Concurrent blockade of aerobic glycolysis and oxidative phosphorylation (OXPHOS) holds great promise in lung cancer therapy yet challenged by tumor cell metabolic plasticity. To address this, we herein grafted dichloroacetic acid into perylenediimide (PDI) skeleton via ionic or covalent bond to create PDIC-AC and PDIC-NAC. Studies demonstrate that ionic bond-driven primary amine positive nitrogen remodeling and mitochondrial localization endow PDIC-AC with significantly stronger inhibitory activity on pyruvate dehydrogenase kinases (PDHKs) than PDIC-NAC. Notably, PDIC-AC targets the Rieske iron-sulfur polypeptide 1 (UQCRFS1) subunit of complex III in mitochondria, triggering electron leakage from the electron transport chain, thereby more efficiently inducing reactive oxygen species (ROS) production relative to PDIC-NAC. Superior PDHKs inhibiting efficacy and ROS generation capacity functionalize PDIC-AC as an efficient inhibitor to block glycolysis and OXPHOS, which not only repolarize macrophages toward anti-tumor M1 phenotype via suppression of lactate production, but also trigger immunogenic cell death via PERK-eIF2α-ATF4-CHOP axis to activate immune response, ultimately reaching effective chemo-immunotherapy against the primary and distant tumors. Overall, this work defines the unambiguous mechanism for PDI-triggered endogenous ROS generation, and meanwhile clarifies small-molecule regulators' energy metabolism intervention mechanism and establishes an innovative chemical bond engineering strategy for energy-targeted chemo-immunotherapy.

ETHNOPHARMACOLOGICAL RELEVANCE:Danshensu (DSS) is one of the water-soluble components extractable from the traditional Chinese medicine Salvia miltiorrhiza Bge., exhibiting pharmacological effects such as promoting blood circulation, dilating coronary arteries, and improving cerebral blood flow. The Danshensu derivative (OZD-1) obtained through the derivatization of DSS is a potential multi-target drug for the central nervous system, however, its mechanism of action against cerebral ischemia-reperfusion injury (CIRI) remains unclear. AIM OF THE STUDY:Systematically investigating the therapeutic potential and mechanisms of action of Danshensu derivative against cerebral ischemia-reperfusion injury. MATERIALS AND METHODS:Rat brain microvascular endothelial cells (RBMVECs) were cultured in vitro to establish an oxygen-glucose deprivation/reoxygenation (OGD/R) injury model. Groups included a control group, an OGD/R model group, and OZD-1 low-dose (12.5 μmol/L), medium-dose (25 μmol/L), and high-dose (50 μmol/L) groups. Cell viability, migration capacity, and vascular lumen formation were assessed using the CCK-8 assay, cell scratch assay, and matrigel matrix gel assay, respectively. In vivo, a transient middle cerebral artery occlusion (tMCAO) model was established in rats. Animals were randomly divided into the sham, model, OZD-1 (35, 70, 140 mg/kg), Edaravone (Eda), and DSS groups. Daily oral administration was performed post-surgery for 14 consecutive days. Tissue pathology staining, behavioral tests, and regional cerebral blood flow imaging assessed brain tissue damage, cognitive function, and ischemic side cerebral blood flow recovery, respectively. Transcriptome sequencing analyzed differential gene expression and pathway enrichment patterns. Western blot detection measured expression levels of proteins related to the PI3K-AKT-CREB signaling pathway, phosphoproteins, downstream apoptosis-related proteins, and CD31, CD34, and VEGFA proteins. RESULTS:In vitro experiments demonstrated that OZD-1 dose-dependently enhanced the viability of RBMVECs following OGD/R injury, significantly improving cell migration and luminal formation capabilities. In vivo studies revealed that compared to the model group, rats in all OZD-1 dosage groups exhibited markedly improved cognitive function, significantly restored cerebral blood flow in the ischemic hemisphere, and substantially reduced pathological brain tissue damage. Transcriptome sequencing results indicated significant enrichment of genes associated with the PI3K-AKT signaling pathway following OZD-1 intervention. Western blot experiments confirmed that OZD-1 significantly upregulates the phosphorylation levels of proteins related to the PI3K-AKT-CREB signaling pathway in OGD/R-injured cells and brain tissue from tMCAO rats, thereby promoting VEGFA-mediated angiogenesis and inhibiting apoptosis. To further verify pathway involvement, in vitro inhibition experiments were performed in RBMVECs using the PI3K inhibitor LY294002 and CREB inhibitor 666-15. These inhibitors abolished the OZD-1-induced upregulation of p-PI3K, p-AKT, and p-CREB, and reversed its protective effects on cell viability, migration, and tube formation. These results confirm that OZD-1 protects vascular endothelial cells directly via activating the PI3K-AKT-CREB pathway. CONCLUSION:OZD-1 exhibits significant neuroprotective effects against CIRI in rats, improving cognitive function, promoting vascular regeneration in ischemic areas, repairing damaged RBMVECs, and reducing apoptosis. Its mechanism of action is associated with the activation of the PI3K-AKT-CREB-VEGFA signaling pathway.

