Heather Wilson

Preclinical evaluation of the novel, orally bioavailable Selective Inhibitor of Nuclear Export (SINE) KPT-335 in spontaneous canine cancer: Results of a phase i study

Background The purpose of this study was to evaluate the activity of Selective Inhibitors of Nuclear Export (SINE) compounds that inhibit the function of the nuclear export protein Exportin 1 (XPO1/CRM1) against canine tumor cell lines and perform a Phase I clinical trial of KPT-335 in dogs with spontaneous cancer to provide a preliminary assessment of biologic activity and tolerability. Methods and Findings Canine tumor cell lines derived from non-Hodgkin lymphoma (NHL), mast cell tumor, melanoma and osteosarcoma exhibited growth inhibition and apoptosis in response to nanomolar concentrations of SINE compounds; NHL cells were particularly sensitive with IC50 concentrations ranging from 2–42 nM. A Phase I clinical trial of KPT-335 was performed in 17 dogs with NHL (naive or relapsed), mast cell tumor or osteosarcoma. The maximum tolerated dose was 1.75 mg/kg given orally twice/week (Monday/Thursday) although biologic activity was observed at 1 mg/kg. Clinical benefit (CB) including partial response to therapy (PR, nu200a=u200a2) and stable disease (SD, nu200a=u200a7) was observed in 9/14 dogs with NHL with a median time to progression (TTP) for responders of 66 days (range 35–256 days). A dose expansion study was performed in 6 dogs with NHL given 1.5 mg/kg KPT-335 Monday/Wednesday/Friday; CB was observed in 4/6 dogs with a median TTP for responders of 83 days (range 35–354 days). Toxicities were primarily gastrointestinal consisting of anorexia, weight loss, vomiting and diarrhea and were manageable with supportive care, dose modulation and administration of low dose prednisone; hepatotoxicity, anorexia and weight loss were the dose limiting toxicities. Conclusions This study provides evidence that the novel orally bioavailable XPO1 inhibitor KPT-335 is safe and exhibits activity in a relevant, spontaneous large animal model of cancer. Data from this study provides critical new information that lays the groundwork for evaluation of SINE compounds in human cancer.

Adoptive T-cell therapy improves treatment of canine non–Hodgkin lymphoma post chemotherapy

Clinical observations reveal that an augmented pace of T-cell recovery after chemotherapy correlates with improved tumor-free survival, suggesting the add-back of T cells after chemotherapy may improve outcomes. To evaluate adoptive immunotherapy treatment for B-lineage non-Hodgkin lymphoma (NHL), we expanded T cells from client-owned canines diagnosed with NHL on artificial antigen presenting cells (aAPC) in the presence of human interleukin (IL)-2 and IL-21. Graded doses of autologous T cells were infused after CHOP chemotherapy and persisted for 49 days, homed to tumor, and significantly improved survival. Serum thymidine kinase changes predicted T-cell engraftment, while anti-tumor effects correlated with neutrophil-to-lymphocyte ratios and granzyme B expression in manufactured T cells. Therefore, chemotherapy can be used to modulate infused T-cell responses to enhance anti-tumor effects. The companion canine model has translational implications for human immunotherapy which can be readily exploited since clinical-grade canine and human T cells are propagated using identical approaches.

Safety of Masitinib Mesylate in Healthy Cats

BACKGROUNDnMasitinib mesylate is a PO-administered tyrosine kinase inhibitor developed both for human and animal diseases with activity against both mutated and wild type forms of the c-kit receptor and platelet-derived growth factor receptors α and β, and is currently registered in Europe for the treatment of mast cell tumors in dogs.nnnHYPOTHESIS/OBJECTIVESnThe objective of this study was to determine if healthy cats can tolerate administration of masitinib without clinically relevant adverse effects.nnnANIMALSnTwenty healthy research colony-specific pathogen-free cats.nnnMETHODSnThis study was a prospective, randomized phase 1 clinical trial. Masitinib was administered PO to 20 healthy cats. Ten cats received 50 mg masitinib every other day for 4 weeks, and 10 cats received 50 mg masitinib daily for 4 weeks.nnnRESULTSnClinically relevant proteinuria was noted in 2/20 (10%) cats (both treated daily), and neutropenia was noted in 3/20 (15%) (seen in both treatment groups). An increase in serum creatinine concentration and adverse gastrointestinal effects were noted in some cats.nnnCONCLUSIONS AND CLINICAL IMPORTANCEnMasitinib mesylate was tolerated in the majority of cats. Long-term administration and pharmacokinetic studies are needed to further assess the use of masitinib in cats.

