Low-dose Combination Research Articles

FOLFOXIRI Resistance Induction and Characterization in Human Colorectal Cancer Cells.

Simple SummaryWe created colorectal cancer in vitro models to study how an induced drug resistance profile can alter cell response and sensitivity to a treatment. By chronically exposing the cells to current first-line treatments (5-FU+folinic acid+oxaliplatin+SN38), resistance to the chemotherapy was obtained. We further investigated the mechanism underlying the acquired chemoresistance and highlighted the main up- and downregulated genes implicated. We also showed that optimized drug combination composed of tyrosine kinase inhibitors overcome chemotherapy-induced resistance.FOLFOXIRI, i.e., the combination of folinic acid, 5-fluorouracil, oxaliplatin, and irinotecan, is a first-line treatment for colorectal carcinoma (CRC), yet non-personalized and aggressive. In this study, to mimic the clinical situation of patients diagnosed with advanced CRC and exposed to a chronic treatment with FOLFOXIRI, we have generated the CRC cell clones chronically treated with FOLFOXIRI. A significant loss in sensitivity to FOLFOXIRI was obtained in all four cell lines, compared to their treatment-naïve calls, as shown in 2D cultures and heterotypic 3D co-cultures. Acquired drug resistance induction was observed through morphometric changes in terms of the organization of the actin filament. Bulk RNA sequencing revealed important upregulation of glucose transporter family 5 (GLUT5) in SW620 resistant cell line, while in the LS174T-resistant cell line, a significant downregulation of protein tyrosine phosphatase receptor S (PTPRS) and oxoglutarate dehydrogenase-like gene (OGDHL). This acquired resistance to FOLFOXIRI was overcome with optimized low-dose synergistic drug combinations (ODCs) acting via the Ras-Raf-MEK-ERK pathway. The ODCs inhibited the cell metabolic activity in SW620 and LS174T 3Dcc, respectively by up to 82%.

Trio-Drug Combination of Sodium Valproate, Baclofen and Thymoquinone Exhibits Synergistic Anticonvulsant Effects in Rats and Neuro-Protective Effects in HEK-293 Cells.

Epilepsy is a chronic brain disorder, with anti-epileptic drugs (AEDs) providing relief from hyper-excitability of neurons, but largely failing to restrain neurodegeneration. We investigated a progressive preclinical trial in rats, whereby the test drugs; sodium valproate (SVP; 150 and 300 mg/kg), baclofen (BFN; 5 and 10 mg/kg), and thymoquinone (THQ; 40 and 80 mg/kg) were administered (i.p, once/day for 15 days) alone, and as low dose combinations, and subsequently tested for antiseizure and neuroprotective potential using electrical stimulation of neurons by Maximal electroshock (MES). The seizure stages were monitored, and hippocampal levels of m-TOR, IL-1β, IL-6 were measured. Hippocampal histopathology was also performed. Invitro and Insilco studies were run to counter-confirm the results from rodent studies. We report the synergistic effect of trio-drug combination; SVP (150 mg/kg), BFN (5 mg/kg) and THQ (40 mg/kg) against generalized seizures. The Insilco results revealed that trio-drug combination binds the Akt active site as a supramolecular complex, which could have served as a delivery system that affects the penetration and the binding to the new target. The potential energy of the ternary complex in the Akt active site after dynamics simulation was found to be −370.426 Kcal/mol, while the supramolecular ternary complex alone was −38.732 Kcal/mol, with a potential energy difference of −331.694 Kcal/mol, which favors the supramolecular ternary complex at Akt active site binding. In addition, the said combination increased cell viability by 267% and reduced morphological changes induced by Pentylenetetrazol (PTZ) in HEK-293 cells, which indicates the neuroprotective property of said combination. To conclude, we are the first to report the anti-convulsant and neuroprotective potential of the trio-drug combination.

Combination therapy of solifenacin and mirabegron vs monotherapy in overactive bladder

