Efficacy Of Combined Treatment Research Articles

Real-world evidence of combined treatment of biologics and exclusive enteral nutrition in patients with ileum-dominant Crohn's disease: A multicenter study

Background & AimsAlthough biologics were prescribed to achieve and maintain clinical remission of active Crohn’s disease (CD), almost half of patients experienced a loss of response or intolerance. Here, we investigated the efficacy of combined treatment of biologics and 16-weeks exclusive enteral nutrition (EEN) in moderate-to-severe CD patients with small intestine lesions. MethodsThis was a real-world, multicenter retrospective study, from October 2016 to March 2023, medical records of patients registered at three IBD centers were reviewed for patients with ileal or ileocolonic CD in moderate-to-severe activity. All patients received treatment of biologics with concomitant 16-week EEN (BioEEN) or biologics alone (Bio). The clinical outcomes and endoscopic outcomes were assessed at week 16 and 52. ResultsThere was no statistically significant difference between Bio (97 patients) and BioEEN group (100 patients) at baseline for demographic and clinical characteristics. Compared to treatment with biologics alone, patients with BioEEN treatment achieved higher rates of clinical response (95.0% vs. 66.0%), clinical remission (87.0% vs. 52.6%), endoscopic response (91.4% vs. 47.4%) including mucosal healing (85.7% vs. 23.7%) at week 16. The superiority of BioEEN sustained in maintenance, with 87.4% (vs. 49.1%) clinical response, 77.8% (vs. 38.6%) clinical remission, 69.2% (vs. 32.6%) endoscopic response and 51.9% (vs. 18.6%) mucosal healing at week 52. ConclusionsCombined treatment of biologics and 16-week EEN was an efficient therapeutic strategy with affirmative effectiveness for small intestine diseases of active CD.

Abstract 2122: Enhanced anticancer efficacy via ROS-dependent ferroptosis: Synergy between surufatinib and cisplatin in small cell lung cancer

Abstract Objective: Small cell lung cancer (SCLC), a highly malignant subset of neuroendocrine carcinoma, accounts for 13% of lung cancer cases and is characterized by its high aggressiveness and poor prognosis. The longstanding first-line treatment, a platinum-etoposide chemotherapy regimen, is increasingly compromised by resistance issues, highlighting the critical need for new therapeutic approaches. Surufatinib, an inhibitor selectively targeting vascular endothelial growth factor receptors (VEGFR) 1, 2, and 3, fibroblast growth factor receptor type 1 (FGFR1), and colony-stimulating factor-1 receptor (CSF-1R), has demonstrated efficacy in a spectrum of solid tumors, including neuroendocrine neoplasm, thyroid cancers and biliary cancers. Recently, ferroptosis, an iron-dependent cell death pathway, offers therapeutic promise, especially by inhibiting the xCT system, effective in pancreatic and lung adenocarcinomas. Yet, surufatinib's potential for inducing ferroptosis in SCLC remains to be investigated. Methods: Cytotoxicity assays were performed on SCLC cell lines to assess the combinatorial effect of surufatinib and cisplatin, revealing a synergistic interaction. Ferroptosis inhibitors were applied to assess the reversal of cytotoxic effects. Mitochondrial morphology, reactive oxygen species (ROS) accumulation, membrane potential dissipation (JC-1 assay), lipid peroxidation (MDA levels), and intracellular iron accumulation (Fe2+ quantification) assays confirmed ferroptosis induction. Western blot analyses were conducted to explore the mechanistic pathways, supplemented by in vivo antitumor efficacy evaluations. Results: In vitro assays revealed that surufatinib, when combined with cisplatin, exerts a pronounced synergistic effect on small cell lung cancer (SCLC) cell lines by inducing ferroptosis. The combination therapy strategically modulated ferroptosis-related pathways: cisplatin effectively decreased the expression of glutathione peroxidase 4 (GPX4), critical for cellular antioxidant defenses, while surufatinib increased the expression of acyl-CoA synthetase long-chain family member 4 (ACSL4), which is essential for the accumulation of lipid peroxides. This dual pathway engagement significantly amplified the ferroptotic response, culminating in heightened antitumor activity. Corroborative in vivo experiments substantiated these findings, validating the combined treatment's efficacy in a living model and underscoring its potential as a robust therapeutic strategy for SCLC. Conclusion: Our findings reveal that surufatinib triggers ferroptosis in small cell lung cancer—a novel discovery. Additionally, it also unveils that surufatinib and cisplatin together enhance anti-cancer efficacy via the ROS-mediated ferroptosis pathway, marking a significant advance in SCLC therapy. Citation Format: Xiaolin Li, Ziting Zhang, Qiyun Tang, Huae Xu. Enhanced anticancer efficacy via ROS-dependent ferroptosis: Synergy between surufatinib and cisplatin in small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2122.

