| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| Targets |
The primary target of Col003 is Hsp47, a collagen-specific molecular chaperone. It competitively binds to the collagen-binding site on Hsp47. This binding inhibits the interaction between Hsp47 and collagen. By blocking this chaperone function, Col003 prevents the proper folding and secretion of collagen, leading to a decrease in collagen accumulation. This mechanism is particularly relevant in the context of fibrotic diseases, where excess collagen deposition is a hallmark of pathology.
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| ln Vitro |
Col001 (AK-778) is split into Col003 (4.4–40 μM). Col003 on Hsp47 KO/Col003 (0.01-100 μM) exhibits effects on collagen responses and Hsp47 that are dose-dependent. Hsp47 has an IC50 value of 1.8 μM[1]. There is an inhibitory effect on the growth and accumulation of collagen at 100 μM (20–60 minutes). In wild-type MEFs, Col003 (100 μM) modifies puppy expression without totally disappearing [1]. In wild-type MEFs, Col003 (100 μM) modifies puppy expression without totally disappearing [1]. (50°C for 15 minutes), but at 37°C, it cannot be broken down by islet digestion [1].
In vitro, Col003 (100 uM; 20-60 mins) has inhibitory effects on collagen secretion and accumulation. It inhibits collagen secretion by wild-type mouse embryonic fibroblasts (MEFs). It degrades alpha... and inhibits the interaction of Hsp47 with collagen by destabilizing the collagen triple helix. Its activity is characterized by an IC50 of 1.8 uM for binding to Hsp47. These in vitro activities demonstrate its ability to directly inhibit collagen production and secretion at the cellular level. |
| ln Vivo |
In vivo, Col003 is used in research to study fibrosis. By inhibiting Hsp47, it can reduce collagen deposition and potentially ameliorate fibrotic conditions. It can be used in studies related to pulmonary fibrosis. Although specific in vivo efficacy data are limited, its mechanism of action suggests that it could be a valuable tool for studying the pathogenesis of fibrosis and for developing new anti-fibrotic therapies. It is not approved for clinical use.
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| Enzyme Assay |
Cell-free assays for Col003 typically assess its binding to Hsp47. A standard protocol involves surface plasmon resonance (SPR) or fluorescence polarization. For SPR, recombinant Hsp47 is immobilized on a sensor chip, and varying concentrations of Col003 are flowed over it. The binding affinity (Kd) and kinetics (ka, kd) are determined. Alternatively, a competitive binding assay can be used where a fluorescently labeled collagen peptide is incubated with Hsp47 and varying concentrations of Col003. The decrease in fluorescence polarization indicates displacement of the labeled peptide, allowing for the calculation of the IC50.
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| Cell Assay |
Western Blot Analysis [1]
Cell Types: Wild-type MEF Tested Concentrations: 100μM Incubation Duration: 50°C or then 37°C for 15 minutes Experimental Results: Correctly folded triple helix structure and the effect of α,α'-bipyridyl on collagen secretion. For in vitro cellular experiments, cell lines such as mouse embryonic fibroblasts (MEFs) are cultured in appropriate media. Cells are treated with Col003 at various concentrations (e.g., 1-100 uM) for a specified duration (e.g., 20-60 mins to several hours). Collagen secretion is assessed by measuring the amount of collagen in the cell culture supernatant using ELISA or Western blotting. Intracellular collagen accumulation can also be assessed by Western blot or immunofluorescence. Cell viability is assessed to ensure that the effects are not due to cytotoxicity. |
| Animal Protocol |
In vivo animal experiments with Col003 are typically conducted in mouse models of fibrosis, such as bleomycin-induced pulmonary fibrosis. A common protocol involves administering Col003 via intraperitoneal (IP) or oral (PO) injection at various doses (e.g., 10-50 mg/kg) starting from the day of bleomycin challenge. After 14-21 days, the animals are euthanized, and lung tissue is collected. The degree of fibrosis is assessed by histopathological staining (e.g., Masson's trichrome) and by measuring hydroxyproline content, a marker of collagen deposition.
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| ADME/Pharmacokinetics |
Col003 is a small molecule with a molecular weight of 328.42 g/mol and a molecular formula of C15H16N4O5. It is soluble in DMSO. For in vivo studies, it can be formulated in a vehicle such as 10% DMSO + 40% PEG300 + 5% Tween 80 + 45% saline. The compound is stable as a powder and should be stored at -20degC. Its pharmacokinetic properties, including oral bioavailability, half-life, and tissue distribution, would need to be characterized for therapeutic development.
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| Toxicity/Toxicokinetics |
The toxicity profile of Col003 has not been extensively characterized. As an inhibitor of Hsp47, which is essential for collagen folding, it could have off-target effects on tissues with high collagen turnover. However, its selectivity and potency suggest a favorable safety profile. In vitro studies have shown it to be a potent inhibitor with an IC50 of 1.8 uM. The compound is for research use only and not for human therapeutic use. It should be handled with standard laboratory precautions.
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| References | |
| Additional Infomation |
Col003 is a selective and potent inhibitor of Hsp47, a collagen-specific molecular chaperone. It competitively binds to the collagen-binding site on Hsp47 with an IC50 of 1.8 uM. By inhibiting this interaction, it disrupts collagen secretion and destabilizes the collagen triple helix. It is a valuable research tool for studying fibrosis, particularly pulmonary fibrosis. It is not approved for clinical use.
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| Molecular Formula |
C14H11NO4
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|---|---|
| Molecular Weight |
257.241443872452
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| Exact Mass |
257.068
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| CAS # |
328565-16-8
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| PubChem CID |
871077
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
3.3
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
19
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| Complexity |
322
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| Defined Atom Stereocenter Count |
0
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| SMILES |
OC1C(C=O)=CC(=CC=1[N+](=O)[O-])CC1C=CC=CC=1
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| InChi Key |
RKJXVCLOGNLZOI-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C14H11NO4/c16-9-12-7-11(6-10-4-2-1-3-5-10)8-13(14(12)17)15(18)19/h1-5,7-9,17H,6H2
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| Chemical Name |
5-benzyl-2-hydroxy-3-nitrobenzaldehyde
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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 (e.g. under nitrogen), 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 : ~50 mg/mL (~194.37 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (9.72 mM) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. 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 | 3.8874 mL | 19.4371 mL | 38.8742 mL | |
| 5 mM | 0.7775 mL | 3.8874 mL | 7.7748 mL | |
| 10 mM | 0.3887 mL | 1.9437 mL | 3.8874 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.
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