3-methyl-4-oxo-3H,4H-imidazo[4,3-c][1,2,4]triazine-8-carboxamide

• ChemBase ID: 72587
• Molecular Formular: C7H7N5O2
• Molecular Mass: 193.16278
• Monoisotopic Mass: 193.05997449
• SMILES: C1(N=Nc2n(C1=O)cnc2C(=O)N)C
• Canonical SMILES: CC1N=Nc2n(C1=O)cnc2C(=O)N
• InChI: InChI=1S/C7H7N5O2/c1-3-7(14)12-2-9-4(5(8)13)6(12)11-10-3/h2-3H,1H3,(H2,8,13)
• InChIKey: HVCFAINNORHGHC-UHFFFAOYSA-N

NAMES AND DATABASE IDS

Names

• IUPAC name: 3-methyl-4-oxo-3H,4H-imidazo[4,3-c][1,2,4]triazine-8-carboxamide
• IUPAC Traditional name: 3-methyl-4-oxo-3H-imidazo[4,3-c][1,2,4]triazine-8-carboxamide
• Synonyms: Temozolomide; Temodar; Temodal; Methazolastone

Database IDs

• CAS Number: 85622-93-1
• PubChem SID: 162037512
• PubChem CID: 46930983

CALCULATED PROPERTIES

• Acid pKa: 10.57281
• H Acceptors: 5
• H Donor: 1
• LogD (pH = 5.5): -0.91549164
• LogD (pH = 7.4): -0.915263
• Log P: -0.91549456
• Molar Refractivity: 46.7048 cm^3
• Polarizability: 16.714983 Å^3
• Polar Surface Area: 102.7 Å^2
• Rotatable Bonds: 1
• Lipinski's Rule of Five: true

PROPERTIES

Physical Property

• Solubility: DMSO

Safety Information

• Storage Condition: -20°C

Product Information

• Salt Data: Free Base

DETAILS

Selleck Chemicals - S1237

Research Area

• Description: Cancer

Biological Activity

• Description: Methazolastone (Temozolomide, Temodar, Temodal) is a DNA damage inducer.
• In Vitro: Methazolastone causes formation of DNA alkali-labile sites which are present in similar amounts and repaired at a similar rate in L-1210 and L-1210/BCNU. In L-1210 but not in L-1210/BCNU methazolastone induces an arrest of cells in SL-G2-M phases. Methazolastone induces a similar amount of DNA ALS which is also repaired at a similar rate in L-1210 and L-1210/BCNU cell lines. Methazolastone induces a similar amount of DNA ALS which is also repaired at a similar rate in L-1210 and L-1210/BCNU cell lines. ^[1] Methazolastone sensitivity of both chemo-sensitive and resistant cells (D54-R and U87-R) is enhanced significantly under hyperoxia. Both Methazolastone and hyperoxia are associated with increased phosphorylation of ERK p44/42 MAPK (Erk1/2), but to a lesser extent in D54-R cells, suggesting that Erk1/2 activity may be involved in regulation of hyperoxia and Methazolastone-mediated cell death. Hyperoxia enhances Methazolastone toxicity in GBM cells by induction of apoptosis, possibly via MAPK-related pathways. ^[2] Methazolastone induces in monocytes the DNA damage response pathways ATM-Chk2 and ATR-Chk1 resulting in p53 activation. ^[3] Chronic Methazolastone exposure results in acquired Methazolastone-resistance and elevates miR-21 expression. ^[4] Methazolastone treatment triggers endoplasmic reticula (ER) stress with increased expression of GADD153 and GRP78 proteins, and deceases pro-caspase 12 protein. Methazolastone induces autophagy through mitochondrial damage- and ER stress-dependent mechanisms to protect glioma cells. ^[5]
• In Vivo: After a daily i.p. dose of 40 mg/kg for 5 consecutive days (days 1-5 after tumor transplant), methazolastone increases life-span by 86% in L-1210 and 22% in L-1210/BCNU. In L-1210/BCNU no effect is seen after 100 μM or 200 μM treatment; only 400 μM methazolastone produced an accumulation of cells in premitotic phase but much less than in L-1210. In L-1210/BCNU the maximum accumulation of cells in SL-G2-M is, after 48 hours-72 hours, approximately 30% as compared to 23% in untreated cells. Cells accumulates in SL-G2-M occurred too when L- 1210 leukemia-bearing mice are treated i.v. with methazola stone (40 mg/kg). No such effect is seen on L-1210/BCNU cells from mice given the same drug dose. ^[1]
• Clinical Trials: Methazolastone plus TPI 287 has entered in a phase II clinical trial in the treatment of melanoma.
• Features: Methazolastone is a second-generation alkylating agent.

Protocol

• Protocol: Cell Assay ^[1]
• Cell Lines: L-1210 and L-1210/BCNU cells
• Concentrations: 0 μM -100 μM
• Incubation Time: l hours
• Methods: L-1210 and L-1210/BCNU cells are seeded at 0.2 × 10₄ cells/mL and incubated for 24 hours. The cultures are treated with Methazolastone for l hours at 37oC, then washed twice in PBS by centrifugation and resuspended in fresh medium. Controls and treated samples are diluted in fresh medium 1:4 at 48 hours and 1:2 at 96 hours. Using these dilutions cell concentrations throughout the experiments are between 3 × 10₅ and 8 × 10₅/mL. Control growth is logarithmic in this range.
• Protocol: Animal Study ^[1]
• Animal Models: DBA/2 mice with L-1210 and L-1210/BCNU cells
• Formulation: 95% ethanol
• Doses: 40 mg/kg
• Administration: Administered via i.v.

References

• References: [1] Catapano CV, et al. Cancer Res. 1987, 47(18), 4884-4889.
• References: [2] Sun S, et al. J Neurooncol. 2012.
• References: [3] Bauer M, et al. PLoS One. 2012, 7(6):e39956.
• References: [4] Wong ST, et al. Anticancer Res. 2012, 32(7), 2835-2841.
• References: [5] Lin CJ, et al. PLoS One. 2012, 7(6), e38706.
• References: [6] Gori JL, et al. Cancer Gene Ther. 2012.

REFERENCES

• Catapano CV, et al. Cancer Res. 1987, 47(18), 4884-4889.
• Sun S, et al. J Neurooncol. 2012.
• Bauer M, et al. PLoS One. 2012, 7(6):e39956.
• Wong ST, et al. Anticancer Res. 2012, 32(7), 2835-2841.
• Lin CJ, et al. PLoS One. 2012, 7(6), e38706.
• Gori JL, et al. Cancer Gene Ther. 2012.