Cobalt protoporphyrin IX is a potent and specific inducer of heme oxygenase-1 (HO-1). Cobalt protoporphyrin IX displays a broad-spectrum antiviral effect against influenza A virus (IAV).

Physicochemical Properties

Molecular Formula	C34H32CON4O4
Molecular Weight	619.58
Exact Mass	619.176
CAS #	14325-03-2
PubChem CID	3000479
Appearance	Brown to black solid powder
Boiling Point	1122ºC at 760mmHg
Flash Point	632.4ºC
Vapour Pressure	0mmHg at 25°C
LogP	2.324
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	8
Heavy Atom Count	43
Complexity	1010
Defined Atom Stereocenter Count	0
SMILES	[Co+2].OC(CCC1=C(C)C2[N-]C1=CC1[N-]C(C=C3N=C(C=C4N=C(C=2)C(C)=C4C=C)C(C)=C3C=C)=C(C)C=1CCC(=O)O)=O
InChi Key	AQTFKGDWFRRIHR-UHFFFAOYSA-L
InChi Code	InChI=1S/C34H34N4O4.Co/c1-7-21-17(3)25-13-26-19(5)23(9-11-33(39)40)31(37-26)16-32-24(10-12-34(41)42)20(6)28(38-32)15-30-22(8-2)18(4)27(36-30)14-29(21)35-25;/h7-8,13-16H,1-2,9-12H2,3-6H3,(H4,35,36,37,38,39,40,41,42);/q;+2/p-2
Chemical Name	3-[18-(2-carboxylatoethyl)-7,12-bis(ethenyl)-3,8,13,17-tetramethylporphyrin-21,23-diid-2-yl]propanoate;cobalt(2+);hydron
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
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	On MDCK cells and RAW264.7 cells, the highest non-toxic concentration (TC0) of cobalt protoporphyrin IX (72 h) is 3.13 μM [1]. In MDCK and RAW264.7 cells, cobalt protoporphyrin IX (2 μM; 0-24 h) raises the levels of intracellular protein HO-1 in a time-dependent way [1]. The antagonistic concentration (IC50) of Cobalt protoporphyrin IX against the influenza viruses Empty Cell A/Fort Monmouth /1/1947, A/TianjinJinnan /15/2009, A/Wuhan /359/1995, A/FujianTongan /196/2009, and BY/FujianXinluo /54/2006 are 0.40 ± 0.16, 0.42 ± 0.15, 0.46 ± 0.19, 0.34 ± 0.05, and 0.64 ± 0.30 μM, respectively [1]. Cobalt protoporphyrin IX (0.25-2 μM; 18 or 24 h) prevents IAV replication by boosting the IFN response; its anti-IAV action is not dependent on HO-1's catalytic role [1]. In order to boost antiviral IFN responses, IRF3 phosphorylation and translocation may be necessary for Cobalt protoporphyrin IX (2 μM; 0–6 h) to induce HO-1 anti-IAV activity [1].
Cell Assay	Western Blot Analysis[1] Cell Types: MDCK cells and RAW264.7 cells Tested Concentrations: 2 μM Incubation Duration: 0, 3, 6, 12 and 24 h Experimental Results: Dramatically increased the intracellular protein levels of HO-1 in a time-dependent manner in MDCK cells and RAW264.7 cells. Real Time qPCR[1] Cell Types: RAW264.7 cells were infected with IAV A/Fort Monmouth/1/1947 (0.2 MOI) Tested Concentrations: 0.25, 0.5, 1 and 2 μM Incubation Duration: 18 h Experimental Results: Enhanced mRNA expressions of IFN-α/β, as well as protein expressions of some ISGs, such as IFN-inducible transmembrane protein 3 (IFITM3), double-stranded RNA-dependent protein kinase (PKR) and 2′-5′-oligoadenylate synthetase 1 (OAS1), in a dose-dependent manner. Western Blot Analysis[1] Cell Types: RAW264.7 cells or RAW264.7 cells infected with IAV A/Fort Monmouth/1/1947 (0.2 MOI) Tested Concentrations: 2 μM Incubation Duration: 0, 2, 4 and 6 h Experimental Results: Increased protein levels of IRF3 and p- IRF3. Promoted the cytoplasmic protein levels of IRF3 and p-IRF3 in RAW264.7 cells followi
References	[1]. heme oxygenase-1 agonist CoPP suppresses influenza virus replication through IRF3-mediated generation of IFN-α/β. Virology. 2019 Feb;528:80-88.
Additional Infomation	See also: cobalt protoporphyrin IX (preferred).

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.6140 mL	8.0700 mL	16.1400 mL
	5 mM	0.3228 mL	1.6140 mL	3.2280 mL
	10 mM	0.1614 mL	0.8070 mL	1.6140 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.