Journal: PLOS Pathogens

Article Title: Kaposi sarcoma-associated herpesvirus cooperates with Epstein-Barr virus to co-transform a small set of human B cells oncogenically

doi: 10.1371/journal.ppat.1013281

Figure Lengend Snippet: (A) Cytokines known to support the growth of B cells were measured in conditioned media from cells infected only with EBV, from non-immortalized and immortalized co-transformed cells, and from established PEL cell lines (KSHV + EBV + or KSHV + EBV - ). (B) Co-transformed cells were grown at a density of 5X10 4 cells/ml in basal medium or in 50% conditioned medium. (*: p < 0.05) (C) Co-transformed cells were grown in complete medium or complete medium supplemented with rapamycin (100 nM) or pacritinib (100 nM) for 48 hours. (**: p < 0.01; ***: p < 0.001).

Article Snippet: Cells were grown in complete medium (RPMI-1640 supplemented with FBS and antibiotics as described previously) or complete medium supplemented with 100nM Rapamycin (Tocris Bioscience, Minneapolis MN) or 100nM Pacritinib (MedChem Express, Monmouth Junction NJ) for 48 hours.

Techniques: Infection, Transformation Assay

Journal: Current Opinion in Hematology

Article Title: IRAK1 and IRAK4 as emerging therapeutic targets in hematologic malignancies

doi: 10.1097/MOH.0000000000000693

Figure Lengend Snippet: Preclinical and clinical interleukin 1 receptor-associated kinase inhibitors

Article Snippet: Pacritinib , CTI Biopharma , FLT3 (IC 50 = 22 nmol/l) , Kinase inhibitor , Myeloproliferative neoplasms , Clinical.

Techniques:

Journal: Drug Discovery Today

Article Title: Calming the cytokine storm of COVID-19 through inhibition of JAK2/STAT3 signaling

doi: 10.1016/j.drudis.2021.10.016

Figure Lengend Snippet: JAK2 inhibitors that are approved by the US FDA or being evaluated in human clinical trials.

Article Snippet: Pacritinib (SB1518) , AbbVie , Primary myelofibrosis (PMF)/post-polycythemia vera MF, post-essential thrombocythemia MF , 3/Recruiting/May 2017 , NCT03165734.

Techniques:

Journal: Oncogene

Article Title: Combined inhibition of JAK2-STAT3 and SMO-GLI1/tGLI1 pathways suppresses breast cancer stem cells, tumor growth, and metastasis

doi: 10.1038/s41388-020-01454-1

Figure Lengend Snippet: (A) IC 50 values calculated from the results of the CellTiter Blue assay. Cell lines were seeded in 96-well plates and treated with test compound for 48 hours. (B) Combination Index (CI) values calculated for various drug combinations and ratios using CompuSyn. Breast cancer cells were seeded in 96-well plates and treated with drug combinations for 48 hours. Only a selection of tested drug ratios is shown. CI<1 indicates synergy, CI=1 indicates addition, and CI>1 indicates antagonism. Synergistic combinations are highlighted in red. (C) Normalized MDA-231 cell viability following combination treatments. Experiments were repeated at least three times to derive averages. Data are presented as mean ± SEM. One-way ANOVA with Tukey’s multiple comparison post hoc test was used to compute p values. TN, triple-negative. CL, claudin-low. LumB, Luminal B. Rux, ruxolitinib. Pac, pacritinib. Vis, vismodegib. Son, sonigedib. TtzmR, trastuzumab-resistant. ***, p < 0.001.

Article Snippet: Pacritinib, ruxolitinib, sonidegib, and vismodegib were obtained from AdooQ Bioscience (Irvine, CA, USA) and stock solutions were prepared using dimethyl sulfoxide (DMSO) for administration in cell culture.

Techniques: CtB Assay, Selection, Comparison

Journal: Oncogene

Article Title: Combined inhibition of JAK2-STAT3 and SMO-GLI1/tGLI1 pathways suppresses breast cancer stem cells, tumor growth, and metastasis