Abstract 7203: Anti-HDGF antibody attenuates the rise of resistance to EGFR tyrosine kinase inhibitor in NSCLC patient-derived xenograft tumors

Abstract Mutant epidermal growth factor receptor (EGFR), a major oncogenic driver in non-small cell lung cancer (NSCLC), is found in the tumors of 15% (western countries) to 50% (Asian countries) of patients. EGFR-targeted therapy using tyrosine kinase inhibitors (TKIs) is a standard of care for those patients who developed metastatic or recurrent diseases. However, despite having an 80% initial response rate, most patients had partial tumor regression and their disease would progress when acquired resistance emerges. Thus, developing more effective treatment is an unmet need. The clinical response of these patients can be recapitulated in mouse models of patient-derived xenograft (PDX) tumor bearing sensitizing EGFR mutations. In this study, treating established PDX tumors with osimertinib, a third generation EGFR TKI, induced partial tumor regression (50% to 70% size reduction from baseline) followed by progression with a median progression-free survival (PFS) of 26 to 28 days. Interestingly, co-administering an antibody against hepatoma-derived growth factor (HDGF) with osimertinib resulted in enhanced tumor regression (98% to 100% size reduction from baseline) which translated into significantly extended animal survival with a median PFS of 103 or 135 days. Immunohistochemical and western blot examination of tumors collected at different treatment timepoints indicated that AKT/mTOR and MAPK pathways were reactivated after initial suppression in TKI monotherapy arm despite persistent EGFR inhibition. Since AKT/mTOR and MAPK pathways are key survival and proliferative signaling pathways, their reactivation can promote TKI tolerance of tumor cells and the emergence of TKI resistance. However, blocking HDGF attenuated the reactivation of these pathways which could partially explain the enhanced efficacy of osimertinib plus anti-HDGF antibody combination treatment seen in this study. Our results suggest tumor-derived HDGF could play a crucial role in the rise of acquired resistance to osimertinib. Identifying the exact cells and molecular pathways utilized by HDGF to promote tumor cell tolerance to TKI will help us understand how TKI sensitive tumor cells survive EGFR blockade and emerge as resistant cells. Citation Format: Cindy Q. Zhou, Ariel Li, Kaoru Ri, Ahmed Sultan, Hening Ren. Anti-HDGF antibody attenuates the rise of resistance to EGFR tyrosine kinase inhibitor in NSCLC patient-derived xenograft tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7203.

Abstract 2048: Integrated tumor models for immune oncology: Using live cell imaging for prediction of treatment efficacy in vitro and in vivo

Abstract Background: The preclinical evaluation of novel cancer treatments demands comprehensive model systems in vitro that provide meaningful data before entering in vivo studies. Here we evaluate the capabilities of live cell imaging systems to evaluate novel immune therapies. Using integrated immune and tumor cell models in vitro we demonstrate, that these model systems can generate reliable data of pharmacodynamic activity of biologicals, small molecules or combinatorial approaches for further preclinical in vivo characterization. Methods: Target tumor cell killing was assessed in vitro with immune cells (T- and NK-cells) and engagers. Tumor cells were transduced with a fluorescent marker to discriminate tumor cells from immune cells. The technology was used to determine inhibition of cell motility (re-invasion) after scratching of tumor cell monolayers. Cells were monitored using the IncuCyte. Dose-response-curves of single treatments and all combinations were generated in parallel. Active therapies were selected for further in vivo validation of immune cell killing. Humanized mice were generated by injection of CD34+ HSC or human immune cell subsets. Immune cell engraftment was monitored by FACS. To analyze the effect biologicals or small molecules, tumor cells were transplanted into these humanized mice. Tumor development and therapeutic effects were monitored by BLI measurements. Results and conclusion: Tumor cell killing by immune cells and monolayer scratch assay in 96 well format were successfully monitored in the IncuCyte. Here, data can be generated over time without the need of new samples at every time point compared with conventional end point measurements. Using antibodies directing immune cells to attack target cells extensive cell killing was observed over time. These data predicted in vivo treatment outcome in mice co-engrafted with human immune cells. After successful humanization of mice, immune cells can be directed to kill target tumor cells. Small molecule combinations were tested in vitro utilizing the metastasis/2D scratch assay. After setup of dose-response curves for two molecules combinatorial treatments were tested. Here we found a synergistic increase in efficacy. These combinations were tested in vivo to evaluate their abilities to inhibit cell motility and distant metastasis. Here we show that the in vitro assays predicted correctly the highest efficacy of combined treatments compared to mono treatments.The IncuCyte System provides data that translate our integrated model systems into in vivo studies. We have shown that activated immune cells can kill target tumor cells in vitro. These data have been validated in vivo using immune cell humanized mice. Further, immune cells, biologicals and small molecule based treatments can be tested either alone or in combination, allowing the preselection of active combinations for further development. Citation Format: Dennis Kobelt, Maria Stecklum, Simone Rhein, Wolfgang Walther, Jens Hoffmann. Integrated tumor models for immune oncology: Using live cell imaging for prediction of treatment efficacy in vitro and in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2048.

