Molecular Pharmaceutics最新文献

The intracellular delivery of full-length antibodiesoffers substantialtherapeutic potential but remains limited by poor cellular uptake,extracellular degradation, and inefficient encapsulation strategies.Here, we report a noninvasive, scalable, and biocompatible nanocarrierplatform based on poly-l-lysine (PLL)-coated polyethyleneglycol-block-poly(lactic-co-glycolicacid) (PEG–PLGA) polymersomes for efficient intracellular antibodydelivery. Coating polymersomes with 30 kDa ε-poly-l-lysine increased antibody encapsulation efficiency to ∼ 80%.It enabled precise modulation of surface charge to a mildly positiveζ-potential (∼+4.5 mV), while maintaining nanoscale dimensions(hydrodynamic diameter ≈ 420 ± 30 nm). The resulting formulationexhibited excellent biocompatibility, preserving >96% cell viabilityin primary human pulmonary fibroblasts. Importantly, PLL-coated polymersomesfacilitated efficient intracellular delivery of full-length antibodiesand preserved their biological function, as demonstrated by robustsuppression of NOD-like receptor family pyrin domain-containing 3(NLRP3)-dependent IL-1β signaling. Upon pulmonary administrationvia aerosolization, polymersomes delivered the antibody efficientlyto lung-resident cells in vivo without detectable acute cytotoxicity.To our knowledge, this work represents the first demonstration ofPLL-coated PEG–PLGA polymersomes enabling intracellular deliveryof full-length antibodies both in vitro and in vivo, establishinga versatile nanoplatform for lung-targeted intracellular antibodytherapeutics.

Residual water in freeze-dried protein formulations playsa criticalrole in molecular relaxation behavior. In a previous study (Kumaret al., Mol Pharmaceutics 2025, 22, 4125–4136), we examinedthe impact of sucrose concentration on native structure retentionin monoclonal antibody (mAb)–sucrose systems. The present workinvestigates how residual water content affects relaxation dynamicsin this formulation system. Water affinity was first assessed viasorption isotherms across a range of water activities. There was anincrease in residual water content as a function of mAb content infreeze-dried formulations. mAb–sucrose interactions were evidentfrom detailed analyses of sorption data. There was a progressive decreasein the glass transition temperature with increasing water content.Dielectric spectroscopy was then employed to characterize the β-and γ-relaxation processes in the glassy state. While the β-relaxationtime decreased with increasing water content, there was no significanteffect on the corresponding activation energy. The effect of watercontent on the γ-relaxation was complex, with an increase inactivation energies as a function of water content. In light of theinfluence of mobility on both physical and chemical stability, theseresults underscore the importance of controlling water content inoptimizing the stability of freeze-dried mAb formulations.

Rapid crystallizationof an active pharmaceutical ingredientcansuppress its observable supersaturation and negate this potentialadvantage of the amorphous form. Excipients added to formulationscan prolong supersaturation, but reduce drug loading and obscure theintrinsic dissolution behavior of the drug in testing. Here, we describespring-and-parachute dissolution behavior in fasted-state simulatedintestinal fluid of an archetypal poorly soluble and strongly crystallizingdrug, griseofulvin (GSF), in the form of excipient-free, pure, amorphousnanopowders generated using a novel, single-step, solvent-free techniqueof organic vapor jet desublimation. Using experiments and modeling,we establish how initial dissolution rate, peak concentration, andarea under the concentration–time curve scale with dose, distinguishingbetween dose regimes in which supersaturation behavior is monotonicand nonmonotonic. The roles of local surface crystallization, particleaggregation, and nonequilibrium crystallization at high supersaturationare elucidated, providing a framework for attaining excipient-freesupersaturation and guiding the development of improved dissolutionmodels.