Introduction and Objectives: Overactive bladder (OAB) with symptoms of frequency, urgency with or without urgency incontinence and nocturia is one of the most bothersome symptom affecting the quality of life of both men and women. Antimuscarinics and Beta 3 agonists both play an important role in reducing the bothersome symptoms. In this study we find the efficacy of low dose combination therapy of solifenacin 5 mg with mirabegron 25 mg vs the monotherapy solifenacin 10 mg or Mirabegron 50 mg.Materials and Methods: It is a prospective study conducted on 150 patients at Madras medical college and were divided into 3 groups. Group A received solifenacin 10 mg/day, Group B received solifenacin 5 mg/day + Mirabegron 25 mg/day and Group C received Mirabegron 50 mg/day. Patients were evaluated for improvement in frequency, urgency, leak episodes, voided volume and improvement in quality of life after a period of 4 weeks and 12 weeks. The three groups were compared using anova test and chi square test and p value was calculated.Results and Observations: Majority of patients of OAB belong to age group > 50 years and majority were females showing female predilection towards OAB. The mean duration of symptoms was 36.01± 7.91 months. Compared with monotherapy, the combination therapy resulted in a significant improvement in reducing the mean number of micturition/24hr from baseline to End of Treatment (EoT). The frequency of urgency episodes and mean number of nocturia episodes/24hr was significantly reduced in patients taking combination therapy as compared to monotherapy. In all the groups a decrease in mean number of incontinence episodes/24hr was observed from baseline to EoT but was not statistically significant. The frequency of antimuscarinic side effects (dry mouth, constipation, urinary retention) had lower incidence in combination therapy compared with solifenacin group. Conclusions: Combination therapy of solifenacin and mirabegron demonstrated significant improvement over monotherapy (solifenacin 10 mg & mirabegron 50 mg) in frequency of micturition, urgency and nocturia episodes without increasing bothersome adverse effects associated with antimuscarinic therapy.

Low-dose aspirin and rivaroxaban combination therapy to overcome aspirin non-sensitivity in patients with vascular disease.

Approximately 20% of vascular patients treated with acetyl salicylic acid (i.e., aspirin) demonstrate less than expected platelet inhibition – putting them at a four-fold increased risk of adverse cardiovascular events. Low-dose rivaroxaban (2.5 mg twice daily) in combination with low-dose aspirin has been shown to reduce adverse cardiovascular and limb events when compared to aspirin alone. In this study, light transmission aggregometry was used to measure arachidonic acid-induced platelet aggregation to evaluate the potential of combining low-dose rivaroxaban and aspirin in attenuating or overcoming aspirin non-sensitivity. In the discovery phase, 83 patients with peripheral arterial disease (PAD) taking 81 mg aspirin daily were recruited from the outpatient vascular surgery clinic at St Michael's Hospital between January to September 2021. 19 (23%) were determined to be non-sensitive to aspirin. After ex-vivo addition of 2.5 mg dosage equivalent of rivaroxaban, aspirin non-sensitivity was overcome in 11 (58%) of these 19 patients. In the validation phase, 58 patients with cardiovascular risk factors who were not previously prescribed aspirin were recruited. In this group, ex-vivo addition of 2.5 mg dosage equivalent of rivaroxaban significantly reduced arachidonic acid-induced platelet aggregation in the presence of aspirin. These results demonstrate the potential for low-dose rivaroxaban to overcome aspirin non-sensitivity in patients with PAD. Further studies are needed to evaluate and confirm these findings.

Metformin Enhances the Anti-Cancer Efficacy of Sorafenib via Suppressing MAPK/ERK/Stat3 Axis in Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) incidence, as well as related mortality, has been steadily increasing in the USA and across the globe, partly due to the lack of effective therapeutic options for advanced HCC. Though sorafenib is considered standard-of-care for advanced HCC, it only improves median survival by a few months when compared to placebo. Sorafenib is also associated with several unpleasant side effects that often lead to early abatement of therapy. Here, we investigate whether a combination regimen including low-dose sorafenib and a non-toxic dose of anti-diabetic drug metformin can achieve effective inhibition of HCC. Indeed, combining metformin with low-dose sorafenib inhibited growth, proliferation, migration, and invasion potential of HCC cells. We observed a 5.3- and 1.9-fold increase in sub-G1 population in the combination treatment compared to sorafenib alone. We found that the combination of metformin enhanced the efficacy of sorafenib and inhibited the MAPK/ERK/Stat3 axis. Our in vivo studies corroborated the in vitro findings, and mice harboring HepG2-derived tumors showed effective tumor reduction upon treatment with low-dose sorafenib and metformin combination. This work sheds light on a therapeutic strategy aiming to augment sorafenib efficacy or dose-de-escalation that may prove beneficial in circumventing sorafenib resistance as well as minimizing related side effects.

Evaluation of the effectiveness of a combination of incoherent broadband light (IPL) and low doses of systemic isotretinoin compared with monotherapy with standard doses of isotretinoin in patients with acne

