Acta Pharmaceutica Sinica B

AI-powered therapeutic aptamer drug discovery: Targeting the CT-domain of CTGF for duchenne muscular dystrophy

Shanshan Yao, Xin Yang, Meishen Ren, Huarui Zhang, Sifan Yu, Tienan Chen, Shijian Ding, Yufei Pan, Luyao Wang, Yuan Ma, Wei Kang, Xiaoling Peng, Jiming Liu, Aiping Lyu, Zongkang Zhang, Yuanyuan Yu, Yixin He, Ge Zhang, Bao-Ting Zhang

Acta Pharmaceutica Sinica B. 2026, 16(7): 4552-4574.

Duchenne muscular dystrophy (DMD), a fatal X-linked disorder, features progressive muscle fibrosis as a key driver of mortality. While CTGF represents a therapeutic target for DMD, its VWC-domain-targeting antibody (FG-3019) failed in clinical trials. Through experimental validation, we identified CT-domain as a superior target domain, as it contributed more fibrosis activity of CTGF than VWC-domain without elevating compensatory TGF-β1 level. Aptamers are synthetic oligonucleotides identified through SELEX, which can specifically bind to flexible protein domains through their unique 3D conformations. Their small molecular size enables effective tissue penetration while maintaining high target specificity, making them ideal CT-domain inhibitors. Nevertheless, conventional SELEX involves time-consuming and inefficient multiple screening rounds. Here, we employed our generative AI model, AptGEN, to rapidly discover a potent CT-domain specific aptamer within 42 days. This chemically modified aptamer (Apc003OA) distributed and remained in muscle tissues for an extended period, whereas FG-3019 could not. Importantly, it demonstrated better fibrosis inhibitory activity in vitro and in mdx mice when compared to FG-3019. Furthermore, Apc003OA demonstrated a favorable safety profile in mdx mice. Within 10 months, we progressed from target domain discovery, aptamer drug discovery, and then obtained both Orphan Drug Designation and Pediatric Rare Disease Designation by US Food and Drug Administration.

Enhanced tumor immunotherapy by nanovesicles derived from engineered DC-like M1 macrophage

Linxia Jiang, Yi Yan, Hongqian Xie, Zhiyi Xu, Xiaoru Guo, Jinyue Li, Xiaoyu Liu, Di Wu, Yuanjun Zhu, Yujie Shi, Xiaoyan Liu, Jian-Cheng Wang

Acta Pharmaceutica Sinica B. 2026, 16(7): 4654-4675.

To explore a therapeutic approach with dual functions of activating cytotoxic CD8⁺ T cells and remodeling immunosuppressive tumor-associated macrophages (TAMs), the engineered macrophage-derived nanovesicles (MAC-PNV) were developed in this study. The MAC-PNV were derived from activated DC-like M1 macrophages after reprogramming macrophages by transcription factors PIB (PU.1, IRF8, and BATF3) and further stimulated with antigenic peptide, lipopolysaccharide (LPS), and interferon-γ (IFN-γ). In vitro results demonstrated that the enriched antigen-presenting complexes and co-stimulatory molecules were displayed on the surface of pro-inflammatory cargo-contained MAC-PNV, which enabled significant activation of CD8⁺ T cells and repolarization of M2 macrophages towards the M1 phenotype. After peritumoral administration, MAC-PNV alone significantly inhibited tumor growth by promoting the activation and intra-tumoral infiltration of CD8⁺ T cells, and remodeling the immunosuppressive tumor microenvironment (TME) in B16-OVA-bearing mouse models. More importantly, MAC-PNV remarkably enhanced the anti-tumor efficacy of low-dose liposomal doxorubicin (DOX-Lipo, 1 mg/kg), along with reducing its dose-limiting toxicities in B16-F10-bearing mouse models. This study highlights that the MAC-PNV would be a potential and effective immunomodulatory enhancer for providing a promising combination strategy with clinical chemotherapeutics.

Targeting bacterial phosphotransferase system to prevent the dissemination of antibiotic resistance genes

Acta Pharmaceutica Sinica B. 2026, 16(7): 4575-4591.

