effectively suppressed. Cancer cell lines harboring PIK3CA mutations are significantly sensitive to CH5132799 ^[1] In human tumor cell lines with PI3K pathway activation by mutation, CH5132799 shows potent antiproliferative activity [HCT116(CRC): IC50 = 0.20 lM, KPL-4(BC):13 IC50 = 0.032 lM, T-47D(BC): IC50 = 0.056 lM, SK-OV-3(Ovarian): IC50 = 0.12 lM]. CH5132799 effectively suppresses phosphorylation of AKT in KPL-4 cells. ^[2]
• In Vivo: CH5132799 shows potent in vivo antitumor activity in several different xenograft models with PIK3CA mutations. CH5132799 overcomes mTORC1 inhibition-mediated Akt activation and regrowth of xenograft tumor by long-term everolimus administration. ^[1] CH5132799 as a clinical candidate that shows excellent oral bioavailability (BA) (101% in mouse), human liver microsomal stability and in vivo antitumor activity in the PC-3 xenograft model (TGI: 101% at 25 mg/kg, po, q.d. × 11 days). CH5132799 exhibits good oral BA in mouse, rat, monkey and dog (F: 54.2–101%, Table 2). In a human breast cancer (KPL-4: PI3Ka H1047R) xenograft model in mice, oral treatment with CH5132799 (12.5 mg/kg, q.d.) shows strong tumor regression. The strong regression is maintained during the 6 week administration, even in the intermittent dosing schedule (q.d., 2 weeks on/1 week off; q.d., 5 days on/2 days off), suggesting that a flexible administration schedule can be applicable in the clinic. ^[2]
• Clinical Trials: CH5132799 is now in Phase 1 clinic trial fro the orally administration in human with advanced solid tumors.

Combination Therapy

• Description: Combination with CH5132799 and trastuzumab, tumors disappears in the trastuzumab-insensitive breast cancer model with the PIK3CA mutation. ^[1]

Protocol

• Protocol: Kinase Assay ^[1]
• PI3K Assay: The E542K, E545K, and H1047R mutants of PI3Kα are prepared with an overlapped extension-polymerase chain reaction. Glutathione S-transferase-tagged PI3Kα mutants and His-tagged p85α are co-expressed with BAC-TO-BAC Baculovirus Expression System. The inhibitory activities of CH5132799 on PI3Kα (p110α/p85α), PI3Kβ(p110β/p85α), PI3Kδ (p110δ/p85α), PI3Kγ (p110γ), PI3KC2α, PI3KC2β, Vps34, and PI3Kα mutants are determined by Adapta Universal Kinase Assay Kit. Time-resolved fluorescence is measured with an EnVision HTS microplate reader. IC50 values are calculated using XLfit.
• Protocol: Cell Assay ^[1]
• Cell Lines: various cancer cell lines belonging to 4 types of cancer—breast, ovarian, prostate, and endometrial cancer—in which the PIK3CA mutation and PTEN deficiency are frequently found and in which RAS/RAF is rarely mutated
• Concentrations: 0 – 10μM
• Incubation Time: 4 days
• Methods: The cell lines are added to the wells of 96-well plates containing 0.076 to 10,000 nM CH5132799 and incubated at 37 °C. After 4 days of incubation, Cell Counting Kit-8 solution is added and, after incubation for several more hours, absorbance at 450 nm is measured with Microplate-Reader iMark. The antiproliferative activity is calculated by the formula (1- T/C) × 100 (%), in which T and C represent absorbance at 450 nm of the cells treated with CH5132799 (T) and that of untreated control cells (C). The IC50 values are calculated by using Microsoft Excel 2007.
• Protocol: Animal Study ^[1]
• Animal Models: A total of 4 × 10^6 to 1.2 × 10^7 cells are injected subcutaneously into the right flank of female BALB-nu/nu mice.
• Formulation: CH5132799 is dissolved in DMSO.
• Doses: 0.39, 0.78, 1.56, 3.13, 6.25, 12.5 and 25 mg/kg
• Administration: Orally administered once a day.

References

• References: [1] Tanaka H, et al, Clin Cancer Res, 2011, 17(10), 3272-3281.
• References: [2] Ohwada J, et al, Bioorg Med Chem Leff, 2011, 21(6), 1767-1772.

REFERENCES

• Tanaka H, et al, Clin Cancer Res, 2011, 17(10), 3272-3281.
• Ohwada J, et al, Bioorg Med Chem Leff, 2011, 21(6), 1767-1772.