Combination Drug Research Articles

Particulates in CGT Guidance Survey Results – Cell and Gene Therapy Industry Knowledge Awareness

The ISCT Particulates Working Group has surveyed the global industry on commonly used regulatory guidance documents pertaining to particulates across various cell and gene therapy (CGT) product-based chemistry and manufacturing controls. Responses were collated from academic and industrially based individuals involved in, or making decisions for, manufacturing, quality control, and analytics. Two surveys covering therapy developers and manufacturing suppliers were created and distributed via email, through focus groups, and via social media platforms. In total, 97 responses were received from CGT professionals at different career levels, which varied from technicians to corporate executives. The responses represent a snapshot of the different CGT industries, with a bias in cell therapy developers, due to the distribution of respondents. Overall, we observed a widespread use of common guidance and legislative documents with specific testing requirements pinned to certain cell and gene therapy product types. Cell-based combination drugs and devices are not well addressed and may require other guidance that were not referenced. These results indicate that CGT particulate risk management should be clarified by collating various particulate guidelines into an easy-to-use guide that brings awareness to the various legislative and guidance documents pertaining to new and novel therapies, their classification, route of administration, and regulatory jurisdiction under which they are handled. The group also recommends clearly outlining the differences and similarities in particulate management regulations across the globe. As the CGT industry is undergoing maturation, the risk analysis methods are also maturing. With such a broad range in product types, routes of administration, source materials, and processing methods, standards of risk analysis and control need to be specific to the risk profile associated with any given drug product. The manufacturing strategy of novel drug types represented under the advanced therapy umbrella will require careful interpretation and implementation of the existing guidance identified in this survey, as not one guidance covers all manufacturing challenges with particulates.

Immune-related and common adverse events with PD-1/PD-L1 inhibitors combined with other anticancer therapy for solid tumors: a systematic review and meta-analysis

AimsThe combination of programmed cell death protein 1 (PD-1)/PD-1 ligand (PD-L1) inhibitors and anticancer therapies has been in the spotlight in recent years. However, the risks associated with these combination therapies are not fully elucidated. The primary objective of this study was to evaluate the relative risk of organ-specific immune-related adverse events (irAEs) and common adverse events (AEs) in patients treated with PD-1/PD-L1 inhibitor-based combination therapies compared to those treated with PD-1/PD-L1 inhibitor monotherapy for solid tumors. Materials and methodsAn electronic database search was performed using ClinicalTrials.gov, Medline, and ASCO/ESMO Annual Meeting Libraries. We included randomized controlled trials designed to assess the safety of combination therapies using PD-1/PD-L1 inhibitors and other anticancer drugs. All the selected clinical studies included solid tumors and provided information on the incidence of non-serious and serious AEs. The quality of evidence was assessed using the Cochrane risk-of-bias tool. A meta-analysis was performed using random effects models to pool the results. ResultsThe primary analysis included 16 relevant clinical studies comprising 4232 patients, of whom 2071 and 2161 patients received PD-1/PD-L1 inhibitor-based combination therapy, and PD-1/PD-L1 inhibitor monotherapy, respectively. Serious organ-specific irAEs were infrequent, even when PD-1/PD-L1 inhibitors were combined with other anticancer drugs. The incidence of serious colitis was significantly higher in the combination therapy group than in the monotherapy group. Among the common AEs associated with PD-1/PD-L1 inhibitors, the incidence of serious pyrexia/fever, non-serious pyrexia/fever, fatigue, nausea, decreased appetite, vomiting, diarrhea, dyspnea, and rash significantly increased in the combination therapy group. In the subgroup analysis based on the modes of action of concomitant anticancer drugs, the combination of PD-1/PD-L1 inhibitors and DNA synthesis inhibitors significantly increased the risk of serious colitis compared to PD-1/PD-L1 inhibitor monotherapy. ConclusionOrgan-specific irAEs occur infrequently when combinations of PD-1/PD-L1 inhibitors and other anticancer drugs are used. However, the risk of serious colitis and certain AEs is higher than that associated with PD-1/PD-L1 inhibitor monotherapy. Vigilant monitoring of AEs and implementation of appropriate clinical management strategies guided by the mode of action of the combination drugs are essential. ProsperoRegistration number CRD42022379088.