Abstract 4552: EGFR inhibitors increase LGR5 expression and enhance potency of LGR5 antibody-drug conjugates targeting colorectal cancer stem cells

Abstract Colorectal cancer (CRC) is the second leading cause of cancer-associated deaths in the United States. One of the primary challenges in treating CRC is therapy resistance thought to be mediated by cancer stem cells (CSCs), a subpopulation of cancer cells with infinite replicative potential that can differentiate to drive tumorigenesis and disease relapse. Antibody-drug conjugates (ADCs) are amongst the fastest growing classes of anticancer drugs and utilize the specificity of monoclonal antibodies (mAbs) to hone cytotoxic payloads to cancer cells. We generated ADCs directed against leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5), a well-defined biomarker of CSCs that is highly overexpressed in CRCs. LGR5 ADCs demonstrated high specificity and efficacy in CRC cells and xenograft models, though tumor relapse after treatment withdrawal was a major obstacle. Interestingly, FDA-approved therapy targeting epidermal growth factor receptor (EGFR) has been shown to increase LGR5 mRNA expression levels in patient-derived models of CRC. This study aims to determine the mechanism of EGFR-mediated regulation of LGR5 expression and evaluate the therapeutic efficacy of combination treatments targeting EGFR and LGR5 for the improved treatment of CRC. To first analyze the effect of EGFR inhibition on LGR5 expression, a panel of CRC cell lines of different KRAS and PI3CKA mutational statuses were treated with FDA-approved EGFR-targeted mAbs or small molecule inhibitors. LGR5 protein expression was shown to be upregulated by EGFR-targeted therapies in a dose- and time-dependent manner, independent of mutation status. Notably, treatment with EGF, MEK1/2 inhibitor trametinib, and EGFR-directed siRNA all resulted in concomitant reduction in EGFR and LGR5 protein levels, suggesting EGFR and LGR5 are co-degraded. Co-immunoprecipitation and immunocytochemistry experiments identified a novel EGFR-LGR5 interaction and co-internalization mechanism in CRC cells, respectively. Furthermore, we showed EGFR-LGR5 interaction is enhanced by treatment with the EGFR-targeting mAb, cetuximab (CTX). To evaluate efficacy of combination therapies targeting EGFR and LGR5, CRC cells were treated with LGR5 ADCs with or without CTX. Importantly, we showed CTX significantly enhanced the potency of LGR5 ADCs incorporating different classes of cytotoxic payloads. These results suggest combining LGR5 ADCs with EGFR inhibitors may be a more effective approach for the treatment of CRC and eliminating CSCs to overcome resistance and relapse. Citation Format: Peyton C. High, Zhengdong Liang, Kendra S. Carmon. EGFR inhibitors increase LGR5 expression and enhance potency of LGR5 antibody-drug conjugates targeting colorectal cancer stem cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4552.

Abstract 5559: The anti-tumor efficacy and immune effects of combining of ceralasertib, an oral ATR kinase inhibitor, with imipridones, in prostate cancer treatment in vitro and in vivo

Abstract Prostate cancer is the most common cancer and the second leading cause of cancer death among men in the United States. There is an estimated 10% to 50% of cases progress to metastatic castration resistant prostate cancer (mCRPC) state within 3 years of diagnosis. mCPRC remains lethal despite therapeutic advances. There is approximately 20% of mCRPC patients present somatic DNA damage repair (DDR) gene mutations. Ceralasertib, formerly known as AZD6738, is a potent and selective orally bioavailable inhibitor of the ataxia tenlangiectasia and Rad3-related (ATR) kinase, which is involved in DNA repair in response to DNA damage and replication stress. Preclinical studies have demonstrated ATRi sensitizing alterations in DNA damage response (DDR) genes. Ceralasertib’s antitumor activity as a monotherapy in treating prostate cancer is moderate. Thus, we combined ceralasertib with imipridones (ONC201 and ONC206), which target mitochondrial caseinolytic protease P (ClpP) and the integrated stress response, resulting in enhanced antitumor efficacy in vitro. Prior data have demonstrated sensitivity of 4 prostate cancer lines to ceralasertib and imipridones monotherapies, synergistic activities with the combination treatments, and immune enhancement effects with the combination treatments when co-cultured with NK92MI cell line. We proceeded to a short term in-vivo study validating the combination treatment efficacy. Preliminary results include cytokine profiling using the Luminex 200 technology to understand treatment induced changes in the tumor microenvironment (TME) from both in vitro and in vivo studies. Our results guide novel clinical trials for effective clinical responses in mCPRC patients. Citation Format: Yutong Linda Xia, Leiqing Zhang, Maryam Ghandali, Maximilian P. Schwermann, Wafik El-Deiry. The anti-tumor efficacy and immune effects of combining of ceralasertib, an oral ATR kinase inhibitor, with imipridones, in prostate cancer treatment in vitro and in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5559.

Repurposing of the Cardiovascular Drug Statin for the Treatment of Cancers: Efficacy of Statin-Dipyridamole Combination Treatment in Melanoma Cell Lines.