BACKGROUND: Long-term experience of clinical use of isotretinoin for acne indicates that it has a good safety and efficacy profile; however, against the background of taking standard doses of isotretinoin, pronounced adverse events that are dose-dependent are noted. Therefore, the search for new combined treatment regimens that reduce the dose of isotretinoin is an urgent task.
 AIMS: The aim of our work was to evaluate the effectiveness of a combination of incoherent broadband light (IPL) and low doses of systemic isotretinoin (0.10.3 mg/kg of body weight per day) compared with monotherapy with standard doses of isotretinoin (0.51 mg/kg of body weight per day) in patients with acne.
 MATERIAL AND METHODS: During the study, the dermatological status of patients was assessed taking into account the dermatological index GSS (Global Severity Scog), dermatological acne index (DIA), IGA scale (Investigator's Global Assessment). For a comparative analysis of the impact of various techniques on the quality of life, a high-valid quality of life index (HQIQ) was used. Dermatological status and quality of life were assessed in patients before and after 6 months of therapy. 12 months after the end of therapy, patients were invited to the clinic to identify delayed relapses.
 RESULTS: Evaluating the overall results of treatment of patients with moderate severity of acne by isotretinone monotherapy and combination therapy with low doses of isotretinoin and broadband incoherent light, the effectiveness of both methods can be noted, however, according to the global severity scale (GSS), clinical remission was significantly more often observed in the combination therapy group compared with the motor therapy group (88 and 74%, respectively, p 0.05). In addition, no relapses were detected after 12 months in the combination therapy group, and adverse events caused by the use of isotretionine were also less frequent.
 CONCLUSION: Thus, the combination therapy of acne with low doses of isotretinoin and broadband incoherent light is more effective.

Abstract 5850: p53 gene expression and nitric oxide levels after artemisinin-caffeine treatment in breast, lungs and liver of DMBA-induced tumorigenesis

Abstract Background: With increasing incidence of cancers globally and limited resources in some affected countries, repurposing existing drugs for reducing tumorigenesis is highly important. Artemisinin and caffeine have potent anti-oxidative and anti-cancer properties, although they are therapies for other diseases. When p53 is activated, it repairs DNA damage, prevents tumorigenesis and induces apoptosis whereas nitric oxide is a pro-apoptotic modulator which can upregulate p53 thereby reducing tumorigenesis. We evaluated the biochemical and p53 gene modulatory effects of doses of artemisinin-caffeine combination on breast, lung and liver tumors in rats induced with DMBA. Methods: After due ethical approval, 30 animals were treated with 40mg/kg single dose of 7,12-dimethylbenzene anthracene (DMBA) as a model for DNA damage and induction of carcinogenesis. Five animals each received low dose artemisinin (4mg/kg), low dose caffeine (25mg/kg), low dose combination of Art + Caff (4+25mg/kg), high dose combination of Art + Caff (8+50 mg/kg) or no treatment. All treatment doses were orally administered daily for two weeks post-DMBA treatment, normal control received saline. Nitric oxide levels were assessed and p53 relative gene expression was determined by primer-specific RT-PCR (GAPDH was used as loading control) and amplicons were resolved by gel electrophoresis. Results: DMBA induced lesions in breast, liver, and lung tissues were evident from histology analysis, compared to untreated group. In all 3 tissues, low dose caffeine and combination of Art + Caff significantly reduced p53 gene expression (p < 0.05), but there was significant increase in of p53 expression in low dose Art and high dose Art + Caff groups, which were similar to DMBA group (p<0.05). There was also decrease in nitric oxide levels in treated groups compared to DMBA group. Conclusions: This study shows a dose-dependent anticancer effects of caffeine and artemisinin combination on p53 gene and nitric oxide regulation and hence can mitigate against tumorigenesis. Citation Format: Titilope M. Dokunmu, Sandra C. Opara, Omiete U. Awani. p53 gene expression and nitric oxide levels after artemisinin-caffeine treatment in breast, lungs and liver of DMBA-induced tumorigenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5850.

Abstract 3095: Breaking the crosstalk of the cellular tumorigenic network in NSCLC by a highly effective drug combination

Abstract Background: Non-small cell lung cancer (NSCLC) is commonly diagnosed at advanced stages with limited treatment options particularly in tumors without targetable driver mutations. Methods: Considering previously published criteria for selecting drug combinations for overcoming drug resistances1, we have established a low dose combination therapy with cabozantinib, afatinib, etoricoxib and plerixafor. Utilizing NSCLC patient-derived xenograft (PDX) models we aimed to use this drug regimen to disrupt paracrine substitutional signaling within the tumorigenic network in order to circumvent pre-existing and adaptive drug resistances. In contrast to daily dosing for respective monotherapies the low dose treatment was paused every five days for two days. Result: All PDX tumors treated, including highly therapy-resistant adeno- and squamous cell carcinomas without identifiable driver mutations, were completely suppressed by this drug regimen leading to an Objective Response Rate of 81% and a Clinical Benefit Rate of 100%. After treatment discontinuation growth suppression remained until the end of the follow-up period with only slight increase in tumor growth. The application and the safety profile of this low dose therapy regimen was well manageable and without pronounced side effects in this pre-clinical setting. Discussion: For the first time we show that simultaneous inhibition of pathways forming the cellular tumorigenic network in NSCLC tumors is highly effective and overcomes drug resistances for monotherapies. All of the drugs used in this combination regimen are approved for different indications and therefore readily available. Conclusion: Our results emphatically encourage a setup for a clinical study in advanced stage NSCLC patients without targetable driver mutations combining the drugs cabozantinib, afatinib, plerixafor and etoricoxib in a low dose treatment regimen as novel therapy strategy. Indication of Source: 1. Langhammer S and Scheerer J. Oncotarget 8, 43555-43570 (2017). Citation Format: Dennis Gürgen, Theresia Conrad, Michael Becker, Susanne Sebens, Christoph Röcken, Jens Hoffmann, Stefan Langhammer. Breaking the crosstalk of the cellular tumorigenic network in NSCLC by a highly effective drug combination [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3095.

