Assay and drug development technologies最新文献

From the steady stream of drug repurposing patent applications published under the Patent Cooperation Treaty (PCT), we have selected fifteen documents that first became available during the first quarter of 2025. As in each installment, some of these claims are truly surprising. Few researchers would have expected that SSRI antidepressants such as sertraline and indatraline could exhibit pronounced anticancer effects. Equally unexpected is the disclosure that sitagliptin, the first antidiabetic agent from the DPP-4 inhibitor class, may be used for the treatment of glioblastoma. Another striking example is the report that artemisinin derivatives, well known for their use against malaria, may induce differentiation in undifferentiated erythroid and myeloid cells in patients with myelodysplastic syndrome. In addition, the compound bucillamine-relatively obscure in Western medicine but long used for the treatment of rheumatoid arthritis in East Asia-has been proposed for potential benefit in organophosphate poisoning. These highlights exemplify the breadth of innovation currently shaping the drug repurposing landscape. The reviewed patent applications originate from a diverse range of jurisdictions, including France, Spain, Greece, Slovenia, South Korea, China, Japan, Canada, and the United States, illustrating the global nature of ongoing research efforts in this field.

This study presents the first validated High-performance liquid chromatography (HPLC) technique for quantifying naphthol AS-E phosphate (NASEP) in bulk drugs and nanoparticle formulation. A C18 HPLC cartridge (250 × 4.6 mm, 5 µm particle size) served as the stationary phase for quantification. The mobile phase consisted of Milli-Q water with 0.1% trifluoroacetic acid (TFA) in pump A and acetonitrile with 0.1% TFA in pump B, with a flow rate ranging from 0.8 to 1.2 mL/min. A 3² factorial design was employed to evaluate the robustness of the proposed method, using mobile phase composition (X₁), flow rate (X₂), and column temperature (X₃) as independent variables and peak area (R₁), retention time (R₂), and percent recovery (R₃) as response variables. The calibration range curve (10-500 µg/mL) was best fitted by quadratic regression. The linearity was reported in the above-mentioned range. The accuracy was 99.952% ± 0.961% at the 75% level, 99.58% ± 1.483% at the 100% level, and 99.789% ± 1.936% at the 125% level. The coefficient of variation was below 2% for both intraday and interday measurements, and the limits of detection and quantification were 0.038 and 0.115 µg/mL, respectively. The NASEP solution was stable (99.04% ± 0.0251%) for 48 h at 8°C. The forced degradation study also revealed that the NASEP solution remained stable in an acidic environment for 48 h at 40°C but degraded at 80°C (p < 0.046) in a time-dependent manner. In contrast, it was unstable in an alkaline medium, independent of temperature, and degraded in the presence of strong oxidizing agents (p < 0.039). Furthermore, NASEP encapsulated in a Gly-Arg-Gly-Asp-Ser pentapeptide and low-molecular-weight heparin functionalized metal-organic framework exhibited sustained drug release at acidic pH 5.4. The proposed NASEP quantification method was validated and is suitable for routine analysis in pharmaceutical formulations.

In this study, we integrated computational and experimental approaches to identify novel therapeutic targets and candidate compounds for gastric cancer (GC). Through in silico analyses, including target prediction, pathway enrichment, and molecular docking, four key proteins-MET, ADORA2A, CDK5R1, and ADORA1-were identified as critical regulators of tumor proliferation, metastasis, and drug resistance pathways (e.g., Semaphorin interactions and NTRK1 signaling). Molecular docking and dynamics simulations revealed strong binding affinities and structural stability between selected compounds and these targets, prioritizing ADORA1 as a promising therapeutic node. To validate these findings, we synthesized compound 3 via a two-step chemical route, yielding a white solid product with 63% overall efficiency. Structural characterization by High-Resolution Mass Spectrometry (HRMS) and ¹H Nuclear Magnetic Resonance (NMR) confirmed its identity. In vitro inhibition assays demonstrated that compound 3 exhibited potent activity against ADORA1, with a mean Half-Maximal Inhibitory Concentration (IC₅₀) of 0.23 nM-approximately twofold more effective than the positive control antagonist Dipropylcyclopentylxanthine (DPCPX) (IC₅₀ = 0.46 nM). These results highlight compound 3 as a promising lead compound for further development in GC therapy, with potential to modulate ADORA1-mediated signaling pathways.

