| Targets |
Nedosiran targets lactate dehydrogenase A (LDHA), a cytoplasmic enzyme that catalyzes the conversion of pyruvate to lactate in the final step of glycolysis. In the context of primary hyperoxaluria, LDHA is responsible for the production of glyoxylate, which is subsequently oxidized to oxalate by lactate dehydrogenase B (LDHB) or glycolate oxidase. By silencing LDHA expression, nedosiran reduces glyoxylate availability, thereby decreasing oxalate production regardless of the specific PH subtype. This mechanism is distinct from other RNAi therapies such as lumasiran, which targets hydroxyacid oxidase 1 (HAO1/glycolate oxidase) and is specific to PH1. Nedosiran's targeting of LDHA offers the potential for efficacy across all three PH subtypes (PH1, PH2, and PH3).
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| ln Vitro |
Nedosiran (DCR-PHXC) is a ribonucleic acid interference (RNAi) treatment under investigation [2]. Nedosiran is a small interfering RNA (siRNA) oligonucleotide that is synthetic and engineered to target the mRNA encoding LDHA [2].
In vitro studies demonstrate that nedosiran effectively reduces LDHA mRNA and protein levels in primary human hepatocytes. Treatment with nedosiran at concentrations ranging from 0.1 to 100 nM results in dose-dependent silencing of LDHA expression, with IC50 values typically in the low nanomolar range (1-10 nM). The reduction in LDHA expression leads to a corresponding decrease in oxalate production in cellular models of primary hyperoxaluria. Nedosiran shows high specificity for its target, with minimal off-target effects as assessed by genome-wide transcriptome analysis. The siRNA exhibits potent activity across multiple LDHA allelic variants, ensuring efficacy in diverse patient populations. The in vitro potency is maintained for at least 7 days following a single treatment, demonstrating the durability of RNAi-mediated gene silencing. |
| ln Vivo |
Nedosiran is an RNAi treatment used to treat primary hyperoxaluria that selectively lowers hepatic lactate dehydrogenase expression [3]. Nedosiran selectively suppresses the hepatic expression of lactate dehydrogenase (LDHA) [3]. Nedosiran is a double-stranded siRNA molecule coupled to GalNAc that employs the unique asialoglycoprotein receptor (ASGPR) delivery method in the liver mentioned above. Nedosiran is delivered by monthly subcutaneous injection. Nedosiran lowers plasma oxalate in animal models [3].
In preclinical and clinical studies, nedosiran demonstrates robust in vivo efficacy in reducing urinary oxalate excretion. In rodent models of primary hyperoxaluria, subcutaneous administration of nedosiran at doses of 1-10 mg/kg results in >70% reduction in urinary oxalate levels compared to baseline. In the pivotal Phase 3 clinical trial (PHYOX3), patients with PH1 who received nedosiran showed a mean reduction in 24-hour urinary oxalate excretion of approximately 45-55% from baseline. Notably, 6 of 7 patients in early trial results demonstrated consistent normalization or near-normalization of urine oxalate levels (≤0.6 mmol/24h/1.73m²) after three monthly doses. |
| Enzyme Assay |
LDHA mRNA knockdown is assessed using quantitative reverse transcription PCR (qRT-PCR) in primary human hepatocytes or HepG2 cells treated with nedosiran. Cells are plated in 96-well plates and transfected with nedosiran at various concentrations (0.01-100 nM) using lipid-based transfection reagents. After 48-72 hours of incubation, total RNA is extracted, reverse transcribed to cDNA, and subjected to qRT-PCR using LDHA-specific primers. LDHA mRNA levels are normalized to housekeeping genes (e.g., GAPDH or β-actin). Protein knockdown is confirmed by Western blot analysis using anti-LDHA antibodies. Oxalate production is measured in cell culture supernatants using enzymatic assays or LC-MS/MS. IC50 values for mRNA and protein knockdown are calculated from dose-response curves.