Metastatic melanoma has a very poor prognosis. Statins, 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGCR) inhibitors, are cholesterol-lowering agents with a potential for cancer treatment. The inhibition of HMGCR by statins, however, induces feedback, which paradoxically upregulates HMGCR expression via sterol regulatory element-binding protein-2 (SREBP2). Dipyridamole, an antiplatelet agent, is known to inhibit SREBP2 upregulation. We aimed to demonstrate the efficacy of statin-dipyridamole combination treatment in both human and spontaneously occurring canine melanoma cell lines. The half maximal inhibitory concentration (IC50) of atorvastatin showed a 68-92% reduction when combined with dipyridamole, compared with that of atorvastatin alone. In some melanoma cell lines, cell proliferation was suppressed to almost zero by the combination treatment (≥3 μM atorvastatin). Finally, the BRAF inhibitor, vemurafenib, further potentiated the effects of the combined statin-dipyridamole treatment in BRAF V600E mutation-bearing human melanoma cell lines. In conclusion, the inexpensive and frequently prescribed statin-dipyridamole combination therapy may lead to new developments in the treatment of melanoma and may potentiate the effects of vemurafenib for the targeted therapy of BRAF V600E-mutation bearing melanoma patients. The concordance between the data from canine and human melanoma cell lines reinforces this possibility.

Lipid Nanocarriers-Enabled Delivery of Antibiotics and Antimicrobial Adjuvants to Overcome Bacterial Biofilms.

The opportunistic bacteria growing in biofilms play a decisive role in the pathogenesis of chronic infectious diseases. Biofilm-dwelling bacteria behave differently than planktonic bacteria and are likely to increase resistance and tolerance to antimicrobial therapeutics. Antimicrobial adjuvants have emerged as a promising strategy to combat antimicrobial resistance (AMR) and restore the efficacy of existing antibiotics. A combination of antibiotics and potential antimicrobial adjuvants, (e.g., extracellular polymeric substance (EPS)-degrading enzymes and quorum sensing inhibitors (QSI) can improve the effects of antibiotics and potentially reduce bacterial resistance). In addition, encapsulation of antimicrobials within nanoparticulate systems can improve their stability and their delivery into biofilms. Lipid nanocarriers (LNCs) have been established as having the potential to improve the efficacy of existing antibiotics in combination with antimicrobial adjuvants. Among them, liquid crystal nanoparticles (LCNPs), liposomes, solid lipid nanoparticles (SLNs), and nanostructured lipid carriers (NLCs) are promising due to their superior properties compared to traditional formulations, including their greater biocompatibility, higher drug loading capacity, drug protection from chemical or enzymatic degradation, controlled drug release, targeted delivery, ease of preparation, and scale-up feasibility. This article reviews the recent advances in developing various LNCs to co-deliver some well-studied antimicrobial adjuvants combined with antibiotics from different classes. The efficacy of various combination treatments is compared against bacterial biofilms, and synergistic therapeutics that deserve further investigation are also highlighted. This review identifies promising LNCs for the delivery of combination therapies that are in recent development. It discusses how LNC-enabled co-delivery of antibiotics and adjuvants can advance current clinical antimicrobial treatments, leading to innovative products, enabling the reuse of antibiotics, and providing opportunities for saving millions of lives from bacterial infections.

Immunotherapy or Chemoimmunotherapy in Older Adults With Advanced Non–Small Cell Lung Cancer

Immune checkpoint inhibitor (ICI) plus chemotherapy combination treatment (ICI-chemotherapy) is now a standard treatment for non-small cell lung cancer (NSCLC) without targetable oncogene alterations, but there are few data on ICI-chemotherapy for patients 75 years and older. To inform the choice of first-line drugs in clinical practice and assess the safety and efficacy of ICI-chemotherapy combination treatment in older adult patients with previously untreated advanced NSCLC. This retrospective cohort study included 58 centers in Japan. The cohort consisted of patients 75 years and older with clinical stage IIIB, IIIC, IV, postoperative or radiotherapy recurrent NSCLC. Patients started first-line systemic therapy between December 2018 and March 2021. Those receiving first-line molecular targeted drugs were excluded. The data were analyzed from February 2022 to October 2022. Systemic therapy. The main outcomes were overall survival (OS), progression-free survival (PFS), and safety. A total of 1245 patients (median [range] age, 78 [75-95] years; 967 [78%] male) with NSCLC were included in the cohort. Programmed death ligand-1 (PD-L1) expression of less than 1% occurred in 268 tumors (22%); 1% to 49% in 387 tumors (31%); 50% and higher in 410 tumors (33%), and unknown expression in 180 tumors (14%). Median OS was 20.0 (95% CI, 17.1-23.6) months for the 354 patients receiving ICI-chemotherapy (28%); 19.8 (95% CI, 16.5-23.8) months for the 425 patients receiving ICI alone (34%); 12.8 (95% CI, 10.7-15.6) months for the 311 patients receiving platinum-doublet chemotherapy (25%); and 9.5 (95% CI, 7.4-13.4) months for the 155 patients receiving single-agent chemotherapy (12%). After propensity score matching, no differences in OS and PFS were found between the patients receiving ICI-chemotherapy vs ICI alone. Each group consisted of 118 patients. For PD-L1 expression of 1% and higher the OS hazard ratio (HR) was 0.98 (95% CI, 0.67-1.42; P = .90), and the PFS HR was 0.92 (95% CI, 0.67-1.25; P = .59). Significance was also not reached when separately analyzed for lower or higher PD-L1 expression (1%-49% or ≥50%). However, grade 3 or higher immune-related adverse events occurred in 86 patients (24.3%) treated with ICI-chemotherapy and 76 (17.9%) with ICI alone (P = .03). In this study, ICI-chemotherapy combination treatment did not improve survival and increased the incidence of grade 3 and higher immune-related adverse events compared with ICI alone in patients 75 years and older. Based on these results, ICI alone may be recommended for older adult patients with PD-L1-positive NSCLC.