Abstract 5631: Immunomodulatory activity of sulindac plus erlotinib for colon cancer prevention in familial adenomatous polyposis

Abstract Familial adenomatous polyposis (FAP) is an autosomal dominant disorder caused by germline mutation in Adenomatous Polyposis Coli (APC)1. Patients with FAP have a nearly 100% lifetime risk of colorectal cancer (CRC) due to the formation of hundreds to thousands of adenomatous polyps in the gastrointestinal tract2. A clinical trial in FAP patients demonstrated that Cox-1/2 inhibitor sulindac (SUL) plus EGFR inhibitor erlotinib (ERL) reduced polyp burden significantly in the colon and small intestine (SI), but toxicity issues raised concerns for durable long-term CRC interception3. In the Apc-mutant polyposis in rat colon (Pirc) rat model of FAP, we determined low-dose regimens of ERL+SUL that were both safe and efficacious4. In the current investigation, the lowest tested dose of dietary SUL (125 ppm continuously) plus ERL (5 mg/kg twice weekly), given at one-half the SUL clinical dose and one-fourth of the ERL clinical dose, administered for 46 weeks respectively, demonstrated good inhibition of tumor burden in the colon (>85% efficacy, p<0.05) and SI (>94% efficacy, p<0.05) compared to untreated controls. Moreover, treatment normalized organ weights and hematocrits that were altered in the untreated AIN controls.Recently, ERL5 and SUL6 were individually demonstrated to increase the recruitment of inflammatory immune cells in mouse models of lung and colon cancer, respectively. However, ERL+SUL in combination was not investigated in the context of immune cell infiltration and polarization. Our preliminary IHC data in the Pirc model indicated increased recruitment of CD8+ T cells in colon polyps by SUL 250 ppm + ERL 42 mg/kg once a week. RT-qPCR and immunoblotting analysis of colon polyps obtained from the low-dose combination of SUL 125 ppm + ERL 5 mg/kg twice weekly demonstrated increased CD4 expression and concurrently decreased oncogenic IL-17 expression and tumor associated macrophage markers (CD163 and CD206) (P<0.05 vs control). Additional immune markers will be identified and validated via CyTOF. We hypothesize that low-dose combination of SUL+ERL can enhance the recruitment of immune cells into early adenomatous colon and SI polyps, providing an effective immunointerception strategy, with potential clinical relevance for FAP patients before or after colectomy. Acknowledgements Research supported by NCI Contract Number 75N91019D00021, Task Order 75N91019F00130. 1.Groden J et al., Cell 1991; 66:589-600.2.Jasperson KW et al., Gastroenterology 2010; 138:2044-2058.3.Samadder NJ et. al., JAMA 2016; 315:1266-75.4.Ulusan AM et. al., Cancer Prev Res (Phila). 2021; 14:325-336.5.Ayeni D et al., J Immunother Cancer 2019; 7:172.6.Yi B et al., Mol Cancer Ther 2021; Apr 20. Citation Format: Chakrapani Tripathi, Roderick H. Dashwood. Immunomodulatory activity of sulindac plus erlotinib for colon cancer prevention in familial adenomatous polyposis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5631.

Dietary Collagen Hydrolysates Retard Estrogen Deficiency-Induced Bone Loss through Blocking Osteoclastic Activation and Enhancing Osteoblastic Matrix Mineralization