Antimicrobial resistance (AMR) poses a significant challenge to public health and human security, with plasmid-mediated horizontal gene transfer (HGT) being a primary driver for its dissemination. Here, we identify indole analogs containing electron-withdrawing groups as a new category of HGT inhibitors. Using indole-3-acetic acid (IAA) as a representative scaffold, we demonstrate that IAA targets glycolytic enolase to deplete phosphoenolpyruvate (PEP) in donor bacteria; this suppresses phosphotransferase system (PTS) activity, blocks PtsI phosphorylation, reduces cAMP synthesis and prevents catabolite repressor protein (CRP) activation, ultimately limiting intracellular ATP availability. Concurrently, IAA attenuates reactive oxygen species (ROS) generation by inhibiting FADH₂ oxidation and riboflavin biosynthesis. The concerted reduction in ATP and ROS arrests conjugative plasmid transfer. Overall, our work suggests the potential of indole analogs as a new class of conjugative transfer inhibitors and highlights that bacterial phosphotransferase system represents a promising target to prevent the propagation of AMR.

Ultrasensitive bioorthogonal probes for selective discrimination of trace H₂S₂/H₂Sn in sulfide-competitive contexts

Xidan Tong, Xiaowei Xu, Jiaxuan Chen, Jinkang Feng, Yixing Li, Yangfei Shi, Zhen Li, Weiwei Guo, Yueqin Zheng

Acta Pharmaceutica Sinica B. 2026, 16(7): 4539-4551.

Hydrogen persulfide/polysulfides (H₂S₂/H₂Sn), as an oxidized derivative of hydrogen sulfide (H₂S), is capable of directly inducing the S-persulfidation of cysteine residues, thereby modulating the activity of relevant enzymes. Owing to its unique reactive properties, H₂S₂/H₂Sn is emerging as a central focus in the study of reactive sulfur species. Therefore, the precise detection of H₂S₂/H₂Sn in vivo is critical for elucidating their roles in redox signaling and cellular regulation. However, conventional probes face challenges such as poor sensitivity, cross-reactivity, and instability. Here, we report a bioorthogonal ether linkage fluorescent probe toolkit (Cyne-1-5) with a cyclooctyne warhead, enabling ultra-sensitive (LOD = 3.3 nmol/L), selective, and real-time tracking of H₂S₂/H₂Sn in living systems. These probes feature rapid activation (>1018-fold fluorescence activation in 5 min), broad spectral coverage (blue to NIR), and exceptional enzymatic stability. Using this toolkit, we uncovered the spontaneous oxidation of H₂S to trace H₂S₂/H₂Sn and demonstrated steric hindrance-driven self-disproportionation of persulfides, where less bulky persulfides efficiently yield H₂S₂/H₂Sn. Furthermore, we achieved the cellular-level visualization of protein S-persulfidation dynamics. This work advances persulfide chemical biology and offers transformative tools for probing H₂S₂/H₂Sn in disease mechanisms and therapeutic development.

Attenuating AAV-triggered innate immunity in the adult mouse nervous system via cGAS-STING pathway inhibition

Yaowei Guo, Jiehao Huang, Yuqi Zhang, Yuxiang Huang, Zeru Li, Yu Xie, Yingqi Cao, Kun Qian, Fan Bai, Wen Li, Wenliang Lei, Gong Chen, Zheng Wu

Acta Pharmaceutica Sinica B. 2026, 16(7): 4389-4409.

While adeno-associated virus (AAV)-mediated gene delivery has emerged as a promising therapeutic modality for neurological disorders, dose-dependent immune responses remain a critical barrier to clinical translation. Here we reveal the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway as a key mediator of innate immune activation following intracranial AAV administration. Through comparative analyses in genetic and pharmacological intervention models, we demonstrate that STING signaling mediates key neuroinflammatory sequelae including glia reactivation, cytotoxic T cell infiltration, and neuronal injury. Mechanistically, microglia serve as the predominant sentinels detecting AAV immunogenicity via cGAS-STING activation. Therapeutic inhibition of this pathway by either microglia depletion or antagonism of STING by small molecules significantly mitigates high-dose AAV9-induced neurotoxicity while enhancing transgene delivery efficacy. Our work delineates a unified mechanistic framework linking AAV-triggered DNA sensing to neuroinflammatory pathology, and provides two clinically actionable approaches to decouple therapeutic gene delivery from detrimental immune activation in nervous system targeted gene therapy.

