2-(4-benzylpiperazin-1-yl)-N'-[(1E)-[2-hydroxy-3-(prop-2-en-1-yl)phenyl]methylidene]acetohydrazide

• ChemBase ID: 73221
• Molecular Formular: C23H28N4O2
• Molecular Mass: 392.49402
• Monoisotopic Mass: 392.22122616
• SMILES: c1(CN2CCN(CC2)CC(=O)N/N=C/c2cccc(c2O)CC=C)ccccc1
• Canonical SMILES: C=CCc1cccc(c1O)/C=N/NC(=O)CN1CCN(CC1)Cc1ccccc1
• InChI: InChI=1S/C23H28N4O2/c1-2-7-20-10-6-11-21(23(20)29)16-24-25-22(28)18-27-14-12-26(13-15-27)17-19-8-4-3-5-9-19/h2-6,8-11,16,29H,1,7,12-15,17-18H2,(H,25,28)/b24-16+
• InChIKey: YQNRVGJCPCNMKT-LFVJCYFKSA-N

NAMES AND DATABASE IDS

Names

• IUPAC name: 2-(4-benzylpiperazin-1-yl)-N'-[(1E)-[2-hydroxy-3-(prop-2-en-1-yl)phenyl]methylidene]acetohydrazide; 2-(4-benzylpiperazin-1-yl)-N'-{[2-hydroxy-3-(prop-2-en-1-yl)phenyl]methylidene}acetohydrazide
• IUPAC Traditional name: 2-(4-benzylpiperazin-1-yl)-N'-[(1E)-[2-hydroxy-3-(prop-2-en-1-yl)phenyl]methylidene]acetohydrazide; 2-(4-benzylpiperazin-1-yl)-N'-{[2-hydroxy-3-(prop-2-en-1-yl)phenyl]methylidene}acetohydrazide
• Synonyms: Procaspase activating compound 1; PAC-1; (4-Benzylpiperazino)acetic acid (3-allyl-2-hyroxybenzylidene)hydrazide; 1-Piperazineacetic acid, 2-(phenylmethyl)-,[[2-hydroxy-3-(2-propenyl)phenyl]methylene]hyrrazide; PAC-1; 4-(Phenylmethyl)-1-piperazineacetic Acid (2E)-2-[[2-Hydroxy-3-(2-propen-1-yl)phenyl]methylene]hydrazide

Database IDs

• CAS Number: 315183-21-2; 1044929-62-5
• MDL Number: MFCD03302441
• PubChem SID: 162038141
• PubChem CID: 6851947

CALCULATED PROPERTIES

• Acid pKa: 8.614495
• H Acceptors: 5
• H Donor: 2
• LogD (pH = 5.5): 1.2224518
• LogD (pH = 7.4): 2.8888998
• Log P: 3.10083
• Molar Refractivity: 117.9249 cm^3
• Polarizability: 44.798073 Å^3
• Polar Surface Area: 68.17 Å^2
• Rotatable Bonds: 8
• Lipinski's Rule of Five: true

PROPERTIES

Physical Property

• Solubility: deionized water: ≤2 mg/mL; DMSO: ≥10 mg/mL
• Apperance: white solid

Safety Information

• Storage Condition: -20°C
• European Hazard Symbols: Nature polluting (N); Harmful (Xn)
• UN Number: 3077
• German water hazard class: 3
• Hazard Class: 9
• Packing Group: 3
• Risk Statements: 22-36/37/38-50/53
• Safety Statements: 26-36/37-60-61
• GHS Pictograms: GHS07; GHS09
• GHS Signal Word: Warning
• GHS Hazard statements: H302-H315-H319-H335-H400
• GHS Precautionary statements: P261-P273-P305 + P351 + P338
• Personal Protective Equipment: dust mask type N95 (US), Eyeshields, Gloves
• RID/ADR: UN 3077 9/PG 3
• Storage Temperature: room temp

Pharmacology Properties

• Target: Caspase

Product Information

• Purity: ≥98% (HPLC)
• Salt Data: Free base
• Empirical Formula (Hill Notation): C23H28N4O2

