Polyinosinic-polycytidylic acid potassium (Poly(I:C) potassium) is a synthetic analog of double-stranded RNA and an agonist of TLR3 and retinoic acid-inducible gene I receptors (RIG-I and MDA5). Polyinosinic-polycytidylic acid potassium could be utilized as a vaccine adjuvant to enhance innate and adaptive immune responses, change the tumor microenvironment, and directly trigger apoptosis in cancer cells.

Physicochemical Properties

Molecular Formula	C19H27KN7O16P2
Molecular Weight	710.50080704689
Exact Mass	710.062
CAS #	31852-29-6
Related CAS #	Polyinosinic-polycytidylic acid;24939-03-5;Polyinosinic-polycytidylic acid sodium;42424-50-0
PubChem CID	137031716
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	10
Hydrogen Bond Acceptor Count	18
Rotatable Bond Count	8
Heavy Atom Count	45
Complexity	1090
Defined Atom Stereocenter Count	8
SMILES	C1=CN(C(=O)N=C1N)[C@H]2[C@@H]([C@@H]([C@H](O2)COP(=O)(O)O)O)O.C1=NC2=C(C(=O)N1)N=CN2[C@H]3[C@@H]([C@@H]([C@H](O3)COP(=O)(O)O)O)O.[K]
InChi Key	KHPUABRWHJGXNJ-VRQAYDGLSA-N
InChi Code	InChI=1S/C10H13N4O8P.C9H14N3O8P.K/c15-6-4(1-21-23(18,19)20)22-10(7(6)16)14-3-13-5-8(14)11-2-12-9(5)17;10-5-1-2-12(9(15)11-5)8-7(14)6(13)4(20-8)3-19-21(16,17)18;/h2-4,6-7,10,15-16H,1H2,(H,11,12,17)(H2,18,19,20);1-2,4,6-8,13-14H,3H2,(H2,10,11,15)(H2,16,17,18);/t4-,6-,7-,10-;4-,6-,7-,8-;/m11./s1
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

Targets	TLR3
ln Vitro	Potassium polyinosinic-polycytidylic acid (20 ng/mL; 24 hours; WM793, WM278, WM239A, WM9, and 1205Lu cells) inhibits the growth activity of cells [1]. Apoptosis in 1205Lu cells is induced by 200 ng/mL of potassium polyinosinic-polycytidylic acid for a 24-hour period [1]. Melanoma cells are induced to express IFN-β when exposed to potassium polyinosinic-polycytidylic acid (5 ng/mL; 24 hours; 1205Lu cells). IFN-β induction by polyinosinic-polycytidylic acid potassium needs IPS-1, RIG-I, and MDA-5, respectively, as demonstrated by the silencing of these two proteins [1]. Potassium polyinosinic-polycytidylic acid potassium was made ready for injection by resuspending it in sterile saline at a concentration of 2 mg/ml, heating it to 50 °C to guarantee total dissolution, and letting it cool naturally to room temperature to guarantee appropriate double-stranded RNA annealing. Potassium polyinonose-polycytidylic acid. I:C Poly Until used, keep stored at -20 °C [3].
ln Vivo	In NOD/SCID immunodeficient mice, polyinosinic-polycytidylic acid potassium (50 µg; iv; on days 3, 6, and 9 following tumor inoculation) exhibits anti-tumor action [1].
Cell Assay	Cell Viability Assay[1] Cell Types: WM793, WM278, WM239A, WM9 and 1205Lu cells Tested Concentrations: 20 ng/mL Incubation Duration: 24 hrs (hours) Experimental Results: Strongly decreased viability from 100% in controls to 20%–50% within 24 hrs (hours). Apoptosis Analysis[1] Cell Types: 1205Lu cells Tested Concentrations: 200 ng/mL Incubation Duration: 24 hrs (hours) Experimental Results: Induced apoptosis in 1205Lu cells. RT-PCR[1] Cell Types: WM793, WM278, WM239A, WM9 and 1205Lu cells Tested Concentrations: 3 ng/mL Incubation Duration: 24 hrs (hours) Experimental Results: Induced IFN-β expression in melanoma cells. Western Blot Analysis[1] Cell Types: 1205Lu cells Tested Concentrations: 5 ng/mL Incubation Duration: 24 hrs (hours) Experimental Results: Revealed active subunits of caspase-9 and caspase-8 in melanoma cells.
Animal Protocol	Animal/Disease Models: NOD/SCID immunodeficient mice (1205Lu cells)[1] Doses: 50 µg Route of Administration: Iv; on days 3, 6, and 9 after tumor inoculation Experimental Results: diminished the size of metastases and the total amount of tumor tissue, and the level of human DNA was 50% lower in mice.
References	[1]. Proapoptotic signaling induced by RIG-I and MDA-5 results in type I interferon-independent apoptosis in human melanoma cells. J Clin Invest. 2009 Aug;119(8):2399-411. [2]. Anticancer function of polyinosinic-polycytidylic acid. Cancer Biol Ther. 2010 Dec 15;10(12):1219-23. [3]. Systemic challenge with the TLR3 agonist poly I:C induces amplified IFNalpha/beta and IL-1beta responses in the diseased brain and exacerbates chronic neurodegeneration. Brain Behav Immun. 2010 Aug;24(6):996-1007.

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	1.4075 mL	7.0373 mL	14.0746 mL
	5 mM	0.2815 mL	1.4075 mL	2.8149 mL
	10 mM	0.1407 mL	0.7037 mL	1.4075 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.