Formulation & Performance — CQ-12

DCL0019 · Updated 20 October 2026

Study overview

CQ-12 is the C12-chain lead from a chloroquine–epoxide lipidoid series. It is used together with a parent ionizable lipid to inhibit downstream autophagic degradation and improve mRNA expression. The evidence below is from the supplied full article and Figures S1–S34 / Tables S1–S5.

Literature results are formulation- and experiment-specific; they are not product release specifications or guaranteed performance.

Five-component formulations and matched controls

30% substitution means replacing 30% of the original ionizable-lipid molar fraction. For SM-102/MC3, 50 mol% becomes 35 mol% parent lipid plus 15 mol% CQ-12. For ALC-0315, 46.3 mol% becomes 32.41 mol% plus 13.89 mol% CQ-12. All six Table S2 recipes sum to 100 mol%.

Formulation	Component order	Molar ratio	Diameter (nm)	PDI	Zeta potential (mV)	Encapsulation (%)	Apparent LNP pKa
SM-102 parent	SM-102 / DSPC / cholesterol / DMG-PEG2000	50:10:38.5:1.5	127.7±1.4	0.141	−0.74±13.4	95.91±0.01	6.77
SM-102 AUDI	SM-102 / CQ-12 / DSPC / cholesterol / DMG-PEG2000	35:15:10:38.5:1.5	123.7±0.4	0.105	0.33±5.50	94.92±0.03	6.57
MC3 parent	MC3 / DSPC / cholesterol / DMG-PEG2000	50:10:38.5:1.5	133.8±0.9	0.153	−0.23±0.17	91.59±0.30	6.30
MC3 AUDI	MC3 / CQ-12 / DSPC / cholesterol / DMG-PEG2000	35:15:10:38.5:1.5	125.3±2.7	0.122	0.58±1.24	88.90±0.25	6.20
ALC-0315 parent	ALC-0315 / DSPC / cholesterol / ALC-0159	46.3:9.4:42.7:1.6	139.0±0.1	0.083	−1.57±3.14	95.62±0.03	6.30
ALC-0315 AUDI	ALC-0315 / CQ-12 / DSPC / cholesterol / ALC-0159	32.41:13.89:9.4:42.7:1.6	131.8±2.4	0.091	0.34±2.24	94.26±1.79	6.07

Source: SI Table S2; Fig. 2G–J and Fig. S19.

pKa was measured by TNS fluorescence and belongs to the formulated nanoparticle, not isolated CQ-12. Table S2 values are reproduced as reported; uncertainty is shown only where supplied. The pKa values for SM-102 AUDI-LNPs in the separate substitution and chain-length screens are 6.47 (S4) and 6.51 (S5), respectively.

Component molecular weights and mass-ratio tables

Calculated input masses per 100 mg total lipid follow mᵢ = 100 × (mol%ᵢ × MWᵢ) / Σ(mol% × MW). These are lipid-only calculations, before purification or recovery losses. DMG-PEG2000 uses the catalog convention 2507.

SM-102 AUDI-LNP

Component	Molecular weight	mol%	Mass / 100 mg total lipid (mg)
SM-102	710.17	35	42.152
CQ-12	504.20	15	12.826
DSPC	790.15	10	13.400
Cholesterol	386.65	38.5	25.245
DMG-PEG2000	2507	1.5	6.377

MC3 AUDI-LNP

Component	Molecular weight	mol%	Mass / 100 mg total lipid (mg)
DLin-MC3-DMA	642.11	35	39.717
CQ-12	504.20	15	13.366
DSPC	790.15	10	13.964
Cholesterol	386.65	38.5	26.307
DMG-PEG2000	2507	1.5	6.646

MC3 calculation uses average molecular weight 642.11 from C43H79NO2; an exact-mass value must not be substituted for average molecular weight. SM-102, DSPC and cholesterol calculation inputs are 710.17, 790.15 and 386.65, respectively. Replace nominal PEG-lipid values with verified batch-average values when available.

ALC-0315 AUDI-LNP

Component	Molecular weight	mol%	Mass calculation
ALC-0315	766.27	32.41	Pending ALC-0159 batch-average MW
CQ-12	504.20	13.89	Pending ALC-0159 batch-average MW
DSPC	790.15	9.4	Pending ALC-0159 batch-average MW
Cholesterol	386.65	42.7	Pending ALC-0159 batch-average MW
ALC-0159	Batch-average value required	1.6	Not calculated

The ALC-0159 batch-average molecular weight is not supplied in the paper, so a falsely precise total-mass recipe is not shown for this formulation. ALC-0159 is the PEG lipid in this branch; DMG-PEG2000 belongs to the SM-102 and MC3 branches.

