Development and validation of simultaneous quantification method for gemcitabine and betulinic acid: augmenting industrial application.

Antitumor drug effect of betulinic acid mediated by polyethylene glycol modified liposomes

− Nanotechnology for cancer therapy is playing a pivotal role in dramatically improving current approaches to cancer detection, diagnosis, and therapy while reducing toxic side effects associated with previous cancer therapy. A wide -spread understanding of these new technologies will lead to develop the more refined design of optimized nanoparticles with improved selectivity, efficacy and safety in the clinical practice of oncology. This review provides an integrated overview of applications and advances of nanotechnology in cancer therapy, based on molecular diagnostics, treatment, monitoring, target drug delivery, approved nanoparticle-based chemotherapeutic agents, and current clinical trials in the development of nanomedicine and ultimately personalized medicine.Key words − Nanotechnology, Nanomedicine, Nanoparticles, Cancer therapy, Personalized therapy Nanotechnology is the interdisciplinary field which is focused on man-made materials up to 100×10 -9 Diagnosism (Grobmyer et al., 2010). Applications of nanotechnology in the screening, diagnosis, and treatment of disease are collectively referred to as “nanomedicine” – an emerging field that has the potential to revolutionize individual and population-based health in the 21st century (Pautler et al., 2010). In recent years, considerable progress and attention have been made in the design and appli-cation of cancer nanotechnology, that is, nanooncology, which is currently the most important chapter of nanomedicine for improving cancer detection, diagnosis, and treatment (Cuenca et al., 2006; Jain, 2010; Misra et al., 2010).Nanobiotechnology plays an important role in the discovery of biomarkers of cancer as well as the development of several drugs for cancer and aids to cancer surgery (Jain, 2007; Jain, 2010). It also provides a unique approach and comprehensive technology against cancer through early diagnosis, prediction, prevention, personalized therapy and medicine for cancer (Misra et al., 2010). The impact of nanobiotechnology on oncology is shown schematically in Figure 1 (Jain, 2005; Jain, 2007; Jain, 2010). This article will focus our attention on cur-rent advances and applications of nanotechnology in cancer therapy for helping with further advancement of developing better cancer drug delivery system that can be applied clin-ically.

ABSTRACTGemcitabine (GEM) is first-line therapy for pancreatic cancer but has limited efficacy in most cases. Nanoparticle-albumin bound (nab)-paclitaxel is becoming first-line therapy for pancreatic cancer, but also has limited efficacy for pancreatic cancer. Our goal was to improve the treatment outcome in patient-like models of pancreatic cancer. We previously established patient-derived orthotopic xenografts (PDOX) pancreatic cancers from two patients. The pancreatic tumor was implanted orthotopically in the pancreatic tail of nude mice to establish the PDOX models. Five weeks after implantation, 50 PDOX mouse models were randomized into five groups of 10 mice for each pancreatic cancer PDOX: untreated control; GEM (100 mg/kg, i.p., once a week for 2 weeks); GEM + nab-PTX (GEM: 100 mg/kg, i.p., once a week for 2 weeks, nab-PTX: 10 mg/kg, i.v., twice a week for 2 weeks); S. typhimurium A1-R (5 × 107 CFU/100 μl, i.v., once a week for 2 weeks); GEM + S. typhimurium A1-R (GEM: 100 mg/kg, i.p., once a week for 2 weeks, S. typhimurium A1-R; 5 × 107 CFU/100 μl, i.v., once a week for 2 weeks). GEM + nab-PTX was significantly more effective than GEM alone in one PDOX model (p = 0.0004), but there was no significant difference in the other PDOX model. The combination of GEM + S. typhimurium A1-R regressed both PDOX models. These results show S. typhimurium A1-R can overcome the ineffectiveness or partial effectiveness of GEM in patient-like models of pancreatic cancer and demonstrate clinical potential for this combination.

