Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

Article Title: Synthesis and Molecular Modeling Studies of N′ -Hydroxyindazolecarboximidamides as Novel Indoleamine 2,3-Dioxygenase 1 (IDO1) Inhibitors

doi: 10.3390/molecules22111936

Figure Lengend Snippet: Structure of IDO1 inhibitors in clinical trials.

Article Snippet: To analyze the hIDO1 inhibitory activities of compounds, we used IDO1 inhibitor screening assay kit and followed the protocol provided from BPS Bioscience (San Diego, CA, USA).

Techniques:

Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

Article Title: Synthesis and Molecular Modeling Studies of N′ -Hydroxyindazolecarboximidamides as Novel Indoleamine 2,3-Dioxygenase 1 (IDO1) Inhibitors

doi: 10.3390/molecules22111936

Figure Lengend Snippet: Rational design of novel IDO1 inhibitor ( 1 ) from the known molecules.

Article Snippet: To analyze the hIDO1 inhibitory activities of compounds, we used IDO1 inhibitor screening assay kit and followed the protocol provided from BPS Bioscience (San Diego, CA, USA).

Techniques:

Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

Article Title: Synthesis and Molecular Modeling Studies of N′ -Hydroxyindazolecarboximidamides as Novel Indoleamine 2,3-Dioxygenase 1 (IDO1) Inhibitors

doi: 10.3390/molecules22111936

Figure Lengend Snippet: Inhibition of tryptophan depletion and kynurenine production through IDO1.

Article Snippet: To analyze the hIDO1 inhibitory activities of compounds, we used IDO1 inhibitor screening assay kit and followed the protocol provided from BPS Bioscience (San Diego, CA, USA).

Techniques: Inhibition

Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

Article Title: Synthesis and Molecular Modeling Studies of N′ -Hydroxyindazolecarboximidamides as Novel Indoleamine 2,3-Dioxygenase 1 (IDO1) Inhibitors

doi: 10.3390/molecules22111936

Figure Lengend Snippet: Docking model 8a against IDO1. ( a ) Binding mode of 8a (pink, ball and stick style) coordinated to heme; ( b ) The surface model of the active site bound to 8a is colored because of hydrophobicity. Hydrogen bonds of inter- and intra-molecular interactions are shown as green dashed lines and interaction residues are represented by the stick model.

Article Snippet: To analyze the hIDO1 inhibitory activities of compounds, we used IDO1 inhibitor screening assay kit and followed the protocol provided from BPS Bioscience (San Diego, CA, USA).

Techniques: Binding Assay

Journal: British Journal of Cancer

Article Title: The therapeutic potential of targeting tryptophan catabolism in cancer

doi: 10.1038/s41416-019-0664-6

Figure Lengend Snippet: Analysing IDO1 inhibitors clinical trial results. a Intra-tumoural penetrance of IDO1 inhibitors (IDO1i) could be hindered due to diminished blood supply to tumours or elevated oncotic pressure in solid tumours (left). Even if IDO1i are able to access tumours, ABC transporters can interfere with their intracellular penetrance (middle). Cellular uptake of IDO1i can lead to AHR activation in various cell types and therefore contribute to cancer progression (right). b Combination of IDOi with other types of therapies can lead to the upregulation of IDO1 expression through various mechanisms (see text). In addition, clinical trials have included patients pre-treated with BRAF inhibitors (BRAFi), which were shown to promote an enrichment of a population of cells with constitutive AHR activity. This might further lead to the induction of IDO1 expression and contribute to cancer progression. c Even if IDO1 inhibition is effectively achieved, IDO2 and TDO2 might compensate for IDO1 blockade. BRAFi and IDO1i can promote AHR activation in various cells present in the tumour microenvironment, which unleashes the intrinsic cancer promoting effects driven by the AHR. Moreover, AHR activation leads to the upregulation of IDO2 and TDO2, which can further activate the AHR and deplete Trp. IDO1i and immune check point blockade (ICB) lead to an increase in cytotoxic TILs, which in turn secrete pro-inflammatory molecules, such as IFN-γ, which also induces IDO2. In addition, the pro-inflammatory microenvironment driven by IDO1i and ICB therapy can also lead to the upregulation of TDO2, via the COX2–PGE 2 –EP4 pathway. d In order to obtain a more robust view of the effects of IDO1 inhibition and improve the outcome of its use in clinical trials, we suggest stratifying patients based on the concentration of Trp, Kyn and Kyn-derived metabolites in plasma and tumours, as well as on the expression of the TCE and AHR activity present in tumours. Once stratified, patients can be treated more accurately with one or more therapies targeting Trp catabolism.