Efficacy and Safety of Combination Treatment of Oral Terbinafine and Itraconazole in Patients with Extensive Superficial Dermatophytes

Background: Dermatophytosis, a prevalent fungal infection, is instigated by species like Trichophyton, Epidermophytonand Microsporum. The current approach for treating dermatophytosis involves a blend of systemic and topical antifungal therapies, applicable to all patients except those with localized, untreated Tinea. Notably, there is a growing incidence of inadequate response to the standard treatment regimen in recent times. Objective: To evaluate the efficacy andsafety of combination treatment of oralterbinafine anditraconazole in patients with extensive superficial dermatophytes. Materials and Methods: This research comprised a randomized clinical trial involving 50 individuals diagnosed with extensive superficial dermatophytosis. Study conducted at the Department of Dermatology and Venereology, Combined Military HospitalCumilla, the study spanned from July 2022 to June 2023. During the six-week duration, patients underwent treatment with terbinafine and itraconazole, with follow-up examinations every two weeks, including relevant investigations. Evaluation of treatment outcomes occurred at both four and six weeks. Statistical analysis was performed using version 25 of the Statistical Package for the Social Sciences (SPSS). Results: This study shows the majority of the participants within the age range of 16-30 years (56.0%) and the average age of the participants is 31.74±11.77 years. In terms of gender, the majority are male (78.0%). The most prevalent diagnosis is Tinea corporis (68.0%), followed by Tinea cruris (36.0%), Tinea faciei (28.0%), and Tinea incognito (12.0%). The majority of participants (85.0%) have had the disease for 12 months or less. The most common sites are the trunk (35.0%), whole body (26.0%), and crural area (24.0%). The majority of participants experience severe itching (56.0%), followed by moderate itching (34.0%), and a smaller percentage report mild itching (10.0%). The majority of participants showed improvement, with percentages increasing from 66.0% after 2 weeks to 84.0% after 4 weeks.The percentage of cured individuals also increased over time, reaching a substantial 90.0% after 6 weeks. Conclusion: The amalgamation of terbinafine and itraconazole demonstrates a superior clinical cure rate compared to use of single systemic antifungals. However, the observed clinical cure rate percentage is lower when juxtaposed with findings from previous studies. Notably, neither combination of systemic antifungals has exhibited efficacy against tinea infections previously treated with formulations containing topical steroids

Biocontrol of Alternaria alternata in cold-stored table grapes using psychrotrophic yeasts and bioactive compounds of natural sources

Alternaria alternata is a common fungal pathogen causing postharvest decay in table grapes. This study addressed the potential of autochthonous yeasts and bioactive compounds of natural sources to act as biocontrol agents (BCAs) against A. alternata in cold-stored table grapes. With this purpose, 19 yeast capable of growing at 0–1 °C were isolated from the surface of Red Globe table grapes. These isolates, along with the pre-isolated strain Metschnikowia pulcherrima RCM2, were evaluated as BCAs in wounded berries. From these results, six yeast isolates were pre-selected to be combined with bioactive compounds of natural sources, like phenolic compounds (PCs) of side streams of wine industry, including bunch stem extract (BSE) (5–25 %), and cane extract (CE) (5–25 %), and functional polysaccharides from shrimp waste such as chitosan (CH) (0.5 %). Then, the biocontrol efficacy of combined treatments beyond individual ones was compared. The results revealed that 4 yeast isolates, namely M. pulcherrima RCM2 and ULA146, and Aureobasidium pullulans FUL14 and FUL18, were the most effective. However, when combined with the natural bioactive compounds, their efficacy against A. alternata did not increase significantly. Notably, ULA146 and FUL18 demonstrated a biocontrol efficacy of 36–37 %, comparable to that of the treatment with commercial doses of SO2, which only showed a 27 % reduction in the lesion diameter. These findings highlight the potential of using psychrotrophic yeasts as BCAs against A. alternata in cold-stored table grapes. Combining these yeast strains with BSE, CE and CH did not increase BCAs efficacy against this pathogen at the concentrations tested. The development of effective biocontrol strategies for A. alternata could contribute to reducing reliance on chemically synthesized fungicides, promoting sustainable practices, aiming to improve the quality and safety of cold-stored table grapes.

