ACS Pharmacology and Translational Science最新文献

Antisense oligonucleotides (ASOs) represent a unique category of therapeutics targeting disease-related RNAs. Since this new therapeutic category emerged, the immediate need to analyze ASOs in clinically relevant biological matrices has led to several methodologies, such as ligand binding assays and imaging techniques. To overcome issues in specificity and provide exact quantitative data for ASOs, a new LC-MS/MS method was developed to analyze in brain tissue a novel 4-10-4 gapmer ASO with the potential for treating Parkinson's disease with phosphorothioated backbone and 2'-O-(2-methoxyethyl) modifications. The sample pretreatment protocol to extract the ASO from brain tissue employed solid phase extraction (SPE) and protein digestion. The LC-MS/MS method was fully optimized, validated and applied to quantify the target ASO in brain tissue samples following an in vivo brain distribution study. The method has a Lower Limit Of Quantification of 1 ng/mg and was applied to the study's samples, demonstrating satisfactory sensitivity and providing valuable information about the ASO's distribution in different brain regions over 45 days.

The mechanisms underlying the onset and progression of chronic pain in COVID-19 patients have been understudied. Using network meta-analysis, we previously demonstrated that alcohol augments COVID-19 symptoms and pathologies possibly by inducing a severe cytokine storm. We and others have also reported that acute alcohol consumption produces analgesic effects, while chronic alcohol consumption results in hyperalgesia and chronic pain. This study aimed to identify the influence of alcohol consumption and COVID-19 on pain. Using publicly available curated gene expression data sets of differentially expressed genes (DEGs) in the prefrontal cortex (PFC) and amygdala of COVID-19 patients, we employed a bioinformatics application, QIAGEN ingenuity pathway analysis (IPA), to identify the key signaling pathways, upstream regulators, and biological functions in these brain areas known to play a role in pain. Canonical pathway analysis revealed activation of the neuropathic pain pathway and signaling pathways involving the cytokine storm, S100 family, IL-6, and neuroinflammation. IPA’s network builder was employed to construct a network map of shared molecules between alcohol and pain-related constructs (discomfort, neuropathic pain, and inflammatory pain). The simulation of alcohol consumption inhibited pain in this network map. To study the influence of COVID-19, we overlaid the DEGs from the PFC and amygdala onto these networks, mimicking alcohol consumption during SARS-CoV-2 infection. Upregulation of molecules in the amygdala and PFC predicted an increase in neuropathic pain, as well as an increase in inflammatory pain in the PFC. Our results suggest that while alcohol consumption directly inhibits pain, the presence of COVID-19 exaggerates impaired cytokine signaling, neuroinflammation, and neuropathic pain signaling in the CNS providing novel insights into the signaling pathways associated with chronic pain of the COVID-19 patients.

To develop novel ^99mTc-labeled ubiquicidin 29-41 derivatives for bacterial infection SPECT imaging aiming at achieving a high target-to-nontarget ratio and lower nontarget organ uptake, a novel 6-hydrazinoicotinamide (HYNIC) ubiquicidin 29-41 derivative (HYNIC-UBI 29-41) was designed and synthesized. It was then radiolabeled with ternary ligands, including TPPTS, PDA, 2,6-PDA, NIC, ISONIC, PSA, 4-PSA, and PES, to obtain eight ^99mTc-labeled HYNIC-UBI 29-41 complexes. All the complexes demonstrated hydrophilicity, exhibited good in vitro stability, and specifically bound Staphylococcus aureus in vitro. Biodistribution studies in mice with bacterial infection demonstrated that [^99mTc]Tc-tricine/TPPTS-HYNIC-UBI 29-41 resulted in increased abscess-to-muscle and abscess-to-blood ratios as well as decreased nontarget organ uptake. Furthermore, it was able to distinguish between bacterial infection and sterile inflammation. Single-photon emission computed tomography (SPECT) imaging studies in mice with bacterial infection revealed visible accumulation at the site of infection, indicating that [^99mTc]Tc-tricine/TPPTS-HYNIC-UBI 29-41 is a potential radiotracer for imaging bacterial infection.

