Therapeutic Formulations Research Articles

Scaling up controlled nucleation in freeze drying: Translating vacuum-induced surface freezing from laboratory to GMP

The freezing step often causes batch inhomogeneity and issues during freeze drying process transfer. The nucleation temperature at which the first ice is formed during freezing differs from vial to vial, and significantly between scales. To solve this issue, Controlled Ice Nucleation techniques can be applied to induce ice nucleation at a defined product temperature across the whole batch. This study describes the application of vacuum-induced surface freezing (VISF) for a therapeutic antibody formulation, including the process transfer from laboratory scale through pilot scale to a GMP line. The VISF method could be successfully implemented on all scales of freeze dryers without equipment adaptation. Some scale-dependent changes in pressure control and degassing were necessary to achieve nucleation in all vials and avoid defects. The resulting lyophilized products were characterized and further analyzed in a stability study. While most critical quality attributes were comparable for product manufactured with and without Controlled Nucleation, the appearance of cakes processed using VISF was much better, which could be linked to different product morphology due to freeze-concentration. The results of this study allow direct comparison of the application of controlled nucleation for an antibody formulation at different scales and confirm the applicability of the technology.

Drugs in Development to Manage Acute Pain.

Acute pain, defined as short-term pain arising from injury or other noxious stimuli, affects patient outcomes, quality of life, and healthcare costs. Safe, effective treatment of acute pain is essential in preventing increased morbidity, mortality, and the transition to chronic pain. In this review, we explore some of the latest therapeutic agents, formulations, combinations, and administration routes of drugs emerging in clinical practice in the USA for the treatment of acute pain. These agents include VX-548 (Suzetrigine), Cebranopadol, AAT-076, Combogesic intravenous (IV), sublingual ketamine, XG004 (naproxen/pregabalin conjugate), and HTX-011 (Zynrelef). We analyze the pharmacodynamics, pharmacokinetics, development status, and clinical implications of these drugs, emphasizing the importance of finding an agent that provides both a strong safety profile and effective relief from acute pain. Our findings show promise but also highlight the need for further large-scale research to allow these drugs to be utilized in a clinical context for patients experiencing acute pain.

Generation of Functional Neurons from Mesenchymal Stem Cells Using Neural Differentiator and Engineered Peptide Hydrogel: Potential Therapeutic Lead for Traumatic Brain Injury.

Traumatic brain injuries (TBIs) cause multifaceted disruption in the neural network, initiate huge inflammation processes, and form glial scars that result in severe damage to the brain. Thus, the treatment of TBI is a challenging task. To address this challenge, a newer and innovative approach is extremely important to develop a successful therapeutic strategy. Toward this aim, we hereby report an extremely effective therapeutic strategy. This interesting approach showcased the development and validation of a combination therapy comprising a neuro-regenerative protective peptide hydrogel (SLNAP) and a potent neuro-regenerative chemical modulator (NCM). It is noteworthy to mention that this hydrogel formulation has injectable nature, which allows it to be applied at focal injury site of brain. Remarkably, our results reveal excellent transdifferentiation of human umbilical cord-derived mesenchymal stem cells (hMSCs) into functional neuron upon treatment with this combination therapeutic formulation. The functionality of regenerated neurons was thoroughly checked through observation of active signals generated from those neurons in electrophysiology recording using patch clamp. Further, we also observed that this strategy not only successfully converts hMSCs into neuron but also spontaneously formed neural stem cells (NSCs) like neurosphere. This work also showcased that this multidomain self-assembling peptide hydrogel emerges as an attractive soft-biomaterial due to its capability of slow and sustained release of the drug at the injury site upon topical application. This resulted in significant regeneration of functional neuron at the injury site. Fascinatingly, we found that this combination therapeutic strategy is highly effective in in vivo brain injury model establishing that this could be a potential and highly effective therapeutic strategy for TBI.

