Combinatorial Chemistry & High Throughput Screening - Volume 28, Issue 7, 2025

Quality control is a system of validated procedures in which many samples, including active pharmaceutical ingredients and final products, are analyzed using standard or validated analytical methods.

Analytical methods used in analyzing active pharmaceutical ingredients or final products in the pharmaceutical industry can be methods registered in pharmacopeias and developed by the company itself. For this reason, published papers related to pharmaceutical analysis attract analysts and researchers' attention. In this study, pharmaceutical analysis and bioanalysis studies carried out between 2015 and 2023 were examined using Google Scholar, and the recent trends were determined for pharmaceutical analysis. Among the published papers performing conventional analytical techniques for pharmaceutical analysis, those applying UV-VIS spectrophotometry method were selected to predict a future perspective in this study. In addition to the data obtained, the current situation of the pharmaceutical industry was considered to correlate with the obtained data for pharmaceutical analysis.

The results were presented with comparative tables and summarizing graphs. Interpreting the results allowed us to determine the trends that pharmaceutical analysis studies will lead in the future. This study can be helpful for researchers working on pharmaceutical analysis in both the industry and academia to predict future trends in pharmaceutical analysis. As a result of the literature research covering the dates 2015-2023, 56% of UV-VIS Spectrophotometric methods are used on pharmaceutical dosage forms, 27% are bulk, 16% are pure, 2% are biological materials, and 0.4% are herbal made from materials. Of these studies, 28% were conducted in the 200-240 nm range, 27% were conducted in the 240-300 nm range, and only 44% were conducted at >300 nm. Interpreting the results allowed us to determine the trends that pharmaceutical analysis studies will lead in the future.

This study can be helpful for researchers working on pharmaceutical analysis in both the industry and academy side to predict future trends for pharmaceutical analysis.

Ballota acetabulosa native to the Mediterranean region, belonging to the Lamiaceae family, holds significance in folk medicine. Externally, it is applied for treating cuts and burns, while internally, it is utilized to alleviate inflammation, suppress cough, and address gastrointestinal issues.

This study aimed to investigate the chemical composition of the essential oil of Ballota acetabulosa and to evaluate the antioxidant capacity of the essential oil, as well as the aqueous and ethanolic extracts of the plant. Essential oil analysis was performed using Gas Chromatography-Mass Spectrometry (GC–MS), while 1,1-Diphenyl-2-Picrylhydrazyl (DPPH) radical scavenging activity and inhibition of lipid peroxidation assays were applied to the essential oil, water, and ethanol extracts of the plant.

Spathulenol was found to be the predominant constituent of the essential oil, comprising 25.03% of the oil. Compared to the control group (Propyl gallate for DPPH, IC50 0.109; BHT for inhibition of lipid peroxidation, IC50 0.133), the essential oil was found to have insignificant antioxidant activity (IC50 value 10.395 mg/mL for DPPH, 1.051 mg/mL for inhibition of lipid peroxidation). Moreover, ethanolic extract (IC50 value 1.583 mg/mL for DPPH, 0.029 mg/mL for inhibition of lipid peroxidation) exerted more antioxidant activity than aqueous extract (IC50 value 1.9017 mg/mL for DPPH, 0.161 mg/mL for inhibition of lipid peroxidation).

Hitherto, this is the earliest report on the composition and activity of the essential oil Ballota acetabulosa. However, further investigation of different antioxidant capacity assays is suggested to highlight potential variations in mechanisms of action and subsequent results. Everything considered, this study advances the comprehension of the chemical composition and possible therapeutic uses of Ballota acetabulosa, highlighting the need for more research into its uses.

ALD is a chronic liver disease caused by chronic excessive alcohol consumption, for which there are no drugs with better efficacy. Ancient literature and modern studies have shown that Massa Medicata Fermentata (MMF) has a hangover effect and ameliorates hepatic inflammation, so we believe that MMF has a potential role in the treatment of alcoholic liver disease.

UPLC-Q-Orbitrap HRMS was used to characterize the chemical constituents in MMF. The database was utilized to collect targets for the components and diseases, and cross-targeting analysis of the targets was performed. PPI, KEGG, GO enrichment analysis and molecular docking were performed using the core cross-targeting information to preliminarily validate the mechanism of action of MMF on disease. Finally, animal validation was carried out using male KM mice of the alcoholic liver injury model.

MMF could play a role in the therapeutic prevention of alcoholic liver disease through the core targets AKT1, TNF, TP53, IL6 and CASP3 to regulate cancer pathways, lipid, and atherosclerosis, targeting IL-17 signaling, TNF signaling pathway, and hepatitis C, which was confirmed by animal pharmacodynamic experiments.