Inflammatory dysregulation is intimately associated with the occurrence and progression of many life-threatening diseases. Accurate detection and timely therapeutic intervention on inflammatory dysregulation are crucial for the effective therapy of inflammation-associated diseases. However, the clinical outcomes of inflammation-involved disorders are still unsatisfactory. Therefore, there is an urgent need to develop innovative anti-inflammatory strategies by integrating emerging technological innovations with traditional therapeutics. Biomedical nanotechnology is one of the promising fields that can potentially transform the diagnosis and treatment of inflammation. In this review, we outline recent advances in biomedical nanotechnology for the diagnosis and treatment of inflammation, with special attention paid to nanosensors and nanoprobes for precise diagnosis of inflammation-related diseases, emerging anti-inflammatory nanotherapeutics, as well as nanotheranostics and combined anti-inflammatory applications. Moreover, the prospects and challenges for clinical translation of nanoprobes and anti-inflammatory nanomedicines are highlighted.

Most BTB-containing E3 ligases homodimerize to recognize a single substrate by engaging multiple degrons, represented by E3 ligase KEAP1 dimer and its substrate NRF2. Inactivating KEAP1 to hinder ubiquitination-dependent NRF2 degradation activates NRF2. While various KEAP1 inhibitors have been reported, all reported inhibitors bind to KEAP1 in a monovalent fashion and activate NRF2 in a lagging manner. Herein, we report a unique bivalent KEAP1 inhibitor, biKEAP1 (3), that engages cellular KEAP1 dimer to directly release sequestered NRF2 protein, leading to an instant NRF2 activation. 3 promotes the nuclear translocation of NRF2, directly suppressing proinflammatory cytokine transcription. Data from in vivo experiments showed that 3, with unprecedented potency, reduced acute inflammatory burden in several acute inflammation models in a timely manner. Our findings demonstrate that the bivalent KEAP1 inhibitor can directly enable sequestered substrate NRF2 to suppress inflammatory transcription response and dampen various acute inflammation injuries.

Background: Cerebral ischemia-reperfusion injury (CIRI) is a phenomenon that pathological injury of ischemic brain tissue is further aggravated after the restoration of blood supply. The complex pathological mechanism of CIRI has led to the failure of multiple neuroprotective agents in clinical studies. Salvianolic acid A (SAA) is a neuroprotective extract from Salvia miltiorrhiza Bge., with significant pharmacological activities in the treatment of brain injury. However, the neuroprotective mechanisms of SAA remain unclear. Purpose: To explore the potential protective effect of SAA on CIRI and its mechanism, and to provide experimental basis for the research of new drugs for CIRI. Study design: A model of transient middle cerebral artery occlusion (tMCAO) in rats was used to simulate clinical CIRI, and the neuroprotective effect of SAA on tMCAO rats was investigated within 14 days after reperfusion. The improvement effects of SAA on cognitive impairment of tMCAO rats were investigated by behavioral tests from days 7-14. Finally, the neuroprotective mechanism of SAA was investigated on day 14. Methods: The neuroprotective effects and mechanism of SAA were investigated by behavioral tests, HE and TUNEL staining, RNA sequence (RNA-seq) analysis and Western blot in tMCAO rats. Results: The brain protective effects of SAA were achieved by alleviating cerebral infarction, cerebral edema, cerebral atrophy and nerve injury in tMCAO rats. Meanwhile, SAA could effectively improve the cognitive impairment and pathological damage of hippocampal tissue, and inhibit cell apoptosis in tMCAO rats. Besides, SAA could provide neuroprotective effects by up-regulating the expression of Bcl-2, inhibiting the activation of Caspase 3, and regulating PKA/CREB/c-Fos signaling pathway. Conclusion: SAA can significantly improve brain injury and cognitive impairment in CIRI rats, and this neuroprotective effect may be achieved through the anti-apoptotic effect and the regulation of PKA/CREB/c-Fos signaling pathway.

