Journal of pharmaceutical sciences最新文献_第5页

Stability of therapeutic monoclonal antibodies during infusion with elastomeric pumps intended for home-based therapy 用于家庭治疗的弹性泵输注过程中治疗性单克隆抗体的稳定性。

IF 3.8 3区医学 Q2 CHEMISTRY, MEDICINAL

Journal of pharmaceutical sciences

Pub Date : 2026-01-10 DOI: 10.1016/j.xphs.2026.104159

Elham Taherian , Mirjam Crul , Reza Nejadnik

The majority of monoclonal antibodies (mAbs) are administered through intravenous infusion, typically in hospitals. A shift towards home-based therapy, initially driven by the COVID-19 pandemic, has been underway in recent years. Elastomeric pumps are devices that can be used for home-based therapy. They have been used for analgesics and antibiotics but not for delicate APIs such as proteins. While there is interest in using elastomeric pumps for administration of mAbs, and a few reports indicate that some hospitals have already started using them, valid stability data are lacking. This study aimed to investigate the stability of two marketed antibodies, Erbitux (cetuximab) and Herzuma (trastuzumab), during infusion using an elastomeric pump. High-performance size exclusion chromatography (HPSEC), flow imaging microscopy (FIM), dynamic light scattering (DLS), UV-VIS spectroscopy, and nanoparticle tracking analysis (NTA) were employed to assess subvisible particles and aggregation. Results show high stability and no degradation in form of aggregation in HPSEC. FIM data revealed that particle counts were the same before and after infusion using the pump. DLS and NTA data exhibited uniform particle sizes across samples, suggesting undetectable aggregation. Overall, findings support the technical feasibility of utilizing this combination of elastomeric pump and conditions for administration of cetuximab and trastuzumab, providing valuable insights into maintaining product stability outside traditional healthcare settings.

大多数单克隆抗体（mab）是通过静脉输注给药的，通常在医院。近年来，人们开始转向以家庭为基础的治疗，最初是由COVID-19大流行推动的。弹性泵是一种可用于家庭治疗的装置。它们已被用于止痛剂和抗生素，但未用于精细的原料药，如蛋白质。虽然人们对使用弹性体泵给药单克隆抗体很感兴趣，而且一些报告表明一些医院已经开始使用它们，但缺乏有效的稳定性数据。本研究旨在调查两种已上市抗体，Erbitux（西妥昔单抗）和Herzuma（曲妥珠单抗）在使用弹性泵输注期间的稳定性。采用高效粒径排除色谱（HPSEC）、流动成像显微镜（FIM）、动态光散射（DLS）、紫外可见光谱（UV-VIS）和纳米颗粒跟踪分析（NTA）来评估亚可见颗粒及其聚集。结果表明，HPSEC具有较高的稳定性和不以聚集形式降解。FIM数据显示，注射泵前后颗粒计数相同。DLS和NTA数据显示样品中颗粒大小均匀，表明无法检测到聚集。总的来说，研究结果支持利用这种弹性泵组合的技术可行性，以及西妥昔单抗和曲妥珠单抗给药的条件，为在传统医疗保健环境之外保持产品稳定性提供了有价值的见解。

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引用次数: 0

Extracellular vesicles from microalgae Pavlova in non-heated condition potently enhance innate immune response as a novel cell-derived immunostimulatory adjuvant 巴甫洛娃微藻细胞外囊泡作为一种新型细胞源性免疫刺激佐剂，在非加热条件下可增强先天免疫应答。

IF 3.8 3区医学 Q2 CHEMISTRY, MEDICINAL

Journal of pharmaceutical sciences

Pub Date : 2026-01-10 DOI: 10.1016/j.xphs.2026.104158

Masaki Morishita , Kotaro Yamada , Toshio Taira , Ken Nakahara , Tomonori Waku , Kotone Minato , Ken-ichi Ogawara