Comparative effectiveness of new pharmacotherapy regimens for mild COVID-19 based on symptom resolution dynamics

the treatment of COVID-19 remains a challenging issue worldwide due to the huge economic losses to society as a result of the disease's widespread prevalence, peculiarities of its course and complications of its development. The purpose of the work was comparative analysis of the effectiveness of new treatment regimens for outpatients with mild COVID-19. 64 medical charts of outpatient diseases with a mild course of COVID-19 were studied. Depending on the treatment regimen used, all patients were divided into two groups that received inhaled ACEs or antiviral therapy with the combination drug nortrelvir/ritonavir. The efficacy of each pharmacotherapeutic alternative would be determined on the basis of retrospective data for the timeframe of normalisation of clinical and laboratory parameters. A rule was established that the set of all clinical and laboratory parameters in the normal range is considered as a clinical endpoint. It was found that the use of ACC treats the disease until the rapid disappearance of clinical symptoms, especially in the first treatment, which works on its rapid therapeutic effect in the initial stages. Nirmatrelvir/ritonavir has also been shown to be effective in reducing symptoms, but its effect is incremental, reaching full relief slightly later than in ACC. Modeling of the dynamics of reaching the clinical point of elimination of all clinical symptoms indicates a pharmacoeconomic advantage of the use of ACC over nirmatrelvir/ritonavir, if the additional costs of treatment do not exceed the cost of three days of treatment.

Administration sequences in single-day chemotherapy regimens for breast cancer: a comprehensive review from a practical perspective.

Breast cancer is one of the most prevalent malignant tumors globally, posing a severe threat to human life and health. Chemotherapy, a cornerstone in the treatment of breast cancer, often overlooks the sequence of drug administration within single-day regimens. This study aims to explore the impact of drug administration order on the efficacy and toxicity of combination chemotherapy protocols for breast cancer. Through a comprehensive review and analysis based on current evidence from evidence-based medicine, we delved into how the order of drug administration affects both efficacy and toxicity. We systematically classified and analyzed commonly used combination drug regimens, providing graded recommendations and a reasoned analysis to offer valuable references for clinical decision-making. Our findings indicate that the sequence of drug administration in complex combination chemotherapy protocols is not arbitrary but necessitates multifaceted considerations. Rational drug sequencing can maximize synergistic effects between drugs, thereby augmenting therapeutic efficacy while effectively mitigating drug-related adverse effects. Additionally, some drug labels and clinical trials have explicitly highlighted the therapeutic benefits of specific drug sequences. This study underscores the importance of considering the sequence of drug administration in clinical practice. It is recommended to prioritize the sequential drug administration according to official drug product labeling, while also considering factors such as the administration sequence from large randomized controlled trials, cell proliferation kinetics specific to cancer types, drug interactions, chronopharmacology, drug irritability, clinical experiences, and patient preferences. By taking these factors into account, the goal is to maximize treatment efficacy and minimize the occurrence of adverse reactions.

Discussion on the evaluation of the therapeutic efficacy of uterine artery blood flow parameters and serum PLGF and sFlt-1 in patients with recurrent spontaneous abortion

ObjectiveTo investigate the effects of different drug treatments on uterine artery blood flow parameters, serum placental growth factor (PLGF), soluble fms-like tyrosine kinase-1 (sFlt-1), and sFlt-1/PLGF in patients with recurrent spontaneous abortion and to explore the predictive value of uterine artery blood flow parameters, serum PLGF, sFlt-1, and sFlt-1/PLGF for pregnancy outcomes.MethodsThis retrospective cohort study included 173 patients who experienced recurrent spontaneous abortion and 100 control patients. Patients with recurrent spontaneous abortion were divided into an aspirin group (75 patients), aspirin combined with low molecular weight heparin (LMWH) group (68 patients), and non-drug group (30 patients) based on different drug treatments. Uterine artery blood flow parameters at gestational weeks 30-31+6 were monitored for the four groups, and serum samples were collected at gestational weeks 30-31+6 to measure the levels of serum PLGF and sFlt-1 and calculate the sFlt-1/PLGF ratio.Results1. Uterine artery blood flow parameters at gestational weeks 30-31+6 were significantly greater in the non-drug group than in the aspirin group, combined drug group, and control group (p<0.05). 2. Serum PLGF levels and the sFlt-1/PLGF ratio at gestational weeks 30-31+6 were significantly lower in the non-drug group than in the aspirin group, combined drug group, and control group, while serum sFlt-1 levels were significantly greater in the non-drug group than in the aspirin group, combined drug group, and control group (p<0.05). 3. Serum PLGF, sFlt-1, and sFlt-1/PLGF had lower diagnostic efficiency for predicting hypertensive disorders during pregnancy than the combined diagnostic efficiency of serum PLGF, sFlt-1, and sFlt-1/PLGF with uterine artery blood flow parameters at gestational weeks 30-31+6.ConclusionAspirin and aspirin combined with LMWH can upregulate serum PLGF and decrease serum sFlt-1 levels in patients with recurrent spontaneous abortion, reduce the miscarriage rate, and significantly improve pregnancy outcomes. The combination of serum PLGF, sFlt-1, sFlt-1/PLGF, and uterine artery blood flow parameters can effectively predict hypertensive disorders during pregnancy.

