Molecular Pharmaceutics最新文献

Among the photosensitizers (PSs) developed in our laboratory,3-(1′-hexyloxy)ethyl-3-devinyl pyropheophorbide-a (HPPH) is undergoing Phase-II multicenterclinical trials for the treatment of head and neck and esophagealcancers. The PS exhibits absorption at 665 nm (in vivo) and showsdesired pharmacokinetics with limited skin phototoxicity in patients,compared to FDA-approved Photofrin. This study determined the effectof HPPH-PDT in vitro and in vivo on human bladder cancer (BCa) tissuegrown in immune-deficient mice. We observed high tumor uptake/retentionof HPPH at 24 h post injection. The anticancer activity of HPPH-PDTwas associated with undesirable induction of COX-2, the key enzymeenhancing the production of prostaglandin E2 (PGE2), a pathway wepreviously found accompanying tumors associated with immune suppressionand tumor progression. Combination of COX-2 inhibition with PDT orwith BCG-immunotherapy showed an improved rate of tumor cures. Hence,HPPH-PDT in combination with COX-2 inhibitors not only reduces theexpression of COX-2, IL-6, VEGF, and PGE2 but also enhances long-termtumor cure.

Synthetic trehaloseglycolipids (TGLs) that modulate the MacrophageInducible C-Type Lectin receptor (Mincle) are attractive targets foradjuvant development. However, natural ligands such as trehalose dimycolate(TDM), as well as existing synthetic mimetics, are limited by poorphysicochemical properties and reactogenicity, motivating the developmentof rationally designed synthetic TGLs. Herein, we investigated thebiophysical properties of amphiphilic TGLs featuring hydrophobic,aromatic moieties attached to the polar trehalose headgroup via ester,amide, or triazole linkages. The formation of homogeneous and colloidallystable aqueous nanodispersions was readily achieved through focusedultrasonication, eliminating the need for cosolvents and/or surfactantsthat might interfere with the self-assembling and biological propertiesof these compounds. These self-assembled particles were characterizedusing DLS, cryo-TEM, and synchrotron small-angle X-ray scatteringto gain insight into their particle morphology and liquid crystallinephase behavior. Additionally, molecular dynamics (MD) simulationswere conducted to evaluate the critical packing parameter (CPP), apredictive descriptor of the supramolecular structures formed by self-assemblingamphiphiles, under conditions that closely mimic experimental environments.The synthetic Mincle ligands formed lyotropic liquid crystals, includingcubosomal or hexagonal phases as well as lamellar phases. In vitro and in vivo testing of these self-assembliesrevealed that all tested TGLs exhibited measurable immunological activity.Analogues forming cubic phases were associated with comparativelystronger cytokine production and enhanced antigen-specific humoraland cell-mediated immune responses within this compound set. Takentogether, the data support a model in which both molecular structureand supramolecular organization contribute to immunostimulatory activity,although these effects cannot be fully decoupled in the present study.Additionally, we developed an in silico model capableof predicting supramolecular assembly, providing insight into theself-assembly tendencies of trehalose-based amphiphiles, and offeringbroader relevance to other biologically active, self-assembling systems.

Noninvasive detection of microglia is crucial for monitoringtheirspatiotemporal dynamics and facilitating early medical interventionduring the prodromal stages of these neuroinflammatory diseases. Colony-stimulatingfactor 1 receptor (CSF1R) is a promising imaging biomarker for microgliain neuroinflammation. However, currently developed CSF1R-targetedprobes lack [¹⁸F]-labeled variants, and their stability,sensitivity, and specificity are all suboptimal to some extent. Inour study, based on pyrrolopyrimidines(a distinct structure of CSF1Rinhibitors), we developed the [¹⁸F]-labeled CSF1R-targetedprobe-¹⁸F-FPPA (6-(4-(2-(2-(2-(fluoro-¹⁸F)ethoxy)ethoxy)ethoxy)phenyl)-N-methyl-N-(3-methylbenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine).¹⁸F-FPPA was synthesizedwith a radiochemical yield of 4–7%, molar activity of 30–70GBq/μmol, and radiochemical purity exceeding 99%, which possessesexcellent physicochemical properties, rapid blood pharmacokinetics(T_1/2α = 2.50 min, T_1/2β = 32.4 min), favorable invitro and in vivo stability, and a clearmetabolic pathway via dual hepatobiliary and renal excretion. A markeduptake of ¹⁸F-FPPA was observed at the LPS injection sitein the LPS group at 30–40 min postinjection. A decreased uptakealso was observed in cold ¹⁹F-FPPA, BLZ945, and PLX3397blocking groups (P < 0.05), and no obvious increasein the group of intraperitoneal injection of cyclosporine A (P > 0.05). The in vivo imaging and immunohistochemicalresults confirmed the specific uptake of ¹⁸F-FPPA by targetingCSF1R on microglia in inflamed tissues. Overall, our findings demonstratedthat ¹⁸F-FPPA had great potential as a promising tracerfor the imaging of neuroinflammatory diseases, while its unique bindingprofile also provides insights into CSF1R-targeted ligand development.

