Synucleozid (NSC 377363) is a novel inhibitor of SNCA mRNA encoding alpha-synuclein. Synucleozid inhibits the translation of α-synuclein protein by selectively targeting the IRE site of the 5'UTR of α-synuclein mRNA and lowering the amount of SNCA mRNA loaded into polyribosomes. It is possible to use synthucleozid in Parkinson's disease research.

Physicochemical Properties

Molecular Formula	C22H21CLN6
Molecular Weight	404.895342588425
Exact Mass	404.15
CAS #	2741856-68-6
Related CAS #	Synucleozid;502139-01-7
PubChem CID	54601217
Appearance	Light yellow to yellow solid powder
Hydrogen Bond Donor Count	7
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	5
Heavy Atom Count	29
Complexity	561
Defined Atom Stereocenter Count	0
SMILES	C(C1=CC=C2C=C(C3C=CC(NC4C=CC(C(N)=N)=CC=4)=CC=3)NC2=C1)(N)=N.Cl
InChi Key	GKVDOKIPCRSEMM-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H20N6.ClH/c23-21(24)14-5-9-18(10-6-14)27-17-7-3-13(4-8-17)19-11-15-1-2-16(22(25)26)12-20(15)28-19;/h1-12,27-28H,(H3,23,24)(H3,25,26);1H
Chemical Name	2-[4-(4-carbamimidoylanilino)phenyl]-1H-indole-6-carboximidamide;hydrochloride
Synonyms	NSC 377363 hydrochloride; Synucleozid hydrochloride
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, 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	Synucleozid (0.25-1 μM; 24 hours) eliminates the cytotoxicity caused by produced fibrils of α-synuclein, which function as seeds and attract endogenous α-synuclein to aggregate[1]. Synucleozid (0-1 μM; 24 hours) inhibits the expression of α-synuclein protein in SH-SY5Y neuroblastoma cells and binds to the A bulge near the base of the IRE hairpin. It also reduces α-synuclein levels in a dose-dependent manner with an IC50 of 500 nM[1]. Similar affinities are seen in the binding of synucleozid (100 nM–100 μM; 24 hours) to both natural and 2-AP-labeled IRE RNA. It reduces 2-AP emission with an EC50 value of 2.7 ± 0.4 μM. Recovery of 2-AP emissions is shown in relation to the concentration of unlabeled SNCA IRE RNA (RNA-0), providing a competitive Kd of 1.5 ± 0.3 μM[1]. In a dose-dependent manner, synucleozid (0.25-1 μM; 24 hours) reduces α-synuclein and other proteins with IREs in their mRNA's UTR, such as APP, PrP, Ferritin, and TfR. Except for the PrP protein, which is evaluated in Neuro-2A cells, all panels are finished in SH-SY5Y cells[1].
Cell Assay	Cell Viability Assay[1] Cell Types: SH-SY5Y neuroblastoma cells Tested Concentrations: 0.25 μM, 0.5 μM, 1 μM Incubation Duration: 24 hrs (hours) Experimental Results: diminished LDH release as a dose-dependent manner. Western Blot Analysis[1] Cell Types: SH-SY5Y neuroblastoma cells Tested Concentrations: 0.25 μM, 0.5 μM, 1 μM Incubation Duration: 24 hrs (hours) Experimental Results: diminished α-synuclein expression as a concentration-dependent manner.
References	[1]. Translation of the intrinsically disordered protein α-synuclein is inhibited by a small molecule targeting its structured mRNA.Proc Natl Acad Sci U S A. 2020 Jan 21;117(3):1457-1467.

Solubility Data

Solubility (In Vitro)

DMSO : ~125 mg/mL (~308.7 mM) H2O : ~2 mg/mL (~5 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (5.14 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 20.8 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: ≥ 2.08 mg/mL (5.14 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 20.8 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.4697 mL	12.3487 mL	24.6975 mL
	5 mM	0.4939 mL	2.4697 mL	4.9395 mL
	10 mM	0.2470 mL	1.2349 mL	2.4697 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.