Journal: Molecular cancer therapeutics

Article Title: DHA-SBT-1214 Taxoid Nanoemulsion and Anti-PD-L1 Antibody Combination Therapy Enhances Anti-Tumor Efficacy in a Syngeneic Pancreatic Adenocarcinoma Model

doi: 10.1158/1535-7163.MCT-18-1046

Figure Lengend Snippet: In vivo efficacy of the PD-L1 antibody in combination to different therapeutic drugs including gemcitabine solution, Abraxane™, paclitaxel in solution, DHA-SBT-1214 in solution and NE-DHA-SBT-1214 against Panc02 induced syngeneic mice tumors. (A)– Graph summarizing all treatment modalities. The values are means ± SD (n=3). Significant differences are indicated as follows: *p < 0.05, and **p< 0.01. (B)– Tumor images taken at the time of harvest from different treatment modalities. (3b-A)– Tumors from mice treated with vehicle; (3b-B)– Three tumors each from PD-L1 (200μg) treated mice; (3b-C, D)–Tumors from Abraxane™ plus IgG or PD-L1 (200μg) treated mice respectively; (3b-E)– Tumors from NE-DHA-SBT-1214 (10mg/kg) plus IgG (200μg) treated mice; (3b-F, G)– Tumors from gemcitabine plus IgG or PD-L1 (200μg) treated mice respectively; (3b-H)– Tumors from NE-DHA-SBT-1214 (10mg/kg) plus PD-L1 (200μg) treated mice; (3b-I, J)– Tumors from NE-DHA-SBT-1214 (25mg/kg) plus IgG or PD-L1 (200μg) treated mice respectively. (C)– Graph for all the tumors from (3B) to show their progression over time.

Article Snippet: In Vivo Single and Combination Therapies Mouse antibody against PD-L1 (10F.9G2) and relevant isotype IgG control was purchased from Bio X Cell.

Techniques: In Vivo

Journal: Molecular cancer therapeutics

Article Title: DHA-SBT-1214 Taxoid Nanoemulsion and Anti-PD-L1 Antibody Combination Therapy Enhances Anti-Tumor Efficacy in a Syngeneic Pancreatic Adenocarcinoma Model

doi: 10.1158/1535-7163.MCT-18-1046

Figure Lengend Snippet: In vivo PD-L1 surface protein expression in response to different therapeutic modalities. (A)– mRNA expression of PD-L1 from different mouse tumor treatment groups analyzed using RT-PCR. Relative gene expression for RT-PCR data was calculated relative to murine β-actin. (B)– Tumor tissue lysate from different treated groups was prepared and protein level of different proteins was analyzed using western blotting. (C)– The bands corresponding to PD-L1 were quantified using Image J software and was normalized relative to band intensities for the corresponding Histone 3 loading controls. The bar represents the mean ± standard deviation of data from at least 3 independent experiments; *p<0.05, **p<0.01.

Article Snippet: In Vivo Single and Combination Therapies Mouse antibody against PD-L1 (10F.9G2) and relevant isotype IgG control was purchased from Bio X Cell.

Techniques: In Vivo, Expressing, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Western Blot, Software, Standard Deviation

Journal: Molecular cancer therapeutics

Article Title: DHA-SBT-1214 Taxoid Nanoemulsion and Anti-PD-L1 Antibody Combination Therapy Enhances Anti-Tumor Efficacy in a Syngeneic Pancreatic Adenocarcinoma Model

doi: 10.1158/1535-7163.MCT-18-1046

Figure Lengend Snippet: Histopathological evaluation of the Panc02-induced tumor tissues collected from control and different combination treated mice (hematoxylin & eosin staining). Significant reduction in tumor stroma observed with combination of NE-DHA-SBT-1214 and anti-PD-L1 treated groups. The images were taken at 63x magnification.

Article Snippet: In Vivo Single and Combination Therapies Mouse antibody against PD-L1 (10F.9G2) and relevant isotype IgG control was purchased from Bio X Cell.