Preparation and dosing basis

Parameter	Reported information
Preparation	Ethanol injection; ethanol lipid phase:aqueous RNA phase = 1:3 v/v.
Aqueous phase	Citrate buffer pH 4.0; buffer concentration not specified.
Mass ratio	The source states lipidoid/RNA = 20:1 w/w. It does not unambiguously define whether the numerator includes all five lipids.
N/P	Not reported. The 20:1 mass ratio is not N/P.
Post-processing	Source describes PBS exchange using a 30 kDa centrifugal ultrafiltration device at 3,000 rpm for 15 min, followed by 0.22 µm filtration and 4 °C storage.
Storage duration	No validated formulation or neat-product shelf life is established.
Characterization	DLS size/PDI/zeta, TEM morphology, RiboGreen encapsulation, and TNS apparent pKa.

Source: Methods, printed p. 17.

Formulation-specific performance and experimental conditions

Endpoint	Finding	Conditions / comparator	Source
Lead selection	CQ12 was the highest-expression C6–C18 analogue at the selected 30% SM-102 substitution.	SM-102 base; HEK293T; luciferase mRNA 200 ng/well; 12 h; n=3; comparator: CQ6/8/10/14/16/18 and other substitution ratios	Fig. 2C–E; SI Tables S4–S5
Luciferase expression	Approximately 5.3-fold higher expression than unmodified LNPs in HEK293T cells. This is not a general in vivo fold change.	AUDI-LNPs SM-102; 200 ng mRNA/well; 12 h; n=3; comparator: Unmodified SM-102 LNPs	Main p. 9; Fig. 4C
Compatibility	CQ12 substitution improved reporter expression for SM-102, MC3 and ALC-0315 parent formulations. No exact cross-formulation fold values are tabulated.	HEK293T; 200 ng luciferase mRNA/well; 12 h; n=3; comparator: Matched parent LNPs and free CQ plus parent LNPs	Fig. 4D; SI Table S2
Cell breadth	Higher reporter expression reported in HEK293T, DC2.4, B16, mouse epidermal keratinocytes and mouse lung fibroblasts.	Luciferase mRNA 200 ng/well; 12 h; n=3; comparator: Matched LNP and CQ+LNP controls	Fig. 4C; SI Fig. S33
EGFP expression	Higher EGFP expression with AUDI-LNPs; Fig. 4F plots relative mean fluorescence intensity, despite caption wording referring to positive cells. No exact positive-cell percentage assigned.	HEK293T; EGFP mRNA 2 µg/mL; 12 h; n=3; comparator: LNPs and free CQ+LNPs	Fig. 4E–F; Methods p. 17
Autophagic flux	Accumulation of dual-color LC3 puncta, increased LC3-II/LC3-I and p62, and TEM observations support inhibited downstream autophagic degradation.	HEK293T; western blot and TEM 6 h; Cy5-RNA/LC3 colocalization 4 h; comparator: LNPs and free CQ+LNPs	Fig. 3C–H; SI Fig. S22
RNA escape imaging	No significant difference at 2 h; higher relative signal outside endo/lysosomal regions at 6 h. An absolute escape percentage is not tabulated.	HEK293T; microscopy; n=4 fields; method uses Cy5-siRNA:mLuc 1:2; comparator: LNPs and free CQ+LNPs	Fig. 4G–I; Methods p. 18
Subcutaneous reporter	Stronger injection-site expression at 12 and 24 h; 2 h comparison is marked nonsignificant.	BALB/c mice; 5 µg luciferase mRNA/mouse; single dorsal subcutaneous injection; imaging 2/12/24 h; n=3; comparator: LNPs and free CQ+LNPs	Fig. 6A–C
Subcutaneous IL-10	Increased IL-10 staining in skin at 12 h after local delivery. Separate skin autophagy-marker assay uses mLuc at 6 h.	BALB/c; 5 µg IL-10 mRNA/mouse; dorsal subcutaneous administration; comparator: LNPs and free CQ+LNPs	Fig. 6D–F
Intratracheal reporter	Higher lung reporter signal after local airway delivery; this is not evidence of lung targeting after intravenous dosing.	C57BL/6; 5 µg luciferase mRNA/mouse; intratracheal instillation; 24 h; n=3; comparator: LNPs and free CQ+LNPs	Main p. 11; Fig. 6G–I
Intravenous reporter	Higher systemic reporter expression with liver/spleen signals reported.	BALB/c; 5 µg luciferase mRNA/mouse; single tail-vein injection; 24 h in SI; n=3; comparator: LNPs and free CQ+LNPs	Fig. 6J–L; SI Fig. S29
Psoriasis model	Reported reductions versus model group: scaling 72.73%, erythema 83.33%, and skin thickness 53.5%; lower inflammatory-marker signals.	IMQ-induced BALB/c; IL-10 mRNA 5 µg/mouse; dorsal subcutaneous treatment on day 1; day-5 tissue assessment; comparator: Model, reporter-mRNA AUDI-LNP, IL-10 LNP, free CQ+IL-10 LNP	Main p. 13; Fig. 7; Methods p. 19
Acute lung injury	Lung wet/dry ratio: model 5.84±0.18, TGF-β AUDI-LNP 4.52±0.25, healthy control 4.23±0.15. Authors report ~22.67% reduction versus model; rounded means give ~22.60%.	LPS-induced C57BL/6; TGF-β mRNA 5 µg/mouse; intratracheal treatment 24 h after LPS; evaluate 48 h after treatment; comparator: Healthy/model, reporter control and TGF-β LNP/CQ+LNP comparators	Main p. 15; Fig. 8G; Methods p. 19
Hyperuricemia model	Lower serum uric acid with Uox-mRNA AUDI-LNPs than the comparator groups; no exact effect size assigned.	C57BL/6; Uox mRNA 5 µg/mouse; IV according to Results; intervention 12 h before adenine/oxonate induction; comparator: Healthy/model, reporter control, Uox LNP and CQ+Uox LNP	Main p. 13 and 18–19; SI Fig. S34
Cell viability	At 300 ng/well the main text reports viability around 80%; at 200 ng/well it remains above 85%. SI Fig. S27 gives a more general no-obvious-cytotoxicity description.	HEK293T dose series; context-specific viability, not neat-chemical toxicology; comparator: LNP and CQ+LNP controls	Main p. 9; Fig. 4B; SI Fig. S27
Acute tolerability	No significant differences in displayed liver/kidney chemistry; no obvious pathology in representative organ images. No chronic or human safety inference.	BALB/c; IV 5 µg luciferase mRNA/mouse; serum at 24 h; n=3. Separate SC/IV IL-10 histology.; comparator: PBS, LNP and free CQ+LNP	SI Figs. S28/S30/S31; Table S3