Safety and Efficacy of Extracorporeal Photopheresis for Acute and Chronic Graft-versus-Host Disease.

Background/Objectives: Despite novel biological agents, steroid-dependent or -refractory graft-versus-host disease (GvHD) remains a severe complication of allogeneic hematopoietic cell transplantation (allo-HCT). Extracorporeal photopheresis (ECP) is an alternative, non-immunosuppressive treatment for patients with acute (aGvHD) or chronic (cGvHD) GvHD. The aim of this study was to investigate the safety and efficacy of ECP in the treatment of acute and chronic GvHD; Methods: We prospectively studied 112 patients with cGvHD who received one or more previous lines of treatment and 28 patients with steroid-dependent or refractory grade II-IV aGvHD post-alloHSCT. Results: In terms of severe aGvHD, most of the patients (19/28) responded to ECP treatment, while the five-year overall survival (OS) was 34%. After adjustment for several confounder factors, the reduction in immunosuppression (p = 0.026) and number of ECP sessions (p < 0.001) were associated with improved OS. Regarding chronic GvHD, only 19 patients failed to respond to ECP treatment; though significantly lower rates of response were presented in patients with visceral involvement (p = 0.037) and earlier post-transplant GVHD diagnosis (p = 0.001). Over a follow-up period of 45.2 [interquartile range (IQR): 5.6-345.1] months, the 5-year cumulative incidence (CI) of cGvHD-related mortality was 21.2% and was significantly reduced in patients with ECP response (p < 0.001), while the 5-year OS was 65.3%. Conclusions: Our results confirm the safety and efficacy of ECP in patients with GvHD and provide sufficient data for further investigation and the best combination drugs needed such that GvHD will not be the major barrier of allo-HCT in the near future.

A heterotypic tumor-on-a-chip platform for user-friendly combinatorial chemotherapeutic testing

BackgroundThree-dimensional (3D) tumor microdevices are promising platform for biomimetic antitumor prediction and high-throughput chemotherapeutic screening and play crucial roles in the exploration of cancer-associated pharmaceutics and therapeutics. Traditional cell manipulation tools (e.g., non-adhesive surfaces and hanging drops) and recent microengineered systems (e.g., microfluidic chips and micropatterned array chips) have progressed in terms of microscale control, substantial tumor production, programmable drug combinations, and throughput analysis. However, establishing a facile 3D tumor microdevice to construct heterotypic tumor-microenvironmental profiles and for throughput, implementable, multi-instrument-compatible analysis of chemotherapies to advance consumer-grade tumor modelling tools is still being explored. ResultsIn this study, we present a facilely operated tumor-on-a-chip platform for massive production of heterotypic 3D tumors and diverse investigations of combinatorial chemotherapy screening. Large quantity of heterotypic tumor generation with high geometric controllability (size difference: 19.6 μm) and operational repeatability (n = 10) was achieved using simple-to-fabricate micropatterned chips. Multiple characteristics of solid tumors, including phenotypic gradients (viability and proliferation) and heterogeneous cellular compositions (multi-cell participation and stroma composition), were reproduced in heterotypic tumors, being more biomimetic than homotypic tumors. We completed the user-friendly analytical evaluation of individual and combinatorial drug therapies, and demonstrated the high applicability of the platform in biomimetic tumor-related large-scale manipulation and on-chip analysis, as well as its high compatibility for off-chip detection. The entire operative process during tumor production and chemotherapy only requires the routine and easy-to-master pipetting manipulation. SignificanceThe establishment of a biomimetic and easy-to-use 3D tumor platform and the large-scale screening-like evaluation of combinatorial chemotherapies based on the usage of the micropatterned chip was achieved in a user-friendly manner. This advancement has significant application potential in the fields of oncology, drug discovery, and tissue engineering, and is expected to be valuable for developing accessible and generalizable tumor-on-a-chip microsystems for exploring cancer therapies.