Assessment of growth and essential oil profiling in leafy coriander (Coriandrum sativum L.) genotypes

Coriander is a versatile plant prized for its dual role as a culinary spice and a medicinal herb. Leafy coriander, an herbaceous plant, is particularly noted for its rich content of bioactive compounds, such as essential oils, flavonoids, and vitamins. The experiment was conducted at the Department of Spices and Plantation Crops, Horticultural College and Research Institute, Tamil Nadu Agricultural University, Coimbatore. A study was conducted using a Randomized Block Design (RBD) with two replications, focusing on critical growth and yield traits. Among the fifteen genotypes evaluated, the CSL1 genotype illustrates superior performance in various growth and yield parameters. Coriander leaves are extensively studied for their antioxidant, anti-inflammatory, and antimicrobial properties, underscoring their significance in nutritional and medicinal contexts. In this research, coriander leaf oil was extracted and analysed using gas chromatography-mass spectrometry (GC-MS) to determine its chemical composition. The GC-MS analysis identified 60 distinct compounds in the essential oil, with the major constituents being 3-dimethylamino acrylonitrile, trans-2-Dodecen-1-ol, Cyclooctane, Decanal, 13-Octadecenal, 9-Decen-1-yl acetate and 2-Propen-1-amine. These results offer valuable insights into the unique chemical profile of coriander leaf oil, contributing to its distinctive aroma and potential applications in flavouring fragrance and therapeutic formulations.

Efficacy of Dairy Protein and Essential Nutrient Density in Formulated Food Products for Treating Severe and Moderate Acute Malnutrition at the Community Level: A Narrative Review.

Community-based management of acute malnutrition (CMAM) relies on a food-based approach. However, a comprehensive assessment of their nutrient composition and its impact on treatment outcomes is currently lacking in the extant literature. This narrative review summarizes recent evidence on the efficacy of formulations that contain dairy protein and maintain the density of essential nutrients (type I and type II) in managing uncomplicated acute malnutrition at the community level. The literature used for the evidence synthesis was identified using a two-stage screening process. An electronic search was run on PubMed and Cochrane Library, followed by a backward snowball search to identify efficacy studies. A total of 26 efficacy studies involving food formulations used to treat uncomplicated severe and moderate acute malnutrition were identified. The review found that while more evidence favours the inclusion of dairy in formulations as efficacious in supporting recovery from malnutrition, ambiguity in the conclusive findings between dairy and non-dairy formulations remains due to the varied percentages of dairy protein in different formulations. The type of protein source used in a formulation matters, but other approaches, including fortification, can aid in maintaining the nutrient density of formulations, thereby improving the chances of recovery. However, the inclusion of high-added sugar in therapeutic formulations exceeding the WHO norms is a concern that warrants more attention. Future clinical research should assess outcomes like lean or fat mass changes to confirm the benefits of using dairy or non-dairy formulations to treat moderate and severe acute malnutrition.

Drug repurposing: An antidiabetic drug Ipragliflozin as Mycobacterium tuberculosis sirtuin-like protein inhibitor that synergizes with anti-tuberculosis drug isoniazid

The surge of drug-resistant Mycobacterium tuberculosis (DR-TB) impedes the World Health Organization's efforts in ending TB and calls for new therapeutic formulations. M. tuberculosis sirtuin-like protein Rv1151c is a bifunctional enzyme with both deacetylation and desuccinylation activities, which plays an important role in M. tuberculosis drug resistance and stress responses. Thus, it appears to be a promising target for the development of new TB therapeutics. In this study, we screened 31,057 ligand compounds from seven compound libraries in silico to identify inhibitors of Rv1151c. Ipragliflozin can bind to Rv1151c and interact stably. Ipragliflozin can change the acylation level of M. tuberculosis by inhibiting Rv1151c and effectively inhibit the growth of M. tuberculosis H37Rv and M. smegmatis. It can potentiate the first-front anti-TB drug isoniazid. As an antidiabetic drug, Ipragliflozin can be potentially included in the regimen to treat diabetes-tuberculosis comorbidity.

