SBIR-STTR Award

Carcinoma of the pancreas, or pancreatic cancer (PC), is the fourth leading cause of cancer-related death in the United States. According to the American Cancer Society, 55,300 new cases are expected in 2016. Despite advances in therapy, the 5-year survival rate is less than 4%. Current treatments of PC include surgery, radiation therapy, chemotherapy, and immunotherapy but they all only slightly prolong survival or relieve symptoms in patients with PC. Gemcitabine (GEM), first line therapy for advanced PC, is only modestly effective with a median survival of about 6 months in randomized clinical trials, The combination of GEM with different anticancer agents does not show significant survival advantage as compared with GEM alone. These limitations in efficacy of available treatments highlight the need for new treatments. Pancreatic inflammation is known to increase the risk of PC. High macrophage infiltration into the tumor mass correlates with the promotion of tumor growth and metastasis development. Triggering receptor expressed on myeloid cells (TREM-1), an inflammation amplifier, plays a role in PC progression. Expression of TREM-1 on tumor-associated macrophages (TAMs), is upregulated in patients with PC and correlates to disease severity. Recently, we demonstrated that a first-in-class TREM-1 inhibitory peptide GF9 in free form and bound to macrophage-targeted lipopeptide complexes that mimic human high density lipoproteins (GF9-HDL) inhibits tumor growth in animal models of PC. We also showed that blockade of TREM-1 inhibits release of cytokines and M-CSF in these animal models. The main hypothesis of this project is that a combination therapy that includes TREM-1 inhibitors and anticancer agents and targets cancer-related inflammation and tumor cells directly can synergistically improve survival of PC patients. We also hypothesize that this effect will be especially pronounced in PC patients with high intratumoral macrophage infiltration. Our preliminary studies strongly support this hypothesis. The long-term objective of the proposed project is to develop a novel combinatorial approach to efficiently target PC. The major goal of the Phase I study is to demonstrate that specific inactivation of TREM-1 with first-in-class inhibitory peptides in combination with GEM or nanoparticle albumin (nab)- bound paclitaxel (nab-PTX), another promising agent that directly targets cancer cells and is widely approved for the treatment of metastatic breast cancer (BC), synergistically suppresses PC tumor progression in animal model system and improves survival. Phase I specific aims are to: 1) evaluate effects and mechanisms of GF9-GEM and GF9-nab-PTX combinations in vitro, and 2) test GF9-GEM and GF9- nab-PTX combinations in two xenograft mouse models of PC. Non-toxic peptide GF9, which employs novel, ligand-independent mechanisms of TREM-1 inhibition, is anticipated to have less severe side effects. In order to increase peptide solubility, bioavailability and targeting to TAMs, we will utilize SignaBlok's proprietary HDL-based nanosystem for macrophage-targeted delivery of water insoluble and poorly water soluble drugs. We will use in vitro macrophage uptake assay to elucidate the molecular mechanisms of a putative receptor-mediated process of targeted delivery of GF9 to macrophages. We will optimize GF9 formulations based upon their stability, GF9 content, and macrophage uptake in vitro. We will use an in vitro cytotoxicity assay and immunoblot analysis to test proliferation of BxPC-3 and AsPC-1 cells as well as expression of phospho-stathmin and alpha-tubulin in the presence of GEM, nab-PTX or their combinations with GF9 formulations. We will use BxPC-3 and AsPC-1 mouse xenograft models to test the ability of GF9- GEM and GF9-nab-PTX combinations to synergistically inhibit tumor progression and promote survival as compared with GF9, GEM, and nab-PTX alone. Free GF9 and GF9-HDL will be tested. Comprehensive histology and immunohistochemistry studies will be performed to analyze angiogenesis, intratumoral macrophage infiltration, and potential non-specific toxicity for organ/tissues. It is anticipated that the proposed research will identify a novel anticancer combination approach that will set the stage for the development of new targeted combination therapies of PC, thereby leading to a higher survival rate of the patients. If successful, the Phase I will be followed in the Phase II by toxicology, absorption/ disposition/ metabolism/ excretion (ADME), pharmacology and chemistry/ manufacturing/ control (CMC) studies, filing an Investigational New Drug (IND) application with the US Food and Drug Administration (FDA) and subsequent evaluation in humans. Final product will be the stable TREM-1- targeted lipopeptide formulation that can be used in combination therapies of PC patients to prolong their survival. Importantly, our recent data demonstrate that blockade of TREM-1 suppresses in vivo progression of not only PC but also non-small cell lung cancer (NSCLC). Thus, successful completion of Phase I will provide the proof of concept of the hypothesis that might be applicable to a variety of inflammation- associated tumors such as NSCLC, BC, colon cancer, and others.