Osteoporosis manifest in postmenopausal women is an osteolytic disease characterized by bone loss, leading to increased susceptibility to bone fractures and frailty. The use of complementary therapies to alleviate postmenopausal osteoporosis is fairly widespread among women. The current study examined that Pangasius hypophthalmus fish skin collagen hydrolysates (fsCH) inhibited ovariectomy (OVX)-induced bone loss by conducting inter-comparative experiments for anti-osteoporotic activity among 206–618 mg/kg fsCH, 2 mg/kg isoflavone, 15 mg/kg glycine–proline–hydroxyproline (GPH) tripeptide, and calcium lactate. Surgical estrogen loss of mice for 8 weeks reduced serum 17β-estradiol levels with uterus atrophy, which was ameliorated by orally administering fsCH or isoflavone to mice. Similar to isoflavone, fsCH containing GPH-enhanced bone mineral density reduced levels of cathepsin K and proton-handling proteins, and elevated collagen 1 level in OVX bones. The treatment with fsCH and isoflavone enhanced the serum levels of collagen synthesis-related procollagen type 1 carboxy/amino-terminal propeptides reduced by OVX, whereas serum levels of osteocalcin and alkaline phosphatase, as well as collagen breakdown-related carboxy/amino-terminal telopeptides of type 1 collagen were reduced in OVX mice treated with fsCH, isoflavone, and calcium lactate. The trabecular bones were newly formed in OVX bones treated with isoflavone and fsCH, but not with calcium lactate. However, a low-dose combination of fsCH and calcium lactate had a beneficial synergy effect on postmenopausal osteoporosis. Furthermore, similar to isoflavone, 15–70 μg/mL fsCH, with its constituents of GPH and dipeptides of glycine–proline and proline–hydroxyproline, enhanced osteogenesis through stimulating differentiation, matrix mineralization, and calcium deposition of MC3T3-E1 osteoblasts. Accordingly, the presence of fsCH may encumber estrogen deficiency-induced bone loss through enhancing osteoclastogenic differentiation and matrix collagen synthesis. Therefore, fsCH may be a natural compound retarding postmenopausal osteoporosis and pathological osteoresorptive disorders.

Albuterol–Budesonide Fixed-Dose Combination Rescue Inhaler for Asthma

BackgroundAs asthma symptoms worsen, patients typically rely on short-acting β2-agonist (SABA) rescue therapy, but SABAs do not address worsening inflammation, which leaves patients at risk for severe asthma exacerbations. The use of a fixed-dose combination of albuterol and budesonide, as compared with albuterol alone, as rescue medication might reduce the risk of severe asthma exacerbation.MethodsWe conducted a multinational, phase 3, double-blind, randomized, event-driven trial to evaluate the efficacy and safety of albuterol–budesonide, as compared with albuterol alone, as rescue medication in patients with uncontrolled moderate-to-severe asthma who were receiving inhaled glucocorticoid-containing maintenance therapies, which were continued throughout the trial. Adults and adolescents (≥12 years of age) were randomly assigned in a 1:1:1 ratio to one of three trial groups: a fixed-dose combination of 180 μg of albuterol and 160 μg of budesonide (with each dose consisting of two actuations of 90 μg and 80 μg, respectively [the higher-dose combination group]), a fixed-dose combination of 180 μg of albuterol and 80 μg of budesonide (with each dose consisting of two actuations of 90 μg and 40 μg, respectively [the lower-dose combination group]), or 180 μg of albuterol (with each dose consisting of two actuations of 90 μg [the albuterol-alone group]). Children 4 to 11 years of age were randomly assigned to only the lower-dose combination group or the albuterol-alone group. The primary efficacy end point was the first event of severe asthma exacerbation in a time-to-event analysis, which was performed in the intention-to-treat population.ResultsA total of 3132 patients underwent randomization, among whom 97% were 12 years of age or older. The risk of severe asthma exacerbation was significantly lower, by 26%, in the higher-dose combination group than in the albuterol-alone group (hazard ratio, 0.74; 95% confidence interval [CI], 0.62 to 0.89; P=0.001). The hazard ratio in the lower-dose combination group, as compared with the albuterol-alone group, was 0.84 (95% CI, 0.71 to 1.00; P=0.052). The incidence of adverse events was similar in the three trial groups.ConclusionsThe risk of severe asthma exacerbation was significantly lower with as-needed use of a fixed-dose combination of 180 μg of albuterol and 160 μg of budesonide than with as-needed use of albuterol alone among patients with uncontrolled moderate-to-severe asthma who were receiving a wide range of inhaled glucocorticoid-containing maintenance therapies. (Funded by Avillion; MANDALA ClinicalTrials.gov number, NCT03769090.)

Synergistic cytostatic effect of bleomycin with cisplatin on Hela cells.