Dual drug co‐encapsulation of bevacizumab and pemetrexed clocked polymeric nanoparticles improves antiproliferative activity and apoptosis induction in liver cancer cells

AbstractNanoparticle (NP) enabled approaches have been employed for chemotherapeutic administration due to their capacity to regulate drug release and reduce side effects. Additionally, these methods can use several drugs concurrently and impede the proliferation of cancer cells that have developed resistance. Bevacizumab (BVZ) and pemetrexed (PEM) have demonstrated encouraging outcomes in the treatment and management of cancer. This work investigates the combined antiproliferative efficacy of BVZ and PEM co‐loaded PLGA‐PEG NPs (BVZ/PEM@PLGA‐PEG NPs) against HepG2 liver cancerous cells. The BVZ/PEM@PLGA‐PEG exhibited a sphere‐shaped and consistent nanosized distribution. In addition, we evaluated the potential mechanisms for inhibiting cell growth and inducing apoptosis using DAPI staining and cell cycle study. The beneficial combined antiproliferative activity and the apoptosis pathway were detected in the HepG2 cells exposed to BVZ/PEM@PLGA‐PEG NPs. Our study determined that the combinational drug treatment of BVZ/PEM@PLGA‐PEG NPs has a significant effect on promoting the effectiveness of liver cancer treatment.

Assessing the cost-effectiveness of replacing antimetabolites with mTOR inhibitors in heart transplant immunosuppression in China: a network meta-analysis-based economic evaluation.

Although several pharmacoeconomic studies have assessed the cost-effectiveness of maintenance immunosuppressive regimens for heart transplant recipients, economic comparisons between various combination drug therapies remain sparse. This study used an economic evaluation based on network meta-analysis to assess the cost-effectiveness of four immunosuppressive regimens for adult heart transplant recipients in China. We conducted a systematic search for clinical trials in PubMed, Embase, Cochrane Library, Web of Science, China National Knowledge Infrastructure (CNKI), Wanfang Data, and VIP database. A validated Markov model was adapted to reflect the Chinese medical landscape. Four maintenance immunosuppression regimens were considered: tacrolimus/mycophenolate mofetil (TAC/MMF), cyclosporine/mycophenolate mofetil (CSA/MMF), everolimus/cyclosporine (EVL/CSA), and sirolimus/tacrolimus (SRL/TAC). The probabilities of health events were derived from a comprehensive literature review. Direct medical costs, adjusted for 2022 values, were from public documents and websites, while utilities for quality-adjusted life-years (QALYs) were taken from previous studies. Primary outcomes were mean lifetime cost, QALYs, and cost-effectiveness, with a willingness-to-pay (WTP) threshold set at three times China's GDP per capita in 2022. Sensitivity analyses were conducted to test the robustness of the results. The base case analysis identified TAC/MMF as the most cost-effective regimen, producing a mean of 6.31 QALYs per patient at a cost of Chinese Yuan (CNY) 534,182.89. Sensitivity analyses consistently reinforced TAC/MMF as the most cost-effective and robust choice. TAC/MMF is the most cost-effective maintenance immunosuppressive regimen for heart transplant recipients within the Chinese health system. The study findings are reinforced by sensitivity analyses, affirming their robustness amid various uncertainties.

Uncovering the Complexity of Perinatal Polysubstance Use Disclosure Patterns on X: Mixed Methods Study.

According to the Morbidity and Mortality Weekly Report, polysubstance use among pregnant women is prevalent, with 38.2% of those who consume alcohol also engaging in the use of one or more additional substances. However, the underlying mechanisms, contexts, and experiences of polysubstance use are unclear. Organic information is abundant on social media such as X (formerly Twitter). Traditional quantitative and qualitative methods, as well as natural language processing techniques, can be jointly used to derive insights into public opinions, sentiments, and clinical and public health policy implications. Based on perinatal polysubstance use (PPU) data that we extracted on X from May 1, 2019, to October 31, 2021, we proposed two primary research questions: (1) What is the overall trend and sentiment of PPU discussions on X? (2) Are there any distinct patterns in the discussion trends of PPU-related tweets? If so, what are the implications for perinatal care and associated public health policies? We used X's application programming interface to extract >6 million raw tweets worldwide containing ≥2 prenatal health- and substance-related keywords provided by our clinical team. After removing all non-English-language tweets, non-US tweets, and US tweets without disclosed geolocations, we obtained 4848 PPU-related US tweets. We then evaluated them using a mixed methods approach. The quantitative analysis applied frequency, trend analysis, and several natural language processing techniques such as sentiment analysis to derive statistics to preview the corpus. To further understand semantics and clinical insights among these tweets, we conducted an in-depth thematic content analysis with a random sample of 500 PPU-related tweets with a satisfying κ score of 0.7748 for intercoder reliability. Our quantitative analysis indicates the overall trends, bigram and trigram patterns, and negative sentiments were more dominant in PPU tweets (2490/4848, 51.36%) than in the non-PPU sample (1323/4848, 27.29%). Paired polysubstance use (4134/4848, 85.27%) was the most common, with the combination alcohol and drugs identified as the most mentioned. From the qualitative analysis, we identified 3 main themes: nonsubstance, single substance, and polysubstance, and 4 subthemes to contextualize the rationale of underlying PPU behaviors: lifestyle, perceptions of others' drug use, legal implications, and public health. This study identified underexplored, emerging, and important topics related to perinatal PPU, with significant stigmas and legal ramifications discussed on X. Overall, public sentiments on PPU were mixed, encompassing negative (2490/4848, 51.36%), positive (1884/4848, 38.86%), and neutral (474/4848, 9.78%) sentiments. The leading substances in PPU were alcohol and drugs, and the normalization of PPU discussed on X is becoming more prevalent. Thus, this study provides valuable insights to further understand the complexity of PPU and its implications for public health practitioners and policy makers to provide proper access and support to individuals with PPU.

