Clinical Trials Conducted by Chand Khanna DVM, PhD and the Animal Cancer Institute.

Inhaled interleukin-2 liposomes immunotherapy: Prospective case series in dogs with pulmonary metastases or primary lung cancers.
Summary: Dogs with pulmonary metastasis and primary lung cancers were treated with aerosols of IL-2. Minimal toxicity was noted in 9 dogs that were treated. Two dogs with metastatic osteosarcoma had regression of pulmonary metastasis for greater than 1 year.
Publication: Khanna C, Hasz DE, Klausner JS, Katsanis, EM, and Anderson PM. Inhaled interleukin-2 liposome immunotherapy in dogs with spontaneous primary lung cancers and cancers metastatic to the lung. Cancer. 79(7): 1409-21, 1997.
Background: Systemic in vivo toxicity of interleukin-2 (IL-2) has been problematic. Anti-neoplastic activity of IL-2 has been modest. We have previously demonstrated the biological activity and safety of aerosols of IL-2 liposomes in normal dogs. We now report objective regression of naturally occurring pulmonary metastases in dogs after one month of nebulized IL-2 liposome therapy.
Methods: Dogs with pulmonary metastases (n=7) and primary lung carcinoma (n=2) were treated with aerosols of IL-2 liposomes. Response to therapy was monitored with serial chest radiographs. Effector populations, collected by broncho-alveolar lavage (BAL) and from heparinized whole blood, were assessed for cell type, immunophenotype, and tumor cytolytic activity. Immunogenicity of human IL-2 and human serum albumin (HSA) in dogs was assessed by immunofluoresence assay (IFA).
Results: Two of four dogs with pulmonary osteosarcoma metastases had complete regression of metastases stable for greater than 12 and greater than 20 months. One of two dogs with lung cancer had stabilization of disease for greater than 8 months; the other had progression. Toxicity was minimal. BAL cell numbers increased greater than 4 fold (p=0.01) and included significantly greater proportions and total numbers of eosinophils (p=0.006) and lymphocytes (p=0.008). Mean BAL effector lytic activity was significantly greater after 15 days of IL-2 liposome inhalation compared to pretreatment activity (p=0.01); however mean BAL lytic activity decreased after 30 days and was no longer significantly greater than pretreatment BAL lytic activity. No allergic reactions were associated with inhaled IL-2 liposome therapy. Canine antibodies against human IL-2 and HSA were detected in all dogs.
Conclusion: Pet dogs with naturally occurring pulmonary metastases and primary lung cancers accepted inhalation treatments easily. Non-toxic and effective treatment of pulmonary metastases of osteosarcoma is possible using nebulized IL-2 liposomes.

L-glutamine prevention of radiation induced mucositis: Prospective placebo blinded randomized trial in dogs receiving megavoltage radiation therapy for nasal cancers
Summary: This prospective randomized trial demonstrated a significant reduction in oral radiation mucositis (ulceration) in pet dogs receiving radiation therapy for nasal tumors when dogs were given a glutamine containing suspension compared to a standard daily mouthwash. The use of the glutamine suspension was not associated with any adverse effects in dogs and did not alter the activity of the radiation therapy in the treated dogs.
Publication: Khanna C, Klausner JS, Walters P, Bell FW, James K, Lund E, Redic K, Anderson PM. L-glutamine versus placebo in the prevention of radiation induced oral mucositis. In Vivo. In progress.
Background: Oral mucositis is a common complication of radiation therapy of the head and neck. Radiation-induced mucositis of the oral cavity results in patient discomfort and may adversely affect treatment outcomes. Mucositis can lead to interruptions or early stoppages in planned radiation treatments and negatively impact on nutritional status. L-glutamine, an important fuel source for gastrointestinal epithelial cells, may decrease the severity of mucositis associated with radiation therapy. Using a relevant animal model, we evaluated of the radio-protective activity of an oral l-glutamine suspension in preventing oral mucositis.
Methods: Pet dogs, receiving radiation therapy for nasal cancers, were entered to a randomized double-blind placebo-controlled trial to evaluate an oral suspension of l-glutamine in the prevention of radiation-induced mucositis. The l-glutamine suspension, or a corn starch placebo, in a sugar-based vehicle was administered to dogs orally (4.0 gm/m2/d of l-glutamine or an equivalent volume of placebo suspension). Dogs were evaluated daily for the severity of mucositis based on a clinical grading score and a food intake score. The severity of mucositis, the duration of mucositis, and the number of interruptions (due to mucositis) in the planned radiation therapy was compared for the dogs receiving the l-glutamine and placebo oral suspension.
Results: A significant decrease in the number of interruptions in radiation therapy was seen in the dogs receiving l-glutamine compared to placebo (p=0.028). Furthermore dogs receiving l-glutamine had a significant decrease in the number of days of severe mucositis compared to the dogs receiving placebo (p=0.05). There were no adverse effects associated with administration of the l-glutamine suspension
Conclusions: Amelioration of oral radiation induced-mucositis is possible using an oral suspension of l-glutamine. Further controlled clinical trials of the radio-protective properties of l-glutamine are warranted.