In vaccine therapy, there is a growing interest in novel immunostimulatory adjuvants derived from food products or dietary supplements with safe profiles. Pavlova is a microalga commonly used as a dietary supplement. Owing to its immune-activating potency and safety profiles, Pavlova offers novel possibilities as an immunostimulatory adjuvant. However, precise control of their proliferation profile and subsequent immune activation when administered to the host is difficult, limiting their practicality as adjuvants. Extracellular vesicles (EVs) are cell-derived nanoparticles containing nucleic acids and proteins. EVs exhibit biological activity without proliferating, indicating their potential in medical applications as novel cell-free materials. However, information on EVs secreted by microalgae, including Pavlova, is limited. In this study, we characterized the EVs from Pavlova as an immunostimulatory adjuvant. Heated Pavlova-derived EVs (hP-EVs) and non-heated Pavlova-derived EVs (n-hP-EVs) were purified from their culture supernatants, with heated Pavlova secreting larger amounts of EVs. These EVs exhibited a particle size of approximately 130–170 nm and a negatively-charged zeta potential. RAW264.7 macrophage cells more actively took up n-hP-EVs than hP-EVs via endocytosis. As an adjuvant activity, n-hP-EVs potently increased the production of the inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), which are indicators of enhanced innate immune response. Moreover, the effect of the storage temperature on the particle size of n-hP-EVs was small, maintaining their ability to activate innate immune responses. Thus, n-hP-EVs demonstrate attractive characteristics for use as novel cell-derived immunostimulatory adjuvants.

在疫苗治疗方面，人们对从安全的食品或膳食补充剂中提取的新型免疫刺激佐剂越来越感兴趣。巴甫洛娃是一种微藻，通常用作膳食补充剂。由于其免疫激活效力和安全性，巴甫洛娃作为免疫刺激佐剂提供了新的可能性。然而，当给药给宿主时，精确控制它们的增殖特征和随后的免疫激活是困难的，限制了它们作为佐剂的实用性。细胞外囊泡（EVs）是细胞衍生的含有核酸和蛋白质的纳米颗粒。电动汽车表现出不增殖的生物活性，表明其作为新型无细胞材料在医学上的应用潜力。然而，关于微藻（包括Pavlova）分泌的电动汽车的信息有限。在本研究中，我们将来自Pavlova的ev定性为免疫刺激佐剂。加热的帕夫洛娃衍生ev （hp - ev）和未加热的帕夫洛娃衍生ev （n- hp - ev）从培养上清中纯化，加热的帕夫洛娃分泌更多的ev。这些电动汽车的粒径约为130-170 nm，具有带负电的zeta电位。RAW264.7巨噬细胞通过内吞作用对n- hp - ev的吸收比hp - ev更积极。作为一种佐剂活性，n- hp - ev可显著增加炎性细胞因子肿瘤坏死因子-α (TNF-α)和白细胞介素-6 （IL-6）的产生，这是增强先天免疫应答的指标。此外，储存温度对n- hp - ev颗粒大小的影响很小，保持了它们激活先天免疫反应的能力。因此，n- hp - ev表现出作为新型细胞源性免疫刺激佐剂的有吸引力的特性。

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引用次数: 0

Overcoming translational barriers in nose-to-brain drug delivery for clinical applications 克服鼻脑给药的翻译障碍，促进临床应用。

IF 3.8 3区医学 Q2 CHEMISTRY, MEDICINAL

Journal of pharmaceutical sciences

Pub Date : 2026-01-08 DOI: 10.1016/j.xphs.2026.104156

Xinyue Zhang , Stephanie Chow , Ho Wan Chan , Shing Fung Chow

Recent research has shown an increasing interest in nose-to-brain drug delivery due to its non-invasive nature and ability to transport therapeutics directly to the central nervous system. This approach offers significant advantages over traditional administration routes, such as the circumvention of the blood-brain barrier and avoidance of first-pass metabolism, thereby enhancing therapeutic efficacy while reducing systemic side effects. Despite promising preclinical findings, nose-to-brain delivery remains underrepresented in the pharmaceutical market, highlighting a critical gap between experimental research and clinical translation. This review critically examines the major challenges confronting nose-to-brain delivery systems, including formulation development, selection of nasal devices, and methodologies for evaluating the nasal biopharmaceutics of drugs during nose-to-brain delivery. Furthermore, strategic recommendations and research priorities are outlined to address these barriers. By identifying and analyzing factors that contribute to the translational failure of nose-to-brain drug delivery systems, it is believed that more effective delivery systems can be developed, ultimately revolutionizing treatment strategies for neurological diseases.

