ICG-001

• Catalog No.: S2662
• CAS Number: 780757-88-2
• M. F.: C33H32N4O4
• M. W.: 548.63158
• Storage: -20°C

SYNONYMS

• IUPAC name: (6S,9aS)-N-benzyl-6-[(4-hydroxyphenyl)methyl]-8-(naphthalen-1-ylmethyl)-4,7-dioxo-octahydro-1H-pyrimido[1,2-a]piperazine-1-carboxamide
• IUPAC Traditional name: (6S,9aS)-N-benzyl-6-[(4-hydroxyphenyl)methyl]-8-(naphthalen-1-ylmethyl)-4,7-dioxo-tetrahydro-2H-pyrimido[1,2-a]piperazine-1-carboxamide

DATABASE IDS

• CAS Number: 780757-88-2

PROPERTIES

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

DETAILS

Description (English)

Research Area

• Description: Cancer

Biological Activity

• Description: ICG-001 specifically binds to CBP with IC50 of 3 μM.
• Targets: CBP
• IC50: 3 μM ^[1]
• In Vitro: ICG-001 has no effect on the related reporter construct, FOPFLASH, which contains mutated TCF sites. After treatment with 25μM of ICG-001 for 8 hours, SW480 cell reduces the steady-state levels of Survivin and Cyclin D1 RNA and protein, both of which can be up-regulated by β-catenin. ICG-001 selectively induces apoptosis in transformed cells but not in normal colon cells, reduces in vitro growth of colon carcinoma cells. ^[1] ICG-001, can phenotypically rescue normal nerve growth factor (NGF) -induced neuronal differentiation and neurite outgrowth in the presenilin-1 mutant cells, emphasizing the importance of the TCF/β-catenin signaling pathway on neurite outgrowth and neuronal differentiation. ^[2] A recent study demonstrates that 5μM ICG-001 inhibits leptin-induced EMT, invasion and tumorsphere formation in MCF7 cells. ^[3]
• In Vivo: Administration of a water-soluble analog of ICG-001 for 9 weeks reduces the formation of colon and small intestinal polyps by 42% as effectively as the nonsteroidal antiinflammatory agent Sulindac, which has consistently demonstrated efficacy in this model. No overt toxicity is detected throughout the course of treatment. In the SW620 nude mouse xenograft model of tumor regression, 150 mg/kg, i.v. of analog demonstrates a dramatic reduction in tumor volume over the 19-day course of treatment, with no mortality or weight loss. ^[1] ICG-001 (5 mg/kg per day) significantly inhibits beta-catenin signaling and attenuates bleomycin-induced lung fibrosis in mice, while concurrently preserving the epithelium. ^[4]

Combination Therapy

• Description: Administration of ICG-001 concurrent with bleomycin prevents fibrosis, and late administration is able to reverse established fibrosis and significantly improve survival. ^[4]

Protocol

• Protocol: Kinase Assay ^[1]
• DUAL-Luciferase Reporter Assay: The Dual-Luciferase Reporter (DLR) Assay System provides an efficient means of performing dual reporter assays. In the DLR^TM Assay, the activities of firefly (Photinus pyralis) and Renilla (Renilla reniformis, also known as sea pansy) luciferases are measured sequentially from a single sample. The firefly luciferase reporter is measured first by adding Luciferase Assay Reagent II (LAR II) to generate a “glow-type” luminescent signal. After quantifying the firefly luminescence, this reaction is quenched, and the Renilla luciferase reaction is initiated by simultaneously adding Stop & Glo® Reagent to the same tube. The Stop & Glo® Reagent also produces a “glow-type” signal from the Renilla luciferase, which decays slowly over the course of the measurement. In the DLR^TM Assay System, both reporters yield linear assays with subattomole (<10-18) sensitivities="" and="" no="" endogenous="" activity="" of="" either="" reporter="" in="" the="" experimental="" host="" cells.="" furthermore,="" the="" integrated="" format="" of="" the="">^TM Assay provides rapid quantitation of both reporters either in transfected cells or in cell-free transcription/translation reactions.
• Protocol: Cell Assay ^[1]
• Cell Lines: Human colon carcinoma cell lines SW480, SW620, and HCT116, normal colonic epithelial cell line CCD-841Co
• Concentrations: ~25 μM
• Incubation Time: 24 hours
• Methods: 1. Prior to starting the assay, prepare the Apo-ONE? Caspase-3/7 Reagent, and mix thoroughly. 2. For best results, empirical determination of the optimal cell number, apoptosis induction treatment and incubation period for the cell culture system may be necessary.3. Use identical cell numbers and volumes for the assay and the negative control samples.4. Do not mix Apo-ONE? Caspase-3/7 Reagent and samples by manual pipetting. Mixing in this manner is unnecessary and may create bubbles that interfere with fluorescence readings or cross-contaminate the samples. Gentle mixing may be performed using a plate shaker.5. Total incubation time for the assay depends upon the amount ofcaspase- 3/7 present in the sample. 6. The Apo-ONE? Caspase-3/7 Reagent is formulated to mediate cellular lysis and support optimal caspase-3/7 activity. In rare instances, the reagent does not affect complete lysis of cultured cells. In such cases, lysis is enhanced by a freeze-thaw cycle. For best results, freeze at –70 °C, then thaw at room temperature. After equilibration, mix to homogeneity and incubate until measurable fluorescence is achieved.
• Protocol: Animal Study ^[1]
• Animal Models: Seven-week-old male C57BL/6J-Apc ^Min/+
• Formulation: Water-soluble analog of ICG-001 is used.
• Doses: 300 mg/kg
• Administration: Water-soluble analog of ICG-001 is treated orally for 9 weeks everyday.

References

• References: [1] Emami KH, et al, Proc Natl Acad Sci USA, 2004, 101(34), 12682-12687.
• References: [2] Teo JL, et al, Proc Natl Acad Sci USA, 2005，102（34），12171-12176.
• References: [3] Yan D, et al, J Biol Chem, 2012, 287(11), 8598-8612.
• References: [4] Henderson WR Jr, et al, Proc Natl Acad Sci USA, 2010, 107(32), 14309-14314.

REFERENCES

• Emami KH, et al, Proc Natl Acad Sci USA, 2004, 101(34), 12682-12687.
• Teo JL, et al, Proc Natl Acad Sci USA, 2005，102（34），12171-12176.
• Yan D, et al, J Biol Chem, 2012, 287(11), 8598-8612.
• Henderson WR Jr, et al, Proc Natl Acad Sci USA, 2010, 107(32), 14309-14314.