Article Snippet: This Eli Lily-developed IDO1 inhibitor has entered Phase 1 clinical trials in late stage and metastatic solid tumours either alone or in combination with anti-PD-L1 therapies (NCT03343613).

Techniques: Activation Assay, Expressing, Clinical Proteomics, Activity Assay, Inhibition, Concentration Assay, Derivative Assay

Journal: British Journal of Cancer

Article Title: The therapeutic potential of targeting tryptophan catabolism in cancer

doi: 10.1038/s41416-019-0664-6

Figure Lengend Snippet: Analytical approaches for quantification expression and activity of tryptophan catabolising enzymes (TCEs) in cancer therapies. The levels of Trp and its metabolites can be measured by chromatographic methods, including high-performance liquid chromatography (HPLC), gas chromatography–mass spectrometry (GC-MS) or liquid chromatography–mass spectrometry (LC-MS). Antibodies detecting Trp, as well as specific metabolites of the Trp degradation pathway, enable enzyme-linked immunosorbent assay (ELISA) measurements. , – Furthermore, matrix-assisted laser desorption/ionisation (MALDI) mass spectrometry (MS) imaging allows visualisation of Trp and Kyn in tissue, , while position-emission tomography (PET) imaging with Trp-derived tracers enables visualisation of Trp uptake in vivo. – In addition, TCE expression can be detected on mRNA and protein levels, however care must be taken when selecting anti-IDO1 antibodies, as many lack selectivity, particularly for immunohistochemistry. The blue area of the circle illustrates methods used for measuring Trp and its metabolites; the yellow area illustrates methods used for detcting the expression of TCE and AHR activity. IF: immunofluorescence; WB: Western blot. PET-imaging example was reproduced under a Creative Commons CC-BY license from. RNA-seq example was reproduced under a Creative Commons CC-BY license from. Western Blot example was provided by the authors.

Article Snippet: This Eli Lily-developed IDO1 inhibitor has entered Phase 1 clinical trials in late stage and metastatic solid tumours either alone or in combination with anti-PD-L1 therapies (NCT03343613).

Techniques: Expressing, Activity Assay, High Performance Liquid Chromatography, Gas Chromatography, Mass Spectrometry, Gas Chromatography-Mass Spectrometry, Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy, Enzyme-linked Immunosorbent Assay, Imaging, Tomography, Derivative Assay, In Vivo, Immunohistochemistry, Immunofluorescence, Western Blot, RNA Sequencing

Journal: Cancers

Article Title: OATD-02 Validates the Benefits of Pharmacological Inhibition of Arginase 1 and 2 in Cancer

doi: 10.3390/cancers14163967

Figure Lengend Snippet: OATD-02 shows superior efficacy in the CT26 tumour model when combined with immune checkpoint inhibitor (anti-PD-L1 antibody) and IDO inhibitor (epacadostat). ( A )—BALB/c mice were inoculated with CT26 cells and upon 24 h dosed with OATD-02 (50 mg/kg, PO, BID) or epacadostat (30 mg/kg, PO, BID) till the end of the experiment; anti-PD-L1 antibody was administered at 2.5 mg/kg (IP, QD at days: 8, 10, 12, 14 and 16); ( B )—final measurements of tumour volumes were taken at day 24 post-inoculation and TGI was calculated (** 0.0042 < p < 0.0060, U Mann–Whitney test).

Article Snippet: The IDO1 inhibitor (epacadostat, INCB024360) was purchased from Shanghai Sunshine Chemical Technology (lot SSC151112) and used for oral gavage as a formulation containing 2% DMSO and 18% PEG400 in sterile saline.

