A2-Iso5-2DC18 is a dihydroimidazole-linked lipid that is a potent mRNA delivery vehicle. A2-Iso5-2DC18 may be utilized in anti-tumor research including B16F10 melanoma.

Physicochemical Properties

Molecular Formula	C47H87N3O2
Molecular Weight	726.21
Exact Mass	725.679
CAS #	2412492-07-8
PubChem CID	146461367
Appearance	Light yellow to yellow liquid（Density：0.95±0.1 g/cm3）
LogP	16.6
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	37
Heavy Atom Count	52
Complexity	868
Defined Atom Stereocenter Count	0
SMILES	CCCCCCCC/C=C\CCCCCCCC1(N(C(N=C1)C(=O)OCC)CCCN2CCCC2)CCCCCCC/C=C\CCCCCCCC
InChi Key	WMYPEEPUVOTFJU-WRBBJXAJSA-N
InChi Code	InChI=1S/C47H87N3O2/c1-4-7-9-11-13-15-17-19-21-23-25-27-29-31-33-38-47(39-34-32-30-28-26-24-22-20-18-16-14-12-10-8-5-2)44-48-45(46(51)52-6-3)50(47)43-37-42-49-40-35-36-41-49/h19-22,44-45H,4-18,23-43H2,1-3H3/b21-19-,22-20-
Chemical Name	ethyl 5,5-bis[(Z)-heptadec-8-enyl]-1-(3-pyrrolidin-1-ylpropyl)-2H-imidazole-2-carboxylate
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: (1). This product requires protection from light (avoid light exposure) during transportation and storage.(2). 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 Vivo	A2-Iso5-2DC18 transfects central antigen-presenting cells (APCs) in the A14 /Cre mRNA mouse model by loading it with either mLuc or Cre-recombinase mRNA LNPs (mCre) at doses of 0.1 mg/kg and 0.5 mg/kg, subcutaneously, once every week for two weeks[1]. In the B16F10 mouse melanoma model, A2-Iso5-2DC18 loaded with OVA mRNA (mOVA) vaccination (15 μg mOVA per animal; sc; twice dosage, once every 5 d) elicits a robust IFN-ɤ production concurrently with a significantly high antigen-specific cytotoxic T lymphocyte (CTL) response[1].
Animal Protocol	Animal/Disease Models: A14/Cre mRNA mouse model (female B6 mice)[1] Doses: Loaded with Cre-recombinase mRNA LNPs (mCre ); 0.1 mg/kg and 0.5 mg/kg Route of Administration: subcutaneous (sc) injection; 3 weeks Experimental Results: Induced protein expression in the local injection site and the draining lymph nodes and transfected central antigen presenting cells (APCs) including macrophages/monocytes (CD11b+) and dendritic cells (CD11c+) in mice. Animal/Disease Models: Ovalbumin (OVA)-expressing B16F10 mouse melanoma model[1] Doses: Loaded with OVA mRNA (mOVA) vaccine; 15 μg mOVA per mouse Route of Administration: subcutaneous (sc) injection; once per week for the first two weeks; 3 weeks continuous observation Experimental Results: Dramatically diminished tumor volume of B16-OVA melanoma and improved overall survival in mice. Increased the number of systemic and tumor-infiltrating antigen-specific T cells dramatically (20–30-fold).
References	[1]. Miao L, et al. Delivery of mRNA vaccines with heterocyclic lipids increases anti-tumor efficacy by STING-mediated immune cell activation. Nat Biotechnol. 2019 Oct;37(10):1174-1185.

Solubility Data

Solubility (In Vitro)

DMSO: 100 mg/mL (137.70 mM）

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (3.44 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. 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. Solubility in Formulation 2: 2.5 mg/mL (3.44 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (3.44 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. 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 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.3770 mL	6.8851 mL	13.7701 mL
	5 mM	0.2754 mL	1.3770 mL	2.7540 mL
	10 mM	0.1377 mL	0.6885 mL	1.3770 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.