Techniques: Control, Staining

Journal: bioRxiv

Article Title: Combination of Antiviral Drugs to Inhibit SARS-CoV-2 Polymerase and Exonuclease as Potential COVID-19 Therapeutics

doi: 10.1101/2021.07.21.453274

Figure Lengend Snippet: The NS5A inhibitors were built and minimized in terms of energy by Density Functional Theory (DFT). Docking was performed using GOLD 2020.2 software with ChemPLP as scoring function. Because of the diversity in NS5A inhibitors’ molecular weights, they were allowed to dock within 8 or 10 Å spheres in the SARS-CoV-2 nsp14 exonuclease active site, constructed in . The NS5A and catalytic amino acid residues are in stick representation. Mg ++ and Zn ++ are represented as green and indigo blue spheres, respectively.

Article Snippet: Recently, using a digital drug development approach combining artificial intelligence and experimental validation to screen drug combinations for potential combination therapy against SARS-CoV-2, it was determined that one of the most effective potential combinations was Remdesivir, Ritonavir and Lopinavir .

Techniques: Functional Assay, Software, Construct

Journal: bioRxiv

Article Title: Combination of Antiviral Drugs to Inhibit SARS-CoV-2 Polymerase and Exonuclease as Potential COVID-19 Therapeutics

doi: 10.1101/2021.07.21.453274

Figure Lengend Snippet: A mixture of 500 nM RNA (sequence shown at the top of the figure) and 50 nM SARS-CoV-2 pre-assembled exonuclease complex (nsp14/nsp10) was incubated in buffer solution at 37 °C for 15 min in the absence (b, g) and presence of varying amounts of Pibrentasvir (c–e) and Ombitasvir (h, i). The RNA (a, f) and the products of the exonuclease reaction (b–e, g–i) were analyzed by MALDI-TOF MS. The signal intensity was normalized to the highest peak. The peak at 8160 Da corresponds to the intact RNA (8157 Da expected). In the absence of Pibrentasvir, exonuclease activity caused nucleotide cleavage from the 3’-end of the RNA as shown by the 7 lower molecular weight fragments corresponding to cleavage of 1–7 nucleotides (b). Under our experimental conditions, approximately 7% intact RNA remained as indicated by the peak at 8167 Da (b). With increasing amounts of Pibrentasvir, exonuclease activity was reduced as shown by the reduced intensities of the fragmentation peaks and increased intact RNA peak (c–e). Similar results were observed with Ombitasvir (f–i).

Article Snippet: Recently, using a digital drug development approach combining artificial intelligence and experimental validation to screen drug combinations for potential combination therapy against SARS-CoV-2, it was determined that one of the most effective potential combinations was Remdesivir, Ritonavir and Lopinavir .

Techniques: Sequencing, Incubation, Activity Assay, Molecular Weight

Journal: bioRxiv

Article Title: Combination of Antiviral Drugs to Inhibit SARS-CoV-2 Polymerase and Exonuclease as Potential COVID-19 Therapeutics

doi: 10.1101/2021.07.21.453274

Figure Lengend Snippet: A mixture of 500 nM RNAs (sequences shown at the top of the figure) and 50 nM SARS-CoV-2 pre-assembled exonuclease complex (nsp14/nsp10) were incubated in buffer solution at 37 °C for 15 min in the absence (b, e) and presence of 20 μM Pibrentasvir (c, f). The intact RNAs (a, d) and the products of the exonuclease reactions (b–f) were analyzed by MALDI-TOF MS. The signal intensity was normalized to the highest peak. In the absence of Pibrentasvir, exonuclease activity caused nucleotide cleavage from the 3’-end of the RNA as shown by the lower molecular weight fragments corresponding to cleavage of 1–7 nucleotides (b, e). When 20 μM Pibrentasvir was added, exonuclease activity was reduced as shown by the reduced intensities of the fragmentation peaks and increased intact RNA peaks (c, f).

Article Snippet: Recently, using a digital drug development approach combining artificial intelligence and experimental validation to screen drug combinations for potential combination therapy against SARS-CoV-2, it was determined that one of the most effective potential combinations was Remdesivir, Ritonavir and Lopinavir .