最近的研究表明，由于其非侵入性和直接将治疗药物运送到中枢神经系统的能力，鼻子到大脑的药物递送越来越受到关注。与传统给药途径相比，该方法具有明显的优势，如绕过血脑屏障和避免首过代谢，从而提高了治疗效果，同时减少了全身副作用。尽管临床前研究结果很有希望，但鼻到脑给药在制药市场上的代表性仍然不足，这凸显了实验研究与临床转化之间的重大差距。这篇综述批判性地探讨了鼻到脑给药系统面临的主要挑战，包括配方开发，鼻装置的选择，以及在给药过程中评估药物的鼻生物制药方法。此外，还概述了解决这些障碍的战略建议和研究重点。通过识别和分析导致鼻到脑系统转化失败的因素，相信可以开发出更有效的传递系统，最终彻底改变神经系统疾病的治疗策略。

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引用次数: 0

A multicompartment stomach physiologically-based pharmacokinetic model capturing gastric reacidification can improve food effect predictions of weak bases 捕获胃再酸化的多室胃生理药代动力学模型可以改善弱碱的食物效应预测。

IF 3.8 3区医学 Q2 CHEMISTRY, MEDICINAL

Journal of pharmaceutical sciences

Pub Date : 2026-01-03 DOI: 10.1016/j.xphs.2026.104155

Lisa Cheng , Po-Chang Chiang , Matthew R Wright , Harvey Wong

Various changes within the gastrointestinal tract following meal intake may significantly impact oral drug absorption. Physiologically-based pharmacokinetic (PBPK) models are often used to perform early predictions of food effects on oral absorption. However, the stomach compartment within these physiological models does not wholly reflect physiology of the fed state. The current approach where the stomach compartment is fixed at pH 5 overlooks the changing gastric pH profile over time after food consumption. We suggest the integration of a multicompartment stomach that represents the various pH phases of the fed stomach to better capture ionizable drug dissolution. Ibuprofen sodium (weak acid) and posaconazole (weak base) were investigated to determine whether a PBPK model that could better capture the varying gastric pH would yield an improvement in food effect predictions. The one-compartment and multicompartment stomach models’ simulations for ibuprofen sodium exposure with and without food consumption were comparable. For posaconazole, the one-compartment stomach model predicted a smaller area under the curve and lower maximum plasma concentration than the multicompartment stomach model’s output in the fed state. Although both models accurately predicted a positive food effect, the magnitude of the simulated fed to fasted fold-changes in posaconazole exposure by the one-compartment stomach model was much less than observed fold-changes. These results suggest that a multicompartment stomach model featuring gastric re-acidification associated with food intake should be incorporated for more accurate food effect predictions for weakly basic compounds.

膳食摄入后胃肠道内的各种变化可能显著影响口服药物的吸收。基于生理的药代动力学（PBPK）模型通常用于对食物对口服吸收的影响进行早期预测。然而，这些生理模型中的胃隔室并不能完全反映进食状态的生理。目前将胃隔室固定在pH 5的方法忽略了食物摄入后胃pH值随时间的变化。我们建议整合一个多室胃，代表不同的pH相喂养胃，以更好地捕获电离药物溶解。研究了布洛芬钠（弱酸）和泊沙康唑（弱碱），以确定能够更好地捕捉胃pH变化的PBPK模型是否会改善食物效应预测。单室胃模型和多室胃模型对布洛芬钠暴露与不进食的模拟具有可比性。对于泊沙康唑，单室胃模型比多室胃模型在进食状态下预测的曲线下面积更小，最大血浆浓度更低。虽然这两个模型都准确地预测了积极的食物效应，但单室胃模型模拟的泊沙康唑暴露从进食到禁食的折叠变化的幅度远小于观察到的折叠变化。这些结果表明，应该将胃再酸化与食物摄入相关的多室胃模型纳入其中，以更准确地预测弱碱性化合物的食物效应。

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引用次数: 0

Explaining the underlying causes of different tabletability classifications of binary mixtures 解释二元混合物表性分类不同的根本原因。

