European Journal of Pharmaceutical Sciences最新文献

{"title":"Therapeutic effect of cyclosporine A-loading TPGS micelles on a mouse model of LPS-induced neuroinflammation.","authors":"Fabiola Guareschi, Carla Fonseca, Soraia Silva, Silvia Pescina, Sara Nicoli, Francesca Buttini, Fabio Sonvico, Ana Fortuna","doi":"10.1016/j.ejps.2024.106994","DOIUrl":"10.1016/j.ejps.2024.106994","url":null,"abstract":"<p><p>Neuroinflammation is an undoubted hallmark of neurodegenerative processes characterized by memory impairment, loss of coordination and muscle strength in diseases such as Alzheimer's disease, Parkinson's disease and multiple sclerosis as well as depressive disorders. Cyclosporine A (CSA) has already been identified as a promising neuroprotective peptide, due to its well-known anti-inflammatory properties. Herein, CSA was encapsulated into α-tocopherol polyethylene glycol 1000 succinate (TPGS) micelles and intranasally administered to a lipopolysaccharide (LPS) induced mouse model of neuroinflammation. After the treatment, mice were subjected to behavioral tests to assess cognitive and motor skills, while the biodistribution of CSA in plasma and olfactory bulb was studied by a new HPLC method validated for precision and accuracy. The results highlighted that in comparison to the classic oral CSA suspension, the intranasal (IN) administration showed significatively better safety and efficiency profiles. Notably, IN administration of CSA micelles showed relevant antidepressive effects and a certain ability to revert LPS-induced motor impairment. This work pointed out that the innovative and noninvasive IN administration of TPGS micelles could represent a safe and effective alternative to the classic oral route to deliver CSA at the Central Nervous System level, where its beneficial activity against neuroinflammation can be exploited.</p>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":" ","pages":"106994"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142863768","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Research on genetic variant characteristics in ADME genes based on whole-exome sequencing in the Han Chinese population.","authors":"Ling Ye, XiangGuang Meng, Yan Zhan, Tong Li, Xin Huang, Hui Qiu, Jianzhu Zhou, Chengxian Guo","doi":"10.1016/j.ejps.2024.106987","DOIUrl":"10.1016/j.ejps.2024.106987","url":null,"abstract":"<p><strong>Background: </strong>Genetic variants in absorption, distribution, metabolism, and excretion (ADME) genes affect drug efficacy and side effects. Whole-exome sequencing (WES) effectively identifies these variants. Findings from Western populations may not apply to the Han Chinese population due to ethnic differences.</p><p><strong>Objective: </strong>This study aimed to investigate genetic variants, metabolic phenotypes, and drug impacts related to ADME genes in the Han Chinese population using WES data.</p><p><strong>Methods: </strong>ADME genes and drug profiles were sourced from multiple databases. WES data were collected from 357 samples at Third Xiangya Hospital and 5,000 samples from the \"HuaBiao Project.\" After WES, fastp was used for quality control; BWA, samtools, and the Picard software were used for comparison, sequencing, and de-duplication; GATK was used for base quality score recalibration; and variant quality score recalibration was carried out after detecting the variants, and the variants were annotated using ANNOVAR. ADME genes and drug profiles were obtained through PharmGKB and other databases, and then all variants in the exonic regions of ADME genes were extracted. The distributional characteristics of these variants, ethnic differences, and metabolic phenotype distribution of important ADME genes, such as CYP2B6, were analyzed. Prediction of deleterious variants of ADME gene employed the ADME gene variant prediction framework, and western blotting and enzyme assays were used to validate the impact of harmful variants.</p><p><strong>Results: </strong>We integrated 604 ADME genes and 972 drugs. There were 33,925 single nucleotide polymorphisms (SNPs) and 484 insertion-deletions (InDels) in 5,357 Han Chinese WES samples; 88.9 % were rare. Of the SNPs, 60.9 % are new functional mutations in enzyme and transporter genes; InDels also affected these genes. We discovered Chinese-specific variants in key ADME genes and ethnic-specific metabolic phenotypes that could affect drug use. Finally, we screened 3,657 potentially harmful mutations; 45.6 % are novel. Western blotting and enzyme activity assays confirmed several harmful mutations significantly reduced CYP2C19 gene expression and function (P < 0.01).