Antisense oligonucleotides (ASOs) represent a unique category of therapeutics targeting disease-related RNAs. Since this new therapeutic category emerged, the immediate need to analyze ASOs in clinically relevant biological matrices has led to several methodologies, such as ligand binding assays and imaging techniques. To overcome issues in specificity and provide exact quantitative data for ASOs, a new LC-MS/MS method was developed to analyze in brain tissue a novel 4–10–4 gapmer ASO with the potential for treating Parkinson’s disease with phosphorothioated backbone and 2′-O-(2-methoxyethyl) modifications. The sample pretreatment protocol to extract the ASO from brain tissue employed solid phase extraction (SPE) and protein digestion. The LC-MS/MS method was fully optimized, validated and applied to quantify the target ASO in brain tissue samples following an in vivo brain distribution study. The method has a Lower Limit Of Quantification of 1 ng/mg and was applied to the study’s samples, demonstrating satisfactory sensitivity and providing valuable information about the ASO’s distribution in different brain regions over 45 days.

Low-density lipoprotein (LDL) is the primary natural carrier of lipids in the bloodstream and plays a central role in the development of atherosclerosis. By leveraging LDL’s natural tendency to accumulate at sites of plaque formation, LDL can be employed as a carrier to selectively deliver the imaging probes to efficiently detect atherosclerotic plaques. In our previous studies, we reported several LDL-based magnetic resonance imaging contrast agents (MRI-CAs) formed by modifying natural LDL (nLDL) or developing LDL-mimetic (synthetic LDL, sLDL) from lipid nanoparticles (LNPs) utilizing chemical reactions on the nanoparticle surface, including preliminary MRI tests. In this study, we report the in vivo biological functionality of these LDLs (both nLDL and sLDL)-based Gd(III)-based contrast agents (GBCAs) by conducting detailed in vivo studies on two types of atherosclerosis murine models, namely, apoE^–/– and LDLr^–/–. We provide more comprehensive MRI data accompanied by ex vivo results, including microscopic analysis of aorta segments for LDL accumulation and whole-body cryoVIZ analysis for biodistribution of the probe. We also tested in vitro cellular internalization of sLDL on two cell lines (RAW 264.7 and THP-1), which are derived from macrophages and monocytes, respectively, in order to observe sLDL uptake by macrophages, which are often present at the vulnerable types of atherosclerotic plaques. In conclusion, our current study demonstrates that modified LDLs─both nLDL and sLDL─facilitate MRI detection of atheroplaques by efficient uptake by macrophages. Taken together with the high loading capacity of Gd(III)-chelate molecules on LDL, especially sLDL, the LDL-based MRI contrast agents reported here hold significant potential for the early detection of atherosclerosis, including vulnerable ones, and should be useful for preventive diagnosis strategies.

Low-density lipoprotein (LDL) is the primary natural carrier of lipids in the bloodstream and plays a central role in the development of atherosclerosis. By leveraging LDL's natural tendency to accumulate at sites of plaque formation, LDL can be employed as a carrier to selectively deliver the imaging probes to efficiently detect atherosclerotic plaques. In our previous studies, we reported several LDL-based magnetic resonance imaging contrast agents (MRI-CAs) formed by modifying natural LDL (nLDL) or developing LDL-mimetic (synthetic LDL, sLDL) from lipid nanoparticles (LNPs) utilizing chemical reactions on the nanoparticle surface, including preliminary MRI tests. In this study, we report the in vivo biological functionality of these LDLs (both nLDL and sLDL)-based Gd(III)-based contrast agents (GBCAs) by conducting detailed in vivo studies on two types of atherosclerosis murine models, namely, apoE ^-/- and LDLr ^-/- . We provide more comprehensive MRI data accompanied by ex vivo results, including microscopic analysis of aorta segments for LDL accumulation and whole-body cryoVIZ analysis for biodistribution of the probe. We also tested in vitro cellular internalization of sLDL on two cell lines (RAW 264.7 and THP-1), which are derived from macrophages and monocytes, respectively, in order to observe sLDL uptake by macrophages, which are often present at the vulnerable types of atherosclerotic plaques. In conclusion, our current study demonstrates that modified LDLs-both nLDL and sLDL-facilitate MRI detection of atheroplaques by efficient uptake by macrophages. Taken together with the high loading capacity of Gd(III)-chelate molecules on LDL, especially sLDL, the LDL-based MRI contrast agents reported here hold significant potential for the early detection of atherosclerosis, including vulnerable ones, and should be useful for preventive diagnosis strategies.