IF 3.8 3区医学 Q2 CHEMISTRY, MEDICINAL

Journal of pharmaceutical sciences

Pub Date : 2026-01-03 DOI: 10.1016/j.xphs.2026.104154

Pradeep Valekar, Ira S. Buckner

A tabletability classification system has been previously proposed to categorize the range of tabletability behaviors of binary mixtures.¹^,² Although the proposed system has been applied in several subsequent studies, the causes of each behavior have not been explained. The main objectives of this study were to investigate why different mixtures exhibit different behaviors and identify the attributes of components that would display specific behaviors in mixtures. To this end, binary mixtures comprising individual components with varied compressibility and compactibility were characterized. The component properties and mixture behaviors were compared to identify trends in the data. It was found that by combining accurate models of the compressibility and compactibility of pure components, the tabletability behavior of their mixtures can be predicted both qualitatively and quantitatively. The mixtures of components with similar tabletability profiles displayed linear behavior. The pronounced differences in both compactibility and tabletability produced negative deviations, while pronounced differences in compressibility and compactibility resulted in positive deviations.

先前已经提出了一个表性分类系统来对二元混合物的表性行为范围进行分类。尽管提出的系统已在随后的几项研究中得到应用，但每种行为的原因尚未得到解释。本研究的主要目的是研究为什么不同的混合物表现出不同的行为，并确定在混合物中表现出特定行为的成分的属性。为此，对具有不同压缩率和压实性的单个组分组成的二元混合物进行了表征。通过比较组分性质和混合行为来确定数据的趋势。研究发现，结合纯组分的压缩性和压实性的精确模型，可以定性和定量地预测其混合物的压实性行为。具有相似表性轮廓的组分的混合物表现出线性行为。紧致性和可表性的显著差异产生负偏差，而压缩性和紧致性的显著差异导致正偏差。

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引用次数: 0

The impact of bile acid sequestrants on octreotide absorption 胆汁酸螯合物对奥曲肽吸收的影响。

IF 3.8 3区医学 Q2 CHEMISTRY, MEDICINAL

Journal of pharmaceutical sciences

Pub Date : 2026-01-03 DOI: 10.1016/j.xphs.2026.104153

Zahraa Al-Tamimi , Mei Feng , Waleed Elballa , Sydney Houser , Michael J. Hageman

Despite significant advancements in peptide drug development, there is still a challenge in formulating and delivering peptide drugs orally. Current oral peptide drugs have very low bioavailability (<1%), which could be attributed, in part, to enzymatic instability, poor membrane permeability/flux, and the sequestration by intestinal colloids composed of bile acid and phospholipids that form bile acid-phospholipid mixed micelles (BAPMM). In this work, we examined the effect of perturbing the BAPMM with bile acid sequestrants (BAS) on the membrane flux and enzymatic stability of octreotide in vitro, and its potential impact on peptide absorption and bioavailability in vivo. Additionally, we tested the effect of adding cyclic E-cadherin peptide (ECP) permeation enhancers on the bioavailability of octreotide. The results suggest that using BAS decreases the bile acid levels and putatively disrupts the micellar structure, leading to a higher concentration of the free peptide to diffuse across the membrane. In vitro bile acid sequestration enhanced the overall peptide flux rate without compromising the improved enzymatic stability. Our in vivo data suggests that using the BAS, colestipol, did not have a significant impact on peptide absorption though. These results highlight the important role of BAPMM on bioaccessible drug concentration, as well as membrane permeation.

尽管多肽药物的开发取得了重大进展，但在口服多肽药物的配方和给药方面仍然存在挑战。目前口服肽类药物的生物利用度非常低(

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引用次数: 0

Application of ion-exchange chromatographic technique using resins to prepare the salts of polar unstable prodrugs and new chemical entities (NCEs) 离子交换色谱技术在树脂制备极性不稳定前药及新化学实体（nce）盐中的应用。

IF 3.8 3区医学 Q2 CHEMISTRY, MEDICINAL

Journal of pharmaceutical sciences

Pub Date : 2026-01-03 DOI: 10.1016/j.xphs.2026.104152

Raja K. Rit, Venkataramasubramanian V., Deepika V., Pandiyaraj B., Umesh Waman Mali, Olivier Venier, Santoshkumar N. Patil