Synergistic effect of curcumin and tamoxifen loaded in pH-responsive gemini surfactant nanoparticles on breast cancer cells

BackgroundDrug combination therapy is preferred over monotherapy in clinical research to improve therapeutic effects. Developing a new nanodelivery system for cancer drugs can reduce side effects and provide several advantages, including matched pharmacokinetics and potential synergistic activity. This study aimed to examine and determine the efficiency of the gemini surfactants (GSs) as a pH-sensitive polymeric carrier and cell-penetrating agent in cancer cells to achieve dual drug delivery and synergistic effects of curcumin (Cur) combined with tamoxifen citrate (TMX) in the treatment of MCF-7 and MDA-MB-231 human BC cell lines.MethodsThe synthesized NPs were self-assembled using a modified nanoprecipitation method. The functional groups and crystalline form of the nanoformulation were examined by Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), differential scanning calorimetry (DSC), and dynamic light scattering (DLS) used to assess zeta potential and particle size, and the morphological analysis determined by transmission electron microscopy (TEM). The anticancer effect was evaluated through an in vitro cytotoxicity MTT assay, flow cytometry analysis, and apoptosis analysis performed for mechanism investigation.ResultsThe tailored NPs were developed with a size of 252.3 ± 24.6 nm and zeta potential of 18.2 ± 4.4 mV capable of crossing the membrane of cancer cells. The drug loading and release efficacy assessment showed that the loading of TMX and Cur were 93.84% ± 1.95% and 90.18% ± 0.56%, respectively. In addition, the drug release was more controlled and slower than the free state. Polymeric nanocarriers improved controlled drug release 72.19 ± 2.72% of Tmx and 55.50 ± 2.86% of Cur were released from the Tmx-Cur-Gs NPs after 72 h at pH = 5.5. This confirms the positive effect of polymeric nanocarriers on the controlled drug release mechanism. moreover, the toxicity test showed that combination-drug delivery was much more greater than single-drug delivery in MCF-7 and MDA-MB-231 cell lines. Cellular imaging showed excellent internalization of TMX-Cur-GS NPs in both MCF-7 and MDA-MB-231 cells and synergistic anticancer effects, with combination indices of 0.561 and 0.353, respectively.ConclusionThe combined drug delivery system had a greater toxic effect on cell lines than single-drug delivery. The synergistic effect of TMX and Cur with decreasing inhibitory concentrations could be a more promising system for BC-targeted therapy using GS NPs.

A new therapeutic strategy for infectious diseases against intracellular multidrug-resistant bacteria

Bacterial infections result in 7,700,000 deaths per year globally, with intracellular bacteria causing repeated and resistant infection. No drug is currently licenced for the treatment of intracellular bacteria. A new screening platform mimicking the host milieu has been established to explore phytochemical antibiotic adjuvants. Previously neglected isoprenylated flavonoids were found to be effective against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). Specifically, the synergistic effect between glabrol and streptomycin against intracellular bacteria was observed for the first time. The glabrol-streptomycin combination targets bacterial inner membrane phospholipids, disrupts arginine biosynthesis, inhibits cell wall proteins and biofilm formation genes (agrA/B/C/D), and promotes ROS production, causing subsequent membrane and wall damage.To enhance the selective uptake of combination drug into infected cells, hyaluronic acid-streptomycin-lipoic acid-glabrol nanoparticles (HSLGS-S) were designed and synthesized to trigger the intracellular delivery of the glabrol-streptomycin combination. Thus, the treatment can be transported into the infected intracellular region and selectively release the glabrol-streptomycin combination to the bacterial at site. The bioactivity of HSLGS-S in clearing intracellular bacteria was 20-fold higher than that of the glabrol-streptomycin combination alone in vitro and 2- to 10-fold higher in vivo.

Dihydroartemisinin Inhibits Epithelial-Mesenchymal Transition Progression in Medullary Thyroid Carcinoma Through the Hippo Signaling Pathway Regulated by Interleukin-6.

