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| ln Vitro |
When the oxygen content falls below 1.3%, the chemical trimonidine, a 2-nitroimidazole, forms a covalent link with cellular macromolecules and serves as an exogenous hypoxia marker. Detection: Using cellular monoclonal antibodies, immunohistochemical detection of pimonidazole can identify hypoxic cells. The detection of cell proliferation is accelerated by commercially available monoclonal antibodies for ischemia cell nuclear repair (PCNA). Spot counting and image analysis tools were utilized to discover pimonidazole hydrochloride-protein adducts, which were then confirmed by immunofluorescence. Hypoxia and cell proliferation were qualitatively evaluated by light microscopy. Pimonidazole hydrochloride (10–100 μM) was used to treat the cells until 90% confluence was reached. Next, the cells were mounted using Prolong Gold with DAPI mounting medium [3 4 hours], washed four times with HBSS, fixed for ten minutes with 10% neutral solution, and then washed three times with buffered saline and formalin buffer. Prior to fixation, tissues were extracted and kept in 400 μM pimonidazole hydrochloride for two hours in order to track in vivo hypoxic signals. Antibodies produced against the protein addition pimonidazole hydrochloride were used for immunostaining [4].
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| ln Vivo |
Pimonidazole is a 2-nitroimidazole that is activated by potentiometric reduction in hypoxic cells and is a persistent adduct with thiol groups in proteins, peptides and amino acids. Furthermore, the amount of pimonidazole hydrochloride found was closely linked to the extent of hypoxia within the tumor in vivo. This handy tool provides the following choices to examine hypoxia levels in the body. 1. Pimonidazole hydrochloride is suspended in 0.9% diluted physiological saline or PBS at a dosage of 30 mg/mL. 2. Inject 60 mg/kg of pimonidazole hydrochloride into several intravenous veins (tail vein). 3. Make internal circulation of the nozzle for 90 minutes, and then euthanize the mice with halothane or high pressure (CO2). NOTE: For animal euthanasia: follow proper IACUC protocols! 4. Kill the mice, take the organs, weigh them, then immediately freeze them, and store them. 5. Pimonidazole hydrochloride dye can be utilized for paraffin-embedded tissues, frozen tissues and cell lines. Organs were embedded in OCT, put in a cryostat, and incubated in 10 μM thick sections. The portions will be extracted and stained on glass slides.
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| References | |
| Additional Infomation |
Pimonidazole Hydrochloride is the hydrochloride salt form of pimonidazole, a hypoxia-selective 2-nitroimidazole with potential radiosensitizing activity. Pimonidazole is reduced in hypoxic environments such as those found in tumors. In hypoxic cells and tumors, reduced pimonidazole binds to thiol-containing proteins and glutathione. The resulting complexes accumulate in hypoxic tumors, deplete radioprotective thiol compounds, and cause the tumor to be more susceptible to radiation treatment.
See also: Pimonidazole (annotation moved to). |
| Molecular Formula |
C11H18N4O3
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|---|---|
| Molecular Weight |
254.28562
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| Exact Mass |
290.115
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| CAS # |
70132-51-3
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| Related CAS # |
Pimonidazole;70132-50-2
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| PubChem CID |
104997
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| Appearance |
White to yellow solid powder
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| Boiling Point |
490.5ºC at 760 mmHg
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| Flash Point |
250.5ºC
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| LogP |
1.901
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
19
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| Complexity |
278
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
SKUVJFHTKRTQQZ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C11H18N4O3.ClH/c16-10(8-13-5-2-1-3-6-13)9-14-7-4-12-11(14)15(17)18;/h4,7,10,16H,1-3,5-6,8-9H2;1H
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| Chemical Name |
1-(2-nitroimidazol-1-yl)-3-piperidin-1-ylpropan-2-ol;hydrochloride
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~250 mg/mL (~859.85 mM)
H2O : ~5 mg/mL (~17.20 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 10 mg/mL (34.39 mM) in 50% PEG300 +50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.9325 mL | 19.6626 mL | 39.3252 mL | |
| 5 mM | 0.7865 mL | 3.9325 mL | 7.8650 mL | |
| 10 mM | 0.3933 mL | 1.9663 mL | 3.9325 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
| NCT Number | Recruitment | interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
| NCT03410420 | Suspended | Drug: Pimonidazole hydrochloride | Aortic Aneurysm, Thoracic | University of Maryland, Baltimore | October 1, 2018 | Phase 1 |
| NCT00374517 | Withdrawn | Drug: Pimonidazole hydrochloride | Refractory Pulmonary Tuberculosis | National Institute of Allergy and Infectious Diseases (NIAID) |
September 5, 2006 | Phase 2 |
| NCT01248637 | Completed | Drug: Pimonidazole hydrochloride | Pancreatic Cancer | University Health Network, Toronto | October 2010 | |
| NCT05702619 | Recruiting | Drug: Optional non-IMP pimonidazole Diagnostic Test: CT-guided Bone Biopsy |
Prostate Cancer Hypoxia |
The Christie NHS Foundation Trust | October 3, 2021 |
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