BackgroundTriterpenoid compounds such as betulinic acid (BA) and oleanolic acid (OA) exhibit considerable pharmacological activities. However, their current production primarily relies on plant extraction and chemical synthesis, methods that are often plagued by low efficiency, complex extraction processes, and environmental concerns. Microbial-based synthesis has emerged as one of the most effective approaches for producing BA and OA.ResultsThis study presented the first identification of filamentous fungal strains efficiently synthesizing both BA and OA. The strain ZBS49 is a newly identified species of Clonostachys isolated from Gleditsia japonica Miq. (showing 99.82% sequence identity), produced 47.7 mg/L of BA. The strain XJ1-1, characterized as Colletotrichum gloeosporioides and isolated from Cannabis sativa L., yielded 65.76 mg/L of OA. After optimizing the culture medium and cultivation conditions, the yields of ZBS49 and XJ1-1 increased to 288.97 and 86.14 mg/L, representing improvements of 506% and 31%, respectively. Furthermore, we discovered that the BA extract of the ZBS49 strain significantly inhibited hepatocellular cancer cells (SMMC-7721 and HepG2) in a dose-dependent manner, with a minimum inhibitory concentration of 70 µM. Genomic analysis of Clonostachys sp. ZBS49 elucidated that the presence of 16 putative genes was related to triterpenoid biosynthesis and 6 distinct terpene biosynthetic gene clusters. Among the 145 CYP450, 5 genes involved in C-28 oxidation were predicted.ConclusionsThis research underscores the effectiveness of filamentous fungi as a biotechnological platform for the efficient production of BA and its derivatives, highlighting their potential applications in cancer therapy. Furthermore, these findings provide valuable genetic resources and establish a robust technical and theoretical framework for utilizing ZBS49 as a microbial platform for the biosynthesis of triterpenoids.Graphical abstract

Introduction: Intravenous (IV) oncology drug administration plays a pivotal role in cancer therapy, enabling swift and precise delivery of therapeutic agents. The preparation process for these drugs is highly complex, necessitating rigorous adherence to safety standards, efficient operational workflows, and adoption of advanced technologies to ensure both patient safety and treatment effectiveness. This review delves into the practices and innovations within pharmacy settings that underpin the preparation of IV oncology drugs, with an emphasis on safety, operational efficiency, technological advancements, and drug stability. Safety: Ensuring safety in the preparation of IV oncology medications is crucial due to the hazardous properties of chemotherapeutic agents. Compliance with standards such as USP <797> and USP <800> is vital to mitigate contamination risks and minimize exposure. Safety measures include the use of personal protective equipment (PPE), engineering controls such as biological safety cabinets (BSCs) and compounding aseptic containment isolators (CACIs), along with comprehensive staff training programs. Past incidents, such as contamination at the New England Compounding Center, illustrate the necessity for robust safety protocols to safeguard both patients and healthcare professionals. Efficiency: Optimizing efficiency in the preparation process is essential to address high demand while maintaining stringent safety and quality standards. Key steps include prescription validation, drug compounding, quality assurance, and dispensing. The adoption of technologies like automated compounding devices (ACDs) and pharmacy management systems has streamlined these processes, significantly reducing errors and preparation times. However, achieving a balance between operational efficiency and uncompromised safety remains a persistent challenge, requiring ongoing staff education, resource optimization, and strict compliance with regulatory guidelines. Innovative Technologies: The integration of cutting-edge technologies, including robotics, automation, and artificial intelligence (AI), has transformed IV drug preparation. Robotic systems offer enhanced precision and minimize human error, while AI and advanced aseptic techniques optimize workflows and dosing accuracy. Future advancements may include the application of 3D printing, nanotechnology, and blockchain to further elevate the safety, efficiency, and traceability of preparation processes. Stability: Preserving the stability of compounded oncology drugs is fundamental to their therapeutic effectiveness and safety. Stability is influenced by variables such as storage conditions, handling methods, and the compatibility of diluents and containers. Adhering to guidelines like USP <797> is crucial for maintaining stability. Innovations in stabilizing agents, packaging, and real-time monitoring systems are advancing the field, helping to overcome stability-related challenges and improving treatment outcomes. Conclusion: This review underscores the paramount importance of safety, efficiency, innovation, and stability in the preparation of IV oncology drugs. Strict adherence to safety standards, streamlined workflows, the adoption of advanced technologies, and meticulous stability management are critical to enhancing pharmacy practices and patient care. Future efforts should prioritize the integration of emerging technologies, standardization of procedures, continuous education for staff, and implementation of real-time monitoring to further progress in this domain. Addressing these challenges with innovative solutions will drive improvements in both patient safety and therapeutic outcomes.