Techniques: Incubation, Activity Assay, Molecular Weight

Journal: bioRxiv

Article Title: Combination of Antiviral Drugs to Inhibit SARS-CoV-2 Polymerase and Exonuclease as Potential COVID-19 Therapeutics

doi: 10.1101/2021.07.21.453274

Figure Lengend Snippet: A mixture of 400 nM RNAs (sequences shown at the top of the figure) and 50 nM SARS-CoV-2 pre-assembled exonuclease complex (nsp14/nsp10) were incubated in buffer solution at 37 °C for 15 min in the absence (b, e) and presence of 20 μM Pibrentasvir (c, f). The intact RNAs (a, d) and the products of the exonuclease reactions (b, c, e, f) were analyzed by MALDI-TOF MS. The signal intensity was normalized to the highest peak. The peak at 9776 Da corresponds to the intact RNA (9773 Da expected) and the peak at 9812 Da corresponds to the intact Remdesivir delayed RNA termination product (9797 Da expected). The small peak at 10158 Da corresponds to mismatched incorporation of an additional G; this is likely due to the low fidelity of SARS-CoV-2 RdRp . In the absence of Pibrentasvir, exonuclease activity caused nucleotide cleavage from the 3’-end of the RNA as shown by the lower molecular weight fragments corresponding to cleavage of 1–11 nucleotides (b, e). When 20 μM Pibrentasvir was added, exonuclease activity was reduced as shown by the reduced intensities of the fragmentation peaks and increased intact RNA peaks (c, f).

Article Snippet: Recently, using a digital drug development approach combining artificial intelligence and experimental validation to screen drug combinations for potential combination therapy against SARS-CoV-2, it was determined that one of the most effective potential combinations was Remdesivir, Ritonavir and Lopinavir .

Techniques: Incubation, Activity Assay, Molecular Weight

Journal: bioRxiv

Article Title: Combination of Antiviral Drugs to Inhibit SARS-CoV-2 Polymerase and Exonuclease as Potential COVID-19 Therapeutics

doi: 10.1101/2021.07.21.453274

Figure Lengend Snippet: A mixture of 500 nM RNAs (sequences shown at the top of the figure) and 50 nM SARS-CoV-2 pre-assembled exonuclease complex (nsp14/nsp10) were incubated in buffer solution at 37 °C for 15 min in the absence (b, e) and presence of 20 μM Pibrentasvir (c, f). The intact RNAs (a, d) and the products of the exonuclease reactions (b-f) were analyzed by MALDI-TOF MS. The signal intensity was normalized to the highest peak. In the absence of Pibrentasvir, exonuclease activity caused nucleotide cleavage from the 3’-end of the natural RNA as shown by the lower molecular weight fragments corresponding to cleavage of 1–8 nucleotides (b). However, for Tfv terminated RNA, only minor cleavage was observed (e). When 20 μM Pibrentasvir was added, exonuclease activity was reduced as shown by the reduced intensities of the fragmentation peaks and increased peak height of the intact RNA (c, f).

Article Snippet: Recently, using a digital drug development approach combining artificial intelligence and experimental validation to screen drug combinations for potential combination therapy against SARS-CoV-2, it was determined that one of the most effective potential combinations was Remdesivir, Ritonavir and Lopinavir .

Techniques: Incubation, Activity Assay, Molecular Weight

Journal: bioRxiv

Article Title: Combination of Antiviral Drugs to Inhibit SARS-CoV-2 Polymerase and Exonuclease as Potential COVID-19 Therapeutics

doi: 10.1101/2021.07.21.453274

Figure Lengend Snippet: Calu-3 cells, at a density of 5 × 10 5 cells/well in 48-well plates, were infected with SARS-CoV-2 at a MOI of 0.1, for 1 h at 37 °C. An inoculum was removed and cells were washed and incubated with fresh DMEM containing 2% FBS and the indicated concentration of Remdesivir (RDV) (A), Sofosbuvir (B), Tenofovir (C), and Favipiravir (D), alone and in combination with the HCV NS5A inhibitors. Supernatants were assessed after 48–72 h. Viral replication in the culture supernatant was measured as PFU/mL by titering in VeroE6 cells. Results are displayed as virus titers. The data represent means ± SEM of three independent experiments.

Article Snippet: Recently, using a digital drug development approach combining artificial intelligence and experimental validation to screen drug combinations for potential combination therapy against SARS-CoV-2, it was determined that one of the most effective potential combinations was Remdesivir, Ritonavir and Lopinavir .

Techniques: Infection, Incubation, Concentration Assay