The aim of this study was to develop a novel CuFe nanozyme-enhanced sensing platform for the ultrafast detection of trace analytes, specifically targeting environmental pollutants and heavy metals. The objectives of the research included evaluation of the platform's sensitivity, selectivity, and real-world applicability for detecting trace analytes in environmental and biological samples. We synthesized the CuFe nanozyme using a co-precipitation method with metal-organic precursors and a reducing agent. The sensing platform was fabricated using conductive electrodes and immobilized nanozymes. The turnover frequency was calculated under optimized conditions (e.g., temperature, pH, and substrate concentration). Equipment utilized included an X-ray diffraction analyzer, transmission electron microscope, electrochemical workstation, and UV-Vis spectrophotometer. This CuFe nanozyme demonstrated a turnover frequency of 125 s^-1, 3.5 times higher than natural peroxidase enzymes, as determined using a colorimetric assay with 3,3',5,5'-Tetramethylbenzidine. The sensing platform exhibited ultrafast detection with a response time of 5 s, determined through real-time monitoring of analyte interaction via the electrochemical method. The detection limit was established at 0.1 nM for target analytes, as measured by the electrochemical method with calibration curves constructed for each analyte in the concentration range of [0.1 nM-X nM]. Importantly, the system was successfully validated in real-world environmental water samples and spiked clinical fluids, showing high recovery rates (98%-102%). The CuFe nanoenzyme-based electrochemical sensing platform demonstrated high accuracy, precision, and recovery in environmental water and spiked biological fluid samples. This study presents a robust, ultrafast nanozyme-based sensing platform with superior sensitivity and selectivity.

Exosomes, nano-sized extracellular vesicles secreted by almost all cell types, have emerged as biologically compatible vehicles for targeted drug delivery, gene therapy, and molecular diagnostics. Their innate ability to traverse biological barriers and deliver diverse cargoes with minimal immunogenicity has catalyzed intense interest in their therapeutic exploitation. However, the intrinsic heterogeneity and limited targeting specificity of native exosomes necessitate advanced engineering strategies to fulfill their clinical potential. This review focuses on the molecular-level nanoengineering of exosomal surfaces to enhance specificity, loading efficiency, and release control of therapeutic payloads. We systematically examine current methodologies, including genetic modification of parental cells, covalent and non-covalent surface conjugation, lipid insertion, click chemistry, and hybrid vesicle fusion. We further detail the quantitative performance of targeting ligands-such as peptides, aptamers, nanobodies, and glycans-in relation to receptor affinity, conjugation efficiency, and biological outcomes. Payload loading techniques, both endogenous and exogenous, are critically analyzed based on loading yield and membrane preservation. Additionally, we highlight disease-specific applications in oncology, neurology, cardiology, and immunotherapy, supported by preclinical and translational case studies. Emerging technologies such as microfluidics, synthetic biology, artificial intelligence-guided modeling, and multi-omics are discussed as integral components of the next generation of precision exosome platforms. Finally, we address key challenges related to scalability, regulatory frameworks, and standardization. This review provides a comprehensive and quantitative framework to guide the design of molecularly engineered exosomes for future translational and clinical success.

Capsaicin (CAP), a bioactive compound from chili peppers, possesses a wide range of therapeutic properties, including antiobesity, anticancer, anti-inflammatory, analgesic, and cardioprotective effects. However, its clinical application has been limited due to poor aqueous solubility, low bioavailability, rapid metabolism, and potential side effects with high doses. Recent advancements in nanotechnology have addressed these challenges by enhancing CAP's solubility, stability, and targeted delivery through innovative nanoformulations. This review provides an in-depth analysis of various nanocarrier systems such as solid lipid nanoparticles, nanostructured lipid carriers, liposomes, polymeric nanoparticles, nanoemulsions, and nanocrystals, all of which have demonstrated improved therapeutic efficacy of CAP in preclinical studies. These nanoformulations not only protect CAP from degradation but also enable controlled release, reduce side effects, and improve patient compliance. The therapeutic potential of CAP-loaded nanocarriers has been investigated in a variety of diseases, including cancer, neurodegenerative disorders, and chronic pain, with promising results. This review highlights the latest innovations in CAP nanotechnology and discusses the future directions for clinical applications, paving the way for more effective CAP-based therapies in modern medicine.