The discovery of new antimicrobial medications has notkept upwith the evolution of bacterial resistance, and infections with drug-resistantbacteria, such as methicillin-resistant Staphylococcus aureus (MRSA), continue to pose a serious threat toglobal public health. Antimicrobial photodynamic therapy (aPDT), asan alternative strategy, produces reactive oxygen species (ROS) underlight activation and kills drug-resistant bacteria through oxidativedamage. Nevertheless, conventional photosensitizers (PSs) are limitedby aggregation-caused quenching, inadequate bacterial selectivity,and off-target cytotoxicity. Quaternary ammonium salts (QASs) canpreferentially accumulate on bacterial membranes through electrostaticinteractions and disrupt the bacterial membranes. The quaternary ammoniumPS PK4, reported here has a triphenylamineunit that enhances ROS production. Under white light irradiation, PK4 produces abundant ROS that oxidativelydamage bacterial membranes, while exhibiting high photostability andminimal dark toxicity. In an MRSA-infected wound model, PK4-mediated phototherapy can reduce inflammation,accelerate wound healing, and enhance collagen deposition withoutsystemic toxicity, providing a promising aPDT strategy for combatingdrug-resistant bacterial infections.

Acute kidney injury (AKI) is a clinical syndrome withhigh incidenceand mortality involving oxidative stress, ferroptosis, and inflammation,yet there are no clinically effective interventions. Herein, HA@PDA@EGCGnanoparticles were constructed using polydopamine (PDA) as carriers,loaded with epigallocatechin gallate (EGCG) via π-π stacking,and further surface-modified with hyaluronic acid (HA) through electrostaticinteraction. The nanoparticles had uniform morphology with a sizeof approximately 50 nm and an EGCG loading capacity of 15.4 ±0.8%. They exhibited prominent broad-spectrum antioxidant activityand hydrogen peroxide-responsive EGCG release, with a cumulative releaserate of 58.2 ± 2.2% within 2 h and close to 80% at 12 h. Cellularuptake and in vivo distribution confirmed the efficient CD44 receptor-mediatedinternalization of the nanoparticles by human kidney-2 (HK-2) cellsas well as renal-targeted accumulation. Reactive oxygen species (ROS)staining, mitochondrial morphology and apoptosis assays showed thatthe nanoparticles effectively scavenged ROS, alleviated mitochondrialdamage, and inhibited cell apoptosis. In a cisplatin-induced in vivoAKI model, HA@PDA@EGCG significantly restored renal function: bloodurea nitrogen and creatinine decreased to 12.3% and 27.2% of modelgroup levels, respectively, while alleviating renal pathological damageand inflammation. Mechanistically, HA@PDA@EGCG inhibited ferroptosisby downregulating acyl-CoA synthetase long-chain family member 4 (ACSL4)to suppress lipid synthesis, chelating ferrous ions, and stabilizingglutathione peroxidase 4 (GPX4) protein. Collectively, HA@PDA@EGCGachieved renal protection by ameliorating oxidative stress, suppressinginflammation and inhibiting ferroptosis through multiple pathways,offering a novel strategy for targeted AKI therapy.

Liposomeshave been extensively studied as carriers for deliveringanticancer agents, and a formulation that encapsulates irinotecan(IRT) is currently available on the market under the name Onyvide.This study examined the molecular state of IRT within liposomes preparedwith ammonium sulfate (AS) or triethylammonium sucrose octasulfate(TEA-SOS) as trapping agents using various analytical techniques.Cryogenic transmission electron microscopy revealed distinct morphologies:solid IRT precipitates in IRT/TEA-SOS liposomes, and dot-like microstructuresin IRT/AS liposomes. Small-angle and wide-angle X-ray scattering (SAXS/WAXS),Raman spectroscopy, and ¹H NMR analyses indicated thatIRT was in a dissolved state within the inner aqueous phase of IRT/ASliposomes. In contrast, it precipitated in an amorphous form withinIRT/TEA-SOS liposomes. In situ NMR measurements,conducted while varying the temperature, revealed a molecular statechange of IRT during the encapsulation process. Detailed analysisof the NMR spectrum, supported by the magic-angle spinning (MAS) technique,enhanced our understanding of the molecular state, including the mobilityand fraction of IRT between monomers and aggregates within the liposome.These differences in molecular states elucidated the distinct releaseprofiles of IRT from each liposome. The solubility of IRT salts withinthe inner aqueous phase of the liposome played a key role in controllingthe release properties of IRT. The findings highlight that the molecularstate of IRT, determined by the choice of trapping agent, significantlyinfluences release behavior and provides valuable insights for therational design of advanced liposomal drug formulations.