Public Health Relevance Statement:
Project Narrative Pancreatic cancer is the fourth leading cause of cancer-related death in the United States, and the 5-year survival rate is less than 4%. Current treatments are substantially ineffective and only slightly prolong survival or relieve symptoms in the cancer patients. The proposed research is expected to result in the development of novel anticancer therapeutic combinations that could substantially improve treatment of this type of cancer, thereby leading to a higher survival rate of the patients.

Project Terms:
absorption; Adenocarcinoma; Adverse effects; Albumins; alpha Tubulin; American Cancer Society; Amplifiers; angiogenesis; Animal Model; Animal Testing; Animals; anti-cancer therapeutic; anticancer activity; Antineoplastic Agents; Apolipoprotein A-I; base; Biological; Biological Assay; Biological Availability; Biological Models; cancer cell; Cancer Etiology; Cancer Patient; cancer therapy; cancer type; Cells; Cessation of life; Chemistry; chemotherapy; Colon Carcinoma; combinatorial; Combined Modality Therapy; comparative; Comparative Study; Complex; Controlled Study; cytokine; cytotoxicity; Data; Development; Dose; drug candidate; Drug Combinations; drug development; Evaluation; Excretory function; Female; Formulation; Future; gemcitabine; Goals; Half-Life; High Density Lipoproteins; Histology; Human; Immunoblot Analysis; Immunohistochemistry; Immunotherapy; improved; In Vitro; in vivo; Infiltration; Inflammation; inhibitor/antagonist; Interleukin-6; Investigational New Drug Application; lead series; Lesion; Ligands; macrophage; Macrophage Colony-Stimulating Factor; malignant breast neoplasm; Malignant neoplasm of pancreas; Malignant Neoplasms; Mediating; Metabolism; Metastatic breast cancer; Molecular; mouse model; Mus; Myeloid Cells; nanoparticle; nanosystems; Neoplasm Metastasis; neoplastic cell; Non-Small-Cell Lung Carcinoma; novel; Nude Mice; oncology; Operative Surgical Procedures; Organ; Oxides; Paclitaxel; Pancreas; Pancreatic carcinoma; Patients; peptide I; Peptides; Pharmaceutical Preparations; Pharmacology; Phase; phase 1 study; Plasma; Play; Process; Radiation therapy; Randomized Clinical Trials; receptor; reduce symptoms; Regimen; Research; Risk; Role; Severity of illness; sex; Site; Solubility; stathmin; Supervision; Survival Analysis; Survival Rate; targeted delivery; Testing; Tissues; Toxic effect; Toxicology; tumor; tumor growth; tumor progression; Tumor Promotion; United States; United States Food and Drug Administration; uptake; Water; Work; Xenograft Model; Xenograft procedure

