To evaluate the interaction of EGCG and kaempferol combinations, the Chou–Talalay method, which was based on the median-effect equation, was used to measure the combination index (CI) values by describing the dose–effect curves of individual compounds and their combinations. Therefore, the EGCG and kaempferol interaction effects were evaluated as CI using CompuSyn software, CompuSyn, Inc., Paramus, NJ, USA [35 (link)]. In the present study, the combinations were composed of two compounds, and their CI values were evaluated by the CompuSyn software using the following formula: CI = (D)1/(DX)1 + (D)2/(DX)2, in which the (DX)1 and (DX)2 are the concentration, respectively, of sample 1 and sample 2 alone in which the cellular antioxidant effect is x%. Similarly, the (D)1 and (D)2 show the respective doses of sample 1 and sample 2, in which their combination antioxidant activity is x%. When CI = 1, the interaction between EGCG and kaempferol was an additive effect; when CI < 1, the interaction between EGCG and kaempferol indicated a synergistic effect, with the smaller CI showing the stronger synergistic effect; when CI > 1, the interaction between EGCG and kaempferol reflected an antagonistic effect [36 (link)].
Compusyn software
Manufactured by ComboSyn
Sourced in United States, United Kingdom
CompuSyn is a software tool designed for the analysis of drug combination effects. It provides mathematical models and statistical analysis for evaluating synergistic, additive, or antagonistic interactions between multiple drugs or compounds.
Automatically generated - may contain errors
Lab products found in correlation
Prism 5, by GraphPad (13 mentions)
Prism 6, by GraphPad (13 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (11 mentions)
Microplate reader, by Thermo Fisher Scientific (8 mentions)
Prism 8, by GraphPad (8 mentions)
Prism software, by GraphPad (8 mentions)
Microplate reader, by Bio-Rad (6 mentions)
Celltiter glo, by Promega (6 mentions)
Celltiter 96 aqueous one solution cell proliferation assay, by Promega (6 mentions)
Microplate reader, by Agilent Technologies (5 mentions)
Annexin v 7 aad apoptosis detection kit, by BD (4 mentions)
Fitc apoptosis detection kit ce, by Immunostep (3 mentions)
Annexin 5 fluorescein isothiocyanate fitc propidium iodide, by Beckman Coulter (3 mentions)
Mtt assay, by Merck Group (3 mentions)
Cell counting kit 8 (cck8), by Keygen Biotech (3 mentions)
Gelcount, by Oxford Optronix (3 mentions)
Srb reagent, by Merck Group (2 mentions)
Cytarabine, by Merck Group (2 mentions)
Compusyn software version 1, by ComboSyn (2 mentions)
Alamar blue, by Thermo Fisher Scientific (2 mentions)
688 protocols using compusyn software
1
Evaluating EGCG-Kaempferol Synergy
2
Synergistic Drug Combination Evaluation
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The stock of momelotinib and sorafenib was prepared by dissolving 20 mg/mL of the mixture in DMSO. The stocks of each drug were stored at −20 °C until use. Using the CompuSyn software to calculate the half-maximal inhibitory concentration (IC50) values of different cell lines as previously reported by Chou TC and Martin N. The calculation method of IC50 is as described in the PC Software and User’s Guide on the ComboSyn Inc. website (http://www.combosyn.com , accessed on 4 May 2020). The effects of momelotinib and sorafenib on cell proliferation were detected using the sulforhodamine B (SRB) assay. The synergistic effect of these two drugs was analyzed using isobolograms of the drug combination, as previously reported by Chou and Talalay [22 (link),23 (link)]. The interaction between the two drugs was also analyzed by the median-effect principle proposed by them. The combination index (CI) was calculated using CompuSyn software. A CI value is less than 1 represented synergism [23 (link)]. Briefly, HCC cells or tumorspheres were seeded in 96-well plates (8 × 105 cells/well) and treated with the drugs (momelotinib or sorafenib alone or in combination) at the 5 μM and 2.5 μM concentration for 48 h, respectively. After respective drug treatments, the relative cell number was estimated by the SRB reagent according to the manufacturer’s protocol (Sigma, Vallejo, CA, USA).
3
Combination Index Analysis of FIPV Inhibition
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Data collected from the plaque reduction assay were analyzed using Compusyn software (ComboSyn, Inc., https://www.combosyn.com/index.html ). Data were excluded according to the protocol defined by Compusyn software if the concentration of GS-441524 and ICZ resulted in the following conditions: 1) plaque reduction of 100%, and 2) plaque reduction of less than 25% (i.e., ineffective against FIPV) (Chou, 2014 (link); ComboSyn Inc., 2017 ). The combination index (CI) was calculated and categorized according to the classification proposed by Cho et al. (Chou, 2006 (link); Chou and Talalay, 1984 (link)).