doi: 10.1038/s41388-020-01454-1

Figure Lengend Snippet: (A) Representative flow cytometry scatter plots of CD44 + /CD24 − MDA-231 cells. (B–E) Combination JAK2 inhibitor (JAK2i) and SMO inhibitor (SMOi) treatments more effectively suppressed the CD44 high /CD24 low MDA-231 population compared to single agent treatments alone as determined by FACS. The combinations of Pacritinib-Sonidegib (B) and Ruxolitinib-Soniedgib (C) were more effective than vehicle or either agent alone while the Pacritinib-Vismodegib (D) combination was not more effective than Pacritinib alone. The combination of Ruxolitinib-Vismodegib (E) did not suppress the CD44 high /CD24 low MDA-231 population. (F-I) Certain JAK2i-SMOi combination treatments synergized to more effectively inhibit BT20 mammosphere formation compared to single agent treatments alone. The combinations of Pacritinib-Sonidegib (F) and Ruxolitinib-Vismodegib (I) were more effective than vehicle or either agent alone while the Pacritinib-Vismodegib (H) combination was not more effective than Pacritinib alone. The combination of Ruxolitinib-Sonidegib (G) did not inhibit mammosphere formation. (J) Pacritinib-Sonidegib combination treatment suppressed formation of BT474-TtzmR mammospheres. Representative BT20 or BT474-TtzmR mammosphere images are shown below the quantified data in each panel. Scale bars represent 200 μm. Data of three or four experimental repeats are presented as mean ± SEM. One-way ANOVA with Tukey’s multiple comparison post hoc test was used to compute p values. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: Pacritinib, ruxolitinib, sonidegib, and vismodegib were obtained from AdooQ Bioscience (Irvine, CA, USA) and stock solutions were prepared using dimethyl sulfoxide (DMSO) for administration in cell culture.

Techniques: Flow Cytometry, Comparison

Journal: Oncogene

Article Title: Combined inhibition of JAK2-STAT3 and SMO-GLI1/tGLI1 pathways suppresses breast cancer stem cells, tumor growth, and metastasis

doi: 10.1038/s41388-020-01454-1

Figure Lengend Snippet: (A) Schema for MFP mouse model. Luciferase-expressing MDA-231 breast cancer cells were implanted into the left inguinal MFP. Tumor growth was assessed via caliper measurements and bioluminescent imaging. Once tumors reached an average of 90 mm 3 , mice were randomized into vehicle, 7.5 mg/kg pacritinib, 20 mg/kg sonidegib, or combination treatment groups (N=9–10) and received three intraperitoneal treatments per week. (B) Growth curve for MFP tumors. (C) Representative bioluminescent images of xenograft-bearing female nude mice 30 days post-inoculation. (D) Quantification of organ-specific metastasis incidence. E) Representative bioluminescent ex vivo organ images are shown. (F-H) Ex vivo lung (F) , bone (G) , and liver (H) bioluminescence was quantified. (I) Average mouse weights over the course of the study. One-way ANOVA with Tukey’s multiple comparison post hoc test was used to compute p values.

Article Snippet: Pacritinib, ruxolitinib, sonidegib, and vismodegib were obtained from AdooQ Bioscience (Irvine, CA, USA) and stock solutions were prepared using dimethyl sulfoxide (DMSO) for administration in cell culture.

Techniques: Luciferase, Expressing, Imaging, Ex Vivo, Comparison

Journal: Oncogene

Article Title: Combined inhibition of JAK2-STAT3 and SMO-GLI1/tGLI1 pathways suppresses breast cancer stem cells, tumor growth, and metastasis

doi: 10.1038/s41388-020-01454-1

Figure Lengend Snippet: (A) Schema for intracardiac mouse model. Luciferase-expressing MDA-231 breast cancer cells were implanted intracardially via the left ventricle of female nude mice and tumor growth was assessed biweekly via bioluminescent imaging. Mice that underwent successful inoculations were randomized into vehicle, 7.5 mg/kg pacritinib, 20 mg/kg sonidegeib, or combination treatment groups (N=8–9) and received three intraperitoneal treatments per week. (B) Representative bioluminescent images of female nude mice 28 days post-inoculation. (C) Pacritinib-sondiegib combination treatment prolonged overall survival of mice bearing MDA-231 xenografts compared to vehicle or either agent alone. Mantel-Cox log-rank test was used to determine p values. (D) Average mouse weights over the course of the study. (E) Representative bioluminescent ex vivo organ images are shown. (F) Quantification of organ-specific metastasis incidence. ( G–J) Ex vivo lung (G) , liver (H) , brain (I) , and (J) bone bioluminescence was quantified. One-way ANOVA with Tukey’s multiple comparison post hoc test was used to compute p values. *, p < 0.05.

Article Snippet: Pacritinib, ruxolitinib, sonidegib, and vismodegib were obtained from AdooQ Bioscience (Irvine, CA, USA) and stock solutions were prepared using dimethyl sulfoxide (DMSO) for administration in cell culture.