The efficacy of bacteriophage/probiotic combination therapy in periodontal treatment

Relevance. Periodontal inflammations result from chronic, persistent infections triggered by dysbiosis-induced shifts within the microbial community. The high prevalence, multifactorial nature and challenging management of periodontal disease create an environment ripe for ongoing advancements in treatment modalities.Study objective: To validate, using clinical and laboratory test data, the efficacy of combined treatment involving bacteriophages and probiotics in managing chronic periodontitis.Materials and methods. A total of 100 patients aged 18 to 45 with periodontal inflammation underwent clinical laboratory examinations before receiving combination treatment. The patients were divided into two groups: Group 1 received conventional treatment based on clinical recommendations, while Group 2 received sequential treatment with a bacteriophage cocktail (targeting 81 phages against A. actinomycetemcomitans, B. licheniformis, B. fragilis, E. cloacae, E. faecalis, K. pneumonia, S. aureus, S. pyogenes, Wolinella spp. among others) followed by a probiotic containing S. Salivarius. Periodontal pocket lavages served as study material. Periodontal pathogens were isolated using real-time PCR, and proinflammatory cytokines were identified via enzyme immunoassay. Statistical analysis was conducted using STATA v.12 software.Results. Following bacteriophage/probiotic combination treatment, positive changes in study indices and a reduction in periodontopathogenic microbiota were observed. Significant decreases were noted in the levels of key periodontal pathogens: P. gingivalis (5.2-fold decrease, p < 0.001), P. intermedia (3.6-fold decrease, p < 0.001; T. forsythia (3.8-fold decrease, p = 0.905), T. denticola 4-fold decrease, p < 0.001. Levels of IL1-β decreased by factors of 4.29 (p < 0.001) and 8.59 (p = 0.02) in severity subgroups, while IL6 levels decreased by factors of 4.39 (p < 0.001) and 5.94 (p = 0.0002). The lowest TNF-α levels (2.13 in mild CGP subgroup and 2.11 in moderate CGP subgroup) were observed post-treatment (p < 0.001; p = 0.0005).Conclusion. The combination treatment involving the selected drugs exhibited notable therapeutic effectiveness in the management of chronic periodontitis. This approach not only avoided the necessity for systemic antibacterial agents but also facilitated prolonged remission by eradicating periodontopathogenic microbiota in patients with chronic periodontitis. The etiopathogenetic nature of this treatment represents a promising paradigm in the management of periodontal inflammation.

Efficacy of combined treatment with pirfenidone and PD-L1 inhibitor in mice bearing ectopic bladder cancer xenograft

To assess the efficacy of pirfenidone combined with PD-L1 inhibitor for treatment of bladder cancer in a mouse model and its effect on tumor immune microenvironment modulation. Forty C57BL/6 mouse models bearing ectopic human bladder cancer xenografts were randomized into control group, PD-L1 inhibitor group, pirfenidone group and combined treatment group (n=10). After successful modeling, PD-L1 inhibitor treatment was administered via intraperitoneal injection at 12.5 mg/kg every 3 days, and oral pirfenidone (500 mg/kg) was given on a daily basis. The survival rate of the mice and tumor growth rate were compared among the 4 groups. The expressions of CD3, CD8, CD45, E-cadherin and N-cadherin in the tumor tissues were detected with immunohistochemistry after the 21-day treatment, and bone marrow-derived suppressor cells (MDSCs) were observed with immunofluorescence staining; serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen (BUN), creatinine (CRE) and lactate dehydrogenase (LDH-L) were analyzed using an automated biochemical analyzer. Treatment with PD-L1 inhibitor and pirfenidone alone both significantly decreased tumor growth rate and tumor volume at 21 days (P < 0.05), but the combined treatment produced an obviously stronger inhibitory effect (P < 0.05). PD-L1 inhibitor and pirfenidone alone significantly increased E- cadherin expression and decreased N-cadherin expression in the tumor tissue (P < 0.05). The two treatments both significantly increased the percentage of CD3+, CD8 and CD45+ T cells and decreased the percentage of Ly-6G+CD11b+MDSCs in the tumor tissue, and these changes were more obvious in the combined treatment group (P < 0.05). No significant differences were found in serum ALT, AST, BUN, CRE or LDH-L levels among the 4 groups (P>0.05). Combined treatment with pirfenidone and PD-L1 inhibitor significantly inhibits the progression of bladder cancer in mice possibly by regulating tumor immune microenvironment and inhibiting epithelial-mesenchymal transition of the tumor cells.