A novel NAMPT activator ameliorates obesity by repressing ACSL1-dependent lipid synthesis

Zhi Jiang, Yi-Xian Li, Dan-Dan Zhao, Yu-Tao Hu, Ling-Zhen Xiao, Yu-Wei Lin, Shu-Min Xu, Jia-Chun Luo, Qingjiang Li, Shi-Liang Huang, Hui-Hao Zhou, Jia-Heng Tan, Shuo-Bin Chen, Zhi-Shu Huang

Acta Pharmaceutica Sinica B. 2026, 16(7): 4478-4490.

Excessive incorporation of long-chain fatty acids (LCFAs) into triglycerides in adipose tissue is a key contributor to obesity and related metabolic disorders, and pharmacologically modulating this process remains challenging. Here, we synthesized a library of β-indoquinazolinone derivatives via palladium-catalyzed oxidative addition complex chemistry and identified compound b2b as a potent and selective anti-obesity candidate. b2b inhibited triglyceride accumulation in adipocytes in vitro and significantly reduced adiposity, body weight, and lipid metabolic disturbances in diet-induced obese mice without observable toxicity. Mechanistic studies revealed that b2b directly activates nicotinamide phosphoribosyltransferase (NAMPT) and elevates intracellular NAD⁺ levels to enhance the NAD⁺-dependent regulatory protein SIRT1 activity. This activation leads to transcriptional repression of acyl-CoA synthetase long-chain family member 1 (ACSL1), thereby inhibiting LCFAs incorporation into triglycerides. These findings demonstrate that pharmacological activation of the NAMPT-NAD⁺-SIRT1 axis by b2b offers a novel strategy for obesity treatment.

Insights into SARS-CoV-2 replication control via targeting the host hijacking function of NSP12

Shirui Fan, Xinyan Long, Wei Zheng, Zhengrui Xiang, Guangjin Liu, Miao Luo, Yongtang Zheng, Ronghua Luo, Zezhou Yu, Yi Luo, Xiaojiang Hao, Duozhi Chen

Acta Pharmaceutica Sinica B. 2026, 16(7): 4410-4425.

Nonstructural protein 12 (NSP12), the RNA-dependent RNA polymerase (RdRp) of SARS-CoV-2, serves as the catalytic core of the viral replication-transcription complex and is pivotal for viral RNA synthesis. Given its essential role, SARS-CoV-2 must precisely regulate intracellular NSP12 levels to support efficient viral replication. However, the mechanisms governing its stability remain poorly understood. Here, we reveal a previously unrecognized viral strategy in which NSP12 hijacks the host chaperone Hsc70 to modulate its own stability. Mechanistically, Hsc70 plays a dual role: mediating NSP12 degradation via chaperone-mediated autophagy (CMA) while also promoting its accumulation. The dynamic balance between these opposing functions determines NSP12 fate. NSP12 can evade CMA-mediated degradation by binding Hsc70 with higher affinity. This disrupts the Hsc70-LAMP2a interaction and shifts Hsc70’s role toward primarily facilitating NSP12 accumulation, thereby enhancing viral replication. We identified Hlyc41 as a potential antiviral agent that disrupts this hijacking mechanism. Hlyc41 competes with NSP12 for binding to the F428 residue of Hsc70, thereby promoting NSP12 degradation and suppressing viral replication. These findings reveal a novel host hijacking mechanism that regulates NSP12 levels and support a promising therapeutic strategy targeting this process.

Discovery of novel tumor-targeting peptide-oncolytic peptide based conjugates (PPCs): A new paradigm for targeted oncolytic-immunotherapy

Acta Pharmaceutica Sinica B. 2026, 16(7): 4491-4523.

Oncolytic peptides, which possess synergistic oncolytic-immunotherapy effects, have exhibited advantages including unique anticancer mechanism, overcoming drug resistance and broad anticancer spectrum in clinic. Nevertheless, conventional oncolytic peptides often suffer from poor stability, limited efficacy, and moderate anticancer selectivity. In this study, the stability-, potency-, and selectivity-guided optimizations were conducted on the first-in-class oncolytic peptide Clip-71. The robust synthetic strategy, in vitro and in vivo anticancer activity, and anticancer mechanism especially immune activation ability were systematically investigated for obtained peptides. The first round of stability-guided optimization identified the mirror-image peptides represented by QY-8, which possessed strikingly high stability, as well as outstanding in vitro and in vivo anticancer activities. Subsequently, to address the limited selectivity and nonspecific distribution of oncolytic peptides, the second round of selectivity-guided and PPCs-strategy based optimization was conducted on QY-8. Strikingly, the novel PPC QY-13, which was obtained through conjugation of the SSTR2-targeting peptide Tyr³-octreotate to QY-8, exhibited most potent anticancer activities both in vitro and in vivo, enhanced immunostimulatory activity, and superior biosafety as well as tumor targeting potency. Collectively, this study not only established promising strategies to significantly improve the anticancer potential of cytotoxic peptides, but also provided the new paradigm for targeted oncolytic-immunotherapy.