Techniques: MANN-WHITNEY

Journal: Journal for Immunotherapy of Cancer

Article Title: Peptide vaccination directed against IDO1-expressing immune cells elicits CD8 + and CD4 + T-cell-mediated antitumor immunity and enhanced anti-PD1 responses

doi: 10.1136/jitc-2020-000605

Figure Lengend Snippet: Growth delay of indoleamine 2,3-dioxygenase 1 (IDO1)-positive CT26 tumors by prophylactic vaccination with major histocompatibility complex (MHC) class I-directed or II-directed IDO1 peptides requires intact adaptive immunity and host IDO1. (A) Western blot analysis of tumors formed by six different mouse tumor-derived cell lines for comparison of IDO1 protein levels (top panel) with β-tubulin as a loading control (bottom panel). Epididymis lysates from wild-type (WT) and Ido1 -/- male BALB/c strain mice were included as positive and negative controls. M designates the molecular weight marker. (B) Growth curves of CT26 tumors in mice administered single doses of five different MHC class I-directed, IDO1 peptide vaccines (EP1–5) 7 days prior to tumor engraftment. Overall responses are plotted as means±SEM together with concurrent results from both untreated and vehicle-treated animals. Growth curves for all groups are represented with gray lines except for the treatment cohort exhibiting the strongest response (EP2) and the untreated cohort which are distinguished with black lines. (n=6 tumors/cohort). (C) Growth curves of CT26 tumors in mice treated with two different MHC class II-directed, IDO1 peptide vaccines (EP6 left , EP7 right ) 7 days prior to tumor engraftment. Overall responses (black lines) are plotted as means±SEM. Concurrent results from both vehicle and EP2 peptide-treated animals (gray lines) are included on each graph for comparison (n=10 tumors/cohort). (D) Growth curves of CT26 tumors in Rag1 –/– mice vaccinated with the MHC class I-directed and II-directed IDO1 peptides EP2 and EP6 7 days prior to tumor engraftment. Overall responses (black lines) are plotted as means±SEM. Concurrent results from vehicle-treated animals (gray lines) are included on each graph for comparison (n≥6 tumors/cohort). (E) Growth curves of CT26 tumors in Ido1 -/- mice vaccinated with EP2 and EP6 7 days prior to tumor engraftment. Overall responses (black lines) are plotted as means±SEM. Concurrent results from vehicle-treated animals (gray lines) are included on each graph for comparison (n=10 tumors/cohort). P values from longitudinal analysis of tumor growth for each peptide vaccine-treated group compared with untreated or vehicle-treated animals are included on each graph.

Article Snippet: The IDO1 enzyme inhibitor epacadostat (ChemieTek Cat# CT-EPAC) was administered by oral gavage, twice a day, at 30 mg/kg in 100 μL of vehicle (3% N, N-diethylacetamide, 10% 1-hydroxypropyl-β-cyclodextrin).

Techniques: Immunopeptidomics, Western Blot, Derivative Assay, Comparison, Control, Molecular Weight, Marker, Vaccines

Journal: Journal for Immunotherapy of Cancer

Article Title: Peptide vaccination directed against IDO1-expressing immune cells elicits CD8 + and CD4 + T-cell-mediated antitumor immunity and enhanced anti-PD1 responses

doi: 10.1136/jitc-2020-000605

Figure Lengend Snippet: Indoleamine 2,3-dioxygenase 1 (IDO1) expression is localized to infiltrating immune cells within CT26 tumors. (A) Confocal images of CT26 tumor sections. ( top row, left to right ) Immunofluoresence staining of sections from wild-type (WT) and Ido1 -/- mice for IDO1 (Cy3, red), and from WT mice for programmed death-ligand 1 (PDL1) (Cy3, red) and the combination of IDO1 (fluorescein isothiocyanate (FITC), green) and PDL1 (Cy3, red). Nuclei were stained on all sections (DAPI, blue). ( bottom row, left to right ) Staining of CT26 tumor sections from WT mice for combinations of IDO1 (Cy3, red) with CD45, CD11b, Gr1 and CD11c (FITC, green). Nuclei were stained on all sections (DAPI, blue). (B) Confocal images of a field of FACS-isolated CD45 + , CD11b + , CD11c + cells from a CT26 tumor in a WT host. ( left to right ) Immunofluoresence staining for nuclei (DAPI, blue), CD11c (FITC, green), IDO1 (Cy3, red), CD19 (Cy5, purple) and the composite image.