Prevention of neointimal hyperplasia after coronary artery bypass graft via local delivery of sirolimus and rosuvastatin: network pharmacology and in vivo validation

BackgroundCoronary artery bypass graft (CABG) is generally used to treat complex coronary artery disease. Treatment success is affected by neointimal hyperplasia (NIH) of graft and anastomotic sites. Although sirolimus and rosuvastatin individually inhibit NIH progression, the efficacy of combination treatment remains unknown.MethodsWe identified cross-targets associated with CABG, sirolimus, and rosuvastatin by using databases including DisGeNET and GeneCards. GO and KEGG pathway enrichment analyses were conducted using R studio, and target proteins were mapped in PPI networks using Metascape and Cytoscape. For in vivo validation, we established a balloon-injured rabbit model by inducing NIH and applied a localized perivascular drug delivery device containing sirolimus and rosuvastatin. The outcomes were evaluated at 1, 2, and 4 weeks post-surgery.ResultsWe identified 115 shared targets between sirolimus and CABG among databases, 23 between rosuvastatin and CABG, and 96 among all three. TNF, AKT1, and MMP9 were identified as shared targets. Network pharmacology predicted the stages of NIH progression and the corresponding signaling pathways linked to sirolimus (acute stage, IL6/STAT3 signaling) and rosuvastatin (chronic stage, Akt/MMP9 signaling). In vivo experiments demonstrated that the combination of sirolimus and rosuvastatin significantly suppressed NIH progression. This combination treatment also markedly decreased the expression of inflammation and Akt signaling pathway-related proteins, which was consistent with the predictions from network pharmacology analysis.ConclusionsSirolimus and rosuvastatin inhibited pro-inflammatory cytokine production during the acute stage and regulated Akt/mTOR/NF-κB/STAT3 signaling in the chronic stage of NIH progression. These potential synergistic mechanisms may optimize treatment strategies to improve long-term patency after CABG.Graphical

Tretinoin improves the anti-cancer response to cyclophosphamide, in a model-selective manner

BackgroundChemotherapy is included in treatment regimens for many solid cancers, but when administered as a single agent it is rarely curative. The addition of immune checkpoint therapy to standard chemotherapy regimens has improved response rates and increased survival in some cancers. However, most patients do not respond to treatment and immune checkpoint therapy can cause severe side effects. Therefore, there is a need for alternative immunomodulatory drugs that enhance chemotherapy.MethodsWe used gene expression data from cyclophosphamide (CY) responders and non-responders to identify existing clinically approved drugs that could phenocopy a chemosensitive tumor microenvironment (TME), and tested combination treatments in multiple murine cancer models.ResultsThe vitamin A derivative tretinoin was the top predicted upstream regulator of response to CY. Tretinoin pre-treatment induced an inflammatory, interferon-associated TME, with increased infiltration of CD8 + T cells, sensitizing the tumor to subsequent chemotherapy. However, while combination treatment significantly improved survival and cure rate in a CD4+ and CD8+ T cell dependent manner in AB1-HA murine mesothelioma, this effect was model-selective, and could not be replicated using other cell lines.ConclusionsDespite the promising data in one model, the inability to validate the efficacy of combination treatment in multiple cancer models deprioritizes tretinoin/cyclophosphamide combination therapy for clinical translation.

Combined effect of Neurotropin® and methylcobalamin on postherpetic neuralgia in mice infected with herpes simplex virus type-1

BackgroundPostherpetic pain (PHP) is difficult to control. Although Neurotropin® (NTP) and methylcobalamin (MCB) are often prescribed to treat the pain, the efficacy of combined treatment for PHP remains imcompletely understood. ObjectiveIn this study, we investigate the combined effects of NTP and MCB on PHP in mice. MethodsNTP and MCB were administered from day 10–29 after herpes simplex virus type-1 (HSV-1) infection. The pain-related responses were evaluated using a paint brush. The expression of neuropathy-related factor (ATF3) and nerve repair factors (GAP-43 and SPRR1A) in the dorsal root ganglion (DRG) and neurons in the skin were evaluated by immunohistochemical staining. Nerve growth factor (NGF) and neurotrophin-3 (NT3) mRNA expression levels were evaluated using real-time PCR. ResultsRepeated treatment with NTP and MCB after the acute phase inhibited PHP. Combined treatment with these drugs inhibited PHP at an earlier stage than either treatment alone. In the DRG of HSV-1-infected mice, MCB, but not NTP, decreased the number of cells expressing ATF3 and increased the number of cells expressing GAP-43- and SPRR1A. In addition, MCB, but not NTP, also increased and recovered non-myelinated neurons decreased in the lesional skin. NTP increased the mRNA levels of NTF3 in keratinocytes, while MCB increased that of NGF in Schwann cells. ConclusionThese results suggest that combined treatment with NTP and MCB is useful for the treatment of PHP. The combined effect may be attributed to the different analgesic mechanisms of these drugs.