Dihydroartemisinin (DHA), an artemisinin derivative, can influence certain malignancies' inflammatory response and growth. This study used Cell Counting Kit-8 and Transwell assays to show that DHA suppressed the growth, migration, and invasion of medullary thyroid cancer cells. Furthermore, the authors used enzyme-linked immunosorbent assay, Western blotting, and immunofluorescence to confirm the expression of the transcriptional coactivators Yes-associated protein (YAP)/transcriptional coactivator with a PDZ-binding domain (TAZ) downstream of the Hippo pathway and changes in the expression of the epithelial-mesenchymal transition (EMT) process markers E-cadherin and N-cadherin. These results demonstrate that DHA effectively reduced the expression of interleukin (IL)-6 in medullary thyroid carcinoma (MTC) cells and hindered the EMT process by regulating the Hippo pathway. This regulation was achieved by promoting YAP phosphorylation and inhibiting YAP/TAZ protein expression. Additional activation of the Hippo pathway by GA-017 alleviated the inhibitory effect of DHA on IL-6. Hippo pathway activation led to an increase in the expression of E-cadherin, a marker of EMT. In conclusion, DHA was demonstrated to regulate the Hippo pathway by inhibiting IL-6 secretion, leading to the inhibition of EMT in MTC. These findings provide a theoretical foundation for further exploration of the anticancer mechanisms of DHA and offer valuable insights into its potential clinical application as a combinatorial drug.

Manufacturing Process Affects Coagulation Kinetics of Ortho-R, an Injectable Chitosan-Platelet-Rich Plasma Biomaterial for Tissue Repair.

Ortho-R (ChitogenX Inc., Kirkland, QC, Canada) is an injectable combination drug-biologic product that is used as an adjunct to augment the standard of care for the surgical repair of soft tissues. The drug product comprises lyophilized chitosan, trehalose and calcium chloride, and it is dissolved in platelet-rich plasma (PRP), a blood-derived biologic, prior to injection at the surgical site where it will coagulate. The first step of the Ortho-R manufacturing process involves dissolving the chitosan in hydrochloric acid. The purpose of this study was to investigate the effect of increasing the amount of acid used to dissolve the chitosan on final drug product performance, more specifically, on the chitosan-PRP coagulation kinetics. Chitosans were solubilized in hydrochloric acid, with concentrations adjusted to obtain between 60% and 95% protonation of the chitosan amino groups. Freeze-dried Ortho-R was solubilized with PRP, and coagulation was assessed using thromboelastography (TEG). The clotted mixtures were observed with histology. Clot reaction time (TEG R) increased and clot maximal amplitude (TEG MA) decreased with protonation levels as pH decreased. Chitosan distribution was homogeneous in chitosan-PRP clots at the lowest protonation levels, but it accumulated toward the surface of the clots at the highest protonation levels as pH decreased. These changes in coagulation kinetics, clot strength and chitosan distribution induced by high protonation of the chitosan amino groups were partially reversed by adding sodium hydroxide to the dissolved chitosan component in order to decrease pH. Careful control of manufacturing processes is critical, and it is important to consider the impact of each manufacturing step on product performance.

Treatment of cardiotoxicity in patients with invasive breast cancer during neoadjuvant chemotherapy

Aim – assessment of the effectiveness of complex therapy to reduce the development of early cardiovascular complications. Materials and methods. Information from medical histories and outpatient follow-up cards of 139 patients with invasive breast cancer was analyzed: in the G.V. Bondar Republican Cancer Center 64 patients (main and control groups); as an additional comparison group, the study included 75 patients of the Samara Regional Clinical Oncology Dispensary. Patients in the control group (33 people) and the Samara Regional Clinical Oncology Dispensary group (75 people) received standard therapy (RUSSCO, recommendations, 2023): doxorubicin 60 mg/m2 IV on day 1 + cyclophosphamide 600 mg/m2 IV on day 1 Day 4 – 4 courses. In 31 patients of the main observation group, in addition to similar drug treatment, in order to prevent the development of early cardiovascular complications, the method was used, which included 7 courses before the start of 1 cycle of chemotherapy + 3 courses of interval normobaric hypoxic therapy (INHT) before the start of each subsequent cycle of chemotherapy. Results. When analyzing the total effectiveness of treatment between the groups of patients, statistically significant differences were obtained: the control and main groups were compared (χ2=5.4, p=0.03), the group of the Samara Regional Clinical Oncology Dispensary group and the main group (χ2=3.45, p=0.076). Conclusions. Hypoxic therapy significantly affects the quality of life of patients. For all indicators reflecting quality of life, traditional treatment was inferior to combination drug therapy with INHT.