Building upon our previous work on [⁶⁸Ga]Ga/[¹⁷⁷Lu]Lu-DOTA-Ibandronate and its demonstrated clinical applicability,this study investigates the biological behavior and translationalpotential of the novel therapeutic radiopharmaceutical terbium-161(¹⁶¹Tb)-labeled DOTA-IBA. The radiolabeling parametersof [¹⁶¹Tb]Tb-DOTA-IBA were first optimized, followed bya systematic assessment of its physicochemical characteristics. Biodistribution,micro-SPECT/CT imaging, and therapeutic efficacy were evaluated ina 4T1 mouse model of bone metastasis to determine pharmacokineticsand bone-targeting capacity. Safety was verified through dose-escalationexperiments. Furthermore, a preliminary clinical evaluation was performedin one volunteer with bone metastases to validate safety and initialtherapeutic efficacy. [¹⁶¹Tb]Tb-DOTA-IBA was obtained withradiochemical purity exceeding 98%, demonstrating excellent bone-targetingselectivity and retention, with a 24 h bone uptake of 30.61 ±4.61% ID/g, which is substantially higher than in soft tissues. Ina patient receiving 370 MBq, marked pain relief was achieved, reflectedby decreased VAS scores, with therapeutic benefit lasting 8–12weeks and no observed toxic reactions. The compound exhibited favorablepharmacokinetics and safety, with no ≥ grade 3 adverse events.Overall, [¹⁶¹Tb]Tb-DOTA-IBA displayed high tumor-to-backgroundratios and excellent pharmacokinetic behavior. Early human data confirmedits safety and therapeutic potential. The complementary use of [⁶⁸Ga]Ga/[¹⁶¹Tb]Tb-DOTA-IBA enables a unified diagnostic-therapeuticplatform for the management of bone metastases, providing an innovative,targeted treatment option capable of improving both quality of lifeand survival in patients with advanced skeletal disease.

Accurate predictionof human pharmacokinetics is essentialforselecting safe starting doses in first-in-human (FiH) trials. Thisstudy compares the predictive accuracy of three methodologies includingstandard physiologically based pharmacokinetic (PBPK) modeling incorporatinganimal verification, high-throughput (HT) PBPK based on in vitro inputs, and machine learning (ML). We evaluated theirperformance on 40 diverse small molecules that entered clinical developmentat Roche between 2003 and 2024. Standard PBPK predictions were assessedprospectively from original reports, while HT-PBPK and ML predictionswere obtained retrospectively. Predicted area under the curve (AUC_inf) and maximum plasma concentration (C_max) for oral administration were compared to observed clinicaldata. While all three approaches demonstrated a useful level of accuracywith at least 49% of predicted AUC_inf and C_max values within 2-fold, standard PBPK was the most precisemethod with 65 and 63% of AUC_inf and C_max within that margin, respectively. This higher precisionlikely results from the integration of expert judgment and animal in vivo data for model refinement, whereas HT-PBPK and MLprovide valuable animal-free, high-throughput alternatives. Ensemblemodels leveraging two or three models were explored, which generallyimproved accuracy in comparison to single models. This highlightsthe complementary nature of mechanistic and ML-based models. Currently,standard PBPK is still the preferred method for FiH dose selectionfor which highest accuracy and mechanistic insight is sought. However,although less accurate, we believe that HT-PBPK and ML are viableanimal-free alternatives with value for early human dose prediction,reducing costs and cycle times.

Purpose: Claudin18.2 (CLDN18.2) has emerged as a significanttherapeutictarget for advanced gastric cancer. In this study, we aimed to developa ¹⁷⁷Lu-radiolabeled anti-CLDN18.2 VHH-Fc (SN-1A01) forradioimmunotherapy in gastric cancer (GC) tumor-bearing models. Methods:Immunohistochemistry (IHC) for CLDN18.2 was performed on gastric lesionsinduced by tumors from 37 patients with GC, as well as in a GC patient-derivedxenograft (GC-PDX) tumor and GC xenograft tumors (N87–18.2and NUCG4). The SN-1A01 was conjugated with DOTA and radiolabeledusing ¹⁷⁷Lu, resulting in the formation of the radioimmunoconjugate[¹⁷⁷Lu]Lu-DOTA–SN–1A01. We established asafe dosage for the application of [¹⁷⁷Lu]Lu-DOTA–SN–1A01and evaluated the therapeutic efficacy of a single dose in GC models.Ki67 expression levels in tumors were detected via IHC following treatmentwith [¹⁷⁷Lu]Lu-DOTA–SN–1A01. Results: IHCanalysis demonstrated moderate-to-high CLDN18.2 expression in 45.9%of GC patient tumor tissues. Additionally, elevated levels of CLDN18.2were detected in GC-PDX and N87–18.2 tumors, whereas lowerexpression was shown in NUGC4. A single dose of [¹⁷⁷Lu]Lu-DOTA–SN–1A01below 300 μCi (11.1 MBq) was well tolerated. [¹⁷⁷Lu]Lu-DOTA–SN–1A01 significantly inhibited tumor growthand increased survival time in both GC-PDX and N87–18.2 modelsacross a dose of 100 and 300 μCi. However, the tumor-suppressiveeffect was comparatively weaker in the NUGC4 model. Reduced Ki67 expressionwas evident in all [¹⁷⁷Lu]Lu-DOTA–SN–1A01-treatedtumors. Conclusion: In preclinical studies, [¹⁷⁷Lu]Lu-DOTA–SN–1A01demonstrated significant antitumor efficacy with acceptable toxicity,which indicated a strong potential for clinical translation in GCtherapy.