Techniques: Luciferase, Expressing, Imaging, Ex Vivo, Comparison

Journal: Oncogene

Article Title: Combined inhibition of JAK2-STAT3 and SMO-GLI1/tGLI1 pathways suppresses breast cancer stem cells, tumor growth, and metastasis

doi: 10.1038/s41388-020-01454-1

Figure Lengend Snippet: (A) Schema for MFP mouse model. Luciferase-expressing BT474-TtzmR breast cancer cells were implanted into the right inguinal MFP. Tumor growth was assessed via caliper measurements and bioluminescent imaging. Once tumors reached an average of 90 mm 3 , mice were randomized into vehicle, 7.5 mg/kg pacritinib, 20 mg/kg sonidegib, or combination treatment groups (N=8–9) and received three intraperitoneal treatments per week. (B) Growth curve for MFP tumors. (C) Representative bioluminescent images of xenograft-bearing female nude mice 50 days post-inoculation. (D) Quantification of organ-specific metastasis incidence. (E) Representative bioluminescent ex vivo organ images are shown. (F–H) Ex vivo lung (F) , bone (G) , and liver (H) bioluminescence was quantified. (I) Average mouse weights over the course of the study. (J) Circulating alanine transaminase (ALT) levels were not significantly increased in any treatment group. Data presented as mean ± SEM. One-way ANVOA with Tukey’s multiple comparison post hoc test was used to compute p values. *, p < 0.05

Article Snippet: Pacritinib, ruxolitinib, sonidegib, and vismodegib were obtained from AdooQ Bioscience (Irvine, CA, USA) and stock solutions were prepared using dimethyl sulfoxide (DMSO) for administration in cell culture.

Techniques: Luciferase, Expressing, Imaging, Ex Vivo, Comparison

Journal: Oncogene

Article Title: Combined inhibition of JAK2-STAT3 and SMO-GLI1/tGLI1 pathways suppresses breast cancer stem cells, tumor growth, and metastasis

doi: 10.1038/s41388-020-01454-1

Figure Lengend Snippet: (A) RT-qPCR analysis of JAK2-STAT3 and SMO-GLI1/tGLI1 target genes following treatment of BT474-TtzmR cells in vitro . (B) Representative IHC images of VEGF-A, mCD31, Ki-67, and p-STAT3 (Y705) of treated BT474-TtzmR mammary fat pad xenografts. Images were taken under 20X magnification. Arrows point to examples of positive staining. (C–E) Quantification of histologic staining of VEGF-A (C) , mCD31 (microvessel density; (D) , % Ki-67 positivity (E) , and % p-STAT3 (Y705) positivity (F) of treated BT474-TtzmR MFP xenografts. (G) Quantification of the number of TUNEL positive cells of mammary fat pad xenografts from each treatment group. (H) Representative images of TUNEL-positive staining of mammary fat pad xenografts from each treatment group. TUNEL, DAPI, and TUNEL-DAPI merged. Images were taken under 10X magnification. For all analyses, at least 5 fields were quantified per tumor section (N=4–5 tumors/group). All scale bars represent 100 μm. Pac, pacritinib. Son, sonidegib. Data presented as mean ± SEM. One-way ANOVA with Tukey’s multiple comparison post hoc test was used to compute p values. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: Pacritinib, ruxolitinib, sonidegib, and vismodegib were obtained from AdooQ Bioscience (Irvine, CA, USA) and stock solutions were prepared using dimethyl sulfoxide (DMSO) for administration in cell culture.

Techniques: Quantitative RT-PCR, In Vitro, Staining, TUNEL Assay, Comparison

Journal: Cancers

Article Title: Novel Methods of Targeting IL-1 Signalling for the Treatment of Breast Cancer Bone Metastasis

doi: 10.3390/cancers14194816

Figure Lengend Snippet: The effects of IL1β overexpression and inhibitors of the IL1β pathway on viability of E0771 and breast cancer cells in vitro. ( A ) Cartoon showing where VX765/VRT043198 (Caspase-1 inhibitors), MLX01 (IL1β secretion inhibitor), Anakinra (IL1R antagonist) and Pacritinib (IRAK1 inhibitor) inhibit the IL1β signalling pathway. ( B , C ) Effects of overexpression of IL1β (IL1B+), overexpression of IL1R1 (IL1R1+), inhibition of IL1β (12 ng/mL MLX01), inhibition of Caspase-1 (25 ng/mL VRT043198), inhibition of IL1R (0.1mg/kg Anakinra) and inhibition of IRAK1 (250 pg/mL Pacritinib) and recombinant IL1β (5 ng/mL) on tumour cell viability, normalised to control. All experiments are from three biological experiments and their graphs represent mean + SD, * p < 0.05, ** p < 0.01.

Article Snippet: VRT043198 (244133-31-1) and Pacritinib (202571) were purchased from Medkoo (Morrisville, NC, USA).