This study serves as a rationale to support MMF in the treatment of ALD and meets the urgent need for clinical treatment of ALD. At the same time, it broadens the scope of clinical application of MMF.

The development of MI following ischemia damage is influenced by oxidative stress. Myocardial Infarction (MI) generates myocardial ischemia injury, which damages the cardiomyocytes. Ischemia builds up to a critical level over time in MI, causing permanent myocardial cell damage or death.

The current study sought to determine whether Prunetin (PRU) could protect against Isoproterenol (ISO)-induced cardiac heart failure in rats by examining cardiac diagnostic markers, lipid peroxidation products, enzymatic and non-enzymatic antioxidant levels, and histological changes.

PRU (20 mg/kg bwt) was orally administered for 19 days to rats, and after the treatment, ISO (85 mg/kg bwt) was subcutaneously administered with an intermission of 24 h for a couple of days to induce myocardial infarction on 20^th and 21^st days. ISO-treated rats exhibited considerable alterations in cardiac-sensitive markers in the serum. The levels of lipid peroxidation markers augmented drastically in the plasma and myocardium. Enzymatic antioxidant levels in erythrocytes and myocardium and the states of non-enzymatic antioxidants were diminished in the plasma and heart tissue of ISO-treated rats. The histopathological examination of heart tissue exhibited cardiac damage in ISO-induced rats.

The oral administration of PRU significantly lowered the levels of lipid peroxidation and biochemical indicators, while significantly improving the antioxidant system function of ISO-interposed rats. In PRU-treated ISO-injected rats, histological examinations revealed suppressed myocardial destruction.

Our research shows that oral pretreatment of PRU prevented ISO-induced oxidative stress in MI.

A highly efficient superior catalyst of Ni (II) and VO (IV) metal complexes supported on MCM-41 has been synthesized and developed for chemoselective oxidation of sulfides to sulfoxides and oxidative coupling of thiols to their corresponding disulfides using H2O2 as a green and efficient procedure. All sulfoxides and disulfides were obtained in short reaction times with excellent yields. The over-oxidation of sulfides or thiols was not observed and all products were synthesized in high purity. These catalysts could be recovered and reused several times without any significant loss in their catalytic activity. Compared to the old catalysts in the literature, these catalysts showed better activity and selectivity for the synthesis of sulfoxide and disulfide derivatives, which shows the novelty of this work.

At first, the mesoporous MCM-41 was synthesized, and further, its surface was modified by (3-chloropropyl)-triethoxysilane (CPTES). Then, the modified MCM-41 (nPrCl-MCM-41) was functionalized by adenine. In the next step, the functionalized MCM-41 (6AP-MCM-41) was used as support for the immobilization of nickel and oxo-vanadium as final catalysts (Ni-6AP-MCM-41 or VO-6AP-MCM-41). The structure and properties of these catalysts have been identified by XRD, SEM, TGA, FT-IR, and AAS spectral analyses. These catalysts were used in the chemoselective oxidation of sulfides and oxidative coupling of thiols.

These complexes catalyzed all reactions well at room temperature. According to the results obtained, the hydroxyl groups of some derivatives, including 2-(methylthio) ethanol or 2,2-(phenylthio) ethanol, remained unchanged during the reaction.

The method has been found to possess the advantages of low cost, high efficiency, high yields, recovery, and reusability for several runs without significant loss in the catalytic activity.

Synovitis, acne, pustulosis, hyperostosis, and osteitis (SAPHO) syndrome is a rare disease that is characterized by autoinflammatory lesions on both bones and skin. The diverse manifestations and limited understanding of its etiology have hindered the diagnosis and treatment of this condition. SAPHO syndrome is also classified as a primary inflammatory osteitis. The onset of osteoarticular involvement in this disease is typically gradual, and the identification of associated biomarkers may be crucial for accurate diagnosis, effective treatment, and a better understanding of its underlying mechanisms.

We enrolled a total of 6 SAPHO patients and 3 healthy volunteers for this study. The miRNA expression profile in circulating exosomes was analyzed using next-generation sequencing. A total of 45 miRNAs were found to be differentially expressed in SAPHO patients. Linear discriminant analysis effect size analysis and Wilcoxon rank-sum test were employed to identify biomarkers based on these differentially expressed miRNAs. Among them, we selected 4 miRNAs as biomarkers for SAPHO syndrome, resulting in an area under the receiver operating characteristic curve of 1.

The differentially expressed miRNAs indicated enrichment in immune system and endocrine system-related KEGG pathways, as well as infectious diseases and cancers. Furthermore, the most significantly enriched molecular functions in GO analysis were protein binding and catalytic activity.