e15002 Background: Bleomycin is a glycopeptide antibiotic with anticancer properties. Clinical data indicate a relatively low efficacy of bleomycin in mono regimen, which is also accompanied by side effects from high doses. That’s why so it is more often used in relatively low doses in combination with other chemotherapy drugs. In particular, in cervical cancer, bleomycin is used in combination with cisplatin in a dose ratio of 0.8:1. Despite good clinical results, it is not known how the two drugs interact in vitro. The aim of this study was to evaluate the effect of combined application of bleomycin and cisplatin on HeLa cells. Methods: HeLa cells were seeded at 3000 cells per well in a 96-well plate (Eppendorf, Germany) and incubated for 24 hours in DMEM medium (Gibco, USA) supplemented with 10% FBS (HyClone, USA) at 37°C in an atmosphere containing 5.0% CO2. After 24 hours, the culture medium was replaced with a medium containing chemotherapy drugs in combination or as standalone agents in a series of two-fold dilutions: 0.03 - 32 µg/mL cisplatin and 0.012 – 12.8 µg/mL bleomycin. After 2 h exposition to studied compounds the medium was replaced and cells were incubated for another 72 hours. At the end of the experiment, the number of living cells was assessed using the MTT test. Viability was determined as the number of living cells in % of the control without the addition of chemotherapy drugs. Results: The combination of cisplatin with bleomycin demonstrated significantly greater efficacy than pure cisplatin in the concentration range from 0.03 µg/ml to 8 µg/ml, and pure bleomycin in the range of 0.012 - 12.8 µg/ml. The difference between the mean viability values for each concentration of pure bleomycin or pure cisplatin and their 0.8:1 mixture was significant at α = 0.05 (df = 14). Observed decrease in the viability of HeLa cells in comparison with pure substances is most pronounced in the range of non-toxic concentrations for both substances in mono regimen, namely 0.03 - 0.5 μg/ml for cisplatin, and at concentrations less than 0.125 μg/ml for bleomycin. Here, the increase in the difference between the dose-response curves grows as the concentration of the studied drugs increases. At cisplatin concentrations of 0.5–32 μg/ml, the difference in HeLa cell viability between pure cisplatin and its combination with bleomycin, on the contrary, decreases linearly with increasing cisplatin concentration, becoming insignificant above 16 μg/ml. Conclusions: The combined use of bleomycin and cisplatin on HeLa culture in a dose ratio of 0.8:1 exhibits synergistic properties that are most pronounced in the concentration ranges of both substances that do not have a cytotoxic effect in mono regimen.

Toxicological Sequelae of Pesticide Combinations Exposure in Buffalo Mesenchymal Stem Cells under <i>In Vitro</i>

The presence of one or more pesticides in a variety of mediums is responsible for their indirect toxicological events leading to cell senescence. In the present investigation, the endeavor was made to see the effect of pesticides Car- Benda-Zim (CBZ) and IMIdacloprid (IMI) alone and in combination with bone marrow-derived Mesenchymal Stem Cells (bMSCs) of buffalo origin. Isolated and cultured bMSCs were exposed to CBZ and IMI alone and in combinations at lower doses. Cells were observed for alterations in cell morphology, oxidative stress, mitochondrial damage and cellular senescence. bMSCs characterized for stem cell surface markers and found to be positive for AP, CD73 and OCT4. bMSCs exposed to IC25, IC12.5 and IC6.25 CBZ and IMI alone and combinations of IC12.5 and IC6.25 of CBZ and IMI. Results revealed significant reduction (p?0.05) in cell viability noticed on microscopic examination along with loss of normal cell morphology and increased in Reactive Oxygen Species (ROS) positive cells, cells with loss of ??m and number of senescent cells in CBZ and IMI treated groups. Lower dose combination groups showed elevated effects when compared with higher dose alone treated groups and control groups. Present findings suggest that CBZ and IMI induced cytotoxicity in bMSCs mediated via ROS production, altered ??m leading to the cell damage and predisposing senescence process. Moreover, the co-existence of CBZ and IMI in a medium has a considerably more toxic effect than their individual effect.

Topiramate and Acetazolamide Combination a Comparative Study between High and Low Dose Profile of Side Effects on Metabolism

Background: Topiramate, with multiple putative anticonvulsant pathways, is a relatively recent antiepileptic drug. The ability to inhibit carbonic anhydrase, a property common with the acetazolamide anticonvulsant, is among them. Aim: The study refers to compare between the use of combinations of topiramate and acetazolamide high and low doses of drugs and which one is safer for the patients. Methods: Twenty stable adult males wistar rats three weeks after acclimatization and uniformly split into two groups (10 rats in each group) and 60 days of treatment: 9 mg/kg/B.W of Topiramate and 30 mg/kg/B.W of Acetazolamide in the high dose (Hd) group and 5 mg/kg B.W of Topiramate and 30 mg/kg B.W of Acetazolamide in the low dose (Ld) group, orally/day in the high dose (Hd) group. Results: No major differences in mean Glutamate pyruvate transaminase, Glutamic oxaloacetic transaminase, Alkaline phosphatase concentration were found in the statistical analysis of the results in all experimental groups compared with each other at the pretreated period, also revealed that intubation of rats in the 60-days Hd induced a substantial (P>0.05) rise in the mean Glutamate Pyruvate Transaminase and Glutamic Oxaloacetic Transaminase serum concentrations relative to the control and Ld classes. Treatment of low-dose rats showed a substantial (P<0.05) decrease in serum alanine aminotransferase, aspartate aminotransferase, blood urea nitrogen and creatinine enzyme concentrations at 30 and 60 days of the trial compared with the T1 group. Conclusion: The study showed that high-dose combinations of topiramate and acetazolamide had the same adverse effect somewhat in low-dose combinations.