Prodrugs are inactive and bio-reversible derivatives of drugs or clinical candidates. Prodrugs and New Chemical Entities (NCEs) often do not show good bioavailability primarily due to poor solubility. One of the strategies to overcome this challenge is to use their salt forms. Conventional salt preparation techniques fail when applied to polar unstable prodrugs and NCEs. Herein, we have developed a simple efficient salt preparation technique using ion exchange resin chromatography. This technique involves the exchange of cations on sulfonic acid-based resin that could be applied successfully to prepare the salts of acidic prodrugs and NCEs. In this current work, phosphate/sulfate/carboxylate-prodrugs were prepared in the form of their inorganic (e.g. Na⁺, K⁺) and organic amine (e.g. erbumine, trizma, meglumine, arginine) salts with excellent control. The resin cartridge was reused for the preparation of different salts without any significant compromise in yields. Moreover, this protocol was employed to quickly switch the counterions for the pH-sensitive prodrugs, which is very useful in pharmaceutical development. This method is cost effective, operationally simple and presents a broad substrate scope. We wish this technique would help organic and pharmaceutical chemists to prepare various salt forms of prodrugs or NCEs in an efficient and controlled manner.

前药是药物或临床候选药物的非活性和生物可逆的衍生物。前药和新化学实体（NCEs）往往表现出良好的生物利用度，主要是由于溶解度差。克服这一挑战的策略之一是使用它们的盐形式。传统的盐制备技术不适用于极性不稳定的前药和nce。在此，我们开发了一种简单高效的离子交换树脂色谱制盐技术。该技术涉及到阳离子在磺酸基树脂上的交换，可以成功地应用于制备酸性前药和nce的盐。在本工作中，磷酸盐/硫酸盐/羧酸-前药以其无机（如Na+， K+）和有机胺（如erbumine， trizma, meglumine，精氨酸）盐的形式制备，具有良好的控制。树脂筒被重复用于制备不同的盐，而没有任何显着的妥协在产量。此外，该方案可用于快速切换ph敏感性前药的反离子，这在药物开发中非常有用。该方法成本低，操作简单，适用范围广。我们希望这项技术能够帮助有机和药物化学家以有效和可控的方式制备各种盐形式的前药或nce。

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引用次数: 0

Preparation and characterization of oral insulin-trimethyl chitosan complex-loaded solid self-nanoemulsifying drug delivery systems (SNEDDS) 口服胰岛素-三甲基壳聚糖复合物负载固体自纳米乳化给药系统的制备与表征。

IF 3.8 3区医学 Q2 CHEMISTRY, MEDICINAL

Journal of pharmaceutical sciences

Pub Date : 2025-12-30 DOI: 10.1016/j.xphs.2025.104149

Richieline S. Cual, Tajmarah Ayyah M. Sandato, Mae Quenie T. Pontanar

Insulin, a pancreatic hormone regulating blood glucose, is available in forms of syringe, pen, or implants; however, these methods presented challenges related to skin adverse reactions and patient compliance. Oral insulin delivery has long been explored due to higher compliance and reduced costs. Similar to natural insulin secretion, but prone to GIT breakdown and low bioavailability. SNEDDS has been investigated to address these in protein formulations. This study formulated Insulin into SNEDDS with trimethyl chitosan and spray-dried for stability, then analyzed for properties and drug release. The study successfully prepared Insulin-TMC SNEDDS with a 36.17±4.02 s emulsification time, forming a Grade B nanoemulsion. Droplet sizes remained nanoscale (134.20–287.20 nm for liquid, 188.77–214.93 nm for solid), preserving characteristics post–spray drying. Stability was confirmed in TMC-free formulations, while TMC variants showed sedimentation over time. TMC-containing Solid-SNEDDS showed moisture content below 5 %, indicating good stability. Insulin release varied with pH—highest at pH 2.5 with 3 mg/mL TMC (105.48 % cumulative percent drug release), while lower releases were seen at pH 6.6 (61.73±1.38 %) and 7.0 (61.73±1.38 %) without TMC. The study supports TMC-based SNEDDS and spray drying as a potential method for oral insulin delivery.