The gut-liver axis in drug-induced liver injury (DILI): Interplay between gut microbiota, intestinal barrier, and immune system

Irene Díaz-Alberola, Daniel E. Di Zeo-Sánchez, Vanesa Garrido-Rodríguez, Carlos López-Gómez, Cristina Rodríguez-Díaz, Raul J. Andrade, M. Isabel Lucena, Marina Villanueva-Paz, Eduardo García-Fuentes

Acta Pharmaceutica Sinica B. 2026, 16(7): 4233-4260.

Drug-induced liver injury (DILI) is an adverse hepatic reaction with a complex etiopathogenesis, involving direct cellular damage by the drug, immune-mediated mechanisms, and host and environmental factors. The gut microbiota has recently been recognized as a key player in human physiopathology. In DILI, animal models and patients have shown significant alterations in gut microbiota composition. Evidence indicates that patients with DILI often present intestinal barrier dysfunction, characterized by increased permeability and the translocation of pathogen-associated molecular patterns (PAMPs) to the liver. This process may contribute to the onset or aggravation of liver injury by triggering harmful immune responses. Furthermore, dysbiosis can alter bacterial metabolite production, thereby affecting intestinal and hepatic homeostasis. The gut microbiota can also modify the efficacy and toxicity of drugs on an individual level, thereby increasing the risk of DILI. This review provides an overview of the current evidence on the mechanisms by which the gut microbiota contributes to inflammation, immune recruitment, and exacerbation of liver damage in DILI. A more in-depth understanding of how the gut microbiota alters intestinal and hepatic homeostasis is essential for advancing our knowledge of DILI pathogenesis, integrating novel biomarkers into clinical practice, and developing microbiota-based interventions.

Acquired pharmacoresistance in temporal lobe epilepsy is driven by Nav1.6-mediated subicular hyperexcitability

Acta Pharmaceutica Sinica B. 2026, 16(7): 4442-4458.

Pharmacoresistance to anti-seizure medications (ASMs) remains a major unmet challenge in temporal lobe epilepsy (TLE), and occurs diversely, as classified to primary or acquired manner. The pathophysiological underpinnings of acquired pharmacoresistance remain elusive. Here, using a hippocampal kindling mouse model, we established that prolonged lamotrigine (LTG) treatment—either during or after kindling—induces broad-spectrum resistance to multiple ASMs, effectively recapitulating clinical patterns of acquired pharmacoresistance. Multimodal interrogation revealed hyperexcitability of subicular pyramidal neurons as a critical factor in pharmacoresistance, characterized by elevated c-Fos expression specifically within the subiculum, as well as hyperexcitability of subicular glutamatergic pyramidal neurons. This hyperexcitability phenotype stemmed from Nav1.6 upregulation, driving both enhanced persistent sodium current (INaP) and a pro-excitatory shift in voltage-dependent activation kinetics of voltage-gated sodium channel (VGSC). Crucially, pharmacological activation of subicular Nav1.6 sufficed to induce acquired pharmacoresistance in pharmaco-responsive mice. Conversely, subiculum-specific Nav1.6 knockdown in pyramidal neurons (but not GABAergic neurons) prevented or reversed pharmacoresistance, while analogous genetic manipulation in the CA1 had no such impact. Chemogenetic inhibition of subicular pyramidal neurons (mimicking ASM effects) restored drug responsiveness, directly implicating compensatory increases in Nav1.6 offsetting ASM’s inhibitory function on subicular excitability in acquired pharmacoresistance. These findings collectively identify Nav1.6 upregulation in subicular pyramidal neurons as a critical driver of acquired pharmacoresistance in TLE, highlighting a novel therapeutic target for refractory epilepsy.