</p>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":" ","pages":"106987"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142821945","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Intradermal versus subcutaneous immunization: Effects of administration route using a lipid-PLGA hybrid nanoparticle tuberculosis vaccine.","authors":"M M Szachniewicz, S J F van den Eeden, K E van Meijgaarden, K L M C Franken, S van Veen, A Geluk, J A Bouwstra, T H M Ottenhoff","doi":"10.1016/j.ejps.2024.106995","DOIUrl":"10.1016/j.ejps.2024.106995","url":null,"abstract":"<p><p>Tuberculosis (TB) remains a significant global health challenge, latently affecting around a quarter of the global population. The sole licensed TB vaccine, Mycobacterium bovis Bacillus Calmette-Guérin (BCG), shows variable efficacy, particularly among adolescents and adults, underscoring the pressing need for more effective vaccination strategies. The administration route is crucial for vaccine efficacy, and administration via the skin, being rich in immune cells, may offer advantages over conventional subcutaneous routes, which lack direct access to abundant antigen-presenting cells. This study compared the immunogenic effects of intradermal versus subcutaneous administration of a candidate TB vaccine delivering a Ag85B-ESAT6-Rv2034 (AER) multiphase fusion recombinant protein, in lipid-poly(D,L-lactic-co-glycolic acid) (lipid-PLGA) nanoparticles in mice. In-depth evaluation of immune responses in splenocytes was performed using 27-marker spectral flow cytometry. Both routes elicited significant T-cell responses. However, intradermal administration uniquely increased polyfunctional CD4⁺ and CD8⁺ T-cells producing IL-2, IFNγ, and TNFα, associated with protection against TB. Additionally, it significantly increased CD69⁺ B-cell counts and induced higher AER-specific antibody titers, particularly IgG2a. These results underscore the superior immunogenic potential of intradermal vaccine administration by effectively inducing immune cells associated with TB protection, highlighting its significance in the development of new vaccine strategies.</p>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":" ","pages":"106995"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142876547","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Psoriatic skin-sensitive degradable mesoporous zinc phosphate microsphere as a photosensitizer in photodynamic therapy for anti-psoriasis.","authors":"Nan Jin, Liqiong Huang, Xin Chen, Zhuhang Liu, Ruotong Chen, Mengting Gao, Tianhui Liu, Jianmin Chen","doi":"10.1016/j.ejps.2024.107000","DOIUrl":"10.1016/j.ejps.2024.107000","url":null,"abstract":"<p><p>Blue light will be a promising alternative for photodynamic therapy in psoriasis, but the photosensitizer in vivo remains unexplored. Mesoporous zinc phosphate microparticle (MZP) was synthesized successfully in this study, as evidenced by XPS, XRD, and nitrogen adsorption experiments. Its psoriatic skin-sensitive property was corroborated by SEM and the higher cumulative release rate of that impregnated with curcumin (Cur) and glycyrrhizic acid (GA), namely Cur-GA-MZP, at pH 5.4, which mimic the acidic environment of the inflammatory psoriatic skin. Moreover, by detecting the decrease of absorbance of 1,3-Diphenylisobenzofuran (DPBF) which had been mixed with MZP irradiated by blue light, MZP is confirmed to reinforce ROS along with irradiation time at pH 5.4, rather than that at pH 7.2. This phenomenon led to a strengthened anti-inflammatory effect of MZP on xylene-induced auricle inflammation, as well as on imiquimod-induced psoriatic-like plaques exists only under blue light irradiation, with the alleviated macroscopic and microscopic appearance, decreased IL-17A levels of skin lesion area as well as spleen index. The mechanism is likely attributed to the NFƙB pathway as well as the MAPK pathway detected by western blot. In sum, MZP, which could dissociate at psoriatic skin condition, will be a promising photosensitizer with the prerequisite of blue light as an effective photodynamic therapy for the management of psoriasis.