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C12-200
Formulation & Performance
Formulation & Performance
Study-specific compositions, preparation conditions and experimental outcomes for C12-200.
Main-article Results were checked through PMC during this pass. The previously reviewed formulation and NHP-method record is retained from the existing live page, with experiment-level restructuring. Fresh primary PDF and supporting-information downloads were blocked; this is not a new complete SI review.
1. Formulations and component amounts
Reported composition and calculated mass are separate. Molecular-weight inputs use catalog values or an explicitly identified reference average; DMG-PEG2000 uses 2507. Edit inputs only when using a verified alternative material.
C12-200 four-component siRNA LNP
mol%Love 2010, main Methods, Formulation of LNPs; previously reviewed live-page record retained
| Component | Molecular weight | Molar parts | Normalized mol% | Amount (µmol) | Weight (mg) |
|---|---|---|---|---|---|
| C12-200 | 50.000 | 59.9559 | 68.1658 | ||
| DSPC | 10.000 | 11.9912 | 9.4748 | ||
| Cholesterol | 38.500 | 46.1661 | 17.8501 | ||
| DMG-PEG2000 | 1.500 | 1.7987 | 4.5093 |
Calculated total: 100.0000 mg lipid. RNA, solvent and process loss are excluded.
Initial lipidoid library screening
Constant weight ratio across the tested library; not the optimized C12-200 molar ratioMain Results, initial in vivo screening and Fig.3
| Component | Molecular weight | Reported molar entry | Calculated mass |
|---|---|---|---|
| Lipidoid | Not confirmed | Not recovered | Not calculated |
| DSPC | 790.15 | Not recovered | Not calculated |
| Cholesterol | 386.65 | Not recovered | Not calculated |
| PEG lipid | Not confirmed | Not recovered | Not calculated |
No complete mass recipe is generated for this formulation while composition or identity-dependent inputs remain unresolved.
DMG-PEG2000 uses 2507 for catalog component calculations. The earlier formulation record gives 2660 for its PEG stock; calculated component weights here therefore use the current catalog material rather than reproducing that historical stock exactly. C12-200 uses the existing catalog molecular-weight value of 1136.931; its structure and salt-form basis have not been independently reverified in this review.
How component weights are calculated
For total lipid mass M (mg), molar parts rᵢ and molecular weight MWᵢ: component mass = M × rᵢ × MWᵢ / Σ(r × MW). Component amount (µmol) = component mass (mg) × 1000 / MWᵢ. Molar parts are normalized only for the displayed percentage. The calculation does not infer N/P or RNA dose.
2. Preparation and experimental setup
| Parameter | Reported procedure / limitation | Source |
|---|---|---|
| Organic phase | C12-200, DSPC, cholesterol and mPEG2000-DMG were dissolved in 90% ethanol. | Main Methods, Formulation of LNPs; retained prior extraction |
| Aqueous phase | siRNA in 10 mM citrate buffer at pH 3. | Main Methods, Formulation of LNPs; retained prior extraction |
| RNA loading | Total lipid:siRNA 7:1 by mass. This refers to all lipid components together, not C12-200 alone. N/P was not reported in the retained formulation record. | Main Methods, Formulation of LNPs; retained prior extraction |
| Mixing | Lipid and siRNA streams were pumped at equivalent volumetric flow rates through a T-junction. Absolute flow rate was not recovered. | Main Methods, Formulation of LNPs; retained prior extraction |
| Buffer exchange | Dialysis against PBS containing 155 mM NaCl, 3 mM Na2HPO4 and 1 mM KH2PO4, pH 7.5. | Main Methods, Formulation of LNPs; retained prior extraction |
| Experiment separation | Cell-screening lipidoid/siRNA complexes used distinct mass ratios; those values are not the four-component in vivo LNP composition. | Main Results, Figs.2–3 |
3. Particle characterization
| Measurement / batch | Reported result | Evidence context |
|---|---|---|
| Particle diameter | Approximately 80 nm | Main Methods; retained prior extraction; LNP value, not neat-lipid specification |
| siRNA entrapment | Approximately 90%, assessed by RiboGreen | Main Methods; retained prior extraction |
| N/P, PDI and apparent pKa | No experiment-specific numerical values recovered in this review | Do not infer N/P from the total-lipid mass ratio or borrow values from later C12-200 studies |
4. Performance by experiment
| Experiment and dose | Readout and comparator | Result and source |
|---|---|---|
