Col003 is a potent and selective inhibitor of Hsp47 that competitively binds to the collagen-binding site of Hsp47 (IC50, 1.8 μM). Col003 inhibits the interaction between Hsp47 and collagen and inhibits collagen secretion by destroying the stability of the collagen triple helix. Col003 may be utilized in related research on pulmonary fibrosis.

Physicochemical Properties

Molecular Formula	C14H11NO4
Molecular Weight	257.241443872452
Exact Mass	257.068
CAS #	328565-16-8
PubChem CID	871077
Appearance	Light yellow to yellow solid powder
LogP	3.3
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	3
Heavy Atom Count	19
Complexity	322
Defined Atom Stereocenter Count	0
SMILES	OC1C(C=O)=CC(=CC=1[N+](=O)[O-])CC1C=CC=CC=1
InChi Key	RKJXVCLOGNLZOI-UHFFFAOYSA-N
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
Chemical Name	5-benzyl-2-hydroxy-3-nitrobenzaldehyde
HS Tariff Code	2934.99.9001
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)

Biological Activity

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].
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.
References	[1]. A small-molecule compound inhibits a collagen-specific molecular chaperone and could represent a potential remedy for fibrosis. J Biol Chem. 2017 Dec 8;292(49):20076-20085.

Solubility Data

Solubility (In Vitro)

DMSO : ~50 mg/mL (~194.37 mM)

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.