The exosomal miRNA profile in SAPHO syndrome exhibited significant changes, suggesting its potential as a candidate biomarker for diagnostic assistance, although further investigation is warranted to elucidate their role in the pathology.

Sepsis is a frequent cause of Acute Lung Injury (ALI), characterized by immune dysregulation and a high mortality rate. The role of cuproptosis, a recently discovered cell death mechanism, in sepsis-associated ALI is still unclear. The study aimed to investigate the regulatory mechanisms and immune characteristics associated with cuproptosis in sepsis-associated ALI, with implications for novel diagnostic and therapeutic approaches.

Data from the GEO database was utilized to conduct a comprehensive analysis of the cuproptosis-related genes (CRGs) in sepsis-associated ALI. Gene enrichment analysis, WGCNA, CIBERSORT algorithm, and consensus clustering were employed to investigate the associations between CRGs and immune cells. A predictive model for sepsis-associated ALI was developed based on key CRGs, and its diagnostic accuracy was assessed. Finally, qPCR was employed to validate alterations in the expression of CRGs in the sepsis-associated ALI cellular model.

A total of 14 CRGs were identified in sepsis-associated ALI. Strong correlations between the CRGs and immune cells were observed, and two different CRG subtypes were identified. The expression of immune-related factors in both the CRG and gene clusters exhibited similarities, suggesting a connection between the subgroups and immune cells. The prediction model effectively forecasted the incidence of sepsis-associated ALI based on the expression of CRGs. Finally, qPCR analysis confirmed that the expressions of CRGs in the sepsis-associated ALI cell model closely matched those identified through bioinformatic analyses.

The study comprehensively evaluated the complex relationship between cuproptosis and sepsis-associated ALI. CRGs were found to be significantly associated with the occurrence, immune characteristics, and biological processes of sepsis-associated ALI. These findings provide valuable new insights into the mechanisms underlying sepsis-associated ALI.

Alzheimer’s Disease (AD) is a highly prevalent form of age-related dementia. However, the underlying mechanisms of AD are largely unexplored.

In this study, bioinformatics analysis was performed to identify the possible therapeutic targets for AD. The GEO database was used to screen the Differentially Expressed Genes (DEGs). Enrichment analysis, protein-protein interaction network, and LASSO model analyses were successfully performed. Furthermore, an ELISA assay was also conducted to determine the expression of principal genes within the AD and control samples.

A total of 416 differentially expressed genes (DEGs) were recognized based on the GSE48350 and GSE28146 datasets. The IL-1β and CXCR4 levels were markedly elevated in the AD samples relative to the control.

The IL-1β and CXCR4 genes were identified as principal AD-related genes that can be targeted for anti-AD therapy.

Identifying cancer-specific biomarkers is a crucial step in the disease screening process at a very early stage of tumor development. In recent years, Quantitative proteomic approaches have gained importance in identifying novel candidate markers in cancer. Gastric cancer has always been known as a life-threatening medical condition with high mortality rates.

The objective of our research is to adapt serum samples from Indian gastric cancer patients to identify and understand the differentially regulated proteins in comparison with healthy individuals.

A total of 30 serum isolates from gastric cancer patients and healthy individuals were obtained and subjected to 2-D Gel electrophoresis, and Tandem LC-MS analysis revealed 12 differentially expressed protein spots. The functional properties of identified proteins were further analyzed using PANTHER and STRING databases.

The differentially expressed protein spots were identified as three candidate proteins: Haptoglobin, Prohibitin, and Apolipoprotein. The protein interaction studies reveal that the haptoglobin fragments were upregulated, and the remaining two prohibitin and Apolipoprotein were down-regulated in gastric cancer patients.

All the proteins identified as biomarkers were found to be involved in regulating cell proliferation and stabilization of oxidative metabolism in the liver; therefore, differential regulation plays a crucial role in gastric cancer progression.

In Lung Adenocarcinoma (LUAD), Qi-deficiency and Phlegm-turbid stagnation (QP) are the most prevalent Traditional Chinese Medicine (TCM) syndrome.

Herein, we collected 90 fecal samples (Healthy individual (H): 30; other syndrome (O): 30; QP: 30) and explored the composition and diversity of gut microbiota in LUAD patients with QP syndrome using 16s-rRNA sequencing. Then, we identified biomarkers for QP syndrome in LUAD patients with Linear Discriminant Analysis (LDA) effect size (LEfSe) and applied logistic regression analysis to construct a diagnostic model evaluated with the area under the receiver operating characteristic curve (AUC) and validated with data from metagenomics.