Deciphering the multi-scale mechanism of herbal phytoconstituents in targeting breast cancer: a computational pharmacological perspective

Breast Cancer (BC) is the most common cause of cancer-associated deaths in females worldwide. Despite advancements in BC treatment driven by extensive characterization of its molecular hallmarks, challenges such as drug resistance, tumor relapse, and metastasis persist. Therefore, there is an urgent need for alternative treatment approaches with multi-modal efficacy to overcome these hurdles. In this context, natural bioactives are increasingly recognized for their pivotal role as anti-cancer compounds. This study focuses on predicting molecular targets for key herbal phytoconstituents—gallic acid, piperine, quercetin, resveratrol, and beta-sitosterol—present in the polyherbal formulation, Krishnadi Churna. Using an in-silico network pharmacology model, key genes were identified and docked against these marker compounds and controls. Mammary carcinoma emerged as the most significant phenotype of the putative targets. Analysis of an online database revealed that out of 135 predicted targets, 134 were mutated in breast cancer patients. Notably, ESR1, CYP19A1, and EGFR were identified as key genes which are known to regulate the BC progression. Docking studies demonstrated that the herbal phytoconstituents had similar or better docking scores than positive controls for these key genes, with convincing protein-ligand interactions confirmed by molecular dynamics simulations, MM/GBSA and free energy landscape (FEL) analysis. Overall, this study highlights the predictive potential of herbal phytoconstituents in targeting BC genes, suggesting their promise as a basis for developing new therapeutic formulations for BC.

Dihydroquercetin nanoparticles nasal gel is a promising formulation for amelioration of Alzheimer’s disease

Dihydroquercetin is a natural flavonoid with anti-inflammatory, antioxidant, and neuroprotective activities. Dihydroquercetin exhibits a great neuroprotector promise in Alzheimer’s disorder via preventing the aggregation of amyloid-beta-peptide-Aβ(1–42). The goal of the study was to create dihydroquercetin-loaded-chitosan nanoparticles (DHQ-CS NPs) loaded to a mucoadhesive, thermosensitive in-situ gel for direct nasal administration to cure Alzheimer’s disorder. Loading drug in chitosan nanoparticles and incorporation into thermosensitive gel enhanced residence time and reduced mucociliary-clearance. Different in-vitro-physicochemical-characteristics of gels and nanoparticles-characterization were used to evaluate the formulations. The therapeutic effectiveness of DHQ-CS NPs gel was evaluated behaviorally, biochemically and histopathologically in Alzheimer’s-rat-model compared to intranasal DHQ gel. The small particles-size was obtained = 235.3 nm of DHQ-CS NPs. The DHQ-CS NPs gel demonstrated a greater release rate compared to the raw DHQ gel. Additionally, the nasal-administration of the DHQ-CS NPs gel showed better In-vivo results compared to DHQ gel, through improvement of memory and learning deficits and also the exploratory behavior and new object memory in streptozotocin induced-Alzheimer rats. Biochemically, the intranasal DHQ-CS NPs gel, showed reduced both Aβ-protein formation and tau protein hyperphosphorylation, inhibition of acetylcholine esterase activity and oxidative stress in the brain with increase of total antioxidants in the brain and serum, compared to DHQ gel. Histopathologically, the DHQ-CS NPs nasal gel produced improvement in the hippocampal and cerebral cortex structures, being comparable to the normal group. Consequently, the intranasal DHQ-CS NPs loaded in-situ gel seems to be a promising therapeutic formulation for Alzheimer’s disease medication.

Reducing the water quenching processes using heavy water in capillary electrophoresis with fluorescence detection

Water, ubiquitous in analytical methods, is renowned for its fluorescence quenching properties, influencing techniques like fluorescence spectrophotometry or techniques with fluorescence detection. This study explores the impact of water (H₂O) substitution for heavy water (D₂O) on the fluorescence behavior of anthraquinones and anthracyclines. Anthraquinones and anthracyclines play crucial roles in pharmacy, serving as essential components in various therapeutic formulations, particularly in cancer treatment and other pharmacological interventions. Capillary electrophoresis (CE) with heavy water as the background electrolyte (BGE) solvent offers superior sensitivity to the separation and detection of these analytes. Experimental results demonstrate the improved detection limits and separation efficiency of selected anthraquinones rhein (RH), aloe-emodin (AE), and anthracyclines doxorubicin (DOX), epirubicin (EPI) and daunorubicine (DAU) in heavy water-based buffers, highlighting the potential of heavy water in advancing analytical chemistry.