DETAILS

Selleck Chemicals - S2738

Research Area

• Description: Cancer

Biological Activity

• Description: PAC-1 is a potent procaspase-3 activator with EC50 of 0.22 μM.
• Targets: Procaspase-3
• IC50: 0.22 μM ^[1]
• In Vitro: PAC-1 activates procaspase-7 in a less efficient manner with EC50 of 4.5 μM. Elevated caspase 3 level in cancer cell lines allows PAC-1 to selectively induce apoptosis in a manner proportional to procaspase-3 concentration with IC50 of 0.35 μM for NCI-H226 cells to ~3.5 μM for UACC-62 cells. PAC-1 induces apoptosis in the primary cancerous cells with IC50 values of 3 nM to 1.41 μM, more potently than in the adjacent noncancerous cells with IC50 of 5.02 μM to 9.98 μM, which is also directly related to the distinct procaspase-3 concentration. ^[1] PAC-1 activates procaspase-3 by chelating zinc ions, thus relieving the zinc-mediated inhibition and allowing procaspase-3 to auto-activate itself to caspase-3. ^[2] PAC-1 is capable to induce cell death in Bax/Bak double-knockout cells and Bcl-2 and Bcl-xL-overexpressing cells with the same efficacy as its wild-type counterpart in a delayed manner. PAC-1 induces cytochrome c release in a caspase-3 independent manner, which subsequently triggers downstream caspase-3 activation and cell death. PAC-1 can not induce cell death and caspase-3 activation in Apaf-1 knockout cells, suggesting that apoptosome formation is essential for caspase-3 activation by PAC-1-mediated cell death. ^[3]
• In Vivo: Administration of PAC-1 at 5 mg with low and steady releasing significantly inhibits the growth of ACHN renal cancer xenograft in mice. Oral administration of PAC-1 (50 or 100 mg/kg) significantly retards tumor growth of NCI-H226 lung cancer xenograft in a dose-dependent manner, and markedly prevents the cancer cells from infiltrating the lung tissue. The in vivo anti-tumor effect of PAC-1 is ascribed to procaspase-3 activation and subsequently apoptosis induction consistent with the activity in vitro. ^[1]
• Features: PAC-1 is the first small molecule known to directly activate procaspase-3 to caspase-3

Protocol

• Protocol: Kinase Assay ^[1]
• In vitro procaspase-3 activation: Procaspase-3 is expressed and purified in Escherichia coli. Various concentrations of PAC-1 are added to 90 μL of a 50 ng/mL of procaspase-3 in caspase assay buffer in a 96-well plate, The plate is incubated for 12 hours at 37 °C. A 10 μL volume of a 2 mM solution of caspase-3 peptidic substrate acetyl Asp-Glu-Val-Asp-p-nitroanilide (Ac-DEVD-pNa) in caspase assay buffer is then added to each well. The plate is read every 2 minutes at 405 nm for 2 hours in a Spectra Max Plus 384 well plate reader. The slope of the linear portion for each well is determined, and the relative increase in activation from untreated control wells is calculated.
• Protocol: Cell Assay ^[1]
• Cell Lines: U-937, HL-60, CRL-1872, ACHN, NCI-H226, Hs888Lu, Hs578Bst, MCF-10A, SK-MEL-5, BT-20, MDA-MB-231, UACC-62, SK-N-SH, B16-F10 , Hs 578t, and PC-12
• Concentrations: Dissolved in DMSO, final concentrations ~100 μM
• Incubation Time: 72 hours
• Methods: Cells are exposed to various concentrations of PAC-1 for 72 hours. Cell death is quantified by the addition of MTS/PMS CellTiter 96 Cell Proliferation Assay reagent. The plates are incubated at 37 °C for approximately 1 hour (until the colored product formed), and the absorbance is measured at 490 nm.
• Protocol: Animal Study ^[1]
• Animal Models: Ovariectomized female athymic BALB/c (nude, nu/nu) mice injected subcutaneously with ACHN cells, male athymic BALB/c nude mice injected subcutaneously with NCI-H226 cells, and male athymic BALB/c–/– mice injected intravenously with NCI-H226 cells
• Formulation: Mixed with cholesterol and pelleted into a 3-mm-diameter 20-mg (total weight) pellet, or dissolved in a mixture of 24:1 vegetable oil/DMSO
• Doses: ~100 mg/kg
• Administration: Pellet implantation subcutaneously or oral gavage

References

• References: [1] Putt KS, et al. Nat Chem Biol, 2006, 2(10), 543-550.
• References: [2] Peterson QP, et al. J Mol Biol, 2009, 388(1), 144-158.
• References: [3] Seervi M, et al. Cell Death Dis, 2011, 2, e207.

Sigma Aldrich - P0115

Biochem/physiol Actions
PAC-1 is a caspase 3 activator. Caspase activation is a key strategy in cancer therapy. Caspase 3 is a key executioner caspase and direct effector of apoptosis. Hallmark of many cancers is the circumvention of signal in the activation of executioner caspases and loss of apoptotic response. EC50 for caspase 3 activation = 0.33 μM and caspase 7= 4.5 μM. The IC50 for apoptosis induction = 0.92 μM. Elevated caspase 3 level in cancerous cell lines and tissues allows PAC-1 to selectively induce apoptosis in tissues.

Toronto Research Chemicals - P132000

PAC-1 is a caspase 3 activator. Caspase activation is a key strategy in cancer therapy. Caspase 3 is a key executioner caspase and direct effector of apoptosis. Hallmark of many cancers is the circumvention of signal in the activation of executioner caspa

REFERENCES

• Putt KS, et al. Nat Chem Biol, 2006, 2(10), 543-550.
• Peterson QP, et al. J Mol Biol, 2009, 388(1), 144-158.
• Seervi M, et al. Cell Death Dis, 2011, 2, e207.
• Slee, E., et al.: J. Biol. Chem., 276, 7320 (2001)
• Pop, C., et al.: Biochemistry, 42, 12311 (2001)
• Li, L., et al.: Science, 305, 1471 (2001)
• Green, D., et al.: J. Clin. Invest., 115, 2610 (2001)
• Satoh, T., et al.: Clin. Cancer Res., 15, 3872 (2001)