Metal organic frameworks in biomedicine: Innovations in drug delivery

Gemcitabine (GEM) is a nucleoside analogue approved as a first line of therapy for pancreatic ductal adenocarcinoma (PDAC). However, rapid metabolism by plasma cytidine deaminase leading to the short half-life, intricate intracellular metabolism, ineffective cell uptake, and swift development of chemoresistance downgrades the clinical efficacy of GEM. ONC201 is a small molecule that inhibits the Akt and ERK pathways and upregulates the TNF-related apoptosis-inducing ligand (TRAIL), which leads to the reversal of both intrinsic and acquired GEM resistance in PDAC treatment. Moreover, the pancreatic cancer cells that were able to bypass apoptosis after treatment of ONC201 get arrested in the G1-phase, which makes them highly sensitive to GEM. To enhance the in vivo stability of GEM, we first synthesized a disulfide bond containing stearate conjugated GEM (lipid-GEM), which makes it sensitive to the redox tumor microenvironment (TME) comprising high glutathione levels. In addition, with the help of colipids 1,2-dioleoyl-glycero-3-phosphocholine (DOPC), cholesterol, and 1,2-distearoyl-glycero-3-phosphoethanolamine-poly(ethylene glycol)-2000 (DSPE-PEG 2000), we were able to synthesize the lipid-GEM conjugate and ONC201 releasing liposomes. A cumulative drug release study confirmed that both ONC201 and GEM showed sustained release from the formulation. Since MUC1 is highly expressed in 70-90% PDAC, we conjugated a MUC1 binding peptide in the liposomes which showed higher cytotoxicity, apoptosis, and cellular internalization by MIA PaCa-2 cells. A biodistribution study further confirmed that the systemic delivery of the liposomes through the tail vein resulted in a higher accumulation of drugs in orthotopic PDAC tumors in NSG mice. The IHC of the excised tumor grafts further confirmed the higher apoptosis and lower metastasis and cell proliferation. Thus, our MUC1 targeting binary drug-releasing liposomal formulation showed higher drug payload, enhanced plasma stability, and accumulation of drugs in the pancreatic orthotopic tumor and thus is a promising therapeutic alternative for the treatment of PDAC.

Gemcitabine (GEM) is a chemotherapeutic drug widely used for treating pancreatic cancer and other cancers. Despite its efficacy, GEM is associated with adverse side effects and tumor resistance, hampering its therapeutic outcomes. To address these challenges, innovative strategies have emerged to enhance GEM delivery. This comprehensive review explores various distribution systems, including polymer-based platforms, liposomes, and inorganic nanoparticles, highlighting their unique characteristics for improving GEM efficacy in cancer treatment. Additionally, we discuss the promising approach of codelivery with genes and present several methods for augmenting GEM's chemotherapeutic properties. Our findings shed light on novel insights and provide recommendations for overcoming the limitations associated with GEM, guiding researchers and clinicians toward more effective cancer therapies.

Chitosan-based Dy2O3/CuFe3O4 bio-nanocomposite development, characterization, and drug release kinetics

Synergistic inhibition of pancreatic adenocarcinoma cell growth by trichostatin A and gemcitabine

F3 peptide functionalized liquid crystalline nanoparticles for delivering Salinomycin against breast cancer

Sulpiride, which has selective dopaminergic blocking activity, is a substituted benzamide antipsychotic drug playing a prominent role in the treatment of schizophrenia, which more selective and primarily blocks dopamine D2 and D3 receptor. This study has two main objectives, firstly; the molecular modeling studies (MD and Docking, ADME) were conducted to define the molecular profile of sulpiride and sulpiridereceptor interactions, another to synthesize polymeric nanoparticles with chitosan, having the advantage of slow/controlled drug release, to improve drug solubility and stability, to enhance utility and reduce toxicity. Molecular dynamic simulation was carried out to determine the conformational change and stability (in water) of the drug and the binding profile of D3 dopamine receptor was determined by molecular docking calculations. The pharmacological properties of the drug were revealed by ADME analysis. The ionic gelation method was used to prepare sulpiride loaded chitosan nanoparticles (CS NPs). The Dynamic Light Scattering (DLS), UV-vis absorption (UV), Scanning Electron Microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy techniques were carried out to characterize the nanoparticles. In vitro cell cytotoxicity experiments examined with MTT assay on mouse fibroblast (L929), human neuroblastoma (SH-SY5Y) and glioblastoma cells (U-87). The statistical evaluations were produced by ANOVA. The residues (ASP-119, PHE-417) of D3 receptor provided a stable docking with the drug, and the important pharmacological values (blood brain barrier, Caco-2 permeability and human oral absorption) were also determined. The average particle size, PdI and zeta potential value of sulpiride- loaded chitosan NPs having a spherical morphology were calculated as 96.93 nm, 0.202 and +7.91 mV. The NPs with 92.8% encapsulation and 28% loading efficiency were found as a slow release profile with 38.49% at the end of the 10th day. Due to the formation of encapsulation, the prominent shifted wave numbers for C-O, S-O, S-N stretching, S-N-H bending of Sulpiride were also identified. Mitochondrial activity of U87, SHSY-5Y and L929 cell line were assayed and evaluated using the SPSS program. To provide more efficient use of Sulpiride having a low bioavailability of the gastrointestinal tract, the nanoparticle formulation with high solubility and bioavailability was designed and synthesized for the first time in this study for the treatment of schizophrenia. In addition to all pharmacological properties of drug, the dopamine blocking activity was also revealed. The toxic effect on different cell lines have also been interpreted.