A multifunctional DNA tetrahedron for imaging, gene therapy, and chemotherapy-phototherapy combination: Binding affinity and anticancer activity

Imaging, silencing cancer-related microRNA, and chemotherapy-phototherapy (CTPT) combination therapy are crucial for cancer diagnosis and drug resistance overcoming. In this study, we designed a multifunctional DNA tetrahedron (MB-MUC1-TD) for the targeted delivery of combined daunorubicin (DAU) + toluidine blue O (TBO). The detection limit of miRNA-21 was determined to be 0.91 nM. The intercalation of DAU and TBO into MB-MUC1-TD was proved by spectroscopic and calorimetric methods. The thermodynamic parameters for the interactions of DAU and/or TBO with MB-MUC1-TD confirmed high drug loading. The first addition of TBO in the ternary system achieved a higher loading of both drugs and a more stable complex structure. Deoxyribonuclease I (DNase I) accelerated the release of DAU and/or TBO loaded in MB-MUC1-TD. Confocal laser scanning microscope demonstrated that MB-MUC1-TD exhibited good imaging ability for miRNA-21 to accurately identify cancer cells, and DAU/TBO was predominantly distributed within the nucleus of cancer cells. In vitro cytotoxicity showed better gene therapy efficacy of MB on MCF-7 cells, better biocompatibility of loaded DAU and TBO on LO2 cells, and stronger synergistic cytotoxicity of DAU + TBO on MCF-7/ADR cells. This study may establish a theoretical foundation for co-loading CTPT combination drugs based on multifunctional DNA nanostructures.

Electrospun patches to deliver combination drug therapy for fungal infections

Fungal infections, though affecting healthcare globally, receive insufficient attention in clinical and academic settings. Invasive fungal infections, particularly caused by combat wounds, have been identified as a critical threat by the US Department of Defense. Monotherapy with traditional antifungals is often insufficient, and so combination therapies are explored to enhance treatment efficacy. However, systemic combination treatments can result in severe adverse effects, suggesting the need for localised delivery systems, such as drug-loaded electrospun patches, to administer antifungals directly to the infection site. This proof-of-concept study hypothesised that dual amorolfine and terbinafine therapy slowly releasing from electrospun patches would be an effective way of eradicating Candida albicans when the patch was applied directly to the fungal colony. The feasibility of creating electrospun materials loaded with amorolfine and terbinafine for combination antifungal therapy was investigated. Electrospinning was used to fabricate polycaprolactone (PCL) patches with varying drug loadings (2.5%, 5%, and 10% w/w) of amorolfine and terbinafine either individually or in combination. The incorporation of both drugs in the fibres was confirmed, with the drugs predominantly in an amorphous state. Results showed that combination therapy patches had a significantly greater and prolonged antifungal effect compared to monotherapy patches, with larger zones of inhibition and sustained efficacy over at least 7 days. This study therefore demonstrates that PCL-based electrospun patches containing amorolfine and terbinafine provide superior antifungal activity against C. albicans compared to monotherapy patches. This approach could lower required drug doses, reducing adverse effects, and enhance patient compliance due to prolonged drug release, leading to more effective antifungal therapy.

Abstract PR-15: Identifying synergistic combinations with KRAS inhibition in PDAC