Techniques: Over Expression, In Vitro, Inhibition, Recombinant

Journal: Cancers

Article Title: Novel Methods of Targeting IL-1 Signalling for the Treatment of Breast Cancer Bone Metastasis

doi: 10.3390/cancers14194816

Figure Lengend Snippet: Inhibiting IL-1 signalling with IL1β inhibitor MLX01, Caspase-1 inhibitor VX765 and IL1R antagonist Anakinra reduced metastatic outgrowth from E0771 cells disseminated in the bone. ( A ) Outline of experimental protocol for administration of PBS (control), MLX01, VX765 and Anakinra in mice following intra-cardiac injection of E0771. ( B ) Representative whole body IVIS images of mice at Day 8 and Day 15 post injection of E0771 cells. ( C ) Numbers of overt metastases per mouse 8-, 15- and 16-days post injection of E0771 cells following daily administration of PBS (control), MLX01, VX765 or Anakinra. ( D ) Total luciferase expression (photons per second (P/S)) of whole mice 8 and 15 days after E0771 cell injection. ( E ) Numbers of mice that developed bone metastases 8, 15 and 16 days after E0771 cells had been disseminated in the bone following daily administration of PBS (control) MLX01, VX765 or Anakinra. ( F ) Size of tumours that grew in the bone as assessed by photons per second (P/S) in luciferase expressing E0771 tumour cells. ( G ) Shows effects of PBS (control), MLX01, VX765 or Anakinra on metastatic outgrowth of tumours following dissemination into the lungs, kidneys and livers. ( H ) Protocol outline for administration of Pacritinib in mice after intra-cardiac injection of E0771. ( I ) Representative images of whole body IVIS imaging 17 days post E0771 injection. ( J ) Numbers of overt metastases per mouse 17 days post injection of E0771 cells and following daily administration of PBS (control), or Pacritinib. ( K ) Total luciferase signal from E0771 tumour cells (photons per second (P/S)) in whole mice 17 days following injection of E0771 cells and administration of PBS (control) or Pacritinib. ( L ) Numbers of mice that developed bone metastases 17 days after E0771 cells had been disseminated in the bone following daily administration of PBS (control) or Pacritinib and size of tumours that grew in the bone as assessed by IVIS imaging in ( M ). ( N ) Shows effects of PBS (control), or Pacritinib on metastatic outgrowth of tumours following dissemination into the lungs, kidneys and livers. *** p < 0.001, **** p < 0.0001.

Article Snippet: VRT043198 (244133-31-1) and Pacritinib (202571) were purchased from Medkoo (Morrisville, NC, USA).

Techniques: Injection, Luciferase, Expressing, Imaging

Journal: Cancers

Article Title: Novel Methods of Targeting IL-1 Signalling for the Treatment of Breast Cancer Bone Metastasis

doi: 10.3390/cancers14194816

Figure Lengend Snippet: Effects of inhibiting IL1β, IL1R1, Caspase-1 and IRAK1 on bone turnover and volume ( A ) Bone volume/tissue volume (BV/TV) % of trabecular bone in the mouse tibia following dissemination of E0771 cells in this site and 13 consecutive daily treatments with PBS (control), MLX01, VX765 or Anakinra (Study A) or ( B ) 15 days 15 consecutive daily treatments with Pacritinib (Study H). ( C ) Bone volume/tissue volume (BV/TV) % of trabecular bone in mouse tibia following orthotopic injection of E0771 and consecutive daily treatments with PBS (control), MLX01, VX765 or Anakinra until primary tumour reached 1000 mm 3 . ( D , E ) ELISA analysis of TRAP5b ( D ) or P1NP ( E ) in mouse serum following dissemination of E0771 cells and 13 consecutive daily treatments with MLX01, VX765 or Anakinra (Study A) or Pacritinib (Study H). Graphs represent mean + SD, * p < 0.05, ** p < 0.01.

Article Snippet: VRT043198 (244133-31-1) and Pacritinib (202571) were purchased from Medkoo (Morrisville, NC, USA).

Techniques: Injection, Enzyme-linked Immunosorbent Assay

Journal: Blood Cancer Journal

Article Title: The oral HDAC inhibitor pracinostat (SB939) is efficacious and synergistic with the JAK2 inhibitor pacritinib (SB1518) in preclinical models of AML

doi: 10.1038/bcj.2012.14

Figure Lengend Snippet: Pracinostat downregulates JAK and FLT3 signaling in JAK2 V617F and FLT-ITD cell lines, and shows synergy in combination with pacritinib. ( a – d ) Western blot analyses from 25 or 50 μg (pFLT3 and FLT3 blots only) of total cell lysate of the indicated cell lines with JAK2 V617F , JAK2, or FLT3-ITD or FLT3 wt are shown. ( a , b ) Cells were treated with the indicated concentrations of pracinostat (SB939) for 24 h. ( c , d ) Cells were treated for 24 h with pracinostat with pacritinib (SB1518) added in the indicated concentrations for the last 2 h of incubation before lysis.