Preliminary Study on the Combination Effect of Clindamycin and Low Dose Trimethoprim-Sulfamethoxazole on Severe Pneumocystis Pneumonia After Renal Transplantation.

ObjectiveEvaluate the effect of the combination of clindamycin with low-dose trimethoprim-sulfamethoxazole (TMP/SMX) regimen on sever Pneumocystis pneumonia (PCP) after renal transplantation.Method20 severe PCP patients after renal transplantation were included in this historical-control, retrospective study. A 10 patients were treated with the standard dose of TMP/SMX (T group), the other 10 patients were treated with the combination of clindamycin and low dose TMP/SMX (CT group).ResultsAlthough there was no significant difference in the hospital survival between the two groups, the CT protocol improved the PaO2/FiO2 ratio more significantly and rapidly after the 6th ICU day (1.51 vs. 0.38, P = 0.014). CT protocol also ameliorated the pulmonary infiltration and the lactate dehydrogenase level more effectively. Moreover, the CT protocol reduced the incidence of pneumomediastinum (0 vs. 50%, P = 0.008), the length of hospital staying (26.5 vs. 39.0 days, P = 0.011) and ICU staying (12.5 vs. 22.5 days, P = 0.008). Furthermore, more thrombocytopenia (9/10 vs. 3/10, P = 0.020) was emerged in the T group than in the CT group. The total adverse reaction rate was much lower in the CT group than in the T group (8/80 vs. 27/80, P < 0.001). Consequently, the dosage of TMP/SMX was reduced in 8 patients, while only 2 patients in the CT group received TMP/SMX decrement (P = 0.023).ConclusionThe current study proposed that clindamycin combined with low-dose TMP/SMX was more effective and safer the than single use of TMP/SMX for severe PCP patients after renal transplantation (NCT 04328688).

Synergistic Effects of Resveratrol Derivatives with Bortezomib on Reducing Viability and Inducing Apoptosis in Multiple Myeloma Cells

Multiple myeloma (MM) is a cancer of plasma cells in the bone marrow characterized by bone lesions, hypercalcemia, anemia, and renal failure. Bortezomib (BTZ), a common treatment for MM, is a proteasome inhibitor that induces apoptosis in MM cells. However, high doses of BTZ can be toxic, signifying a need for a synergistic drug combination. Resveratrol (RES) is a phenolic compound found in grapes shown to inhibit MM cell growth and induce apoptosis in these cells. We hypothesize that combining BTZ with a RES derivative at certain concentrations is synergistic in reducing viability and inducing apoptosis in MM cells. To address this hypothesis, we performed viability assays to assess the effects of increasing concentrations of RES derivatives and BTZ on human MM cell viability over time (24, 48, 72 hours) and western blots to assess the compounds’ effects on inducing apoptosis in MM cells following 24 hour treatment. We found that BTZ as well as RES and its derivatives pinostilbene (PIN) and piceatannol (PIC) decreased MM cell viability in a dose‐ and time‐dependent manner and increased expression of cleaved proapoptotic proteins PARP1 and caspase‐3 in a dose‐dependent manner. We screened different combinations of RES derivatives with BTZ for synergistic effects on MM viability using CompuSyn and selected the combination of 5 nM BTZ and 5 μM PIN for further assessment as a synergistic treatment. In conclusion, a combination of low doses of BTZ and PIN synergistically decreases MM cell viability, possibly through inducing apoptosis.

PH-sensitive liposomes for colonic co-delivery of mesalazine and curcumin for the treatment of ulcerative colitis

The ineffectiveness of the current management of ulcerative colitis (UC) demands novel pharmaceutical techniques and advance therapeutic strategies to facilitate safe and efficacious treatment of the disease. In this quest, mesalazine (MZ), a 5-aminosalicylate with anti-inflammatory and potent antioxidant activities, along with curcumin (CM), a natural anti-inflammatory and antioxidant polyphenol, were found to produce a synergistic efficacy to efficiently alleviate UC. In the present study, we have formulated pH-sensitive liposomes (LS) containing a low-dose combination of MZ and CM as a potential therapy for UC. It was hypothesized that the synergistic action of the two drugs, pH-dependent colon-specific delivery, and use of LS as a nano-technological drug carrier system could diminish drug-associated side effects, reduce the dose and provide effective treatment strategy for UC. The guinea pig model of UC was used to evaluate the in vivo efficacy of pH sensitive MZ-CM co-loaded LS and found to be more efficacious than single-drug LS or drug solution. Furthermore, both drugs exhibited high antioxidant activity and mitigated the oxidative stress present along UC; resulting in an effective treatment of UC. In conclusion, eudragit-S100 coated MZ-CM liposomes is a promising treatment strategy for the effective oral therapy of UC.