Article Snippet: The IDO1 enzyme inhibitor epacadostat (ChemieTek Cat# CT-EPAC) was administered by oral gavage, twice a day, at 30 mg/kg in 100 μL of vehicle (3% N, N-diethylacetamide, 10% 1-hydroxypropyl-β-cyclodextrin).

Techniques: Expressing, Staining, Isolation

Journal: Journal for Immunotherapy of Cancer

Article Title: Peptide vaccination directed against IDO1-expressing immune cells elicits CD8 + and CD4 + T-cell-mediated antitumor immunity and enhanced anti-PD1 responses

doi: 10.1136/jitc-2020-000605

Figure Lengend Snippet: Major histocompatibility complex (MHC) class I-directed and II-directed indoleamine 2,3-dioxygenase 1 (IDO1) peptides cooperate together but not with IDO1 enzyme inhibition. (A) Growth curves of CT26 tumors in wild-type (WT) mice immunized with the MHC class I-directed and II-directed peptides EP2 and EP6 either separately or together beginning 7 days after tumor engraftment. Responses to adjuvant alone, individual peptides (gray lines) and combined peptides (black lines) are plotted as means±SEM (n=8 tumors/cohort). (B) Evaluation of EP2 and EP6 both separately and in combination in Ido1 -/- mice as described in A (n=8 tumors/cohort). (C) Growth curves of CT26 tumors in WT mice treated with EP2 and epacadostat either separately or together beginning 7 days after tumor engraftment. Responses to adjuvant alone, EP2 and epacadostat individually (gray lines), and combined treatment (black lines) are plotted as means±SEM (n=10 tumors/cohort). (D) Evaluation of EP6 and epacadostat both separately and in combination as described in B (n≥9 tumors/cohort). P values for longitudinal tumor growth comparisons between combined and individual peptide vaccine-treated groups are included on each graph.

Article Snippet: The IDO1 enzyme inhibitor epacadostat (ChemieTek Cat# CT-EPAC) was administered by oral gavage, twice a day, at 30 mg/kg in 100 μL of vehicle (3% N, N-diethylacetamide, 10% 1-hydroxypropyl-β-cyclodextrin).

Techniques: Immunopeptidomics, Enzyme Inhibition Assay, Adjuvant

Journal: Journal for Immunotherapy of Cancer

Article Title: Peptide vaccination directed against IDO1-expressing immune cells elicits CD8 + and CD4 + T-cell-mediated antitumor immunity and enhanced anti-PD1 responses

doi: 10.1136/jitc-2020-000605

Figure Lengend Snippet: Programmed cell death protein 1-binding antibody (anti-PD1 antibody) cooperativity with major histocompatibility complex (MHC) class I-directed and II-directed indoleamine 2,3-dioxygenase 1 (IDO1) peptides compared with epacadostat. (A) Growth curves of CT26 tumors in wild-type (WT) mice receiving the MHC class I-directed and II-directed peptides EP2 and EP6 either separately or together with or without the anti-PD1 antibody beginning 7 days after tumor engraftment. The experiment is divided between two graphs for clarity. ( top left) Responses to adjuvant alone, individual peptides or anti-PD1 alone (gray lines), and the combined peptides (black lines), are plotted as means±SEM (n=10 tumors/cohort). ( bottom left ) Responses to adjuvant alone, anti-PD1 alone or with the individual peptides (gray lines), and anti-PD1 with the combined peptides (black lines) are plotted as means±SEM (n=10 tumors/cohort). (B) Growth curves of CT26 tumors in WT mice treated with the combination of EP2+EP6 or the IDO1 inhibitor epacadostat either without or with anti-PD1 antibody beginning 7 days after tumor engraftment. ( left side) Responses to adjuvant alone, epacadostat, anti-PD1 or EP2+EP6 individually (gray lines), and combinations of epacadostat or EP2+EP6 with anti-PD1 (black lines) are plotted as means±SEM (n=10 tumors/cohort). P values for longitudinal tumor growth comparisons between the anti-PD1 and other treatment groups are included on each graph. P values from additional pairwise determinations are shown in . ( right sides (all )) Individual growth curves for each treatment condition (X-axis is set at −100 on the Y-axis). In groups with complete responders (CRs), the number of animals represented is indicated on the graph.