Enhancing skin aging parameter assessment in clinical trials: AI-Driven analysis of ultrasound images

BackgroundHigh-frequency ultrasound (HFUS) imaging offers valuable insights into skin aging parameters in clinical trials. However, manual measurements are time-consuming and operator-dependent, necessitating the exploration of automated approaches. ObjectiveThis study aimed to develop, train, and validate an artificial intelligence model for automated HFUS image parameter acquisition in dermatological assessments. A secondary goal was its application in evaluating data from an anti-aging skin therapy clinical trial. MethodsSupervised machine learning algorithms were used to predict target variables from ultrasound images. The best models were selected through independent validation. A training dataset of 144 ultrasound skin images and a validation set of 40 images were employed. A clinical trial involved 34 female subjects with aging skin signs, randomized into two groups: (1) cosmetic formulation plus therapeutic unfocused ultrasound (TUS) and (2) placebo formulation plus TUS. HFUS imaging was conducted before and after a 4-week and 8-week study period. ResultsOur algorithms successfully predicted echogenicity and thickness variables, showing competitive performance. In the clinical trial, the treatment group exhibited a significant improvement in skin echogenicity, while no significant changes were observed in the placebo group. ConclusionThis study demonstrates the feasibility of automating the prediction of bioimage variables for anti-aging strategy development. Mathematical algorithms reduced HFUS image evaluations to minutes, expediting clinical trial progress.

An Advanced Mechanically Active Osteoarthritis-on-Chip Model to Test Injectable Therapeutic Formulations: The SYN321 Case Study.

Current treatments for osteoarthritis (OA) often fail to address the underlying pathophysiology and may have systemic side effects, particularly associated with long-term use of non-steroidal anti-inflammatory drugs (NSAIDs). Thus, researchers are currently directing their efforts toward innovative polymer-drug combinations, such as mixtures of hyaluronic acid viscoelastic hydrogels and NSAIDs like diclofenac, to ensure sustained release of the NSAID within the joint following intra-articular injection. However, the progress of novel injectable therapies for OA is hindered by the absence of preclinical models that accurately represent the pathology of the disease. The uBeat® MultiCompress platform is here presented as a novel approach for studying anti-OA injectable therapeutics on human mechanically-damaged OA cartilage microtissues, in a physiologically relevant environment. This platform can accommodate injectable therapeutic formulations and is successfully tested with SYN321, a novel diclofenac-sodium hyaluronate conjugate under development as a treatment for knee OA. Results indicate the platform's effectiveness in evaluating therapeutic potential, showing downregulation of inflammatory markers and reduction in matrix degradation in OA cartilage micro-tissues treated with SYN321. The uBeat® MultiCompress platform thus represents a valuable tool for OA research, offering a bridge between traditional in vitro studies and potential clinical applications, with implications for future drug discovery.

In vitro anticancer efficacy of Calendula Officinalis extract-loaded chitosan nanoparticles against gastric and colon cancer cells

Objective This study assessed the anticancer activities of Calendula officinalis-loaded chitosan nanoparticles in gastric and colon cancer cells compared with fibroblast cells and examined the balance between ROS and antioxidants. Methods Considering this information, we synthesized Calendula officinalis-loaded chitosan nanoparticles (CO-CSNPs) via the ionic gelation method. Their characterizations were carried out with ZetaSizer, UV-Vis, FTIR and SEM devices including size, morphology and surface zeta potential analysis, loading capacity, encapsulation efficiency, in vitro drug release, and chemical interactions. The anticancer activities of CO, CSNPs, and CO-CSNPs were tested against AGS, Caco-2, and normal NIH-3T3 cells using an XTT assay. The anticancer effects were evaluated using DAPI staining, scratch assay, reactive oxygen species (ROS) detection and the CUPRAC method on cellular and non-cellular processes that promote anticancer mechanisms. Results Results showed that CO and CO-CNPs exhibited anticancer activity against AGS and Caco-2. Further, the formulation of CO with CSNPs enhanced the anticancer activity of CO while having no cytotoxicity on NIH-3T3. DAPI staining, scratch assay, ROS, and CUPRAC method confirmed the anticancer activity of CO and CO-CSNPs, which resulted in a reduction in the number of apoptotic cells, inhibited migration, triggered apoptotic pathway via ROS, and higher antioxidant activity. Conclusions The results of the study indicate that CO-CSNPs are a promising therapeutic formulation for gastric and colon cancer treatment. We consider that this study will lead to the investigation of molecular mechanisms of CO-CSNPs in cancer treatment and their investigation in clinical studies.