Carcinoma of the pancreas, or pancreatic cancer (PC), is the third leading cause of cancer-related death in the US. Despite recent advances in the current treatments that include surgery, radiation therapy, chemo- and immunotherapy, the 5-year survival rate is as low as 9%. The long-term goal of this project is to develop a first-in-class, efficient and well tolerable therapy for PC to be used standalone or with standard chemo- and/or immune checkpoint blockade (ICB) treatments as induction and/or maintenance therapy. In PC patients, overexpression of TREM-1 correlates with poor survival, implicating TREM-1 as a new target. Current TREM-1 blockers all attempt to block binding of uncertain ligand(s) to TREM-1. To reduce risk of failure in the clinic, we developed a ligand-independent TREM-1 inhibitory peptide GF9 that can be formulated into macrophage-specific lipopeptide complexes (LPC) to improve its half-life and targeting. In Phase I, we showed that: 1) In suppressing tumor growth and improving survival, TREM-1 blockade using GF9-LPC in PC xenografts is as effective as a standard chemo: gemcitabine (GEM)+nab-paclitaxel (PTX) combo, and 2) addition of GF9-LPC to GEM+nab-PTX sensitizes the tumor to chemo and triples survival rate of mice. Mechanistically, in PC xenografts, GF9-LPC reduces tumor-associated macrophage (TAM) infiltration and serum level of CSF1. As shown by others, in mice with hepatocellular carcinoma, blocking TREM-1+ TAMs by GF9 reverses immunosuppression and overcomes anti-PDL1 resistance. The goal of this project is to further develop GF9 therapy for PC to be used as an induction/maintenance therapy alone or with first-line standard chemo treatments (GEM+nab-PTX) and/or ICB (anti-PD1/PDL1). Phase II aims are to: 1) generate and test rationally designed manufacturing friendly GF9 sequence- based formulations with favorable pharmacokinetic profile and high efficacy in vivo and select the lead (sub-aim – develop an assay to analyze GF9 in blood in PK studies), 2) test the lead in combination with GEM+nab-PTX in xenograft and syngeneic mouse models of PC, 3) test the lead in combination with anti- PD1/PDL1 in syngeneic mouse models of PC, and 4) test the lead in the non-clinical toxicology studies. Histology/IHC studies will be performed to analyze intratumoral macrophage infiltration as well as angiogenesis, tumor cell proliferation and death. Cytokines including CSF-1 will be analyzed. Follow-up Phase IIb will include other administration and combination (eg, radiation+GF9; anti-CSF-1R+ GF9) regimen, TOX, ADME, CMC and other IND-enabling studies. Final manufacturing friendly product will represent safe and stable PC therapy. Its anticipated safety is supported by good tolerability of SignaBlok's GF9 sequence-based formulations by long term-treated healthy, cancer and arthritic mice. Prototypes of SignaBlok's LPC were safe and well tolerated in clinical trials. TREM-1 blockade using peptide LR12 by SignaBlok's top competitor (Inotrem, France) was safe and well tolerated in healthy and septic subjects.

Public Health Relevance Statement:
Project Narrative Pancreatic cancer is the third leading cause of cancer-related death in the United States, and the 5-year survival rate is less than 9%. Current treatments are substantially ineffective and only slightly prolong survival or relieve symptoms in the cancer patients. The proposed research is expected to result in the development of novel, first-in-class therapeutic combinations that could substantially improve treatment of this type of cancer, thereby leading to a higher survival rate of the patients and higher quality of their life.

NIH Spending Category:
Biotechnology; Cancer; Digestive Diseases; Immunotherapy; Lung; Lung Cancer; Orphan Drug; Pancreatic Cancer; Rare Diseases

Project Terms:
angiogenesis; Animal Cancer Model; Animals; anti-PD-1; anti-PD-L1; Arthritis; base; Base Sequence; Binding; Biological Assay; Biophysics; Blood; Cancer Etiology; Cancer Patient; cancer therapy; cancer type; Canis familiaris; Cell Death; Cell Proliferation; Cells; Cessation of life; Clinic; Clinical Trials; Combined Modality Therapy; Complex; CSF1 gene; cytokine; Data; design; Development; Disease; Dose; Drug Kinetics; Drug Targeting; Failure; follow-up; Formulation; France; gemcitabine; Goals; Half-Life; Histology; Human; immune checkpoint blockade; Immunosuppression; Immunotherapy; improved; In Vitro; in vivo; Infiltration; inhibitor/antagonist; innovation; Lead; Life; Ligands; macrophage; Macrophage Colony-Stimulating Factor; Maintenance Therapy; Malignant neoplasm of pancreas; Malignant Neoplasms; mouse model; Mus; Neoadjuvant Therapy; Neoplasm Metastasis; neoplastic cell; Non-Small-Cell Lung Carcinoma; novel; novel therapeutics; Operative Surgical Procedures; overexpression; Paclitaxel; pancreatic cancer model; pancreatic cancer patients; Pancreatic carcinoma; Pathology; Patients; PD-1/PD-L1; Peptides; Phase; Primary carcinoma of the liver cells; protein aminoacid sequence; prototype; Radiation; Radiation therapy; Rattus; reduce symptoms; Regimen; Research; Resistance; Risk; Safety; septic; Serum; Structure; Survival Rate; targeted treatment; Testing; Therapeutic; Toxicology; tumor; tumor growth; Tumor Volume; Tumor-associated macrophages; United States; Xenograft procedure