Minnelide synergizes with conventional chemotherapy by targeting both cancer and associated stroma components in pancreatic cancer

Addition of nab-paclitaxel to gemcitabine offers a survival benefit of only 6 weeks over gemcitabine alone at a cost of increased toxicity in PDAC. The goal of the present study is to evaluate the efficacy of Minnelide, a water-soluble prodrug of triptolide, in combination with the standard of care regimen for chemotherapy with the added advantage of reducing the doses of these drugs to minimize toxicity. Pancreatic cancer cell lines were implanted subcutaneously or orthotopically in athymic nude or C57BL/6J mice. Subsequently, animals were randomized and received saline or minnelide or full dose chemotherapy or low dose chemotherapy or minnelide in combination with low dose chemotherapy. Our results show that a combination of low doses of Minnelide with Gemcitabine + nab-paclitaxel significantly inhibited tumor progression and increased the survival of tumor-bearing mice in comparison with conventional chemotherapy alone. Moreover, combination therapy significantly reduced cancer-related morbidity by decreasing ascites and metastasis and effectively targeted both cancer and the associated stroma. In vitro studies with a combination of low doses of triptolide and paclitaxel significantly decreased the cell viability, increased apoptosis and led to significantly increased M-phase cell cycle arrest in various pancreatic cancer cell lines as compared to either drug alone. Our results show that Minnelide synergizes with conventional chemotherapy leading to a significant reduction in the doses of these toxic drugs, all the while achieving better efficacy in the treatment of PDAC. This combination effectively targeted both the cancer and the associated stromal components of pancreatic cancer.

Effect of Diclofenac and Andrographolide Combination on Carrageenan-Induced Paw Edema and Hyperalgesia in Rats.

Studies into drug combination at low doses are a promising approach to the management of pain and inflammation. The aim of this study was to evaluate the anti-edema and anti-hyperalgesic effects of a combination of diclofenac and andrographolide. Male Sprague-Dawley rats were first treated with diclofenac or andrographolide alone (3–100 mg/kg), as well as a combination of the 2 drugs. Carrageenan was then injected into the right hind paw of rats, and changes in paw volume and sensitivity to mechanical (von Frey) and thermal (Hargreaves test) stimuli measured. Results showed drug combination produced synergistic effects at reducing paw edema especially at lower doses, with a Loewe synergy score of 13.02 ± 8.75 in SynergyFinder and a combination index of .41 ± .18 after isobolographic analysis. Again synergy scores for decreasing response to 1.0 and 3.6 g force application of von Frey filaments after drug combination were 10.127 ± 5.68 and 8.554 ± 6.53, respectively, in SynergyFinder. Synergistic effects were also seen after drug combination in the Hargreaves test with a synergy score of 5.136 ± 16.38. In conclusion, combination of diclofenac with andrographolide showed better pharmacologic effects after carrageenan injection and was more synergistic at low-dose combinations.

Solifenacin and Mirabegron Monotherapies Versus Combination Therapy in Overactive Bladder: A Prospective Observational Study

Overactive bladder (OAB) is a common medical condition, especially in the elderly population, that affects the quality of life. The primary treatment of OAB is a combination of behavioural measures and widely used anti-muscarinic drug therapy like solifenacin, which brings about significant side effects, thereby affecting the quality of life adversely. Mirabegron, a recently approved drug for treatment of OAB is a β3 - adrenergic agonist, which relaxes the detrusor muscle. With this background, the current study was done to compare the efficacy and safety of solifenacin, mirabegron and combination of low doses of solifenacin and mirabegron on various parameters of OAB. A total no. of 70 patients with OAB symptoms coming to urology OPD were included in this study and they were divided into 3 groups - Group 1: 24 patients receiving solifenacin 10mg/day, Group 2: 23 patients receiving solifenacin 5mg/day plus mirabegron 25mg/day, Group 3: 23 patients receiving mirabegron 50 mg/day. Patients were assessed for change in mean numbers of micturition, urgency, incontinence per 24 hour and quality of life using Patient Perception of Bladder Control (PPBC) Questionnaire. They were followed up after 4 weeks and 12 weeks and change in the above parameters from baseline were noted. Patients receiving combination therapy achieved a significant reduction in urgency, frequency, nocturia and urge incontinence and decrease in side effects when compared with solifenacin and mirabegron alone. Combination therapy with solifenacin and mirabegron significantly improved quality of life and reduced side effects like dry mouth, blurred vision, constipation in OAB patients in comparison to solifenacin and mirabegron monotherapy.