</p>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":" ","pages":"107000"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142893247","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Editorial: Pulmonary drug delivery.","authors":"Carsten Ehrhardt, Francesca Buttini, Imran Saleem, Regina Scherließ, Ayca Yıldız-Peköz","doi":"10.1016/j.ejps.2024.106988","DOIUrl":"10.1016/j.ejps.2024.106988","url":null,"abstract":"","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":" ","pages":"106988"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142822020","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Developing a method for the study of perfusion effects in topical product pharmacokinetics.","authors":"Saara K Luna, M Alice Maciel Tabosa, Ting Chean Khoo, Conor L Evans","doi":"10.1016/j.ejps.2024.106975","DOIUrl":"10.1016/j.ejps.2024.106975","url":null,"abstract":"<p><p>Topical drug products are delivered to skin structures to treat numerous skin diseases. Due to the complexities of the skin environment and barrier, topical drug pharmacokinetics are difficult to determine, especially in vivo, as most pharmacokinetic assessment methods can only be performed ex vivo. Notably, in vivo conditions include perfusion via dermal capillaries, which influences topical drug uptake by acting as a clearance route and a \"sink\" driving permeation through the skin. In this study, we develop a method to examine the effects of perfusion on topical drug uptake in vivo using stimulated Raman scattering (SRS) microscopy, a chemically-specific imaging modality ideal for visualizing topical drug permeation over time. In this pilot study, we imaged the in vivo and ex vivo uptake of tazarotene in 70/30 v/v Transcutol:EtOH in paired mouse ear skin, comparing the effects of perfusion on tazarotene concentration (linearly proportional to SRS signal intensity) over time and pharmacokinetic parameters (T_max, C_max, AUC) in lipid-rich and lipid-poor regions in the stratum corneum and sebaceous glands. Obvious variations in SRS signal-time trends and statistically significant differences in stratum corneum pharmacokinetic parameters comparing uptake in lipid-rich and lipid-poor regions in vivo and ex vivo indicated slowed tazarotene flux through ex vivo skin in the absence of perfusion. The observed permeation differences in lipid-rich and lipid-poor regions in perfused and non-perfused skin reflects increased tazarotene permeation rate and removal in the presence of perfusion (in vivo) and decreased permeation rate and lack of elimination route in the absence of perfusion (ex vivo). Our method is demonstrated to be effective in assessing in vivo perfusion effects on topical drug uptake, promoting a better understanding of the influence of perfusion on topical drug delivery.</p>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":" ","pages":"106975"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142778899","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Antitumor activity of selenopsammaplin A analog (SPA-10091-HCl) via histone methyltransferase DOT1L degradation and apoptosis induction in castration-resistant prostate cancer cells.","authors":"Jaeho Han, Sehun Yang, Woong Sub Byun, Sung Chul Jang, Eun Seo Bae, Hyeung-Geun Park, Sang Kook Lee","doi":"10.1016/j.ejps.2024.106991","DOIUrl":"10.1016/j.ejps.2024.106991","url":null,"abstract":"<p><p>Castrate-resistant prostate cancer (CRPC) is one of the most difficult cancers in men and is characterized by a poor prognosis and a high risk of metastasis. The overexpression of the disruptor of telomeric silencing 1-like (DOT1L), which is a specific methyltransferase for histone H3 at lysine residue 79 (H3K79), has been related to poor outcomes in patients with CRPC. Therefore, targeting DOT1L is considered a potential therapeutic approach to overcome the significant medical challenges of CRPC. In our previous study, we designed selenopsammaplin A (SPA) analogs as non-nucleoside DOT1L inhibitors to suppress human breast cancer cell proliferation and metastasis. However, the antitumor activity and the precise underlying mechanism of SPA analogs in PC cells remain unclear. Herein, we administered SPA-10091-HCl, a DOT1L-targeting degrader, to effectively hinder the growth and DOT1L-mediated H3K79 methylation in CRPC (PC3 and DU145) cells. Mechanistically, SPA-10091-HCl selectively degrades DOT1L protein through the nuclear ubiquitin-proteasome pathway, thereby suppressing H3K79 methylation in CRPC cells. SPA-10091-HCl inhibits CRPC cell proliferation, migration, and invasion, with the E-cadherin expression upregulation and N-cadherin and vimentin expression downregulation. Additionally, prolonged SPA-10091-HCl treatment induced apoptosis by regulating apoptosis-associated protein expressions, including Poly (ADP-ribose) polymerase (PARP), caspase-3, caspase-9, and Bcl-2. Moreover, SPA-10091-HCl effectively inhibited tumor growth in the PC3 cells-implanted xenograft mouse model without any overt toxicity. These results indicate SPA-10091-HCl as a potential candidate for further development as a chemotherapeutic agent against CRPC.