Bioflavonoid Daidzein: Therapeutic Insights, Formulation Advances, and Future Directions.

Bioflavonoids, are a diverse group of phytonutrients that are widely distributed in fruits, vegetables, grains, teas, and certain medicinal herbs. They are characterized by their antioxidant properties and play essential roles in plant biology, such as providing color to fruits and flowers, protecting plants from environmental stresses. Daidzein, a bioflavonoid classified under natural products, is sourced from plants like soybeans and legumes. It exists in forms such as glycosides and aglycones, with equol and trihydroxy isoflavone being key metabolites formed by gut bacteria. Known for its wide-ranging therapeutic potential, daidzein has shown effects on cardiovascular health, cancer, diabetes, skin conditions, osteoporosis, and neurodegenerative disorders. Its mechanisms include interaction with estrogen receptors, antioxidative and anti-inflammatory properties, and modulation of apoptosis and cell cycles. Recent advances in formulation technologies aimed at enhancing daidzein's bioavailability and efficacy are critically evaluated, including nanoparticle-based delivery systems and encapsulation strategies. Researchers have developed advanced formulations like nanoparticles and liposomes to enhance daidzein's solubility, stability, bioavailability, and targeted delivery. Considered a promising nutraceutical, daidzein warrants further exploration into its molecular actions and safety profile to fully realize its clinical potential. This review offers a succinct overview encompassing therapeutic benefits, chemical characteristics, historical uses, toxicology insights, recent advancements in delivery systems, and future directions for daidzein research.

Hemoglobin-PEG Interactions Probed by Small-Angle X-ray Scattering: Insights for Crystallization and Diagnostics Applications.

Protein-protein interactions, controlling protein aggregation in the solution phase, are crucial for the formulation of protein therapeutics and the use of proteins in diagnostic applications. Additives in the solution phase are factors that may enhance the protein's conformational stability or induce crystallization. Protein-PEG interactions do not always stabilize the native protein structure. Structural information is needed to validate excipients for protein stabilization in the development of protein therapeutics or use proteins in diagnostic assays. The present study investigates the impact of polyethylene glycol (PEG) molecular weight and concentration on the spatial structure of human hemoglobin (Hb) at neutral pH. Small-angle X-ray scattering (SAXS) in combination with size-exclusion chromatography is employed to characterize the Hb structure in solution without and with the addition of PEG. Our results evidence that human hemoglobin maintains a tetrameric conformation at neutral pH. The dummy atom model, reconstructed from the SAXS data, aligns closely with the known crystallographic structure of methemoglobin (metHb) from the Protein Data Bank. We established that the addition of short-chain PEG600, at concentrations of up to 10% (w/v), acts as a stabilizer for hemoglobin, preserving its spatial structure without significant alterations. By contrast, 5% (w/v) PEG with higher molecular weights of 2000 and 4000 leads to a slight reduction in the maximum particle dimension (Dmax), while the radius of gyration (Rg) remains essentially unchanged. This implies a reduced hydration shell around the protein due to the dehydrating effect of longer PEG chains. At a concentration of 10% (w/v), PEG2000 interacts with Hb to form a complex that does not distort the protein's spatial configuration. The obtained results provide a deeper understanding of PEG's influence on the Hb structure in solution and broader knowledge regarding protein-PEG interactions.

Application of Nanotechnology and Phytochemicals in Anticancer Therapy.

Cancer is well recognized as a leading cause of mortality. Although surgery tends to be the primary treatment option for many solid cancers, cancer surgery is still a risk factor for metastatic diseases and recurrence. For this reason, a variety of medications has been adopted for the postsurgical care of patients with cancer. However, conventional medicines have shown major challenges such as drug resistance, a high level of drug toxicity, and different drug responses, due to tumor heterogeneity. Nanotechnology-based therapeutic formulations could effectively overcome the challenges faced by conventional treatment methods. In particular, the combined use of nanomedicine with natural phytochemicals can enhance tumor targeting and increase the efficacy of anticancer agents with better solubility and bioavailability and reduced side effects. However, there is limited evidence in relation to the application of phytochemicals in cancer treatment, particularly focusing on nanotechnology. Therefore, in this review, first, we introduce the drug carriers used in advanced nanotechnology and their strengths and limitations. Second, we provide an update on well-studied nanotechnology-based anticancer therapies related to the carcinogenesis process, including signaling pathways related to transforming growth factor-β (TGF-β), mitogen-activated protein kinase (MAPK), phosphatidylinositol 3 kinase (PI3K), Wnt, poly(ADP-ribose) polymerase (PARP), Notch, and Hedgehog (HH). Third, we introduce approved nanomedicines currently available for anticancer therapy. Fourth, we discuss the potential roles of natural phytochemicals as anticancer drugs. Fifth, we also discuss the synergistic effect of nanocarriers and phytochemicals in anticancer therapy.

