Abstract

A simple, accurate, and stability-indicating reverse-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the simultaneous quantification of Amlodipine Besylate and Lisinopril in fixed-dose combination (FDC) formulations. Chromatographic separation was achieved using a Phenomenex C18 column (250 × 4.6 mm, 5 µm) with a mobile phase comprising acetonitrile and phosphate buffer (pH 3.5) in the ratio of 60:40 v/v, at a flow rate of 1.0 mL/min. Detection was carried out at 210 nm, with retention times of 3.2 min for Lisinopril and 5.8 min for Amlodipine. The method was validated in accordance with ICH Q2(R1) guidelines for linearity, precision, accuracy, robustness, specificity, limit of detection (LOD), and limit of quantitation (LOQ). Linearity was observed in the range of 2–12 µg/mL for Amlodipine and 5–30 µg/mL for Lisinopril with correlation coefficients >0.999. Recovery studies demonstrated accuracy between 98–102%, while %RSD values for intra- and inter-day precision remained below 2%. Beyond pharmaceutical quality control, this validated RP-HPLC platform reflects broader innovations in clinical microbiology diagnostics, as robust, stability-indicating assays form the foundation for reliable drug monitoring, therapeutic validation, and quality assurance—critical components in improving patient care and addressing challenges such as antimicrobial resistance.

Abstract

Drug designing and improving the physicochemical properties of existing APIs (active pharmaceutical ingredients) are the two major challenges of the pharmaceutical industry. The physicochemical properties of the API can be improved by obtaining their multicomponent compounds (binary, ternary, etc) using suitable coformers, which can be obtained in various forms (cocrystals, molecular salts, hydrates/solvates, polymorphs and combination of these). As structure reflects the function, complete three-dimensional structural elucidation using single crystal X-ray diffraction (SCXRD) technique shed light on properties and hence on the applications of the modified APIs. In our research work, moretha fifty binary compounds of the API, gallic acid were prepared successfully, out of which fifteen compounds were published in international peer reviewed journals. Coformers, which are “Generally recognized as safe” (GRAS) compounds, were selected with the help of CSD data base facility using hydrogen bonding and supramolecular synthon approaches. The novel compounds were subjected to initial confirmation characterization techniques (PXRD, FTIR, TGA, and UV-Vis spectroscopy), and then structural studies were carried out using SCXRD experiment. The contribution of inter/intramolecular hydrogen bond interactions, lone pair⋯π and π⋯π stacking interactions, supramolecular ring frameworks in the formation of multicomponent compounds were explored from the structural analysis. The intermolecular interactions were quantified and 3D energy framework analysis were performed through Hirshfeld surface analysis. The chemical and physical reactive parameters were determined from DFT study. Further, molecular docking studies were also carried out to explore the possible molecular interactions with the targeted bacteria, viruses and proteins. Almost all binary crystal forms of gallic acid were successfully obtained in our research, and we are now progressing toward the preparation of higher-order (ternary, quaternary, and quinary) compounds. It has been demonstrated that combining the API with isomers of coformer compounds leads to the formation of binary crystals in various forms with different stoichiometry. Thus, research on multicomponent crystals of gallic acid provides the pharmaceutical industry with the opportunity to harness its desirable properties, thereby expanding its potential medicinal applications.

Abstract

Background and aim: The suppression of the ovaries could be related to COVID-19, as they are often suppressed in acute illnesses to ensure the normal functioning of other systems and organs. In this context, some patients with COVID-19 were found to have inappropriately high levels of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) [1,2]. In the immune response against SARS-CoV-2, interleukin-6 is released along with other cytokines such as interleukin-8 and tumor necrosis factor alpha, which trigger a pro-coagulant state that is unfavorable for the blastocyst or fetus in a normal human uterus. In addition, high rates of antiphospholipid antibodies (aPLs)  have been found in severe COVID-19 patients, although their association with intrahospital vascular thrombosis or worse outcome was not clearly proven [3], and also in patients with subsequent post-COVID syndrome, which could affect pregnancy and fertility. For diagnosis of antiphospholipid syndrome (APS), vascular thrombosis should be accompanied with the presence of aPLs. The aim of this study was to investigate the presence of aPLs in severe COVID-19 female patients and to establish their association with the actual vascular thrombosis, thrombosis in personal history and pregnancy complications.

