RGLS4326 (RG4326) is a first-in-class microRNA-17 (miR-17) oligonucleotide inhibitor. RGLS4326 may be utilized in the study of autosomal dominant polycystic kidney disease (ADPKD). RGLS4326 acts on HeLa cells and inhibits the function of miR-17 with EC50 of 28.3 nM.

Physicochemical Properties

Molecular Formula	C95H115F3N32O51P8S8
Molecular Weight	3082.42
CAS #	2229964-07-0
Appearance	White to off-white solid powder
SMILES	S=P(O)(OC[C@@]12[C@H](C)O[C@@]([H])([C@H](N3C=NC4C(NC(N)=NC3=4)=O)O1)[C@@H]2OP(=O)(OC[C@@H]1[C@H]([C@H]([C@H](N2C(N=C(C=C2)N)=O)O1)OC)OP(O)(OC[C@@H]1[C@H]([C@H]([C@H](N2C=NC3=C(N)N=CN=C23)O1)F)OP(O)(OC[C@@H]1[C@H]([C@H]([C@H](N2C(N=C(C=C2)N)=O)O1)F)OP(O)(OC[C@@H]1[C@H]([C@H]([C@H](N2C=CC(NC2=O)=O)O1)F)OP(O)(OC[C@@H]1[C@H]([C@H]([C@H](N2C=CC(NC2=O)=O)O1)OC)OP(O)(OC[C@@]12[C@H](C)O[C@@]([H])([C@H](N3C=CC(NC3=O)=O)O1)[C@@H]2OP(O)(OC[C@@]12[C@H](C)O[C@@]([H])([C@H](N3C=NC4C(NC(N)=NC3=4)=O)O1)[C@@H]2O)=S)=S)=S)=S)=S)=S)S)O[C@H]1[C@]2([H])[C@H](N3C=NC4=C(N)N=CN=C34)O[C@@]1(CO)[C@H](C)O2
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 (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

Targets	MicroRNA[1]
ln Vitro	The miR-17 seed sequence is completely complementary to RGLS4326, a single-stranded, chemically altered 9-nt short oligonucleotide. In ADPKD, RGLS4326 suppresses the miR-17 family's pathogenic activity [1]. According to miR-17 PD-Sig, RGLS4326 administration reduces miR-17 function in renal collecting duct cells in culture, with an EC50 value of 77.2 ± 20.2 nM [1]. Human autosomal dominant polycystic kidney disease (ADPKD) cysts are inhibited from growing by RGLS4326 [1].
ln Vivo	RGLS4326 distributes to renal tubules and cysts more selectively. With a half-life of less than four hours in wild-type mice and a Tmax of less than one hour and a Cmax of 8.5 μg/mL, RGLS4326 (single 30 mg/kg subcutaneous injection) is quickly absorbed into plasma [1]. The direct miR-17 target genes Pkd1 and Pkd2 are likewise upregulated in expression when RGLS4326 is administered in vivo[1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: Primary cysts in 3D Matrigel Tested Concentrations: 5, 20, 100, and 300 nM Incubation Duration: 9 days Experimental Results: diminished in cyst epithelial cell proliferation.
Animal Protocol	Animal/Disease Models: Pkd2-KO mice[1] Doses: 20 mg/kg Route of Administration: SC injection Experimental Results: Compared to non-cystic control kidneys, polycystic kidneys of PBS-treated Pkd2-KO mice exhibit an age-dependent progressive decline in miR-17 PD-Sig, indicative of increasing miR-17 activity with disease progression. Administration of RGLS4326 reversed this decline in miR-17 PD-Sig, indicating a sustained functional inhibition of miR-17.
References	[1]. Discovery and preclinical evaluation of anti-miR-17 oligonucleotide RGLS4326 for the treatment of polycystic kidney disease. Nat Commun. 2019 Sep 12;10(1):4148.

Solubility Data

Solubility (In Vitro)

H2O : ≥ 100 mg/mL (32.44 mM)

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	0.3244 mL	1.6221 mL	3.2442 mL
	5 mM	0.0649 mL	0.3244 mL	0.6488 mL
	10 mM	0.0324 mL	0.1622 mL	0.3244 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.