Efficacy and Safety of Combination Treatment of Oral Terbinafine and Itraconazole in Patients with Extensive Superficial Dermatophytes

Efficacy and Safety of Combination Treatment of Oral Terbinafine and Itraconazole in Patients with Extensive Superficial Dermatophytes

Bortezomib in Combination with Physachenolide C Reduces the Tumorigenic Properties of KRASmut/P53mut Lung Cancer Cells by Inhibiting c-FLIP

Background: Defects in apoptosis regulation are one of the classical features of cancer cells, often associated with more aggressiveness and failure to therapeutic options. We investigated the combinatorial antitumor effects of a natural product, physachenolide C (PCC) and bortezomib, in KRASmut/P53mut lung cancer cells and xenograft mice models. Methods: The in vitro anticancer effects of the bortezomib and PCC combination were investigated using cell viability, migration, and invasion assays in 344SQ, H23, and H358 cell lines. Furthermore, the effects of combination treatment on the critical parameters of cellular metabolism, including extracellular acidification rate (ECAR) and mitochondrial oxidative phosphorylation based on the oxygen consumption rate of cancer cells were assessed using Seahorse assay. Finally, the antitumor effect of the bortezomib (1 mg/kg) and PCC (10 mg/kg) combination was evaluated using xenograft mice models. Results: Our data showed that the bortezomib–PCC combination was more effective in reducing the viability of lung cancer cells in comparison with the individual treatments. Similarly, the combination treatment showed a significant inhibition of cell migration and invasion of cancer cells. Additionally, the key anti-apoptotic protein c-FLIP was significantly inhibited along with a substantial reduction in the key parameters of cellular metabolism in cancer cells. Notably, the bortezomib or PCC inhibited the tumor growth compared to the control group, the tumor growth inhibition was much more effective when bortezomib was combined with PCC in tumor xenograft mice models. Conclusion: These findings demonstrate that PCC sensitizes cancer cells to bortezomib, potentially improving the antitumor effects against KRASmut/P53mut lung cancer cells, with an enhanced efficacy of combination treatments without causing significant side effects.

Combination therapy of high-dose intravenous anakinra and baricitinib in patients with critical COVID-19: Promising results from retrospective observational study

IntroductionIn this study, we aimed to evaluate the safety and efficacy of combination treatment of high-dose intravenous anakinra and baricitinib in patients with critically ill COVID-19. Material and methodsThis retrospective observational study was conducted in a tertiary center with diagnosis of COVID-19 patients.Study population consisted of patients with positive polymerase chain reaction and computer tomography findings compatible with COVID-19 as well as critical illness. ResultsData of 15 patients in combination group and 43 patients in control group were evaluated and included into the study. Overall mortality was 46.7 % (n = 7) in combination arm and 69.8 % (n = 30) in control group although it was not statistically significant (p = 0.1). Similarly, need of intubation was also lower in combination arm (46.7 %) compared to control group (69.8 %), it was not significantly different (p = 0.1). ICU admission was significantly lower in combination (46.7 %, n = 7) arm than control group (76.7 %, n = 33) (p = 0.03, Odds ratio [OR]:4.7). Development of severe infection (20 %, n = 3 vs 25 %, n = 9/36), pulmonary embolism (6.7 %, n = 1 vs 0), myocardial infarction (6.7 %, n = 1 vs 2.6 %, n = 1/38) and pneumothorax (13.3 %, n = 2 vs 2.6 %, n = 1/38) were not different between two groups (p = 0.7, p = 0.3, p = 0.5 and p = 0.2). In multivariable analysis only cHIS score was associated with high mortality (p = 0.018, OR:2.8, [95 % confidence interval: 1.2–6.6]). In survival analysis, mortality rate was significantly lower in combination arm than control group (Log-Rank:p = 0.04). ConclusionCombination therapy of high-dose anakinra and baricitinib may be an adequate treatment option in patients with COVID-19 who had critical disease and has acceptable safety profile.

Synergistic and Additive Effects of Herbal Medicines in Combination with Chemotherapeutics: A Scoping Review.

Natural products are increasingly gaining interest as potential new drug candidates for cancer treatment. Herbal formula, which are combinations of several herbs, are primarily used in East Asia and have a long history of use that continues today. Recently, research exploring the combination of herbal formulas and chemotherapy for cancer treatment has been on the rise. This study reviewed research on the co-administration of herbal formulas and chemotherapy for cancer treatment. The databases PubMed, Embase, and Cochrane Library were used for article searches. The following keywords were employed: "Antineoplastic agents," "Chemotherapy," "Phytotherapy," "Herbal medicine," "Drug synergism," and "Synergistic effect." The selection process focused on studies that investigated the synergistic interaction between herbal formulas and chemotherapeutic agents. Among the 30 studies included, 25 herbal formulas and 7 chemotherapies were used. The chemotherapy agents co-administered included cisplatin, 5-fluorouracil, docetaxel, doxorubicin, oxaliplatin, irinotecan, and gemcitabine. The types of cancer most frequently studied were lung, breast, and colon cancers. Most studies evaluating the anticancer efficacy of combined herbal formula and chemotherapy treatment were conducted in vitro or in vivo. Most studies reported synergistic effects on cytotoxicity, apoptosis, and tumor growth inhibition. These effects were found to be associated with cell cycle arrest, anti-angiogenesis, and gene expression regulation. Further studies leading to clinical trials are required. Clinical experiences in East Asian countries could provide insights for future research.