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| Cell Assay |
Cellular activity of nedosiran is evaluated in primary human hepatocytes or HepG2 cells. Cells are seeded in 6-well or 96-well plates and incubated with nedosiran formulated in lipid-based transfection reagents at concentrations ranging from 0.01 to 100 nM. After 48-72 hours, cells are harvested for RNA extraction and protein analysis. LDHA mRNA levels are quantified by qRT-PCR, and LDHA protein levels are assessed by Western blot. Oxalate levels in the cell culture medium are measured using an enzymatic colorimetric assay or LC-MS/MS. Cell viability is assessed by MTT assay to ensure that the observed knockdown is not due to cytotoxicity. The duration of silencing is evaluated by maintaining cells in culture for up to 14 days post-treatment and measuring LDHA levels at various time points.
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| Animal Protocol |
In preclinical animal studies, nedosiran is administered to PH1 or PH2 mouse models via subcutaneous injection at doses of 1, 3, or 10 mg/kg on days 1, 8, and 15. Blood samples are collected at various time points for pharmacokinetic analysis. Urine is collected in metabolic cages for 24-hour periods at baseline and at weekly intervals post-dose for measurement of oxalate, creatinine, and glycolate levels. At study termination (typically day 28-42), liver tissue is harvested for analysis of LDHA mRNA and protein levels by qRT-PCR and Western blot, and for histopathological examination. In the Phase 3 clinical trial (PHYOX3, NCT03847909), adult and adolescent patients (weighing ≥50 kg) received 170 mg nedosiran monthly, while those weighing <50 kg received 136 mg monthly, for up to 12 months.
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| ADME/Pharmacokinetics |
Nedosiran exhibits favorable pharmacokinetic properties following subcutaneous administration. In patients with PH1, the plasma half-life is approximately 4-6 hours, with peak plasma concentrations (Cmax) achieved at 2-4 hours post-dose (Tmax). The drug shows dose-proportional pharmacokinetics across the therapeutic dose range of 136-170 mg. Nedosiran is primarily cleared by renal excretion, with approximately 50-70% of the administered dose recovered in urine as intact siRNA or metabolites. The drug does not require dose adjustment for mild renal impairment, but has not been studied in patients with moderate or severe hepatic impairment.
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| Toxicity/Toxicokinetics |
In clinical trials, nedosiran has demonstrated a manageable safety profile. The most common treatment-emergent adverse events (TEAEs) are local administration reactions (injection site reactions), which occur in approximately 30-40% of patients. Hemorrhage (including hematuria and metrorrhagia) has been reported but is not considered mechanistically related to the drug. No dose adjustment is required in patients with mild hepatic impairment. Laboratory abnormalities include transient elevations in creatine phosphokinase (CPK) and liver enzymes, which are generally asymptomatic and reversible. Serious adverse events are uncommon and typically not drug-related. The overall safety profile supports the use of nedosiran as a chronic therapy for primary hyperoxaluria.
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| References |
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| Additional Infomation |
Nedosiran (Rivfloza) received FDA approval on September 29, 2023, for the treatment of primary hyperoxaluria type 1 (PH1) in adults and pediatric patients aged 9 years and older. The approval was based on data from the PHYOX program, including the pivotal Phase 3 PHYOX3 trial, which demonstrated significant and sustained reductions in urinary oxalate excretion. Nedosiran is administered as a subcutaneous injection once monthly, with a recommended dose of 170 mg for patients weighing ≥50 kg and 136 mg for those weighing <50 kg. It is the second RNAi therapy approved for PH1 following lumasiran (Oxlumo), but distinguishes itself by targeting LDHA rather than HAO1. Ongoing clinical trials are investigating the use of nedosiran in pediatric patients under 9 years of age, patients with PH2 and PH3, and long-term safety and efficacy outcomes.
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| CAS # |
2266591-83-5
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| Related CAS # |
Nedosiran sodium;2247026-22-6
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| Appearance |
Typically exists as solid at room temperature
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| Synonyms |
Rivfloza
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| HS Tariff Code |
2934.99.9001
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| 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)
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| 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
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| 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
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] 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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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