<h3>Background</h3> Microfluidics chip-based approaches (MF) can achieve unique transport properties through laminar flows and vastly increased surface-to-volume ratios, which have been extensively utilized to prepare smaller and homogeneous lipid nanoparticles (LNPs). LNPs have shown potential to carry highly insoluble medicines, especially betulinic acid (BA) which has significant antitumour activity but is difficult to administer for cancer therapy due to its poor water solubility. In this study, we investigated the parameters involved in the continuous production of LNPs encapsulating BA (BA-LNPs) by MF, and the possibility of improving antitumour efficacy and reducing toxicities was described. <h3>Methods</h3> Briefly, a three-inlet MF system was developed and used to produce the LNPs. EggPC, cholesterol and BA (45:37:18% molar) were dissolved in ethanol. The lipid solution was then loaded into a 1 mL glass syringe and injected into the centre inlet channel, while phosphate-buffered saline (PBS; pH 6.5) was loaded into two 5 mL glass syringes and introduced into the two side inlet channels to establish hydrodynamic focusing. To investigate the physicochemical and biological properties of fabricated liposome at different shear forces, the total flow rate (TFR) is varied from 0.3 to 0.8 mL/min. <h3>Results</h3> The TFR has a very small effect on particle size distribution, but an increase in TFR causes a progressive decrease in liposome size. The percentage of encapsulating efficiency (EE) was 77–92% and there were no significant changes when BA-LNPs were stored at 4° C. The inhibitory rate of BA-LNPs was significantly higher compared to free BA in vitro. Immunohistochemical analysis showed many damaged tumor cells after BA-LNPs were injected for 15 days. The survival of mice treated with BA-LNPs was apparently prolonged compared to mice treated with free BA. <h3>Conclusions</h3> This result indicates that much stronger antitumour effects were induced by BA-LNP administration, which is most likely due to the relatively small particle sizes produced by MF and which are suitable for intracellular transportation.

Background &amp; Aims: The new therapy for pancreatic cancer which has the poorest prognosis among malignant tumors is high demand nowadays. The overall 5-year survival rate in all stages of this type of cancer is less than 5%, even with the multimodality therapy, including surgery, chemotherapy and radiotherapy. Although gemcitabine (GEM) is a key drug in the treatment of advanced pancreatic cancer, the therapeutic effect is limited because of drug resistance. Heat shock protein 90 (HSP90) is known to be overexpressed in several types of cancer cells, and its inhibition has shown promise in the treatment of solid malignancies. Src kinases work as a signal switch for multiple molecular signal transduction pathways. The objective of this study is to evaluate an antitumor effect of three kinds of small molecular weight compounds (antimetabolite Gemcitabine, HSP90 inhibitor NVP-AUY922, and Src kinase inhibitor Dasatinib) on tumor growth and chemotaxis in pancreatic cancer cell lines. Methods: Pancreatic cancer cell lines were treated with each of the 3 compounds to examine the effect on proliferation assay, apoptosis assay, Western blotting, and chemotaxis assay. Results: Of these compounds, HSP90 inhibitor inhibited cell proliferation in pancreatic cancer cells best in a dose- and time-dependent manner. Growth inhibitory effect of HSP90 inhibitor for pancreatic cancer cells were highest of the three. Further, HSP90 inhibitor inhibited the growth of gemcitabine sensitive and resistant pancreatic cancer cell lines. With flowcytometry and immunostaining, HSP90 inhibitor had higher apoptosis-inducing effect than gemcitabine. In the chemotaxis assay, HSP90 inhibitor inhibited the migration of pancreatic cancer cell best in the three compounds. In western blotting, HSP90 inhibitor suppressed the expression of multiple receptor tyrosine kinase proteins as well as the signal transduction in PI3K-Akt and RAS-MAPK pathways. Conclusion: In this study, HSP90 inhibitor was the best of 3 small molecular compounds and has proven effective in gemcitabine sensitive and resistant pancreatic cancer. The HSP90 inhibitor is useful as an antitumor drug by inhibiting proliferation and chemotaxis of pancreatic cancer cells through multiple kinase pathways. Citation Format: Masahiro Yamamura, Akira Yamauchi, Naoki Katase, Masakiyo Sakaguchi, Yosuke Katata, Hiroaki Tanioka, Makoto Okawaki, Takeshi Nagasaka, Yoshiyuki Yamaguchi. Comparing effects of small molecular weight compounds on proliferation and chemotaxis of pancreatic cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2188.

P3-080 - Evaluation of Chemotherapy with Cisplatin–Gemcitabine after Failure of Gemcitabine Monotherapy for Unresectable or Recurrent Biliary Tract Cancer