Doxorubicin vs Dactinomycin containing induction chemotherapy: Prospective placebo blinded randomized trial in dogs receiving multi-agent chemotherapy for lymphoma.
Summary: Dogs receiving Doxorubicin compared to Dactinomycin as part of a multi-agent protocol were compared. Median time to first remission was not significantly different between the 2 groups. However, median duration of remission and median survival time was significantly longer for dogs treated with Doxorubicin. There were no significant differences in toxicity noted between the 2 groups
Publication: Khanna C, Lund E, Redic K, Hayden D, Bell FW, Goullaud R, Klausner JS. A randomized trial of doxorubicin versus dactinomycin in the treatment of canine lymphosarcoma. J Am Vet Med Assoc. 213(7):985-90, 1998.
Objective: To compare efficacy and toxicity of 2 multi-agent chemotherapeutic protocols, similar in all respects except that one incorporated dactinomycin and the other incorporated doxorubicin, for treatment of dogs with malignant lymphoma.
Design: Randomized double blind controlled trial.
Animals: 45 dogs with malignant lymphoma.
Methods: Dogs were randomly assigned to a doxorubicin or dactinomycin treatment group. Time to first remission, duration of first remission, survival time, and prevalence of toxicoses, particularly number of episodes of dose-limiting neutropenia and gastrointestinal toxicoses were compared between groups.
Results: 37 dogs received at least 1 dose of doxorubicin (21 dogs) or dactinomycin (16 dogs). Median time to first remission was not significantly different between groups, but median duration of first remission and median survival time were significantly longer for dogs in the doxorubicin treatment group than for dogs in the dactinomycin treatment group. Numbers of dogs that died, numbers of episodes of dose-limiting neutropenia, and numbers of episodes of gastrointestinal toxicoses were not significantly different between groups.
Conclusions: A multi-agent chemotherapeutic protocol incorporating doxorubicin was significantly more effective in dogs with malignant lymphoma than a similar protocol incorporating dactinomycin. Despite the lower cost and lack of cardiotoxicity, dactinomycin is not an equivalent substitute for doxorubicin in the initial treatment of dogs with malignant lymphoma.