Article Snippet: The IDO1 enzyme inhibitor epacadostat (ChemieTek Cat# CT-EPAC) was administered by oral gavage, twice a day, at 30 mg/kg in 100 μL of vehicle (3% N, N-diethylacetamide, 10% 1-hydroxypropyl-β-cyclodextrin).

Techniques: Binding Assay, Immunopeptidomics, Adjuvant

Journal: Journal for Immunotherapy of Cancer

Article Title: Peptide vaccination directed against IDO1-expressing immune cells elicits CD8 + and CD4 + T-cell-mediated antitumor immunity and enhanced anti-PD1 responses

doi: 10.1136/jitc-2020-000605

Figure Lengend Snippet: Major histocompatibility complex (MHC) class I-directed and II-directed indoleamine 2,3-dioxygenase 1 (IDO1) peptide administration does not reduce the proportional representation of IDO1-expressing tumor infiltrating immune cells but does reduce their expression of IDO1. (A) Confocal images of CT26 tumor sections from wild-type (WT) mice administered either adjuvant alone, EP2+EP6 IDO1 peptides or antiprogrammed cell death protein 1 (anti-PD1) antibody as noted. Immunofluoresence staining for CD11b (fluorescein isothiocyanate (FITC), green), IDO1 (Cy3, red) and nuclei (DAPI, blue) was performed on all sections. (B) Quantitative comparison of the proportional representation of the CD11b hi CD11c hi (IDO1-expressing) subset following administration of adjuvant alone, anti-PD1 and the individual EP2 or EP6 peptides alone or combined identified by flow cytometry as shown in from ( left ) within the total population of viable cells from dissociated tumors and ( right ) within the CD45 + population of tumor-infiltrating immune cells. Graphed as means±SEM with significance determined by one-way analysis of variance (ANOVA) with Tukey’s multiple comparison test. (C) Confocal images of a field of FACS-isolated CD45 + , CD11b + , CD11c + cells from CT26 tumors obtained from an Ido1 -/- host or from WT hosts administered either adjuvant alone, EP2, EP6 or EP2+EP6 IDO1 peptides, or anti-PD1 antibody as noted. Immunofluoresence staining for IDO1 (Cy3, red) and nuclei (DAPI, blue) was evaluated on all sections. (D) Quantitative comparison of IDO1 (Cy3)/nuclei (DAPI) ( left ) and CD11b (FITC)/nuclei (DAPI) ( right ) staining per field from FACS-isolated CD45 + , CD11b + , CD11c + cells from treatment groups described in (D). Graphed as means±SEM with significance determined by one-way ANOVA with Tukey’s multiple comparison test. ns, not significant. *P<0.05;**p<0.01; ***p<0.001, ****p<0.0001.

Article Snippet: The IDO1 enzyme inhibitor epacadostat (ChemieTek Cat# CT-EPAC) was administered by oral gavage, twice a day, at 30 mg/kg in 100 μL of vehicle (3% N, N-diethylacetamide, 10% 1-hydroxypropyl-β-cyclodextrin).

Techniques: Immunopeptidomics, Expressing, Adjuvant, Staining, Comparison, Flow Cytometry, Isolation