The α diversity and β diversity revealed that the microbiota community structure in LUAD patients with QP syndrome was different from that with healthy individuals and LUAD patients with other syndromes. At the phylum level, the QP group had more abundance of Bacteroidetes and less Proteobacteria than the O group. At the genus level, the abundance of 4 genera (Bacteroides, Parabacteroides, Prevotella, and Flavonifractor) was different between the QP group and O group. Moreover, LEfSe indicated that those 4 genera might be the biomarkers for LUAD patients with QP syndrome. Then, we used those 4 genera to develop a diagnostic model. The AUC based on 16s-rRNA sequencing and metagenomics was 0.989 and 1, respectively.

A diagnostic model was developed, which would be an available tool for the clinical diagnosis of LUAD with QP syndrome.

The aim of this study was to reveal the hepatic cell landscape and function in the progression of NAFLD to NASH.

Non-Alcoholic Steatohepatitis (NASH) is the progressive form and turning point of Non-Alcoholic Fatty Liver Disease (NAFLD), which severely causes irreversible cirrhosis as well as hepatocellular carcinoma. The mechanism underlying the progression of NAFLD to NASH has not been revealed. Unraveling the mechanism of action of NAFLD-NASH is an important goal in improving the survival of patients with liver disease.

The aim of this study is to discover heterogeneous hepatic cells during the progression of NAFLD to NASH.

Single-nucleus RNA-seq (snRNA-seq) data containing NASH in NAFLD samples were obtained from the Gene Expression Omnibus (GEO) database. Cell types in liver tissues from NASH and NAFLD were identified after dimensionality reduction analysis, cluster analysis, and cell annotation. The cell pathways in which differences existed were identified by analyzing metabolic pathways in Hepatic cells. We also identified cell subpopulations in Hepatic cells. The developmental trajectories of Hepatic cells were characterized by pseudotime trajectory analysis. Single-cell regulatory network inference and clustering analysis identified key transcription factors and gene regulatory networks in Hepatic cells. Moreover, cell communication analysis determined the potential interactions between Hepatic cells and immune cells, and heapatic stellate cells.

Seven cell types were identified in NAFLD and NASH. The proportion of Hepatic cells was lower in NASH and showed greater energy metabolism and glucose metabolism activity. Hepatic cells exhibited heterogeneity, showing two cell subpopulations, Hepatic cells 1 and Hepatic cells 2. Dysregulation of lipid metabolism in Hepatic Cell 2 resulted in lipid accumulation in the liver, which might be involved in the progression of NAFLD. Four key transcription factors, BHLHE40, NFEL2L, RUNX1, and INF4A, were primarily found in Hepatic cells 2. The transcription factors within the hepatic cells 2 subpopulation mainly regulated genes related to lipid metabolism, energy metabolism, and inflammatory response. The cell communication analysis showed that hepatocyte interactions with immune cells were associated with inflammatory responses, while interactions with hepatic astrocytes were associated with liver injury and hepatocyte fibrosis.

The hepatic cells 2 might promote the progression of NAFLD to NASH by regulating metabolic activity, which might contribute to liver injury through inflammation.

Omacetaxine, a semisynthetic form of Homoharringtonine (HHT), was approved for the treatment of Chronic Myeloid Leukemia (CML). Previously, we have published the synthesis of this natural alkaloid and three of its derivatives: Deoxyharringtonine (DHT), Deoxyhomoharringtonine (DHHT), and Bis(demethyl)-deoxyharringtonine (BDHT), and reported its refractory activity against the HL-60/RV+ cells over-expressing P-glycoprotein 1 (MDR1).

In this study, we have explored the extent of this resistance by first expanding the panel of established cell lines and using a panel of 21 leukemia patient-derived primary cells.

Herein, we have reported consistent resistance to HTT of K562-derived cells and to mitoxantrone of MES-SA/MX2-derived cells; all of them have been found to overexpress MDR1, while we have found U87MG-ABCG2 and H69AR cells to be very sensitive to HTT. In contrast, DHT, DHHT, and BDHT seemingly overcame this resistance due to the changes made to the acyl chain of HTT, rendering the derivatives less susceptible to efflux. Surprisingly, the leukemia primary cells were very sensitive to HHT and its derivatives with low nanomolar potencies, followed by a new class of CDC7 kinase inhibitors, the anthracycline class of topoisomerase inhibitors, the DNA intercalator actinomycin-D, and the vinca alkaloid class of microtubule inhibitors. The mechanism of cell death induced by HTT and DHHT was found to be mediated via caspase 3 cleavage, leading to apoptosis.

Taken together, our results confirm that HHT is a substrate for MDR1. It opens the door to a new opportunity to clinically evaluate HHT and its derivatives for the treatment of AML and other cancers.