RENAL EVALUATION IN THE HEALTHY GREEN IGUANA (IGUANA IGUANA): ASSESSMENT OF PLASMA BIOCHEMISTRY, GLOMERULAR FILTRATION RATE, AND ENDOSCOPIC BIOPSY

Abstract Plasma biochemistry, iohexol clearance, endoscopic renal evaluation, and biopsy were performed in 23 clinically healthy 2-yr-old green iguanas (Iguana iguana). Mean (±SD) values for packed cell volume (30 ± 3%), total protein (62 ± 7 g/L, 6.2 ± 0.7 g/dl), albumin (25 ± 2 g/L, 2.5 ± 0.2 g/dl), globulin (37 ± 6 g/L, 3.7 ± 0.6 g/ dl), total calcium (3.0 ± 0.2 mmol/L, 12.0 ± 0.7 mg/dl), ionized calcium (1.38 ± 0.1 mmol/L), phosphorus (1.32 ± 0.28 mmol/L, 4.1 ± 0.9 mg/dl), uric acid (222 ± 100 μmol/L, 3.8 ± 1.7 mg/dl), sodium (148 ± 3 mmol/L or mEq/ L), and potassium (2.6 ± 0.4 mmol/L or mEq/L) were considered within normal limits. Values for urea were low (<1.4 mmol/L, <4 mg/dl) with 70% of samples below the detectable analyzer range. After the i.v. injection of 75 mg/ kg iohexol into the caudal (ventral coccygeal or tail) vein, serial blood collections were performed over 32 hr. Iohexol assays by high-performance liquid chromatography produced plasma iohexol clearance graphs for each lizard. A three-compartment model was used to fit area under the curve values and to obtain the glomerular filtration rate (GFR) using regression analysis. The mean GFR (SD) was 16.56 ± 3.90 ml/kg/hr, with a 95% confidence interval of 14.78–18.34 ml/kg/hr. Bilateral endoscopic renal evaluation and biopsy provided tissue samples of excellent diagnostic quality, which correlated with tissue harvested at necropsy and evaluated histologically. None of the 23 animals demonstrated any adverse effects of iohexol clearance or endoscopy. Recommended diagnostics for the evaluation of renal function and disease in the green iguana include plasma biochemical profiles, iohexol clearance, endoscopic examination, and renal biopsy.

Dose-escalating vinblastine for the treatment of canine mast cell tumour

The purpose of this study was to evaluate the short-term adverse events (AEs) in dogs with mast cell tumours (MCT) receiving prednisone and dose-escalating vinblastine (VBL). Twenty-four dogs were treated with intravenous VBL starting at 2 mg m(-2) and then escalating in weekly increments to 2.33, 2.67 and 3 mg m(-2). AEs were graded using a standardized scoring system. No dogs receiving 2 or 2.33 mg m(-2) experienced grade 3 or 4 AEs. Among the dogs, 9.5 and 5.9% had grade 3 or 4 AEs at dosages of 2.67 and 3 mg m(-2), respectively. Serious AEs included neutropaenia (n = 3) and vomiting (n = 1), only one of which required hospitalization. These data indicate that VBL chemotherapy may be safe to administer at higher than the traditional 2 mg m(-2) dosage for dogs with MCT. Randomized prospective trials are necessary to establish whether dose escalation will translate into improved response rates when compared with the standard 2 mg m(-2) dosage.