To develop novel ^99mTc-labeled ubiquicidin 29-41 derivatives for bacterial infection SPECT imaging aiming at achieving a high target-to-nontarget ratio and lower nontarget organ uptake, a novel 6-hydrazinoicotinamide (HYNIC) ubiquicidin 29-41 derivative (HYNIC-UBI 29-41) was designed and synthesized. It was then radiolabeled with ternary ligands, including TPPTS, PDA, 2,6-PDA, NIC, ISONIC, PSA, 4-PSA, and PES, to obtain eight ^99mTc-labeled HYNIC-UBI 29-41 complexes. All the complexes demonstrated hydrophilicity, exhibited good in vitro stability, and specifically bound Staphylococcus aureus in vitro. Biodistribution studies in mice with bacterial infection demonstrated that [^99mTc]Tc-tricine/TPPTS-HYNIC-UBI 29-41 resulted in increased abscess-to-muscle and abscess-to-blood ratios as well as decreased nontarget organ uptake. Furthermore, it was able to distinguish between bacterial infection and sterile inflammation. Single-photon emission computed tomography (SPECT) imaging studies in mice with bacterial infection revealed visible accumulation at the site of infection, indicating that [^99mTc]Tc-tricine/TPPTS-HYNIC-UBI 29-41 is a potential radiotracer for imaging bacterial infection.

The mechanisms underlying the onset and progression of chronic pain in COVID-19 patients have been understudied. Using network meta-analysis, we previously demonstrated that alcohol augments COVID-19 symptoms and pathologies possibly by inducing a severe cytokine storm. We and others have also reported that acute alcohol consumption produces analgesic effects, while chronic alcohol consumption results in hyperalgesia and chronic pain. This study aimed to identify the influence of alcohol consumption and COVID-19 on pain. Using publicly available curated gene expression data sets of differentially expressed genes (DEGs) in the prefrontal cortex (PFC) and amygdala of COVID-19 patients, we employed a bioinformatics application, QIAGEN ingenuity pathway analysis (IPA), to identify the key signaling pathways, upstream regulators, and biological functions in these brain areas known to play a role in pain. Canonical pathway analysis revealed activation of the neuropathic pain pathway and signaling pathways involving the cytokine storm, S100 family, IL-6, and neuroinflammation. IPA's network builder was employed to construct a network map of shared molecules between alcohol and pain-related constructs (discomfort, neuropathic pain, and inflammatory pain). The simulation of alcohol consumption inhibited pain in this network map. To study the influence of COVID-19, we overlaid the DEGs from the PFC and amygdala onto these networks, mimicking alcohol consumption during SARS-CoV-2 infection. Upregulation of molecules in the amygdala and PFC predicted an increase in neuropathic pain, as well as an increase in inflammatory pain in the PFC. Our results suggest that while alcohol consumption directly inhibits pain, the presence of COVID-19 exaggerates impaired cytokine signaling, neuroinflammation, and neuropathic pain signaling in the CNS providing novel insights into the signaling pathways associated with chronic pain of the COVID-19 patients.

Glioblastoma multiforme (GBM) is highly aggressive, necessitating new therapies. Photoactivated chemotherapy (PACT) offers a promising approach by activating prodrugs with visible light at the tumor site. This study evaluated the anticancer activity of ruthenium-based PACT compounds in U-87MG glioblastoma cells and their safety in SH-SY5Y neuron-like cells. The compound [3](PF₆)₂ showed promising light-activated anticancer effects in U-87MG cells, while [1](PF₆)₂ was inactive, and [2](PF₆)₂ was nonactivated. Interestingly, in SH-SY5Y cells, light-activated [3](PF₆)₂ increased cell proliferation, similar to donepezil, without causing cell death. Increased Ca²⁺ uptake was observed, possibly via interaction with the AMPA receptor, as suggested by docking studies. These findings suggest ruthenium-based PACT compounds may serve as potential treatments for GBM, effectively attacking cancer cells while preserving healthy neuronal cells.

Glioblastoma multiforme (GBM) is highly aggressive, necessitating new therapies. Photoactivated chemotherapy (PACT) offers a promising approach by activating prodrugs with visible light at the tumor site. This study evaluated the anticancer activity of ruthenium-based PACT compounds in U-87MG glioblastoma cells and their safety in SH-SY5Y neuron-like cells. The compound [3](PF₆)₂ showed promising light-activated anticancer effects in U-87MG cells, while [1](PF₆)₂ was inactive, and [2](PF₆)₂ was nonactivated. Interestingly, in SH-SY5Y cells, light-activated [3](PF₆)₂ increased cell proliferation, similar to donepezil, without causing cell death. Increased Ca²⁺ uptake was observed, possibly via interaction with the AMPA receptor, as suggested by docking studies. These findings suggest ruthenium-based PACT compounds may serve as potential treatments for GBM, effectively attacking cancer cells while preserving healthy neuronal cells.