Thehigh expression of transferrin receptors on the surface ofcancer cells makes them a potential target for drug internalization,which has become a golden opportunity in targeted cancer therapy.Carvacrol, a naturally occurring phenolic compound present in essentialoils, has attracted considerable attention for its pronounced anticanceractivity against diverse human malignancies. Nevertheless, its therapeuticapplication in vivo is constrained by intrinsic physicochemicaland pharmacokinetic limitations, including poor water solubility,suboptimal bioavailability, and rapid systemic elimination. To addressthese challenges, the present study highlights the fabrication oftransferrin-targeted TPGS-conjugated carvacrol (CVC)-loaded polydopaminenanoparticles (CVC-PDA-TPGS-Tf NPs), with exploration of the antitumorefficacy of nanocarrier-mediated carvacrol, deploying a murine triple-negativebreast cancer (TNBC) tumor xenograft model and a human TNBC three-dimensional(3D) spheroid model, as 3D cell culture technologies more thoroughlybear a resemblance to in vivo cell environments toaccommodate improved precision in drug discovery. This work demonstratesthe successful creation of nanoconjugate CVC-PDA-TPGS-Tf NPs (∼112nm in size) and their thorough physicochemical characterization aftersynthesis. Intraperitoneal administration of CVC-PDA-TPGS-Tf NPs substantiallyinhibited tumor growth in the 4T1 xenograft model. Carvacrol effectivelydisintegrates MDA-MB-231 spheroids, illustrating a proportionatelyhigher uptake of carvacrol nanoconjugates by these spheroids. Themolecular docking studies also pinpointed the interaction of potentialcancer biotargets with carvacrol in estrogen- and progesterone-receptor-positivebreast cancer cells, in addition to its mentioned therapeutic efficiencyagainst TNBC. In conclusion, these findings support CVC-PDA-TPGS-Tfas a promising platform for TNBC and potentially hormone receptor-positivebreast cancers, warranting further mechanistic and translational studies.

Dueto its high specificity and peak expression in hepatocellularcarcinoma (HCC), Glypican-3 (GPC3) is a heavily researched target.While current GPC3-targeted therapies demonstrate remarkable efficacy,traditional GPC3 assessment methods are limited by time-dependentand spatial heterogeneity, invasiveness, and prolonged waiting periods,highlighting an urgent need for new GPC3 evaluation strategies. Inthis study, first, we selected MHCC97H as a GPC3-high expressing HCCcell line (named GPC3^high MHCC97H) and constructed a GPC3-lowexpressing MHCC97H cell line (named GPC3^low MHCC97H) usinglentiviral transfection technology. Second, a novel radiopharmaceuticalprecursor, CS03, was synthesized. Molecular docking analysis revealedthat CS03 exhibits the potential to bind to multiple amino acid residuesof GPC3, indicating its promise as a targeting molecule for GPC3.Therefore, we labeled it with Ga-68 to synthesize [⁶⁸Ga]Ga-CS03.The radiolabeling yield of [⁶⁸Ga]Ga-CS03 exceeded 80%.After purification, the radiochemical purity of [⁶⁸Ga]Ga-CS03was greater than 98%, with a specific activity of (3.00 ± 0.22)× 10⁶ MBq/mmol. [⁶⁸Ga]Ga-CS03 exhibitedexcellent in vitro stability, rapid blood clearance(T_1/2α = 0.93 ± 0.49 min and T_1/2β = 17.22 ± 1.42 min), and good in vitro affinity (36.30 ± 5.17 nM). Micro-PET/CT imagingof HCC models revealed significant [⁶⁸Ga]Ga-CS03 radiotraceraccumulation in GPC3^high MHCC97H tumors 0.5 h postinjection.In contrast, minimal or no significant accumulation was detected inGPC3^low MHCC97H tumors and blocking group tumors, indicatingthe excellent in vivo affinity and specificity of[⁶⁸Ga]Ga-CS03. [⁶⁸Ga]Ga-CS03 showed rapid tumorclearance, with a significant decrease in tumor radiotracer distributionby 1 h, a finding corroborated by subsequent biodistribution studies.Furthermore, micro-PET/CT images and biodistribution results indicatedthat [⁶⁸Ga]Ga-CS03 was cleared by the kidneys, which explainsits rapid blood clearance. Finally, we collected tumor tissues fromthe animal models after PET imaging and performed hematoxylin andeosin (H&E) staining and GPC3 immunofluorescence (IF) staining.We found that the fluorescence intensity of IF staining was positivelycorrelated with the radiotracer distribution in the tumor sites onPET images, which further validated the specificity of [⁶⁸Ga]Ga-CS03 for GPC3 binding. In conclusion, we developed [⁶⁸Ga]Ga-CS03, a novel GPC3-targeted PET radiotracer. Extensive in vitro and in vivo evaluations demonstratedits favorable stability, high affinity, and specificity. These promisingpreclinical findings highlight its potential for the noninvasive assessmentof GPC3 expression in HCC.