Ethnopharmacological relevance: Tiepishihu Xiyangshen granules (TXG) is a traditional Chinese medicine formula composed of Panax quinquefolius L, Dendrobium officinale Kimura & Migo and Ganoderma lucidum (Curtis) P. Karst. It has long been used as a nutritional supplement and an immune enhancer in China. However, the immunomodulatory effects and the underlying mechanisms of TXG have not been clarified.Aim of the study: This study aims to investigate the immunomodulatory effects of TXG and clarify the underlying mechanism.Materials and method: TXG was administered by gavage for 18 days. From the 15th day, the immunosuppression model was induced by intraperitoneally injecting 80 mg/kg CTX for 3 days. The immune regulatory effects of TXG on immune organs were verified by calculating the organ index and observing the spleen tissue structure through HE staining. The effects of TXG on immune cells were examined by recording the PBWC, the prolifer-ation rate of lymphocyte and the T lymphocyte phenotype. The effects of TXG on immune molecules were measured by detecting serum hemolysin and the content of cytokines. In parallel, kit was utilized to detect its antioxidant capacity. RNA seq and Western blot were used to analyze the possible immune regulation mechanism of TXG. HPLC and UPLC-Q-TOF-MS were used to identify the chemical components in TXG. Results: At the level of immune organs, TXG effectively reduced the adverse reaction to the body and the sub-stantial damage to the spleen after chemotherapy by improving the spleen damage. At the level of immune molecules, TXG upregulated the expression of cytokines and antibodies. At the level of immune cells, TXG antagonized bone marrow suppression by increasing the PBWC of immunosuppressed mice. Meanwhile, TXG upregulated the ratio of CD4+/CD8+ lymphocytes and ameliorated the proliferation of T and B lymphocytes. And the mechanism of TXG to improve immunity might be through TLR4/MAPKs and PI3K/AKT/FOXO3a signaling pathways. Conclusion: The results of this study confirmed that TXG has prominent immunomodulatory activities, and the immunity regulations of TXG may be achieved by regulating TLR4/MAPKs and PI3K/AKT/FOXO3a signal pathways.

Cabazitaxel (CTX) has distinct therapeutic merits for advanced and metastatic cancer. However, the present clinical formulation (Jevtana®) has several defects, especially for undesirable tumor-targeting and serious side effects, greatly limiting the therapeutic efficacy. Small-molecule prodrug-based nanoassemblies integrate the advantages of both prodrug strategy and nanotechnology, emerging as a promising treatment modality. Herein, disulfide bonds with different lengths were employed as linkages to elaborately synthesize three redox-sensitive stearyl alcohol (SAT)-CTX prodrug-based nanoassemblies (SAC NPs, SBC NPs and SGC NPs) for seeking optimal chemotherapeutical treatment. All the prodrug-based nanoassemblies exhibited impressive drug-loading efficiency, superior self-assembly capability and excellent colloidal stability. Interestingly, the drug release behaviors of three prodrug-nanoassemblies in the same reductive environment were different owing to tiny changes in the carbon chain length of disulfide bonds, resulting in disparate cytotoxicity effects, pharmacokinetic outcomes and in vivo antitumor efficacies. Among them, SAC NPs displayed rapid drug release, excellent cytotoxicity, long blood circulation and enhanced tumor accumulation, thus showing strong tumor inhibition in the 4T1-bearing mouse model. Our study shed light on the vital role of connecting bonds in designing high-efficiency, low-toxicity prodrug nanoassemblies.

