ACS Pharmacology and Translational Science最新文献

The Lands cycle is a fundamental process for the continuous renewal of phospholipids in cell membranes, directly influencing their fluidity and functionality. This cycle is particularly active in tissues such as the nervous and immune systems and is crucial for cellular homeostasis. It is implicated in the development of inflammatory, neurodegenerative, and cancerous diseases. The present review discusses the biochemical regulation of the Lands cycle─focusing on phospholipase A2 (PLA₂) and lysophospholipid acyltransferase (LPCAT)─and its impact on lipid metabolism, cell signaling, and disease. Dysregulation of this cycle has been linked to pathological conditions, including oncogenesis and hepatotoxicity. This suggests that modulation of the cycle may have an effect on inflammatory responses and tumor resistance. Advances in the fields of lipidomics and computational modeling have resulted in a more comprehensive understanding of the Lands cycle, thereby emphasizing its potential as a therapeutic target.

Overactive epidermal growth factor receptor (EGFR) signaling is often involved in driving different types of carcinomas. It is a well-studied target for targeted therapies, with both monoclonal antibodies and kinase inhibitors available for clinical use. Even though these drugs show a clinical benefit, most patients develop resistance over time. The development of new therapeutic modalities is therefore highly motivated. Herein, we describe a new type of drug candidate targeting EGFR, a so-called affibody-based drug conjugate. It consists of an EGFR-targeting affibody molecule, Z_EGFR, expressed as a fusion to an albumin-binding domain for half-life extension, and coupled with the potent cytotoxic drug DM1 via a maleimidocaproyl linker. The resulting drug conjugate Z_EGFR-ABD-mcDM1, showed strong binding to recombinant EGFR and EGFR-expressing cells. It was found to be highly potent in killing EGFR-expressing A431 cells with an IC₅₀ of 3.4 nM. In vivo, it showed moderate uptake in A431-derived xenografts with high EGFR expression. Collectively, the results from this study, demonstrate a potent and EGFR-specific drug candidate that holds promise for further development.

Screening diverse chemical structure compounds is an essential task in modern drug discovery. It provides different opportunities to avoid patent invasion, avoid potential toxicity observed in similar compounds, and execute new potential pharmacological functions. G protein-coupled receptors (GPCRs) span an important family of membrane proteins that play a central role in signal transduction and serve as significant drug targets. A prototypical class A GPCR is the β₂-adrenergic receptor (β₂AR), which is widely targeted by agonists to treat respiratory diseases. Although various β₂AR agonists are currently available on the market, there is still an urgent demand for further optimizing drug safety, efficacy, and selectivity. Here, we combine machine learning (ML) methods with other computational methods to efficiently screen agonists against β₂AR from a large compound library, composited of 19 million molecules. Verified by cellular functional assays, we identified several extremely potent agonists showing EC₅₀ values in the range of 0.2–20 nM with new chemical structures, of which the structure is diverse from previous reported molecules. Our machine learning computational approaches provide new possibilities to design novel drug candidates for GPCR.

Hypertension is a major cardiovascular risk factor that perturbs neurohumoral regulation, yet its integrated effects on circadian sleep–wake organization and neurocardiac coupling remain unclear. We examined whether the severity of hypertension induces internal circadian misalignment and neurocardiac desynchronization. Male Wistar rats were assigned to the control, high-fructose (HF; mild hypertension), and DOCA-salt (severe hypertension) groups. Across 24 h, we evaluated neurohumoral markers (melatonin, norepinephrine, angiotensin II, vasopressin, and corticosterone), calcium, cardiovascular function (blood pressure, ECG, HRV, and echocardiography), sleep–wake behavior (EEG/EMG), and molecular oscillations of Bmal1, Per1, CACNA1C, and ANP in the SCN and heart. Temporal allostatic load and causal network inference were applied to assess the systemic strain. Both hypertensive models established new blood pressure set points, with HF rats stabilizing at mild hypertension levels and DOCA-salt rats stabilizing at severe hypertension levels. Both exhibited increased neurohumoral load, autonomic imbalance, and ECG/HRV alterations, while DOCA-salt rats showed marked melatonin suppression, sustained elevations of norepinephrine, AVP, corticosterone, and calcium and pronounced NREM–REM fragmentation. Cardiac Bmal1 and Per1 were phase-shifted, CACNA1C was upregulated, and ANP was downregulated, while SCN rhythms were preserved, indicating peripheral desynchronization. Allostatic load analysis revealed an early and persistent burden in DOCA-salt rats and delayed but significant increases in HF rats. Causal network modeling demonstrated a progressive loss of melatonin’s upstream regulation, replaced by neurohumoral dominance, indicating potential pathways in the treatment of hypertension-induced sleep–wake disturbances. These findings indicate that the severity of hypertension reorganizes systemic temporal architecture, amplifying circadian misalignment and SCN–heart decoupling, highlighting the need for stage-specific chronotherapeutic strategies.