Co-delivery of Liposomal Ketoconazole and Bevacizumab for Synergistical Inhibition of Angiogenesis Against Endometrial Cancer.

In this study, we designed a novel formulation based on liposomes for the co-delivery of cancer-derived exosome inhibitor (ketoconazole, Keto) and angiogenesis inhibitor (bevacizumab, mAb). The designed Combo-Lipo formulation was systematically characterized, exhibiting a uniform average particle size of 100nm, as well as excellent serum and long-term physical stabilities. The cell viability assay revealed that Combo-Lipo treatment significantly reduced the viability of cancer cells compared to free drugs. Moreover, liposomes effectively inhibited angiogenic mediators and reduced tumor immune suppressive factors. The Combo-Lipo formulation demonstrated potent downregulation of angiogenic factors and synergistic effects in suppressing their production. Furthermore, liposomes inhibited tumor-associated macrophages (TAMs), leading to decreased expression of tumor-promoting factors. Together, these findings highlighted the promising characteristics of Combo-Lipo as a therapeutic formulation, including optimal particle size, serum stability, and potent anti-cancer effects, as well as inhibition of angiogenic mediators and TAMs toward treating endometrial cancer.

Enhanced Solubility and Increased Bioavailability with Engineered Nanocrystals

Abstract: The exploration of nanocrystal technology is currently receiving significant attention in various fields, including therapeutic formulation, clinical formulation, in-vivo and in-vitro correlation research, and related investigations. The domain of nanocrystals in pharmaceutical delivery has received significant interest as a potential solution for the difficulties associated with medications that have low solubility. The nanocrystals demonstrate promise in improving solubility and bioavailability, presenting a potential resolution to significant challenges. Significantly, nanocrystals have exhibited efficacy in the context of oral administration, showcasing prompt absorption due to their quick breakdown, hence fitting with the requirements of medications that necessitate fast commencement of action. In addition, the adaptability of drug nanocrystals encompasses several methods of administration, including oral, parenteral, ophthalmic, cutaneous, pulmonary, and targeted delivery modalities. The observed consistency can be ascribed to the increased solubility of nanocrystals of the medicine, which effectively counteracts the influence of food on the absorption of the drug. Surface modification tactics have a significant influence on insoluble medicines by enhancing hydrophilicity and reducing plasma protein adsorption on the crystal surface. The surface properties of nanocrystals are modified through the utilization of specific surfactants and polymers, which are subsequently incorporated into polymer solutions via high-pressure homogenization procedures. This article encompasses an examination of the drug distribution mechanism, the nanocrystal formulation technology, the therapeutic applications, the potential future developments, and the challenges associated with the solubility and bioavailability of tailored nanocrystals, as discussed in this article. Consequently, it possesses the capacity to provide guidance for future investigations pertaining to nanocrystal technology.