Journal: Journal for Immunotherapy of Cancer

Article Title: Peptide vaccination directed against IDO1-expressing immune cells elicits CD8 + and CD4 + T-cell-mediated antitumor immunity and enhanced anti-PD1 responses

doi: 10.1136/jitc-2020-000605

Figure Lengend Snippet: Major histocompatibility complex (MHC) class I-directed and II-directed responses are predominantly mediated by CD8 + and CD4 + T cells, respectively. (A) Flow cytometry characterization of splenic T-cell populations from a complete responder (CR) mouse (bottom panels) compared with a naïve mouse (top panels). CD4 + CD8 - (left panels) and CD8 + CD4 - (right panels) gated populations assessed for levels of CD44 (X-axis) and CD62L (Y-axis). (B) Quantitative comparison of the proportional representation of CD44 hi CD62L hi (activated) populations of CD4 + and CD8 + T cells in the spleens of CR mice relative to naïve mice identified by flow cytometry analysis as shown in (E). Graphed as means±SEM with significance determined by two-tailed Student’s t-test (n≥3 mice/group). (C) Quantitative comparison of the proportional representation of CD44 hi CD62L lo (memory) populations as described for B. (D) Schematic illustration of the basic strategy for adoptive transfer of T cells from IDO1 peptide(s)+programmed cell death protein 1-binding antibody (anti-PD1 antibody)-treated, CR mice to treatment naïve, tumor-bearing mice. (E) Growth curves of CT26 tumors in WT mice that received T cells isolated 10 days following tumor rechallenge from a CR mouse previously treated with anti-PD1 antibody and the MHC class I-directed IDO1 peptide EP2. Responses to PBS (gray line) and T cells (black line) are plotted as means±SEM (n=8 tumors/cohort). (F) Growth curves of CT26 tumors in WT mice that received flow cytometry sorted T-cell subsets isolated 10 days following tumor rechallenge from a CR mouse treated with anti-PD1 and EP2. Responses to PBS, CD4 + , CD8 + (black lines), and combined CD4 + and CD8 + T cells (gray line) are plotted as means±SEM (n=4 tumors/cohort). (G) Growth curves of CT26 tumors in WT mice that received flow cytometry sorted T-cell subsets isolated 10 days following tumor rechallenge from CR mice treated with either anti-PD1 and the MHC class II-directed IDO1 peptide EP6 or anti-PD1 and EP2. Responses to PBS, CD4 + and CD8 + T cells from EP6+anti-PD1 (black lines) or EP2+anti-PD1 (gray lines) treated donors are plotted as means±SEM (n=6 tumors/cohort). (H) Growth curves of CT26 tumors in WT mice that received flow cytometry sorted T-cell subsets isolated 10 days following tumor rechallenge from a CR mouse treated with anti-PD1 and EP2+EP6. Responses to PBS, CD4 + , CD8 + (black lines) and combined CD4 + and CD8 + T cells (gray line) are plotted as means±SEM (n=6 tumors/cohort). (I) As described above in G except the recipient mice were Ido1 -/- (n=6 tumors/cohort). P values for longitudinal tumor growth comparisons between the PBS (no cells) and different T-cell adoptive transfer cohorts are included on graphs E–I. P values for comparisons between the combined and individual CD4 + and CD8 + adoptive transfer cohorts are also included for graphs H and I. *P<0.05;**p<0.01; ***p<0.001.

Article Snippet: The IDO1 enzyme inhibitor epacadostat (ChemieTek Cat# CT-EPAC) was administered by oral gavage, twice a day, at 30 mg/kg in 100 μL of vehicle (3% N, N-diethylacetamide, 10% 1-hydroxypropyl-β-cyclodextrin).

Techniques: Immunopeptidomics, Flow Cytometry, Comparison, Two Tailed Test, Adoptive Transfer Assay, Binding Assay, Isolation

Journal: PLoS ONE

Article Title: Inhibition of immune checkpoints PD-1, CTLA-4, and IDO1 coordinately induces immune-mediated liver injury in mice

doi: 10.1371/journal.pone.0217276

Figure Lengend Snippet: Pharmacokinetics (PK)/pharmacodynamics (PD) in (A) wild-type and (B) PD-1 -/- mice. Mice were administered epacadostat at doses of 100, 300, and 600 mg/kg/dose by oral gavage at t = 0 h and t = 6 h, and total plasma concentrations of epacadostat and kynurenine were measured by LC-MS/MS. Epacadostat concentrations were below the limit of quantification at 24 h for the 100 mg/kg BID dose. All values are reported as mean ± SD. n = 3 (WT) and 3 ( PD-1 -/- ). PD: pharmacodynamics, PK: pharmacokinetics, BID: bis in die (twice daily).