</p>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":" ","pages":"106991"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142846213","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pyrrolidine SS13 induces oxidative stress and autophagy-mediated cell death in colorectal cancer cells.","authors":"Natalia Nosalova, Monika Majirska, Alexandra Keselakova, Miroslava Martinkova, Dominika Fabianova, Andrej Mirossay, Martina Bago Pilatova, Martin Kello","doi":"10.1016/j.ejps.2024.106982","DOIUrl":"10.1016/j.ejps.2024.106982","url":null,"abstract":"<p><strong>Introduction: </strong>Pyrrolidines, nitrogenous organic compounds, are among the most intensively studied agents because of their antibacterial, antiviral, neurological, and promising antitumor effects. Moreover, many medicinal drugs contain pyrrolidine moiety such as sunitinib (anticancer drug), telaprevir and ombitasvir (antiviral drugs) or ramipril (antihypertensive drug).</p><p><strong>Rationale of the study: </strong>Based on the pro-apoptotic effect of pyrrolidine SS13, this study focuses on the pro-oxidative properties of the tested pyrrolidine SS13 on colorectal cancer cells to deepen the understanding of its mechanisms of action.</p><p><strong>Research hypothesis: </strong>We hypothesize that SS13 induces oxidative stress and autophagy activation in HCT116 and Caco-2 cell lines, thus contributing to antiproliferative effects.</p><p><strong>Methods: </strong>Flow cytometry, western blot, fluorescence microscopy and qRT-PCR were used to evaluate the effect of pyrrolidine SS13.</p><p><strong>Conclusion and future directions: </strong>Pyrrolidine SS13 induced oxidative stress through the accumulation of reactive oxygen and nitrogen species in both cell lines and the modulation of both superoxide dismutase isoenzymes (SOD1, SOD2). Oxidative stress was also associated with the activation of DNA damage response system and modulation of stress/survival pathways. We demonstrated for the first time that pyrrolidine SS13 is involved in the induction of autophagy accompanied by increased levels of autophagic markers (p-AMPK, p-ULK, LC3I/II and ATG7) and a significant decrease in p62 protein levels in both cell lines. Finally, chloroquine, an inhibitor of autophagy, enhanced cell survival and suppressed the cytotoxic effect of SS13 in HCT116 and Caco-2 cells, indicating that SS13 contributes to autophagy-mediated cell death. Taken together, our results suggest that oxidative stress and autophagy participate in the antiproliferative effect of pyrrolidine SS13 on colorectal cancer cells. Further research using primary cell cultures obtained from different animal tissues as well as performing in vivo experiments is needed to understand these processes in detail and to investigate the potential therapeutic application of new pyrrolidine derivatives.</p>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":" ","pages":"106982"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142791300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploration of solubilisation effects facilitated by the combination of Soluplus® with ionic surfactants.","authors":"Justus Johann Lange, Lukas Enzner, Martin Kuentz, Patrick J O'Dwyer, Wiebke Saal, Brendan T Griffin, Nicole Wyttenbach","doi":"10.1016/j.ejps.2024.106957","DOIUrl":"10.1016/j.ejps.2024.106957","url":null,"abstract":"<p><p>Preclinical testing of new drug candidates frequently necessitates high-dose solution formulations to support robust testing in rodent models. This study aimed to expand the range of high solubilisation capacity formulations by exploring the solubilisation effects of the polymeric surfactant Soluplus® in combination with ionic surfactants. The interactions between Soluplus® and three ionic surfactants, sodium dodecyl sulphate, dioctyl