| Initial in vivo lipidoid screen C57BL/6 mice; single intravenous tail-vein administration; anti-FVII siRNA 3 mg/kg; 12 purified lipidoids; n=3 or 4 | Readout: 72 h Comparator: Other library members and serum FVII baseline | The lead C12-200 emerged from a fixed-weight-composition screen. This initial screen is distinct from the optimized low-dose C12-200 experiments.Main Results, initial in vivo screen and Fig.3 |
| Low-dose Factor VII response C57BL/6 mice; intravenous anti-FVII siRNA in C12-200 LNP | Readout: Assay timing is experiment-specific; 48/72 h is not pooled into one time point Comparator: LNP01 reference; nontargeting siRNA control at 1 mg/kg | Activity reported below 0.01 mg/kg siRNA; more than two orders of magnitude greater dose potency than the earlier LNP01 system. No exact ED50 is inferred from the abstract claim.Main Abstract and Results, Fig.3 |
| Duration of Factor VII suppression C57BL/6 mice; single intravenous administration of 0.1 or 1 mg/kg anti-FVII siRNA; n=3 | Readout: 24 h to approximately 35 days Comparator: Predose/baseline serum Factor VII | Complete silencing at 24 h; Factor VII protein returned to baseline by approximately 20 days at 0.1 mg/kg and 35 days at 1 mg/kg.Main Results, Fig.4 |
| Five-target liver silencing Mouse liver; FVII, ApoB, PCSK9, XBP1 and SORT1 siRNAs; single intravenous administration; 0.2 mg/kg per siRNA, totaling 1 mg/kg; n=5 | Readout: 72 h Comparator: Saline and control-siRNA groups | More than 65% silencing for each of the five target mRNAs. The reported 0.2 mg/kg is per target and must not be presented as the total mixture dose.Main Results, Fig.5 |
| Cellular uptake mechanism HeLa cells; fluorescent siRNA/C12-200; dextran, transferrin and cholera toxin B trafficking markers; EIPA and cytochalasin D inhibition experiments | Readout: Membrane actin ruffling observed after 15 min; uptake-assay timings depend on experiment Comparator: Marker colocalization and inhibitor controls | Dextran colocalization, actin changes and inhibitor-sensitive uptake support a macropinocytosis-associated route in this cell model. They do not establish an exclusive uptake route in vivo.Main Results, Fig.6 |
| Hepatic TTR silencing in nonhuman primates Cynomolgus monkeys; anti-TTR siRNA 0.03, 0.1 or 0.3 mg/kg; 15-minute cephalic-vein infusion; n=3 per dose | Readout: 48 h liver biopsy Comparator: PBS and irrelevant-siRNA controls | Dose-dependent and sequence-specific hepatic TTR mRNA knockdown. Exact percentage values are not transcribed from the inaccessible figure in this pass.Main Results, nonhuman-primate TTR experiment; NHP methods retained from prior extraction |
| Reported mouse tolerability observations C12-200 siRNA LNP at 1 mg/kg in the reported mouse experiments | Readout: Study-specific observations Comparator: PBS/control measurements | No reported indication of toxicity or body-weight change; liver chemistry results were assessed in the paper. This limited observation does not establish long-term or clinical safety.Main Results, Table1 and Fig.4 discussion |
Mechanistic evidence
C12-200 is an aminoalcohol lipidoid identified through an epoxide-based combinatorial library. Cell experiments support macropinocytosis-associated uptake; hepatic siRNA knockdown demonstrates functional delivery. These observations are separate from direct quantitative measurements of endosomal escape, which are not recovered here.
5. Interpretation and unresolved details
- The 2010 primary evidence concerns siRNA silencing, not mRNA expression or CRISPR delivery. Later uses require their own sources.
- Initial library screening used constant component weight proportions. Do not describe that entire screen as the optimized 50:10:38.5:1.5 mol% formulation.
- The five-siRNA mixture delivers 0.2 mg/kg of each siRNA, or 1 mg/kg total.
- Mouse Factor VII efficacy, HeLa-cell uptake experiments and monkey TTR outcomes concern different biological models.
- High monkey knockdown is reported qualitatively here; no percentage is invented from an unavailable figure.
- The nonhuman-primate experiment is cited by its Results section; Fig. 6 of the main article concerns cellular uptake.
Results apply to the stated LNP composition, cargo, process, dose, route and model. LNP experimental measurements are not batch-release specifications for the isolated lipid.
6. References and data access
Love KT, et al. Lipid-like materials for low-dose, in vivo gene silencing. Proc Natl Acad Sci U S A. 2010;107(5):1864–1869.
Download experiment table (CSV) · Open printable Formulation & Performance guide
Revision FP-20261024-b3 · Source review: 24 October 2026 · DC Chemicals