AZD5069

Our findings confirm the involvement of the CXCL8/CXCR2-axis in promoting pro-tumorigenic effects in TC cells, further demonstrating its immunotherapeutic significance in human cancer.

Interestingly, treatment with AZD-5069, a CXCR2 inhibitor, attenuates invasion and proliferation of tumor spheroids by blocking direct contact with neutrophils. Our findings also show that CXCR2 inhibition reduces neutrophil migration toward tumor spheroids. These results shed new light on the tumor-promoting mechanisms of human neutrophils and the tumor-suppressive mechanisms of CXCR2 inhibition in pancreatic cancer and may aid in the design and optimization of novel immunotherapeutic strategies based on neutrophils.

We conjugate a chemically modified small-molecule inhibitor of MDSC-surface receptor CXCR2 (AZD5069) with polyethylene glycol (PEG) and chemically graft AZD5069-PEG constructs onto amphiphilic polysaccharide derivatives to engineer CXCR2-homing nanoparticles (CXCR2-NP). Encapsulation of JAK2/STAT3i Ruxolitinib (CXCR2-NP Ruxo ) resulted in more durable attenuation in STAT3-regulated arginase activity from PMN-MDSCs and induction of cytolytic T-cell activity vs. free Ruxolitinib in-vitro and in-vivo . Cell-specific delivery of payloads via CXCR2-homing immunonanoparticles represents a novel strategy to disrupt MDSC-mediated immunosuppression and invigorate antitumor immunity in PDAC.

P1/2, N=340, Active, not recruiting, AstraZeneca | Phase classification: P1b/2 --> P1/2 | Trial completion date: Dec 2023 --> Mar 2025

Our findings reveal that blocking GCMSCs-derived IL-8/CXCR2 pathway decreasing PD-L1 expression and lactate production, improving antitumor efficacy of anti-PD-1 immunotherapy, may be of value for the treatment of advanced gastric carcinoma.

P1/2, N=30, Terminated, Institute of Cancer Research, United Kingdom | N=86 --> 30 | Trial completion date: Oct 2023 --> Nov 2022 | Active, not recruiting --> Terminated | Trial primary completion date: Jan 2023 --> Sep 2022; Discontinuation of production of IMP

Conversely, CXCR2 inhibition with AZD5069 led to reduced CD4 proliferation and PD-1 expression.Summary/ This study further highlights the importance of immune evasion in mediating AML relapse following AHSCT. This study further highlights the importance of immune evasion in mediating AML relapse following AHSCT. The chemokines CXCL2 and CXCL8 were identified as potential mediators of this process and may provide prospective targets to enhance the GvL effect to prevent or treat post-AHSCT relapse. Diagram Description automatically generated Chemokine, Acute myeloid leukemia, Allogeneic hematopoietic stem cell transplant

In this study, we analyzed the immune contexture of advanced HCC patients before and after pembrolizumab treatment to investigate the role of myeloid IL-8 in ICB-resistance mechanism and aim to intervene the IL-8 pathway through inhibition of its receptor by a clinical in-use CXCR2 antagonist...Therapeutic efficacy of CXCR2 pathway inhibition by AZD5069 in potentiating ICB response was evaluated in our ICB-resistant orthotopic mouse model which is generated by serial in vivo passaging of anti-PD-L1 residual tumor... We demonstrated the importance of myeloid IL-8/CXCR2 pathway in ICB-resistance from our advanced HCC cohort which paved way for IL-8 to become a novel prognostic target for immunotherapy. Blocking CXCR2 could reduce MDSC trafficking and overcome ICB-resistance in our pre-clinical HCC model, suggesting a promising combination regimen in future development.

The 2nd dose cohort opened to recruitment in September 2022. Exploratory studies (blood; pre- & on-treatment tumor and non-malignant liver biopsies) include biomarkers of CXCR2 inhibition (blood); proof-of-mechanism (tumor: expression of CXCR2, PD-L1, PD-1, CD8, CD4, CD66b, CD69); proof-of-mechanism (blood: ctDNA); drug-induced changes of mRNA expression, CXCR2 ligands & signalling pathway genes, T-cell and myeloid cell pathways, neutrophil-associated genes; predictive biomarkers (blood and tumour) include biomarkers of the CXCR2/PD-L1 immune axis; aberrant CXCR2 signalling pathways; proliferation biomarkers and CD10 (neutrophils), CD68 (macrophages), CD103 (T-regs); tumour mutational status.