Article Snippet: The IDO1 inhibitor epacadostat (Chemscene, Monmouth Junction, NJ) was administered twice daily (6 hours between each dose) as an oral gavage formulation with 0.5% HPMC/0.25% Tween 20 in a water base at 100, 300 or 600 mg/kg/dose.

Techniques: Drug discovery, Clinical Proteomics, Liquid Chromatography with Mass Spectroscopy

Journal: PLoS ONE

Article Title: Inhibition of immune checkpoints PD-1, CTLA-4, and IDO1 coordinately induces immune-mediated liver injury in mice

doi: 10.1371/journal.pone.0217276

Figure Lengend Snippet: Mice were treated with 300 mg/kg BID epacadostat and/or 9D9 for 4 weeks (see methods). (A) Representative H&E images and quantification of liver (B) single cell necrosis and (C) periportal infiltration. (D) GLDH. (E) Quantification of T cells by flow cytometry. Necrosis, infiltration and GLDH data were analyzed with a Kruskal-Wallis test and Dunn’s multiple comparisons test. Flow cytometry data was analyzed with an ordinary one-way ANOVA with Tukey’s multiple comparisons test. Arrows indicate immune cell accumulation. Arrowheads indicate a necrotic cell. Scale bar = 150 μm. All data are reported as mean ± SEM *P<0.05, ***P<0.001, n = 6 per group. GLDH: glutamate dehydrogenase.

Article Snippet: The IDO1 inhibitor epacadostat (Chemscene, Monmouth Junction, NJ) was administered twice daily (6 hours between each dose) as an oral gavage formulation with 0.5% HPMC/0.25% Tween 20 in a water base at 100, 300 or 600 mg/kg/dose.

Techniques: Flow Cytometry

Journal: PLoS ONE

Article Title: Inhibition of immune checkpoints PD-1, CTLA-4, and IDO1 coordinately induces immune-mediated liver injury in mice

doi: 10.1371/journal.pone.0217276

Figure Lengend Snippet: PD-1 -/- mice were treated with 9D9, epacadostat (A, C panels are 300 mg/kg BID only, B includes 300 and 600 mg/kg BID), or in combination for 2 or 6 weeks. (A) Representative H&E images and (B) quantification of single cell necrosis. (C) GLDH levels through 6 weeks. Arrows indicate immune cell accumulation. Arrowheads indicate a necrotic cell. Scale bar = 150 μm. All data (n = 6 per group) are reported as mean ± SEM. Necrosis data was analyzed with a Kruskal-Wallis test and Dunn’s multiple comparisons test. GLDH data was analyzed with a repeated measures two-way ANOVA and Tukey’s multiple comparisons test. a P < 0.05 9D9 + epacadostat vs vehicle, 9D9, b P < 0.05 9D9 + epacadostat vs epacadostat, c P < 0.05 epacadostat vs vehicle, 9D9.

Article Snippet: The IDO1 inhibitor epacadostat (Chemscene, Monmouth Junction, NJ) was administered twice daily (6 hours between each dose) as an oral gavage formulation with 0.5% HPMC/0.25% Tween 20 in a water base at 100, 300 or 600 mg/kg/dose.

Techniques:

Journal: PLoS ONE

Article Title: Inhibition of immune checkpoints PD-1, CTLA-4, and IDO1 coordinately induces immune-mediated liver injury in mice

doi: 10.1371/journal.pone.0217276

Figure Lengend Snippet: PD-1 -/- mice were treated with 9D9, lower dose (300 mg/kg BID) epacadostat or in combination for 2 or 6 weeks. (A) Representative images of the mid-zonal/centrilobular region of the liver in the vehicle and combination groups. Semi-quantitative analysis of immunohistochemical detection of CD4, CD8α or Foxp3 at (B) 2 and (C) 6 weeks of treatment. (D) Statistical measure of the strength of association (gamma value) between liver hepatocyte necrosis score with GLDH, immune infiltrates, and immunohistochemistry scores. All data (n = 6 per group) are reported as mean ± SEM. Immune cell scores were analyzed with a Kruskal-Wallis test and Dunn’s multiple comparisons test. Scale bar = 300 μm. *P<0.05, **P<0.01, ***P<0.0001. p: periportal, mc: mid-zonal/centrilobular.