Stroke is an intricate oxidative and inflammatory response resulting from cerebral ischemia followed by reperfusion injury. The complex pathophysiology of stroke poses challenges for treatment. Peroxisome proliferated-activated receptor (PPAR) expression in the rat hippocampus is markedly elevated post cerebral ischemia/reperfusion injury (I/R injury). Hence, Saroglitazar, a dual PPAR-α/γ agonist, was investigated against cerebral I/R injury in rats. Male Sprague Dawley rats were subjected to bilateral common carotid artery occlusion for 30 min and reperfusion for 3 days. During the reperfusion, animals were treated with vehicle or Saroglitazar once a day for 3 days. The behavioral parameters were assessed, and animals were sacrificed to measure oxidative markers (Malondialdehyde, Superoxide dismutase, catalase, and reduced glutathione), inflammatory markers (interleukin-6, tumor necrosis factor-α, nuclear factor kappa-light chain enhancer of activated B cells [NF-κB], and high mobility group box 1 protein [HMGB-1]), infarction, and histopathology changes. Following I/R injury, antioxidant enzymes were reduced, while nitric oxide and inflammatory markers were increased in the disease group. In the rat hippocampus, these changes led to neurobehavioral impairment and cerebral infarction. Saroglitazar improved the levels of antioxidants and reduced inflammation. 2,3,5-triphenyltetrazolium chloride staining and histopathological analysis revealed the neuroprotective effect of Saroglitazar in the hippocampus region. The neuroprotective effects of Saroglitazar were attributed to its activation of both PPAR-α and PPAR-γ. It improved antioxidant levels and inhibited pro-inflammatory cytokines by suppressing the HMGB-1/NF-κB signaling pathway. These findings underscore the potential of Saroglitazar in mitigating cerebral I/R injury.

High-throughput screening (HTS) approaches incorporating multiplexed, cell-based assays are increasingly common for identifying novel modulators of complex biological processes. To enable the discovery of small molecule modulators of mRNA processing, we developed a multiplexed, bead-based, high-throughput QuantiGene assay, leveraging the Luminex platform that is capable of simultaneously quantifying transcript levels of multiple independent target genes within a single HTS campaign. To address plate variability and potential screening artifacts, a pragmatic hit-calling pipeline was implemented utilizing both plate- and well-based normalization strategies. This dual-normalization approach reduced false negatives and produced consistent hit confirmation rates. Application of this methodology led to the identification of unique compounds selectively modulating individual target genes. Strikingly, among the three exemplary genes, only 5% of primary actives demonstrated activity across all three target genes, underscoring the assay's capacity for detecting selective mRNA modulators. Chemical motif analysis of confirmed actives recovered known RNA privileged scaffolds as well as novel scaffolds that are uniquely enriched for individual targets screened. Validation screening using an orthogonal, four-point concentration-response real-time PCR (qPCR) assay in a disease-relevant cell line demonstrated high validation rates, supporting the robustness and translational relevance of this multiplexed HTS platform. These findings establish a scalable and reliable strategy for identifying selective small molecule modulators of mRNA processing, with broad applicability in early drug discovery.

Berberine (BER) is an antioxidant, anti-inflammatory, and antitumor agent, while paclitaxel (PTX) is a widely used synthetic chemotherapeutic agent for breast cancer. Several reports have suggested the use of a PTX and BER combination for the effective treatment of breast cancer, and many pharmaceutical scientists are working to develop a novel drug delivery system incorporating this combination. However, the literature lacks a reliable simultaneous estimation method for this combination. Therefore, in the present study, we aimed to develop a robust reversed-phase high-performance liquid chromatography method for the simultaneous estimation of PTX and BER in free drug form in liposomal formulation. The method employed a C₁₈ column (250 × 4.6 mm, 5 µm) with acetonitrile and water (70:30, v/v) as the mobile phase at a flow rate of 0.8 mL/min and detection at 250 nm. Retention times were 2.84 and 5.62 min for PTX and BER, respectively. Theoretical plates >2000 were demonstrated, and peak tailing of <2 in validation as per ICH Q2(R2) was observed. In the 10-50 ppm range, linearity was found with R² values of 0.9979 (PTX) and 0.9975 (BER). Furthermore, the method achieved within acceptable limits precision (<2% relative standard deviation) and accuracy (90%-110%). Robustness assessments checked method reliability in small variations. In addition, using the method effectively, entrapment efficiencies of 85.27 ± 1.74% and 78.62 ± 2.38% were obtained for PTX and BER in liposomal formulations. Moreover, in vitro release studies revealed 98.83 ± 2.94% (PTX) and 96.56 ± 1.92% (BER) release over 24 h. The validated method was precise, accurate, and reliable, making it suitable for application in drug formulation analysis.

Here, we briefly discuss 17 PCT patent applications related to drug repurposing that were published during the final quarter of 2024. As always, we have chosen the documents based on significance only, which gave us a wide range of applicants, active agents, and geographic representation. Applicants are not only from the United States and China but also from Japan, Germany, the United Kingdom, Canada, and even remote places such as Algeria and Iceland. Significantly, the contribution from Iceland is how to induce therapeutic hypothermia-but with entacapone, used in Parkinson's disease. The UK company, HealX, presents data showing that Angelman syndrome, an intractable neurodevelopmental disorder, could be treated with the common NSAID sulindac. Gliclazide, an old antidiabetic, could potentially treat schizophrenia, as inventors from Chinese Ping An-Shionogi report. These are only a few highlights from the stream of documents that have been issued towards the end of 2024.