P1b/2, N=340, Active, not recruiting, AstraZeneca | Trial completion date: Sep 2022 --> Dec 2023

We then chemically modified a surface receptor inhibitor of CXCR2 AZD5069 by conjugating it with polyethylene glycol to enhance aqueous solubility. Finally, co-culture of splenocyte-derived murine CD8+ T-cells with J774 treated with Ruxolitinib-NPCXCR2 showed significant improvement in T-cell IFN-γ release compared with free Ruxolitinib. Compartment-specific delivery of payloads targeting tolerogenic signaling in gMDSC via NPCXCR2 may represent a novel strategy to mitigate immunosuppression in PDAC.

Neutrophils were targeted in BPN mice with AZD5069 (CXCR2 inhibitor) +/- anti-PD1 antibody or vehicle control from 85 days post Cre induction (DPI)...Metastasis is reduced by neutrophil inhibition which synergises with anti PD1 therapy, supporting an actionable phenotype. The NAS identifies a poor prognostic patient group and future work will see if neutrophil targeted therapy could benefit these patients.

Conclusions The combination of AZD5069 and ENZA is tolerable and has antitumor activity in mCRPC. We provide clinical evidence for direct targeting of myeloid chemotaxis as a therapeutic strategy in mCRPC.

CXCR2 inhibition abrogated doxorubicin-mediated TGF-β upregulation in 3D in vitro TNBC coculture with PBMCs and eliminated drug resistance in TNBC mammospheres, suggesting a vital role for CXCR2 in TNBC doxorubicin-resistance via TGF-β signaling regulation. Moreover, CXCR2 inhibition improved the efficacy of the immunotherapeutic drug "atezolizumab" where the combined inhibition of CXCR2 and PDL1 in TNBC in vitro coculture showed an additive effect in cytotoxicity. Altogether, the current study suggests CXCR2 inhibitors as a promising approach to improve TNBC treatment if used in combination with chemotherapy and/or immunotherapy.

Inhibition of CXCR2 using small molecule antagonist AZD5069 reversed this behavior, limiting the neutrophil responses to the BrM and retarding the reciprocal tumor development. We further demonstrate that abrogation of NETs formation using Neutrophil Elastase Inhibitor (NEI) significantly decreases the influx of neutrophils towards BrM but not to their parental tumor, suggesting that CXCR2 activation could be used by the brain metastatic tumors as a mechanism to program the tumor-infiltrating TANs into a pro-NETotic state, so as to assume a unique spatial distribution that assists in the subsequent migration and invasion of the metastatic tumor cells. This new perspective indicates that CXCR2 is a critical target for suppressing neutrophilic inflammation in brain metastasis.

In this study we employ Imaging Mass Cytometry (IMC) to determine how co-therapy of anti-PD1 (Biolegend) and a small molecule inhibitor of the neutrophil chemokine receptor CXCR2 (AZD5069 – AstraZeneca) remodels the tumour microenvironment (TME) in a murine model of NASH-HCC... IMC allows for the identification of multiple cell types in a spatial context and quantification of changes in cell-cell interactions within the TME. In this study, IMC demonstrated that combined antiPD1 and CXCR2 inhibitor therapy in NASH-HCC encouraged the formation of cytotoxic immune hubs containing locally proliferating neutrophils, CD8+ T cells and antigen presenting cells. We then propose that the manipulation of neutrophils may have therapeutic potential in NASH-HCC.

P1b/2, N=340, Active, not recruiting, AstraZeneca | Trial completion date: Sep 2021 --> Sep 2022

Here, we investigate the potential use of AZD5069 in combination with atezolizumab (anti-PD-L1) in an ex-vivo TNBC model.   CXCR2 mRNA expression was analyzed using RealTime PCR in 65 BC biopsies and controls.  These data highlight the role of CXCR2 inhibition as a cutting edge approach to boost immunotherapy and combat chemo-resistance in TNBC.

Therapeutic efficacy of CXCR2 blockade was conducted in an orthotopic mouse model using AZD5069 (100 or 150 mg/kg) which is a CXCR2 antagonist currently undergoing clinical trials and in combination with anti-PD-L1 antibody (10F.9G2)...Conclusions Our data demonstrated the intricate link between IL-8/CXCR2 axis and MDSC trafficking to TME, providing insight into the immunosuppression mechanism in HCC. Targeting IL-8/CXCR2 chemotaxis pathway may potentiate ICB responsiveness, serving as a novel potential therapeutic option for effectively combined immunotherapy in liver cancer.

In vivo, administration of a CXCR1/2-targeted pepducin (x1/2pal-i3), or CXCR2-specific antagonist (AZD5069), in combination with IR to PTEN-deficient xenografts attenuated tumour growth and progression compared to control or IR alone. Post-mortem analysis confirmed that x1/2pal-i3 administration attenuated IR-induced CXCL signalling and anti-apoptotic protein expression. Interventions targeting CXC chemokine signalling may provide an effective strategy to combine with RT in locally advanced prostate cancer patients with known presence of PTEN-deficient foci.