Polyethylene glycol (PEG) is widely utilized in pharmaceuticalsand cosmetics as a safe and less- or nonimmunogenic synthesized biocompatiblepolymer. However, the increasing detection of pre-existing anti-PEGantibodies in untreated individuals has raised concerns that dailyexposure to PEG derivatives through cosmetics could induce immuneresponses against PEG. We have shown that daily topical applicationsof this type of cosmetic product cause anti-PEG IgM production innormal mice, and the efficacy of PEGylated therapeutics was suppressedby this induced anti-PEG IgM. In this study, the mechanism underlyinganti-PEG IgM production caused by topical applications of a cosmeticproduct containing PEG derivatives, which are yet to be identified,was investigated in mice. Daily applications of such a cosmetic productto the compromised skin of mice elicited anti-PEG IgM production.Splenectomy, performed before the topical applications, markedly suppressedthe anti-PEG response. In addition, when splenectomy was carried outafter 14 days of daily topical applications, there was a remarkabledecline in the anti-PEG IgM response that had already been induced.In this study, however, splenectomy completely diminished neitheranti-PEG IgM production nor pre-existing anti-PEG IgM titers. Althoughimmunodeficient C.B-17/Icr-scid/scid (SCID) mice caused no anti-PEGIgM production, the adoptive transfer of splenocytes obtained fromnormal mice to SCID mice restored the anti-PEG response to daily topicalapplications of this cosmetic product. Furthermore, anti-PEG IgM productionwas observed in T cell-deficient BALB/cAJcl-nu/nu (BALB/c-nu) mice.In the lymph node-deficient ALY/NscJcl-aly/aly (aly/aly) mice, dailytopical applications of this cosmetic product also caused anti-PEGIgM production, but the titer was lower than that in normal mice.The results of this study show the spleen to be the primary contributor,while lymph nodes play a partial role, in the production of anti-PEGIgM via daily topical applications of this cosmetic product in a Tcell-independent manner in mice. This study provides mechanistic insightinto how daily topical exposure to PEG could lead to the formationof pre-existing anti-PEG IgM, which in turn would alter the pharmacokineticsand therapeutic efficacy of PEGylated therapeutics.

Oral insulin delivery is limited by the intestinal mucusbarrierand intracellular lysosomal degradation within epithelial cells. Toaddress these sequential barriers, we engineered insulin-loaded zwitterioniccharge-switching nanoparticles (INS/Bet-CS-SA NPs) through the rationalintegration of betaine-modified chitosan and sodium alginate. Thisdesign leverages the pH-responsive surface charge transition of thenanocomplex, allowing the formulation to remain compact and positivelycharged under gastric conditions while undergoing swelling and surfacetransformation in intestinal media. The optimized nanoparticles exhibiteda high insulin loading capacity (71.71%) and a sustained release profile(86.03% within 12 h under simulated gastrointestinal conditions).Especially, the surface zeta potential of INS/Bet-CS-SA NPs shiftedfrom +15.73 ± 3.17 mV at pH 1.2 to −24.22 ± 3.47mV at pH 6.8 and −25.68 ± 3.93 mV at pH 7.4, confirmingthe intended charge-switching behavior. This zwitterionic functionalsurface markedly enhanced mucus penetration and transepithelial transport,yielding apparent permeability coefficients 25.4-fold higher thanthat of free insulin. Cellular uptake and trafficking analyses suggestedcaveolae-associated endocytosis and reduced lysosomal retention playedan essential role, which might be caused by betaine-associated membraneperturbation and chitosan’s proton buffering capacity. In atype 2 diabetic rat model, orally administered INS/Bet-CS-SA NPs induceda sustained hypoglycemic response and achieved a relative bioavailabilityof 25.36%. The prolonged pharmacodynamic effect was additionally supportedby a semimechanistic pharmacokinetic/pharmacodynamic model. Overall,this work provides a zwitterionic functional nanoplatform capableof overcoming multiple gastrointestinal barriers as a promising strategyfor the oral delivery of insulin.