Substitution and chain-length screens

Dataset	Condition	Diameter (nm)	PDI	Encapsulation (%)	Apparent pKa
S4 substitution screen	30% replacement; 35:15:10:38.5:1.5	123.00±1.10	0.105±0.017	93.86±1.94	6.47
S4 substitution screen	75% replacement; 12.5:37.5:10:38.5:1.5	189.50±11.00	0.258±0.016	76.12±5.50	6.32
S4 substitution screen	100% replacement; CQ12:DSPC:Chol:PEG 50:10:38.5:1.5	207.57±12.02	0.266±0.032	71.94±6.00	6.15
S5 chain-length screen	C12 at 30% replacement	121.13±2.40	0.112±0.013	93.50±0.66	6.51

Source: SI Tables S4–S5; values are separate experiments, not pooled with S2.

In S4 the printed 1% recipe (45:0.5:10:38.5:1.5) sums to 95.5 and the 10% recipe (40:5:10:38.5:1.5) sums to 95. These two source entries require clarification and are excluded from mass calculations. The selected 30% recipe is consistent across Fig. 2E and S2/S4/S5.

Structure and analytical identity

The C12 epoxide ring-opening product is attached to the exocyclic quinoline amino nitrogen, giving an N-linked 2-hydroxydodecyl group. The terminal diethylamino group is retained. Two stereocenters are unspecified. The source depicts the neutral parent; no salt stoichiometry or counterion is assigned.

Identity item	Evidence
Formula	C30H50ClN3O, calculated from the structure in main Fig. 2D and SI Figs. S1/S4.
Average molecular weight	504.20, calculated from the verified neutral structure.
HRMS [M+H]+	Calculated 504.3721; observed 504.3695 (SI Table S1, Fig. S4 and Methods).
NMR / chromatography	1H NMR in CDCl3: SI Fig. S11; HPLC chromatogram and PDA spectra: SI Fig. S3. No commercial-batch purity is inferred from these figures.
Synthesis	Chloroquine and 1,2-epoxydodecane ring opening; purified product yield 42% (Methods p. 17).
SMILES	`CCCCCCCCCCC(O)CN(c1ccnc2cc(Cl)ccc12)C(C)CCCN(CC)CC`

Source: Main Fig. 2A/D; Methods p.17; SI Figs. S1/S3/S4/S11 and Table S1.

Interpretation limits

The local intratracheal results do not demonstrate lung targeting after systemic dosing. The 5.3-fold value is the HEK293T reporter comparison, not a universal animal or cargo effect. Inhibition of autophagy may be unsuitable where autophagic clearance is therapeutically important; the authors identify chronic dosing and safety-window studies as future work.

The article has internal timing/label differences: Fig. 3C is described as 6 h in text/methods and 12 h in the caption; Fig. 4F plots relative MFI despite a positive-cell caption; source viability statements differ at the highest dose. These are retained as limits rather than silently harmonized. Raw Source Data were not supplied, so graph-only exact values are not assigned.

References

Zhou T-Q, Sun W, Yang H, Zhang G, Naeem A, Weng Y, Zhu M, Wei Z-Z, Huang Y, Liang X-J. Autolysosomal degradation-inhibitory nanoparticles for enhanced mRNA escape and cytokine therapy. Cell Biomaterials. Article in press (2026), article 100593. doi:10.1016/j.celbio.2026.100593. Article and Supplementary Information.

The supplied PDF carries a 2026 article-in-press citation and a future issue footer (volume 3, 18 May 2027). The DOI is the stable reference used here. Supplementary pages cited above use the printed SI numbering.