4
Quantifying Synergistic Drug Interactions
5
Synergistic Nanoparticle Combination Therapy
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Compusyn software program (version 1.0), based on the Chou–Talalay method, was employed to determine the nature of interaction between the two-/three-agents. This method utilizes a multiple drug-effect equation derived from enzyme kinetics model in which the output is represented as combination indexes (CI). Compusyn software defines synergy as CI value less than 1. Based on CI values, the extent of synergism/antagonism may be determined [55] (link). In brief, CI values between 0.9 and 0.85 would suggest a slight synergy, whereas those in the range of 0.7 to 0.3 are indicative of clear synergistic interactions between the drugs/agents. On the other hand, CI values in the range of 0.9–1.1 suggest a near additive effect and values more than 1.1 indicate antagonism. We have determined CI values for all two-and three-drug-loaded NPs (1a , 1b , 1c and 2 ). For detailed in vitro studies, the dose was selected considering both the therapeutic efficacy and extent of synergism. The synergistic doses of 5 and 10 μg/mL for CS-SHMP-CQA-NPs showed below IC50 value and were used for all in vitro experiments. Doses of free Cur, Quer and Asp were chosen according to their entrapment efficiencies in CS-SHMP-CQA-NPs (10 μg/mL).
6
Synergistic Drug Combination Analysis
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Raw absorbance values from MTT assays were normalized to their respective vehicle control wells and represented as a % decrease in absorbance (control = 0 effect, 2% decrease in absorbance =.02 effect). These values were entered into the CompuSyn software (ComboSyn, Paramus, NJ) as “effect” along with the concentrations of both JNK-IN-8 and lapatinib used to obtain that value. Effect values for increasing concentrations of each drug alone were used to calculate the median-effect, which was then used to determine whether the effect caused by a combination concentration was synergistic [25 (link)]. Combination Index plots and values were generated by the CompuSyn software.
7
Synergistic Drug Combination Analysis
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Using MTT assay data, the combination index (CI) was calculated by the Chou-Talalay method as described by Chou et al. using CompuSyn software (CompuSyn, Inc., Paramus, NJ, USA) [42 (link)]. The dose-effect curves for single and cotreatment were generated and the CI for every dose and the corresponding effect, fraction affected (Fa) were calculated. The resultant CI values reflect the potential interactions between two drugs. CI < 1 indicates synergism, CI = 1 indicates an additive effect and CI > 1 indicates antagonism. The Dose-reduction index (DRI ) was calculated from the DRI equation and algorithm using CompuSyn software, (DRI50) values represent the magnitude of dose reduction obtained for the 50% growth inhibitory effect in Amy/Sor cotreatment compared to each drug alone that causes the same growth inhibition effect.
8
Evaluating Synergistic Chemotherapeutic Effects
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100,000 cells were seeded in 96-well plates, treated for 48 h with chemotherapeutic agents at scalar concentrations (from 1 pmol/L to 1 mmol/L), then stained with neutral red solution as described [44 ]. The IC50, i.e. the concentration of each drug that decreased the cell viability by 50%, was calculated with the CompuSyn software (http://www.combosyn.com ). The Rf was calculated by dividing the IC50 of each drug in HMM cells for the IC50 of each drug in HMC. The synergistic, additive or antagonistic effect of the different concentrations of chemotherapeutic drugs in the presence of 1 μmol/L zoledronic acid was calculated with the CI equation of Chou-Talalay [45 (link)], using the CompuSyn software.
9
Synergistic Antioxidant Activity Evaluation
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To determine the synergistic, additive or antagonistic effect of the antioxidant activity between HT and the strawberry puree, data were analyzed using CompuSyn software (CompuSyn Inc., Paramus, NJ, USA). This program is based on Chou-Talalay’s multiple drug effect equations [26 (link)], which defines the theoretical basis for the synergistic, additive and antagonistic effects between components in a mixture by the combination index (CI). CompuSyn software defines synergy as CI values less than 1, CI = 1 additive, and CI > 1 antagonism [28 ].
10
Synergy Analysis of Niclosamide and DDP
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To evaluate whether the antitumor effects of niclosamide combined with DDP were synergistic, additive or antagonistic, combination index (CI) value for drug synergy was calculated using the CompuSyn software (Version 2.1, ComboSyn, Inc., Paramus, NJ, USA) as previously described (14 (link)). Using data obtained from CCK-8 assays and CompuSyn software, the dose-effect curves for single agents and their combinations were generated, and the CI values for each dose and the corresponding effect level, referred to as the fraction affected (Fa; the fraction of cells inhibited following drug exposure, for example 0.5 when cell growth is inhibited by 50%), were calculated. CI values <1 indicated a synergistic effect, values equal to 1 indicated an additive effect and values >1 indicated an antagonistic effect. Then, to provide a visual illustration of drug interactions, the Fa-CI plot was constructed by simulating CI values over a range of Fa levels from 0.1 to 0.95 (18 (link),19 (link)).
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