QUALITATIVE ANALYSIS OF MAHISHADRAVAKA WITH SPECIAL REFERENCE TO LCMS STUDY

Introduction: Mahishadravaka, a renowned Ayurvedic proprietary medicine, has been historically used for its therapeutic properties. It is the arka preparation (distillation process) of Mahisha bone (bone of a buffalo), along with other herbal medicines. It is traditionally used for Vata vyadhies (diseases caused by Vata). Materials and Method: In this study, we conducted a comprehensive LCMS analysis and vitamin D3 and Calcium analyses of Mahishadravaka. Identification and characterisation of bioactive compounds were also aimed at. Result: Our findings reveal a diverse array of compounds with promising pharmacological activities, including Anti-inflammatory, antioxidant, anti-microbial, and anticancer properties. This article provides an overview of the identified compounds and their reported medicinal benefits, shedding light on the potential mechanisms underlying the therapeutic efficacy of Mahishadravaka. Conclusion: The LCMS of Mahishadravaka unveiled a rich array of bioactive compounds with significant pharmacological potential. The elucidation of the bioactive constituents paves the way for further research and development of novel therapeutic formulations based on Ayurvedic principles. These compounds encompass a broad spectrum of therapeutic activities, validating the traditional use of Mahishadravaka in Ayurvedic medicine. The findings from this study provide a scientific basis for the therapeutic efficacy of Mahishadravaka and underscore its potential for developing novel therapeutic interventions targeting various diseases.

Synergistic cryoprotective effect of deaeration and polysorbate 80 on IgG denaturation during thin-film freeze-drying

Polysorbate 80 (PS80) is employed as a cryoprotectant in biotherapeutic formulations to preserve biologics from denaturation during the freezing process. However, excessive quantities of PS80 can lead to stickiness in dry powder, thereby reducing production yields and altering the physical and aerodynamic characteristics of the powder. Additionally, degradation byproducts of PS80 pose a risk of destabilizing proteins in therapeutic formulations. Our study reveals the synergistic cryoprotective effect of combining deaeration with PS80. PS80 plays a protective role by competitively binding to ice/water and air/water interfaces, as well as binding directly to IgG molecules. Deaeration increases the cryoprotective effect of PS80 by reducing the surface area of air microbubbles, thereby reducing the required amount of PS80 for surface coverage and limiting available adsorption sites for IgG molecules, which allows the decrease in the necessary concentration of PS80 required to minimize IgG denaturation during the thin-film freeze-drying (TFFD) process. Even at a low concentration of 0.0025 % w/v, PS80 prevents the aggregation of IgG during the freezing step of TFFD but does not eliminate IgG denaturation during the drying step. PS80 concentrations ranging from 0.0025 % to 0.0400 % w/v exhibit no impact on the specific surface area (SSA) and aerodynamic properties (i.e., fine particle fraction (FPF), mass median aerodynamic diameter (MMAD)) of IgG dry powders. These IgG TFFD powders exhibit a fine particle fraction exceeding 70 % and a median mass aerodynamic diameter of approximately 2.3–2.5 μm, with an SSA of around 20 m2/g.

Targeted Chemo-Phototherapy in Red Light with Novel Doxorubicin and Iron(III) Complex-Functionalized Gold Nanoconjugate (Dox-Fe@FA-AuNPs).

The anticancer efficacy of doxorubicin, an anthracycline-based and FDA-approved chemotherapeutic drug, is significantly hindered by acquired chemoresistance and severe side effects despite its potent anticancer properties. To overcome these challenges, we developed an innovative therapeutic formulation that integrates targeted chemotherapy and phototherapy within a single platform using gold nanoparticles (AuNPs). This novel nanoconjugate, designated as Dox-Fe@FA-AuNPs, is co-functionalized with folic acid, doxorubicin, and an iron(III)-phenolate/carboxylate complex, enabling cancer-specific drug activation. Here, we report the synthesis, characterization, and comprehensive physico-chemical and biological evaluations of Dox-Fe@FA-AuNPs. The nanoconjugate exhibited excellent solubility, stability, and enhanced cellular uptake in folate receptor-positive cancer cells. The nanoconjugate was potently cytotoxic against HeLa and MDA-MB-231 cancer cells (HeLa: 105.5±16.52 μg mL-1; MDA-MB-231: 112.0±12.31 μg mL-1; MDA-MB-231 (3D): 156.31±19.35 μg mL-1) while less cytotoxic to the folate(-) cancer cells (MCF-7, A549 and HepG2). The cytotoxicity was attributed to the pH-dependent release of doxorubicin, which preferentially occurs in the acidic tumor microenvironment. Additionally, under red light irradiation, the nanoconjugate generated ROS, inducing caspase-3/7-dependent apoptosis with a photo-index (PI) >50, and inhibited cancer cell migration. Our findings underscore the potential of Dox-Fe@FA-AuNPs as a highly effective and sustainable platform for targeted chemo-phototherapy.