胰岛素是一种调节血糖的胰腺激素，可以通过注射器、笔或植入物获得；然而，这些方法提出了与皮肤不良反应和患者依从性相关的挑战。长期以来，人们一直在探索口服胰岛素，因为它具有更高的依从性和更低的成本。类似于天然胰岛素分泌，但易于GIT分解和低生物利用度。已经研究了SNEDDS以解决蛋白质配方中的这些问题。本研究采用三甲基壳聚糖将胰岛素配制成SNEDDS，喷雾干燥稳定性，并对胰岛素的性质和释放度进行分析。本研究成功制备了胰岛素- tmc SNEDDS，乳化时间为36.17±4.02s，形成了B级纳米乳。液滴尺寸保持纳米级（液体为134.20 ~ 287.20 nm，固体为188.77 ~ 214.93 nm），保持喷雾干燥后的特性。在不含TMC的配方中，稳定性得到了证实，而TMC变体随着时间的推移会出现沉淀。含tmc的Solid-SNEDDS含水率低于5%，稳定性好。胰岛素释放量随pH值的变化而变化，当TMC浓度为3 mg/mL时，pH值为2.5时释放量最高（累计释放量为105.48%），而不含TMC时，pH值为6.6（61.73±1.38%）和7.0（61.73±1.38%）时释放量较低。该研究支持基于tmc的SNEDDS和喷雾干燥作为口服胰岛素递送的潜在方法。

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引用次数: 0

Interaction of isoniazid derivatives active against drug-resistant tuberculosis with models of the lung surfactant and of the Mycobacterium tuberculosis cell wall 抗耐药结核的异烟肼衍生物与肺表面活性剂和结核分枝杆菌细胞壁模型的相互作用。

IF 3.8 3区医学 Q2 CHEMISTRY, MEDICINAL

Journal of pharmaceutical sciences

Pub Date : 2025-12-30 DOI: 10.1016/j.xphs.2025.104151

Weronika Śliżewska , Filomena Martins , Rodrigo F.M. de Almeida , Joaquim T. Marquês

Tuberculosis is one of the most important causes of death in the world. The emergence and increased prominence of multidrug-resistant strains of Mycobacterium tuberculosis (Mtb), non-susceptible to currently available therapies, has toughened the fight to eradicate this disease. This study focuses on further investigating the therapeutic potential of promising antitubercular compounds, namely, isoniazid (INH), and three INH derivatives, N'-decanoylisonicotinohydrazide (INH-C10), N'-(E)-(4-phenoxybenzylidene)isonicotinohydrazide (N34) and N’-(4-phenoxybenzyl)isonicotinohydrazide (N34red). INH-C10 and N34 have been selected due to their high selectivity index against the Mtb mutant bearing the primary mutation responsible for INH drug resistance. In opposition, N34red, which differs from N34 only in the saturation of the N′ = C bond, exhibits a poor selectivity index. Moreover, INH-C10 and N34 interact with human serum albumin and model lipid membranes mimicking the plasma membrane of human cells, showing their promising potential. In the current study, the interaction of these compounds with models of the lung surfactant (LS) and of the mycolic acid (MA)-enriched Mtb cell wall was assessed, in order to further explore their ability to interact with and cross the various biological barriers to be encountered on their way to the molecular target inside Mtb. We show that all the INH derivatives were able to interact with both the LS and the mycolic acid-enriched cell wall models. INH-C10 and N34 had a smaller impact than N34red on the pulmonary surfactant model. On the other hand, INH-C10 promoted the most extensive perturbation of the MA-enriched cell wall model, which correlates well with the previously shown ability of this compound to incorporate into and disturb gel-phase lipid bilayers. This indicates that INH-C10 may penetrate a MA rich barrier more easily, reaching higher intracellular levels, and increase its permeability. These traits contribute to explain the high antimicrobial activity of this derivative against the most common drug-resistant Mtb mutant.