Methods: In 35 female patients hospitalized with severe COVID-19 pneumonia from March to May 2022, vascular thrombosis and pregnancy complications were recorded during the entire hospital stay and in their medical history. Females were also tested for nine types of aPLs at 4 time points (admission, exacerbation and discharge from hospital and at 3-month follow-up): anticardiolipin (aCL), anti-β2-glycoproteinI (anti-β2GPI) and antiphosphatidylserine/prothrombin (aPS/PT) of the IgM/IgG/IgA isotypes. Because we used anticoagulants at therapeutic doses, we could not use the LA test, even with neutralizers, as these are only effective at low and typically preventive LMWH doses (0.8–1.0 U/mL); instead, the aPS/PT test was performed. Positive values were defined as those above the 99th percentile of the healthy control population. Statistical analysis was performed using SPSS 21 (NY, USA IBM Corp. Released 2021, IBM SPSS Statistics for Windows).

Results: During hospitalization, aPLs were tested positive at least once in 51.4% of females. All 5.7% of patients that passed away were tested aPLs negative at admission and worsening (one had pulmonary artery thrombosis and one microthrombosis). Personal anamnesis (PA) for thromboembolism was verified for 4 patients, at admission all of them tested aPLs negative, however transition to aPLs positivity at discharge (as the disease subsided) was detected in 75%  (all with deep vein thrombosis in PA) due to newly- positive aCL IgG at discharge seen in 75% added with positive aCL IgM in 50%. At follow-up,  one female remained positive for aCL IgG, one negativized and one one did not appear.  None of the women in our cohort were pregnant, and none of them became pregnant while at the hospital and in the follow-up. In their personal history, two women had abortions after 10th week of gestation, but none of them tested positive for aPLs at admission or follow-up.

Conclusion: The incidence of APS in our cohort was 2.8%, whereas in general population it ranges from 0.001% to 0.002%. This high incidence was mostly due to deep vein thrombosis in womens personal history. Therefore, it is advised that all females with personal anamnesis of deep vein thrombosis in the past who had suffered severe COVID-19 and who plan to conceive should be tested for aPLs once and in positive cases retested 3 months later, in order to rule out the APS diagnosis and order appropriate therapy during pregnancy. Also, suppression of the ovaries related to COVID-19 in the post-COVID syndrome could be the reason for infertility [4].

Abstract

Tumor-derived exosomes (TDEs) are key mediators of intercellular communication in cancer, transferring oncogenic cargo: proteins, nucleic acids, and metabolites—to recipient cells within local and distant microenvironments. By disseminating carcinogenic materials, TDEs drive immunosuppression, inflammation, angiogenesis, treatment resistance, epithelial- mesenchymal transition (EMT), and metastasis. Critically, TDEs can also induce malignant transformation in otherwise normal cells, highlighting their central role in cancer progression. Consequently, eliminating TDEs represents a promising strategy to augment current cancer therapies and curb metastatic spread. However, existing approaches to block TDE release or uptake remain ineffective and often incur off-target toxicities. Given the structural and biochemical similarities between TDEs and viruses—including size, membrane composition, and biogenesis—we propose adapting virus-engineered methodolo- gies to develop vaccines targeting TDEs (anti-TDE vaccines). This strategy involves genetically engineered tumor cells to generate modified, immunogenic TDEs that are incapable of binding to or entering recipient cells. Specifically, we advocate for: Surface Engineering: Decorating TDE membranes with foreign antigens to provoke potent, durable immune responses against TDEs, enabling their systemic clearance. Gene Downregulation: Suppressing expression of specific transmembrane proteins (e.g., tetraspanins, integrins) in parent tumor cells to generate TDEs that lack functional ligands required for target cell attachment and internalization.

Further, we refer TDEs as local viruses (loruses) to reflect their endogenous origin and their involvement in the trafficking of pathological information. Engineered loruses could serve as effective vaccines, priming the immune system to recognize and eliminate natural TDEs while avoiding unintended toxicity. This approach merges insights from onco-immunology and virology to overcome the challenges of TDE-mediated pathogenesis and open new avenues for cancer immunotherapy.