Photodynamic therapy (PDT), extensively explored as a non-invasive and spatio-temporal therapeutic modality for cancer treatment, encounters challenges related to the brief half-life and limited diffusion range of singlet oxygen. Lipid peroxides, formed through the oxidation of polyunsaturated fatty acids by singlet oxygen, exhibit prolonged half-life and potent cytotoxicity. Herein, we employed small molecule co-assembly technology to create nanoassemblies of pyropheophorbide a (PPa) and docosahexaenoic acid (DHA) to bolster PDT. DHA, an essential polyunsaturated fatty acid, co-assembled with PPa to generate nanoparticles (PPa@DHA NPs) without the need for additional excipients. To enhance the stability of these nanoassemblies, we introduced 20% DSPE-PEG2k as a stabilizing agent, leading to the formation of PPa@DHA PEG2k NPs. Upon laser irradiation, PPa-produced singlet oxygen swiftly oxidized DHA, resulting in the generation of cytotoxic lipid peroxides. This process significantly augmented the therapeutic efficiency of PDT. Consequently, tumor growth was markedly suppressed, attributed to the sensitizing and amplifying impact of DHA on PDT in a 4T1 tumor-bearing mouse model. In summary, this molecule-engineered nanoassembly introduces an innovative co-delivery approach to enhance PDT with polyunsaturated fatty acids.

随着生物纳米技术在药物递送领域的深入研究和广泛应用,研究人员设计和构建多种功能各异的纳米药物递送系统用于抗肿瘤药物的高效递送.其中,将前药策略与纳米递药技术进行有机整合的前药纳米组装体已经逐渐成为纳米药物递送系统中一个非常重要的领域.前药设计的关键在于药物分子结构的合理修饰和前药在靶部位的选择性高效激活.近年来,研究发现肿瘤微环境智能响应型前药纳米组装体能够在靶部位选择性快速释药,其已成为癌症诊疗相关研究的重要平台.首先,对肿瘤氧化还原微环境和常用的氧化还原敏感化学桥连进行介绍;其次,分别介绍肿瘤氧化还原微环境智能响应型聚合物大分子前药纳米组装体和小分子前药纳米组装体;最后,对前药纳米组装体优缺点和整个纳米药物递送系统的临床转化前景进行总结和分析.通过对以上内容进行综述,以期为肿瘤微环境智能响应型前药纳米系统的设计与构建提供参考.

The Keap1-Nrf2-ARE pathway regulates the constitutive and inducible transcription of various genes that encode detoxification enzymes, antioxidant proteins and anti-inflammatory proteins and has pivotal roles in the defence against cellular oxidative stress. In this study, we investigated the therapeutic potential of CPUY192018, a potent small-molecule inhibitor of the Keap1-Nrf2 protein-protein interaction (PPI), in renal inflammation. In human proximal tubular epithelial HK-2 cells, CPUY192018 treatment significantly increased Nrf2 protein level and Nrf2 nuclear translocation, which enhanced Nrf2-ARE transcription capacity and the downstream protein content in a Nrf2 dependent manner. In lipopolysaccharide (LPS)-challenged human HK-2 cells, CPUY192018 exhibited cytoprotective effects by enhancing the Nrf2-ARE regulated antioxidant system and diminished the LPS-induced inflammatory response by hindering the ROS-mediated activation of the NF-κB pathway. In the LPS-induced mouse model of chronic renal inflammation, by activating Nrf2, CPUY192018 treatment balanced renal oxidative stress and suppressed inflammatory responses. Hence, administration of CPUY192018 reduced kidney damage and ameliorated pathological alterations of the glomerulus. Taken together, our study suggested that small-molecule Keap1-Nrf2 PPI inhibitors can activate the Nrf2-based cytoprotective system and protect the kidney from inflammatory injury, raising a potential application of Keap1-Nrf2 PPI inhibitors in the treatment of inflammatory kidney disorders.

目的 制备法莫替丁生物黏附型中空微球,并对其进行初步评价.方法 采用溶剂-扩散挥发法制备法莫替丁生物黏附型中空微球,以微球包封率、收率及漂浮率为指标通过正交试验优化处方工艺,以优化处方制备的微球进行离体黏附性试验、体外溶出实验,并进行释药机理的探讨.结果 法莫替丁生物黏附型中空微球的最优处方及工艺为:乙基纤维素质量浓度为27.0 g.L-1,法莫替丁质量浓度为4.8g·L-1,无水乙醇与无水乙醚体积比为17∶3.最优处方下制得微球外观圆整,粒径分布较均匀,在扫描电镜下横切面可见明显中空结构,平均包封率和离体黏附率分别为82.0％和98.0％,在人工胃液中12h漂浮率可达87.0％.12h体外释放90％以上.结论 该法所制中空微球在人工胃液中具有良好的漂浮、黏附及缓释特性,体外释药符合Higuchi模型.