结核病是世界上最重要的死亡原因之一。对现有疗法不敏感的耐多药结核分枝杆菌（Mtb）菌株的出现和日益突出，加强了根除这一疾病的斗争。本研究的重点是进一步研究有前景的抗结核化合物，即异烟肼（INH）及其三个INH衍生物N'-癸烷基异烟碱肼（INH- c10）， N'-(E)-（4-苯氧苄基）异烟碱肼（N34）和N'-（4-苯氧苄基）异烟碱肼（N34red）的治疗潜力。选择INH- c10和N34是因为它们对携带INH耐药主要突变的Mtb突变体具有高选择性指数。相反，N34red与N34的区别仅在于N' = C键的饱和度，其选择性指数较差。此外，INH-C10和N34与人血清白蛋白和模拟人细胞膜的模型脂质膜相互作用，显示出其良好的潜力。在本研究中，我们评估了这些化合物与肺表面活性物质（LS）和富含霉菌酸（MA）的结核分枝杆菌细胞壁模型的相互作用，以进一步探索它们与结核分枝杆菌内部分子靶点到达过程中遇到的各种生物屏障相互作用和跨越的能力。我们发现所有的INH衍生物都能与LS和富含霉菌酸的细胞壁模型相互作用。INH-C10和N34对肺表面活性物质模型的影响小于N34red。另一方面，INH-C10促进了对富含ma的细胞壁模型的最广泛的扰动，这与先前显示的该化合物融入并干扰凝胶相脂质双分子层的能力密切相关。这表明INH-C10可能更容易穿透富含MA的屏障，达到更高的细胞内水平，并增加其通透性。这些特征有助于解释该衍生物对最常见的耐药结核分枝杆菌突变体具有较高的抗菌活性。

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引用次数: 0

Experimental determination of extinction coefficients by sedimentation velocity analytical ultracentrifugation for accurate quantification of recombinant adeno-associated virus 用沉降速度分析超离心法测定重组腺相关病毒消光系数的精确定量。

IF 3.8 3区医学 Q2 CHEMISTRY, MEDICINAL

Journal of pharmaceutical sciences

Pub Date : 2025-12-30 DOI: 10.1016/j.xphs.2025.104150

Yuki Yamaguchi , Takahiro Maruno , Takaaki Kurinomaru , Risa Shibuya , Mitsuko Fukuhara , Yasuo Tsunaka , Susumu Uchiyama

Recombinant adeno-associated virus (rAAV) is widely used as a gene delivery vector. Sedimentation velocity analytical ultracentrifugation (SV–AUC) is the gold standard for quantifying the ratio of full particles (FPs) to the sum of empty particles (EPs) and FPs (F/E ratio) of rAAVs. Here, we experimentally determined the molar extinction coefficients (ε) and mass extinction coefficients of highly purified FPs and EPs of AAV serotypes 2, 5, 6, 8, and 9 using SV–AUC with interference and multi-wavelength absorbance detection. At 230 nm, the difference in ε between EPs and FPs was the smallest, although the ε of FPs remained 1.2-fold higher than that of EPs. Expectedly, the differences in ε between FPs and EPs were almost identical across serotypes with the same genome length and increased linearly in a genome length-dependent manner, although both sets of ε differed across serotypes. Consequently, accurate quantification of F/E ratio requires the use of distinct ε values for EPs and FPs. The ε per base of single-stranded DNA was independent of serotype and genome length, allowing estimation of the ε of FPs from that of EPs. Coupling these ε values with SV–AUC enables the determination of absolute rAAV concentrations. This study provides practical guidance for accurate absorbance-based rAAV quantification.

重组腺相关病毒（rAAV）作为一种基因传递载体被广泛应用。沉降速度分析超离心（SV-AUC）是定量测定raav中满颗粒（FPs）与空颗粒（EPs）和FPs （F/E比）之比的金标准。本研究采用干涉和多波长吸光度检测的SV-AUC技术，测定了AAV血清型2、5、6、8和9的高纯度FPs和EPs的摩尔消光系数（ε）和质量消光系数。在230 nm处，EPs和FPs的ε值差异最小，但FPs的ε值仍然是EPs的1.2倍。出乎意料的是，在相同基因组长度的血清型中，FPs和EPs之间的ε差异几乎相同，并且以基因组长度依赖的方式线性增加，尽管两组ε在血清型中存在差异。因此，准确量化F/E比率需要对EPs和FPs使用不同的ε值。单链DNA的每碱基ε与血清型和基因组长度无关，可以通过EPs的ε来估计FPs的ε。将这些ε值与SV-AUC耦合，可以确定rAAV的绝对浓度。本研究为基于吸光度的rAAV准确定量提供了实用指导。

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