sodium succinate, and sodium oleate, with a primary focus on solubility enhancement were investigated over a range of ionic surfactant concentrations. The solubilisation profiles for seven model drugs were obtained, and the vehicles were characterised by their visual characteristics, dynamic light scattering, and viscosity measurements. The solubilisation profiles were non-linear, indicating the formation of different colloidal species with individual solubilisation strengths depending on surfactant type and concentration, demonstrating substantial solubility enhancement. For certain drugs more than additive solubilisation, facilitated by synergistic interactions between Soluplus® and the ionic surfactants, was obtained. Overall, the solubility increase provided by the excipient combinations resulted in non-linear and drug specific solubilisation profiles. The non-linearities observed were reflected in visual observations of the vehicles appearance, DLS and viscosity measurements, which collectively indicated a change in polymer aggregation with increasing concentration of anionic surfactant. This investigation highlights that already low quantities of ionic surfactants introduced to Soluplus® may substantially enhance solubility, which offers a promising approach for further exploration in preclinical drug development where more conventional solubilising formulation strategies may fall short.</p>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":" ","pages":"106957"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142647190","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dual pH-responsive CRISPR/Cas9 ribonucleoprotein xenopeptide complexes for genome editing.","authors":"Xianjin Luo, Janin Germer, Tobias Burghardt, Melina Grau, Yi Lin, Miriam Höhn, Ulrich Lächelt, Ernst Wagner","doi":"10.1016/j.ejps.2024.106983","DOIUrl":"10.1016/j.ejps.2024.106983","url":null,"abstract":"<p><p>Clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR associated (Cas) protein has been proved as a powerful tool for the treatment of genetic diseases. The Cas9 protein, when combined with single-guide RNA (sgRNA), forms a Cas9/sgRNA ribonucleoprotein (RNP) capable of targeting and editing the genome. However, the limited availability of effective carriers has restricted the broader application of CRISPR/Cas9 RNP. In this study, we evaluated dual pH-responsive amphiphilic xenopeptides (XPs) for delivering CRISPR/Cas9 RNP. These artificial lipo-XPs contain apolar cationizable lipoamino fatty acid (LAF) and polar cationizable oligoaminoethylene acid units such as succinoyl-tetraethylenepentamine (Stp) in various ratios and U-shaped topologies. The carriers were screened for functional Cas9/sgRNA RNP delivery in four different reporter cell lines, including a Duchenne muscular dystrophy (DMD) exon skipping reporter cell model. Significantly enhanced cellular uptake into HeLa cells, effective endosomal disruption in HeLa gal8-mRuby3 cells, and potent genome editing by several Cas9/sgRNA RNP complexes was observed in four different cell lines in the 5 nM sgRNA range. Comparing Cas9/sgRNA RNP complexes with Cas9 mRNA/sgRNA polyplexes in the DMD reporter cell model demonstrated similar splice site editing and high exon skipping of the two different molecular Cas9 modalities. Based on these studies, analogues of two potent U1 LAF₂-Stp and LAF₄-Stp₂ structures were deployed, tuning the amphiphilicity of the polar Stp group by replacement with the six oligoamino acids dmGtp, chGtp, dGtp, Htp, Stt, or GEIPA. The most potent LAF₂-Stp analogues (containing dGtp, chGtp or GEIPA) demonstrated further enhanced gene editing efficiency with EC50 values of 1 nM in the DMD exon skipping reporter cell line. Notably, the EC50 of LAF₂-dGtp reached 0.51 nM even upon serum incubation. Another carrier (LAF₄-GEIPA₂) complexing Cas9/sgRNA RNP and donor DNA, facilitated up to 43 % of homology-directed repair (HDR) in HeLa eGFPd2 cells visualized by the switch from green fluorescent protein (eGFP) to blue fluorescent protein (BFP). This study presents a delivery system tunable for Cas9 RNP complexes or Cas9 RNP/donor DNA polyplexes, offering an effective and easily applicable strategy for gene editing.</p>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":" ","pages":"106983"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142794380","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}