Article Snippet: Pracinostat (SB939) as hydrochloride salt and pacritinib (SB1518) as citrate salt were synthesized by SAI Advantium Pharma Ltd (Hyderabad, India).

Techniques: Western Blot, Incubation, Lysis

Journal: Blood Cancer Journal

Article Title: The oral HDAC inhibitor pracinostat (SB939) is efficacious and synergistic with the JAK2 inhibitor pacritinib (SB1518) in preclinical models of AML

doi: 10.1038/bcj.2012.14

Figure Lengend Snippet: In vitro combination of pracinostat (SB939) and pacritinib (SB1518)

Article Snippet: Pracinostat (SB939) as hydrochloride salt and pacritinib (SB1518) as citrate salt were synthesized by SAI Advantium Pharma Ltd (Hyderabad, India).

Techniques: In Vitro, Sequencing

Journal: Blood Cancer Journal

Article Title: The oral HDAC inhibitor pracinostat (SB939) is efficacious and synergistic with the JAK2 inhibitor pacritinib (SB1518) in preclinical models of AML

doi: 10.1038/bcj.2012.14

Figure Lengend Snippet: Pracinostat combined with pacritinib is efficacious and synergistic in vivo in two different models of human AML. Female SCID-beige mice ( n =12 per group) were inoculated with 5 × 10 6 SET-2 cells and treatment was started on d33 post-implantation ( a , b ). For the combination group, both pacritinib (SB1518) and pracinostat (SB939) were administered in half the volume (as a 20 mg/kg solution) every other day; when both compounds were given simultaneously, all other doses were given at 10 mg/kg, at least 8 h apart for 19 days (d0–d18). There was one non-treatment-related death in the vehicle group on study d11 (gavaging error). Female SCID mice ( n =12 per group) were inoculated with 5 × 10 6 Molm-13 cells ( c , d ). Treatment was started on d11, animals were dosed every other day for 8 days, in contrast to ( a ), animals were dosed with pacritinib only daily (in the evenings), pracinostat dosing remained unchanged, all dosing was done in a 10 mg/kg solution. On the last day, mice were killed and excised tumors were weighed. The doses showing significant TGI versus vehicle (on the last day of the study) using analysis of variance/Dunnet's post test, are indicated with ** P <0.01 or *** P <0.001. ( e ) Tumors from the MOLM-13 efficacy study ( c , d ) were harvested on the last day 3 h post pacritinib dosing and immediately lysed. Western blot analyses for pFLT3, FLT3, pSTAT5 and β-actin are shown from the lysates of three randomly selected tumors from each treatment group of ( c ).

Article Snippet: Pracinostat (SB939) as hydrochloride salt and pacritinib (SB1518) as citrate salt were synthesized by SAI Advantium Pharma Ltd (Hyderabad, India).

Techniques: In Vivo, Western Blot

Journal: Blood Cancer Journal

Article Title: The oral HDAC inhibitor pracinostat (SB939) is efficacious and synergistic with the JAK2 inhibitor pacritinib (SB1518) in preclinical models of AML

doi: 10.1038/bcj.2012.14

Figure Lengend Snippet: Pharmacokinetic analysis of pracinostat and pacritinib administered in combination. Female BALB/c nude mice were dosed with 150 mg/kg pacritinib (SB1518) plus 75 mg/kg pracinostat (SB939) simultaneously either once (‘day 1') or for a total of 14 days (i.e., 7 doses of SB939, administered every other day in the morning, plus SB1518 for 14 times, every day morning and evening, ‘day 14'). Plasma was collected at 10, 30 and 60 min post-dose on d1/d14 ( n =5) as well as 4, 8 and 24 h post-dose ( n =3). Pharmacokinetic parameters were calculated by a non-compartmental method, using WinNonlin software, and compared with values obtained in previous experiments in the same mouse strain. * Indicates that values were extrapolated from BALB/c nude mice dosed with 50 mg/kg pacritinib or pracinostat. Results for pracinostat are shown in ( a ), and for pacritinib in ( b ).

Article Snippet: Pracinostat (SB939) as hydrochloride salt and pacritinib (SB1518) as citrate salt were synthesized by SAI Advantium Pharma Ltd (Hyderabad, India).

Techniques: Clinical Proteomics, Software