title: Defining the Pharmacodynamic Profile and Therapeutic Index of NHS-IL12 Immunocytokine in Dogs with Malignant Melanoma.
abstract: BACKGROUND
  Interleukin (IL)-12 is a pro-inflammatory cytokine that mediates T-helper type 1 responses and cytotoxic T-cell activation, contributing to its utility as anti-cancer agent. Systemic administration of IL-12 often results in unacceptable toxicity; therefore, strategies to direct delivery of IL-12 to tumors are under investigation. The objective of this study was to assist the preclinical development of NHS-IL12, an immunocytokine consisting of an antibody, which targets necrotic tumor regions, linked to IL-12. Specifically this study sought to evaluate the safety, serum pharmacokinetics, anti-tumor activity, and immune modulation of NHS-IL12 in dogs with naturally occurring cancers.
  
  
  RESULTS
  A rapid dose-escalation study of NHS-IL12 administered subcutaneously to dogs with melanoma was conducted through the Comparative Oncology Trials Consortium (COTC). Eleven dogs were enrolled in four dose-escalation cohorts; thereafter, an additional seven dogs were treated at the defined tolerable dose of 0.8 mg/m2. The expanded cohort at this fixed dose (ten dogs in total) was accrued for further pharmacokinetics and pharmacodynamics assessment. NHS-IL12 levels, serum cytokine concentrations, and peripheral blood mononuclear cell characterization (post-treatment) and draining lymph node immune profiling, and tumor biopsies (pre- and post-treatment) were collected. Adverse events included thrombocytopenia, liver enzymopathies, fever, and vasculitis. Correlation between interferon (IFN)-γ induction, adverse events, and NHS-IL12 exposure (maximum concentration and area under the concentration-time curve) were dose-dependent. Serum IL-10 levels and intratumoral CD8+ populations increased after treatment. Partial responses, according to Response Evaluation Criteria in Solid Tumors (RECIST) criteria, were observed in two dogs treated with NHS-IL12 0.8 mg/m2 and 1.6 mg/m2.
  
  
  CONCLUSIONS
  NHS-IL12 was administered safely to dogs with melanoma and both immunologic and clinical activity was observed. This study successfully defined a narrow therapeutic window for systemic delivery of NHS-IL12 via the subcutaneous route. Results will inform the design and implementation of first-in-human clinical trials of NHS-IL12 in cancer patients.
authors.affiliation: Comparative Oncology Program, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland, United States of America.
authors.forename: Melissa
authors.initials: M
authors.lastname: Paoloni
authors.affiliation: Comparative Oncology Program, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland, United States of America.
authors.forename: Christina
authors.initials: C
authors.lastname: Mazcko
authors.affiliation: College of Veterinary Medicine, University of Missouri-Columbia, Columbia, Missouri, United States of America.
authors.forename: Kimberly
authors.initials: K
authors.lastname: Selting
authors.affiliation: College of Veterinary Medicine and Biological Sciences, Colorado State University, Fort Collins, Colorado, United States of America.
authors.forename: Susan
authors.initials: S
authors.lastname: Lana
authors.affiliation: School of Veterinary Medicine, Tufts University, North Grafton, Massachusetts, United States of America.
authors.forename: Lisa
authors.initials: L
authors.lastname: Barber
authors.affiliation: College of Veterinary Medicine, University of Tennessee, Knoxville, Tennessee, United States of America.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Phillips
authors.affiliation: School of Veterinary Medicine, University of California Davis, Davis, California, United States of America.
authors.forename: Katherine
authors.initials: K
authors.lastname: Skorupski
authors.affiliation: School of Veterinary Medicine, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
authors.forename: David
authors.initials: D
authors.lastname: Vail
authors.affiliation: College of Veterinary Medicine, Texas A&M University, College Station, Texas, United States of America.
authors.forename: Heather
authors.initials: H
authors.lastname: Wilson
authors.affiliation: College of Veterinary Medicine and Biological Sciences, Colorado State University, Fort Collins, Colorado, United States of America.
authors.forename: Barbara
authors.initials: B
authors.lastname: Biller
authors.affiliation: College of Veterinary Medicine and Biological Sciences, Colorado State University, Fort Collins, Colorado, United States of America.
authors.forename: Anne
authors.initials: A
authors.lastname: Avery
authors.affiliation: College of Veterinary Medicine, Michigan State University, East Lansing, Michigan, United States of America.
authors.forename: Matti
authors.initials: M
authors.lastname: Kiupel
authors.affiliation: Comparative Oncology Program, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland, United States of America.
authors.forename: Amy
authors.initials: A
authors.lastname: LeBlanc
authors.affiliation: EMD-Serono Research and Development Institute, Billerica, Massachusetts, United States of America.
authors.forename: Anna
authors.initials: A
authors.lastname: Bernhardt
authors.affiliation: EMD-Serono Research and Development Institute, Billerica, Massachusetts, United States of America.
authors.forename: Beatrice
authors.initials: B
authors.lastname: Brunkhorst
authors.affiliation: EMD-Serono Research and Development Institute, Billerica, Massachusetts, United States of America.
authors.forename: Robert
authors.initials: R
authors.lastname: Tighe
authors.affiliation: Comparative Oncology Program, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland, United States of America.
authors.forename: Chand
authors.initials: C
authors.lastname: Khanna
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000494:pharmacology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D016207:Cytokines / Q000097:blood; D004283:Dog Diseases / Q000097:blood / Q000188:drug therapy* / Q000473:pathology*; D004285:Dogs; D005260:Female; D007074:Immunoglobulin G / Q000008:administration & dosage / Q000494:pharmacology*; D007155:Immunologic Factors / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000494:pharmacology*; D016130:Immunophenotyping; D055104:Infusions, Subcutaneous; D018664:Interleukin-12 / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000494:pharmacology*; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000378:metabolism; D008198:Lymph Nodes / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology / Q000378:metabolism; D008297:Male; D008545:Melanoma / Q000662:veterinary*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000494:pharmacology*; D016896:Treatment Outcome
chemical_list: D000970:Antineoplastic Agents; D016207:Cytokines; D007074:Immunoglobulin G; D007155:Immunologic Factors; C000594734:NHS-IL12 immunocytokine; D011993:Recombinant Fusion Proteins; D018664:Interleukin-12