UBA5-IN-1 (compound 8.5) is a selective inhibitor of UBA5 with IC50 of 4.0 μM. UBA5-IN-1 suppresses the proliferation/growth of Sk-Luci6 cancer cells that highly express UBA5.

Physicochemical Properties

Molecular Formula	C26H40F6N10O11S2ZN
Molecular Weight	912.16
Exact Mass	910.151
CAS #	1831169-11-9
PubChem CID	102004328
Appearance	White to off-white solid powder
Hydrogen Bond Donor Count	7
Hydrogen Bond Acceptor Count	24
Rotatable Bond Count	8
Heavy Atom Count	56
Complexity	914
Defined Atom Stereocenter Count	4
SMILES	C(F)(F)(F)S([O-])(=O)=O.C(N12CCN3CCN4CCN(CC1)[Zn+2]234)(=O)CCCCCNC([C@@H]1[C@H]([C@@H](O)[C@H](N2C=NC3=C(N=CN=C23)N)O1)O)=O
InChi Key	DLPRYIVGTKSVTR-WAYDLRBYSA-L
InChi Code	InChI=1S/C24H40N10O5.2CHF3O3S.Zn/c25-21-17-22(31-14-30-21)34(15-32-17)24-19(37)18(36)20(39-24)23(38)29-5-3-1-2-4-16(35)33-12-10-27-8-6-26-7-9-28-11-13-33;2*2-1(3,4)8(5,6)7;/h14-15,18-20,24,26-28,36-37H,1-13H2,(H,29,38)(H2,25,30,31);2*(H,5,6,7);/q;;;+2/p-2/t18-,19+,20-,24+;;;/m0.../s1
Chemical Name	zinc;(2S,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxy-N-[6-oxo-6-(1,4,7,10-tetrazacyclododec-1-yl)hexyl]oxolane-2-carboxamide;trifluoromethanesulfonate
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	IC50: 4.0 μM (UBA5), 78.5 μM (UAE), 66.8 μM (NAE)[1]
ln Vitro	UBA5-IN-1 (0.1-1000 μM) inhibits UAE and NAE with IC50 values of 78.5 and 66.8 μM, respectively, and exhibits a dose-dependent inhibition effect to UBA5 with an IC50 value of 4.0 μM[1]. In terms of ATP, UBA5 is non-competitively inhibited by UBA5-IN-1 (0–5 μM)[1]. The effects of UBA5-IN-1 (10 μM) on ATP binding to human kinase are negligible[1]. The proliferation of Sk-Luci6 cancer cells with high UBA5 expression levels is affected by UBA5-IN-1 (0–50 μM; 72 h)[1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: Sk-Luci6 cancer cells with high expression levels of UBA5 Tested Concentrations: 0-50 μM Incubation Duration: 0-72 h Experimental Results: demonstrated selective anti-proliferative activity for Sk-Luci6 cancer cells which expressed higher UBA5 protein levels, but demonstrated no anti-proliferative activity to MRC9 lung fibroblasts and A549 carcinoma cells.
References	[1]. A selective inhibitor of the UFM1-activating enzyme, UBA5. Bioorg Med Chem Lett. 2016 Sep 15;26(18):4542-4547.

Solubility Data

Solubility (In Vitro)

DMF: 100 mg/mL (109.63 mM) DMSO: 50 mg/mL (54.81 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 1.25 mg/mL (1.37 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 12.5 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: ≥ 1.25 mg/mL (1.37 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 12.5 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: ≥ 1.25 mg/mL (1.37 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 12.5 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 4: ≥ 1.25 mg/mL (1.37 mM) (saturation unknown) in 10% DMF 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 5: ≥ 1.25 mg/mL (1.37 mM) (saturation unknown) in 10% DMF 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. 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 6: ≥ 1.25 mg/mL (1.37 mM) (saturation unknown) in 10% DMF 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.0963 mL	5.4815 mL	10.9630 mL
	5 mM	0.2193 mL	1.0963 mL	2.1926 mL
	10 mM	0.1096 mL	0.5481 mL	1.0963 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.