Predictors of outcome in dogs with subcutaneous or intramuscular hemangiosarcoma

OBJECTIVEnTo identify prognostic factors in a large group of dogs with subcutaneous or intramuscular hemangiosarcoma (HSA) or both. Design-Multi-institutional retrospective cohort study. Animals-71 dogs with subcutaneous or intramuscular HSA.nnnPROCEDURESnMedical records of affected dogs were reviewed. The following factors were evaluated for an association with outcome: dog age and sex, clinical signs, anemia, thrombocytopenia, neutrophilia, tumor stage at diagnosis, achievement of complete excision, intramuscular involvement, presence of gross disease, tumor recurrence, and treatment.nnnRESULTSnOf the 71 cases identified, 16 (29%) had intramuscular tumor involvement. For all dogs, median time to tumor progression and overall survival time (OST) were 116 and 172 days, respectively; 25% survived to 1 year. Univariate analysis identified presence of clinical signs or metastasis at diagnosis, dog age, tumor size, use of any surgery, and presence of gross disease as predictors of time to tumor progression and OST. There was no significant difference in survival time between dogs with respect to type of HSA. Multivariate analysis confirmed that adequate local tumor control, tumor diameter ≤ 4 cm, presence of metastasis at diagnosis, and presence of gross disease were significantly associated with OST.nnnCONCLUSIONS AND CLINICAL RELEVANCEnSubcutaneous and intramuscular HSA remains a heterogeneous group of tumors that generally carries a poor prognosis. Adequate local control of smaller tumors with no associated clinical signs or metastasis may provide the best chance of long-term survival.

Feline Alimentary Lymphoma: Demystifying the Enigma

Alimentary lymphoma is one of the most commonly diagnosed neoplasms of the cat. The incidence of this disease has increased significantly over the past 15 years during the post-feline leukemia era. Despite the common prevalence of this disease, appropriate diagnosis and treatment can be challenging. There are two main forms of feline alimentary lymphoma: the small-cell (lymphocytic, well-differentiated, low-grade) lymphoma variety and the large-cell (lymphoblastic, high-grade) lymphoma variety. These two diseases are related; however, each presents its own diagnostic and therapeutic challenges. Additionally, it can be difficult to differentiate these malignancies from other nonneoplastic diseases such as inflammatory bowel disease and other chronic inflammatory conditions of the gastrointestinal tract. The purpose of this article is to tackle the challenges of this allusive disease with a step-by-step approach to diagnosis, staging, and therapy.

Is cancer a stem cell disease? Theory, evidence and implications

Cells of muticellular organisms form part of a specialized society that cooperates to promote survival of the organism. In this, cell division, proliferation and differentiation are strictly controlled and a balance exists between normal cell birth and cell death. Inextricably linked to this fundamental concept is the role of the adult stem cell (ASC) whose progeny and microenvironment make up the anatomical structure that coordinates normal homeostatic production of functional mature cells. ASCs have been best characterized in the haematopoietic system but exist in all major organ systems. These cells are characterized by a capacity for selfrenewal, being undifferentiated but capable of multilineage differentiation, slowly cycling cells but clonogenic, and capable of asymmetric division. Further, ASC reside in particular ‘ niche ’ environments that support an appropriate spatiotemporal dialogue between ASC and microenvironmental cells in order to fulfi l the lifelong demands for normal differentiated cells. 1

Defining the Pharmacodynamic Profile and Therapeutic Index of NHS-IL12 Immunocytokine in Dogs with Malignant Melanoma

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. Methodology/Principal Findings 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/Significance 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.

Evaluation of Spray BIO-Max DIM-P in Dogs for Oral Bioavailability and in Nu/nu Mice Bearing Orthotopic/Metastatic Lung Tumor Models for Anticancer Activity

ABSTRACTPurposeIn an effort to prepare an oral dosage form for poorly bioavailable anti-cancer agents, we have incorporated spray drying using a customized spray gun generating enteric coated Self-emulsifying drug delivery systems. The objective of this study was to design and evaluate pharmacokinetics and pharmacodynamic characteristics of Spray BIO-Max DIM-P (SB DIM-P).MethodsSB DIM-P was prepared and optimized based on physico-chemical characteristics using design of experiment (DOE–Vr 8.0) software. Pharmacokinetic parameters in dogs and rats were evaluated and analyzed using Winonlin. Anti-tumor activity was carried out in orthotopic and metastatic lung tumor models using size M capsules in mice.ResultsBased on the optimization using DOE analysis of SB DIM-P characteristics, formulations were selected for further investigation. Pharmacokinetic studies showed a 30% increase in oral bioavailability in rats and ~2.9 times more bioavailability of SB DIM-P compare to solution in dogs. SB DIM-P showed ~20–25% more tumor volume/weight reduction in H1650 metastatic tumor model and ~25–30% tumor volume/weight reduction in A549 orthotopic tumor model compared to DIM-P solution.ConclusionsOur studies demonstrate the potential application of spray dried enteric coated self-emulsifying delivery system (SB DIM-P) to enhances oral absorption and efficacy of DIM-P in lung tumor models.