IGF-I inhibition plus chemotherapy vs chemotherapy alone in osteosarcoma: enhanced tumor apoptosis
Summary: OncoLAR® is an agent that blocks the production of insulin-like growth factor (IGF-I), a hormone that is believed to enhance the survival of cancer cells. This study evaluated the effects of OncoLAR® in canine patients being treated with chemotherapy for osteosarcoma. A comparison was made in this group of study patients to osteosarcoma patients receiving chemotherapy but not OncoLAR®. Results showed significant suppression of IGF-I in patients treated with OncoLAR®, but there was no improvement in cancer cell destruction or patient survival time using this agent.
Publication: Khanna C , Prehn J, Yeung C, Caylor J, Bose S, Cassaday RJacob S, Helman L. Randomized trial of OncoLAR® (SMS-201 995pa LAR) and carboplatin versus carbplatin alone in dogs with naturally occurring osteogenic sarcoma. Clinical Cancer Res In Press, 7/1/02.
Background: Osteosarcoma is the most common primary tumor of bone. Several lines of evidence suggest the role of the insulin-like growth factor - growth hormone pathway in the biology of this cancer. Insulin like growth factor has been demonstrated to act as a survival signal for cancer cells, and as such has been hypothesized to be a mechanism of resistance against chemotherapy-induced death. OncoLAR® is a novel long acting analog of somatostatin that results in growth hormone blockade from the pituitary and therein suppression of serum insulin-like growth hormone release from the liver. The use of OncoLAR® pediatric osteosarcoma patients resulted in approximately fifty percent suppression in serum IGF-I levels with limited toxicity.
Methods: To determine if insulin-like growth factor suppression will decrease chemotherapy resistance by eliminating an important survival signal to osteosarcoma cells we conducted a randomized, blinded, placebo-controlled pre-clinical study in pet dogs with naturally occurring osteosarcoma. The trial compared primary tumor necrosis and apoptosis and survival of pet dogs receiving OncoLAR® and carboplatin chemotherapy compared to a placebo and carboplatin.
Results: demonstrated suppression of serum insulin-like growth factor levels by approximately 46% without toxicity in dogs receiving OncoLAR®. No differences in primary tumor necrosis, apoptosis, tumor insulin-like growth factor mRNA expression, or survival were seen between the two treatment groups.
Conclusions: Results suggest that suppression of insulin-like growth factor levels by the extent and/or duration achieved in the trial was not sufficient to improve chemotherapy related anti-tumor effects in pet dogs with osteosarcoma.

Thalidomide antiangiogenic therapy in dogs :Prospective case series in dogs with measurable malignant cancers
Summary: This study evaluated the effects of thalidomide in 7 canine patients with a variety of tumors. No objective response was seen in these patients. No adverse effects were noted associated with Thalidomide treatment.
Publication: Jankowski M, Fulton L, Sheafor S, Prescott D, Khanna C. Ongoing evaluation of single agent thalidomide in dogs with measurable cancer. Proc Vet Cancer Soc 1999.
Background: Angiogenesis is essential for cancer progressin and metastasis. Inhibition of angiogenesis may be an effective treatment for cancers. The anticancer activity of thalidomide has been demonstrated in early human clinical trials. The objectives of this study are: 1) to determine the safety and efficacy of oral thalidomide against measurable canine cancers, 2) to define an optimal biological dose for thalidomide in dogs, and 3) to define tumor types sensitive to antiangiogenic therapy with thalidomide.
Methods: This is a single agent, phase I/II clinical study. Entry requirements include measurable and histologically confirmed diease, body weight> 10 kg, no chemotherapy within 10 days of treatment, and informed owner consent. Three escalating dose cohorts (30 cases/cohort) have been defined: 3.3-6.5, 6.6-13, 13.3-26 mg/kg QD. Physical examinations, CBC, serum biochemistry, and tumor response will be evaluated at day 0 and every 30 days thereafter. Urine and plasma samples collected before and during thalidomide therapy will be assayed for markers of antiangiogenic activity including, basic fibroblast growth factor, vascular endothelial growth factor and interleukin-8.
Results: Seven cases to date have been evaluated. All dogs had progressive disease prior to study entry. No side effects to thalidomide have been noted in the first dose cohort.

 SD-Stable Disease; PD-Progressive Disease

Conclusions: Thalidomide appears to be well tolerated in the current dose cohort. No objective tumor responses have been documented at this time. Additional entry of cases with smaller tumor burden may allow antiangiogenic activity to be exerted before disease progression. Surrogate markers of systemic antiangiogenic activity, measured in plasma and urine, may be helpful in defining optimal biologic doses.

ACI-002c: Thrombospondin-I antiangiogenic therapy in cats: Prospective case series in cats with measurable malignant cancers
Summary: This prospective study evaluated the effects of thrombospondin-1 peptides in 10 feline patients with various cancers. No objective response was seen in these patients. No adverse effects were noted.

ACI-004c: Thrombospondin-I antiangiogenic therapy in dogs: Prospective case series in dogs with measurable malignant cancers
Summary: This prospective study evaluated the effects of thrombospondin-I peptides in canine patients with various cancers. No adverse effects were noted. Approximately 40% of these patients showed improvements identified by stable disease unexpected for the biology of that given cancer, partial remissions, or complete remissions. Abstract submitted to the Amercian Society of Clinical Oncology Annual Meeting 2002