Article Snippet: The IDO1 inhibitor epacadostat (Chemscene, Monmouth Junction, NJ) was administered twice daily (6 hours between each dose) as an oral gavage formulation with 0.5% HPMC/0.25% Tween 20 in a water base at 100, 300 or 600 mg/kg/dose.

Techniques: Immunohistochemical staining, Immunohistochemistry

Journal: PLoS ONE

Article Title: Inhibition of immune checkpoints PD-1, CTLA-4, and IDO1 coordinately induces immune-mediated liver injury in mice

doi: 10.1371/journal.pone.0217276

Figure Lengend Snippet: Mice were treated with anti-mouse CTLA-4 (9D9), lower dose (300 mg/kg BID) epacadostat or in combination for 6 weeks (A–H) or 3 weeks (I). Single cell suspensions of the whole liver were analyzed for (A) CD45+ cells, (B) CD11b+F4/80+ macrophages, (C) Gr1+CD11b+ myeloid derived suppressor cells (MDSC)s, (D) CD49b+ natural kill (NK) cells, (E) CD3+ T cells, (F) CD3+CD8+ T cells (G) CD3+CD4+ T cells, (H) Foxp3+CD4+ T cells, and (I) Ki67+ T cell subsets by flow cytometry. All data are reported as mean ± SEM. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, n = 6 per group, one-way ANOVA with Tukey’s multiple comparisons test (A–H) or n = 12 per group, unpaired t-test (I).

Article Snippet: The IDO1 inhibitor epacadostat (Chemscene, Monmouth Junction, NJ) was administered twice daily (6 hours between each dose) as an oral gavage formulation with 0.5% HPMC/0.25% Tween 20 in a water base at 100, 300 or 600 mg/kg/dose.

Techniques: Derivative Assay, Flow Cytometry

Journal: PLoS ONE

Article Title: Inhibition of immune checkpoints PD-1, CTLA-4, and IDO1 coordinately induces immune-mediated liver injury in mice

doi: 10.1371/journal.pone.0217276

Figure Lengend Snippet: Mice were treated with anti-mouse CTLA-4 (9D9), 300 mg/kg BID epacadostat or in combination for 6 weeks (A-E) or with 600 mg/kg BID epacadostat or in combination for 2 weeks (F-G). Linear regression analysis of (A) leukocytes (CD45+ cells), (B) macrophages, (C) total T cells and (D) CD4+ and (E) CD8+ T cells as defined in with single cell necrosis. (F) The top activated (> 1.5 z-score) toxicity pathways significantly modulated by 9D9 + epacadostat treatment compared with vehicle control, sorted by p-value and (G) the top five significantly regulated canonical pathways relative to vehicle identified by ingenuity pathway analysis (sorted by combination vs vehicle ascending p-values). There were no significant changes, and thus no activation of these canonical pathways, in the 9D9 vs vehicle group. PRR: pattern recognition receptor, ICOS: inducible T-cell costimulator ICOSL: inducible T-cell costimulatory ligand.

Article Snippet: The IDO1 inhibitor epacadostat (Chemscene, Monmouth Junction, NJ) was administered twice daily (6 hours between each dose) as an oral gavage formulation with 0.5% HPMC/0.25% Tween 20 in a water base at 100, 300 or 600 mg/kg/dose.

Techniques: Control, Activation Assay

Journal: The Febs Journal

Article Title: Pharmacological validation of TDO as a target for Parkinson’s disease

doi: 10.1111/febs.15721

Figure Lengend Snippet: Potency (IC 50 ) in n m of TDO inhibitors NTRC 3531‐0 and LM10 in biochemical and cell‐based assays for human (h) and mouse (m) TDO or IDO1. Confidence intervals and number of experimental replicates (n) are given within brackets

Article Snippet: The IDO1 inhibitor epacadostat, used as a reference inhibitor in the biochemical and cell‐based assays for IDO1, was purchased at MedKoo (cat. no. 206461).

Techniques: Inhibition