The primary goal of antiplatelet therapy is to inhibit platelet aggregation without increasing the risk of bleeding. Treatment resistance and recurrence of thrombotic events are common, underscoring the need to identify new molecules with antiplatelet activity. In this research, we synthesized and characterized spiro-hydroquinone derivatives substituted with various aliphatic chain lengths (1–9 carbons) and evaluated the effect of these modifications on platelet activation. The structure–activity relationship study revealed that increasing the aliphatic chain length did not enhance antiplatelet activity; instead, it increased cytotoxicity and negatively affected solubility. Notably, the shortest molecule, SD3A, inhibits mitochondrial function and acts selectively on collagen-mediated activation, resulting in reduced thrombus formation without affecting coagulation, thereby representing a low risk of bleeding in vitro. These results identify ortho-carbonylhydroquinone spiro derivatives, specifically SD3A, as a promising antiplatelet molecule, demonstrating an optimal combination of low cytotoxicity and pathway-selective activity against collagen.

Nociceptive pain, resulting from tissue injury or inflammation, affects a large portion of the global population. This type of pain is commonly treated by small molecules that are associated with a variety of drawbacks, including addiction and potential liver or kidney damage, highlighting the need for new therapeutic strategies. Here, we report the design, synthesis, and characterization of EG01449 (12h), a quinoline-based neuropilin-1 (NRP1) antagonist with analgesic effects in vascular endothelial growth factor (VEGF)-induced pain models. Neuropilin-1 is a critical coreceptor mediating VEGF signaling. In models of VEGF-induced pain, the VEGFA₁₆₅a isoform increases currents through voltage-gated sodium and calcium channels in dorsal root ganglia sensory neurons. Notably, this effect was mitigated upon the inhibition of NRP1 by 12h, while 12h alone showed no discernible impact on sodium currents. Compound 12h also attenuated sensitivity to mechanical stimuli and cold-induced allodynia. Unlike the previously reported NRP1-targeting compounds that may activate intracellular signaling, 12h did not activate p38 mitogen-activated protein kinase and exhibited a purely inhibitory pharmacological profile. Structural comparison using X-ray crystallography revealed an additional hydrogen bond that contributes to the increased stabilization of the 12h/NRP1 complex. These findings demonstrate that the NRP1 inhibitor 12h elicits an antinociceptive effect and highlight the impact of subtle structural modifications on biological outcomes. NRP1 antagonism thus represents a promising new modality for the treatment of chronic pain conditions.

This study evaluates the potential of a ¹⁷⁷Lu-labeled GRPR-targeting antagonist as a radiotherapeutic agent for tumors expressing the gastrin-releasing peptide receptor (GRPR). The therapeutic effect of the radioligand was investigated both as a monotherapy and in combination with the mTOR inhibitor everolimus. The GRPR antagonist, LF1 (AAZTA⁵-Pip-d-Phe-Gln-Trp-Ala-Val-Gly-His-Sta-Leu-NH₂), was synthesized using the chelator AAZTA⁵ linked via a 4-amino-1-carboxymethylpiperidine (Pip) spacer and radiolabeled with lutetium-177. The preclinical evaluation included assessments of binding kinetics, blood and organ clearance, plasma protein binding, and metabolic stability. SPECT/CT imaging and biodistribution studies were performed in mice bearing PC3 xenograft tumors. To assess its therapeutic efficacy, PC-3-mice were treated with [¹⁷⁷Lu]Lu-LF1 either alone or following everolimus pretreatment. [¹⁷⁷Lu]Lu-LF1 showed high binding affinity (K_d = 0.12 ± 0.01 nM) and favorable pharmacokinetics, including rapid blood clearance and low plasma protein binding (2–3% at 5 and 15 min p.i.). Although subject to enzymatic degradation, the radioligand demonstrated high, sustained, and specific tumor uptake (42 ± 5.0% IA/g at 1 h and 3.9 ± 1.1% IA/g at 144 h p.i.). Pancreatic uptake cleared quickly, allowing for high-contrast SPECT/CT imaging. Therapeutically, tumors treated with 93 MBq of [¹⁷⁷Lu]Lu-LF1 grew more slowly than those treated with 41 MBq. The combination of everolimus and [¹⁷⁷Lu]Lu-LF1 resulted in significant tumor growth inhibition, compared to the relevant monotherapies with either [¹⁷⁷Lu]Lu-LF1 or everolimus. [¹⁷⁷Lu]Lu-LF1 shows promise as a therapeutic radioligand for GRPR-positive prostate cancer, offering high tumor uptake and rapid clearance from nontarget tissues. Mice bearing PC3 xenograft tumors were well tolerated and demonstrated enhanced therapeutic efficacy when combined with everolimus.