Abstract Background: To date, targeted therapies have largely failed in eliciting sustained responses in advanced pancreatic ductal adenocarcinoma (PDAC). KRAS-targeted therapies have the potential to drastically transform clinical management. Emerging data indicate that only a subset of patients respond, and that acquired resistance is common. Here, we used a combination of in vivo CRISPR activation (CRISPRa) screening and in vitro/preclinical models to identify synergistic combinations with KRAS inhibition (KRASi) in pancreatic and lung cancer. Material and Methods: We used our CRISPRa-competent PPKS (P53(F/F), Kras(LSL-G12D/+), R26(LSL-SAM)) model of adenocarcinoma to conduct targeted screening for drivers of resistance to MRTX133. Our library consisted of ∼450 frequently mutated and amplified putative oncogenes across human adenocarcinoma based on TCGA data. Mice were treated with 1e8 TU of lentiviral library through nasal instillation. After 6 weeks, treatment was initiated with 3 mg/kg MRTX1133 daily, 5 days/week (i.p.). Mice were sacrificed after 4 weeks of treatment and the guide distribution determined through bioinformatic analysis. Genes statistically significantly enriched (FDR&amp;lt;0.05) in the treatment group were determined. Cell lines with acquired resistance to MRTX1133 derived from the KPC PDAC model and the KPP lung adenocarcinoma (LUAD) model were generated using long-term, low-dose treatment. Resistance was confirmed with MTT drug response assays. In vitro combinatorial drug response was determined using a 72h, 96-well MTT assay, and clonogenic response using a 2 week, 384-well FACS assay. Mice were treated with 20 mg/kg MRTX1133 and/or 20mg/kg Temuterkib once daily, 7 days/week (i.p.) for 2 weeks. Results: We performed targeted in vivo CRISPRa screening for drivers of resistance to KRASi in an autochthons model of lung adenocarcinoma (LUAD) using a library targeting 452 genes commonly amplified or mutated in cancer. We identified a set of genes that were specifically enriched in at least 50% of tumors treated with MRTX1133, including MAPK1/ERK2. We also generated 16 cell lines with acquired resistance to MRTX1133, derived from KPC PDAC or KPP LUAD tumors. Transcriptomic analysis identified a large set of genes differentially expressed in cell with acquired resistance relative to controls. GSEA indicated that reactivation of the MAPK cascade as a common trait in resistant cells. To test if ERK inhibition synergizes with KRASi, we quantified the combinatorial response to MRTX1133 and Temuterkib, a specific inhibitor of ERK1 and ERK2, in a set of KPC and KPP derived cell lines, revealing significant synergy and reduced clonogenic potential in a majority of cell lines. A majority of cell lines with acquired resistance to MRTX1133 remained sensitive to Temuterkib. Finally, we tested combinatorial treatment of MRTX1133 and Temuterkib in a syngenetic subcutaneous model of PDAC, which showed at minimum additive effect. In conclusion we have identified ERK inhibition as a promising combination with KRAS inhibition in KRAS-mutant PDAC and LUAD. Citation Format: Fredrik I Thege, Amber Hoskins, Sonja M Woermann, Anirban Maitra. Identifying synergistic combinations with KRAS inhibition in PDAC [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr PR-15.

Topical psoriasis therapy: An innovative classic

Introduction. Psoriasis affects various aspects of the quality of life of patients, causing psycho-emotional stress, anxiety and depression, therefore treatment of patients in terms of long-term control is the main goal of therapy.Aim. To study the effectiveness of drugs containing calcipotriol and betamethasone in the treatment of plaque psoriasis of smooth skin, scalp, palmoplantar mild to moderate severity.Material and methods. We observed 59 patients and 32 patients with moderate plaque psoriasis (PASI &gt; 10 and &lt; 20 points). In the groups, patients were randomized into subgroups with mild and moderate severity of psoriasis.Results. In patients with mild and moderate plaque psoriasis, after 4 weeks there was a decrease in the PASI index by 89.1% and 64.1%, respectively, the DLQI index decreased by 53.8% and 61.9%, the sPGA index by 71.8% and 64.9% for mild and moderate degrees, respectively. In patients with mild and moderate palmoplantar psoriasis, after 4 weeks there was a decrease in the PPASI index by 64.2% and 62.1%, respectively, the DLQI index decreased by 56.4% and 56.7%, and the sPGA index by 63.4 % and 66.7% for mild and moderate degrees, respectively. In patients with mild to moderate psoriasis of the scalp, after 4 weeks there was a decrease in the PSSI index by 75.1% and 72.6%, the DLQI index decreased by 64.6% and 69.6%, and the sPGA index by 62.3% and 67.6% for mild and moderate degrees, respectively.Conclusion. A 52-week follow-up demonstrated the effectiveness and safety of therapy with combination drugs (Daivobet® and Xamiol®) in patients with plaque psoriasis, including scalp psoriasis and palmoplantar psoriasis. It also showed a pronounced positive effect on the quality of life of patients, including in terms of long-term control of the disease.

The Synergistic Effect of Baloxavir and Neuraminidase Inhibitors against Influenza Viruses In Vitro.

Influenza viruses remain a major threat to human health. Four classes of drugs have been approved for the prevention and treatment of influenza infections. Oseltamivir, a neuraminidase inhibitor, is a first-line anti-influenza drug, and baloxavir is part of the newest generation of anti-influenza drugs that targets the viral polymerase. The emergence of drug resistance has reduced the efficacy of established antiviral drugs. Combination therapy is one of the options for controlling drug resistance and enhancing therapeutical efficacies. Here, we evaluate the antiviral effects of baloxavir combined with neuraminidase inhibitors (NAIs) against wild-type influenza viruses, as well as influenza viruses with drug-resistance mutations. The combination of baloxavir with NAIs led to significant synergistic effects; however, the combination of baloxavir with laninamivir failed to result in a synergistic effect on influenza B viruses. Considering the rapid emergence of drug resistance to baloxavir, we believe that these results will be beneficial for combined drug use against influenza.