Tumor-associated macrophages (TAMs) critically shape the multiple myeloma (MM) microenvironment, yet the molecular programs linking immune signaling to MM dissemination remain unclear. Here, we identify a TAM-derived IL6-STAT3-PIM2-cMyc-FN1 axis that governs cell adhesion and epithelial-mesenchymal transition (EMT) in MM. Proviral Integration Site for Moloney murine leukemia virus 2 (PIM2) acts as a central effector by transcriptionally suppressing fibronectin 1 (FN1) via stabilization of c-Myc, thereby reducing MM-stromal adhesion and promoting migratory capacity. IL6-family cytokines secreted by M2-like TAMs activate STAT3 to induce PIM2 expression, forming a feed-forward loop that reinforces the EMT-like phenotype. Functional assays confirm that PIM2 knockdown restores FN1, increases adhesion, and impairs cell migration, while the dual silencing of FN1 reverses these effects. Analysis of patient biopsies and xenograft models revealed a reciprocal pattern of PIM2 and FN1 expression. These findings delineate a TAM-controlled signaling circuit that integrates inflammatory cues with adhesion loss and invasive behavior, highlighting the IL6-STAT3-PIM2-cMyc-FN1 axis as a potential target in MM therapy.

Gangliosides are sialic acid-enriched glycosphingolipids that play a vital role in regulating multiple signaling pathways during cancer progression. The diversity in their cell- and tissue-specific expression and dysregulations in cancer cells contributes to the unique pathophysiology of triple-negative breast cancer (TNBC). In this study, we follow up on our previously established hydrogel-mediated localized delivery of a combination of docetaxel (DTX) and carboplatin (CPT) (DTX-CPT-Gel therapy) that ensured effective tumor regression in multiple murine syngeneic and xenograft tumor models. Here, we demonstrate that DTX-CPT-Gel therapy downregulates GM3/GD3/GM1 gangliosides by targeting different ganglioside metabolic genes at the transcriptional and translational levels. DTX-CPT-Gel therapy-mediated alterations in ganglioside metabolism affect the activity of key growth factor receptor-mediated signaling pathways, including the epidermal growth factor receptor (EGFR) and cMET/hepatic growth factor receptor (HGFR), which positively impact tumor mitigation. Our work on DTX-CPT-Gel therapy, in continuum, highlights the potential of this therapy for TNBC treatment by intercepting multiple lipid-mediated signaling pathways and reinforces GD3 synthase/ST8SIA1 as a promising target for TNBC therapy.

The noncompetitive lysine-specific demethylase 1 (LSD1) inhibitors SP-2509 and SP-2577 are N′-(1-phenylethylidene)benzohydrazides that display potent activity in Ewing sarcoma. They block transcriptional regulation of the causative oncogenic fusion protein EWSR1::FLI1 and cause cell death. However, SP-2509 and SP-2577 are the only LSD1 inhibitors active in Ewing sarcoma; other LSD1 inhibitors have little effect. Studies from our group and others suggest that SP-2509 activity may result from off-target activity affecting the mitochondria. Here, we identified potential off-target mechanisms of N′-(1-phenylethylidene)benzohydrazides using an unbiased approach, cellular thermal shift assay coupled to mass spectrometry. Interestingly, this revealed significant destabilization of the electron transport chain complex III protein ubiquinol-cytochrome c reductase (UQCRFS1). We find that UQCRFS1 destabilization is likely linked to impaired iron–sulfur (Fe–S) cofactor binding and that SP-2509 broadly destabilizes cellular Fe–S proteins. Using both chemical and genetic tools, we show that SP-2509 mediated cell death is LSD1 independent and instead requires a N′-(2-hydroxybenzylidene)benzohydrazide. Our studies suggest that this core moiety alters iron metabolism in the cell. Importantly, we also find that the reversal of EWSR1::FLI1 transcriptional regulation by SP-2509 is independent of LSD1 inhibition. This unique activity is instead associated with the N′-(2-hydroxybenzylidene)benzohydrazide core and destabilization of Fe–S proteins. These findings reveal a novel mechanism of action for this class of compounds and raise additional questions regarding how EWSR1::FLI1 transcriptional regulation is linked to Fe–S biogenesis, the precise mechanisms of cell death, the biological features of susceptible cancer cells, and strategies for clinical translation.