Tetsuya Taura

Targeting Attenuated Interferon-α to Myeloma Cells with a CD38 Antibody Induces Potent Tumor Regression with Reduced Off-Target Activity.

Interferon-α (IFNα) has been prescribed to effectively treat multiple myeloma (MM) and other malignancies for decades. Its use has waned in recent years, however, due to significant toxicity and a narrow therapeutic index (TI). We sought to improve IFNα’s TI by, first, attaching it to an anti-CD38 antibody, thereby directly targeting it to MM cells, and, second, by introducing an attenuating mutation into the IFNα portion of the fusion protein rendering it relatively inactive on normal, CD38 negative cells. This anti-CD38-IFNα(attenuated) immunocytokine, or CD38-Attenukine™, exhibits 10,000-fold increased specificity for CD38 positive cells in vitro compared to native IFNα and, significantly, is ~6,000-fold less toxic to normal bone marrow cells in vitro than native IFNα. Moreover, the attenuating mutation significantly decreases IFNα biomarker activity in cynomolgus macaques indicating that this approach may yield a better safety profile in humans than native IFNα or a non-attenuated IFNα immunocytokine. In human xenograft MM tumor models, anti-CD38-IFNα(attenuated) exerts potent anti-tumor activity in mice, inducing complete tumor regression in most cases. Furthermore, anti-CD38-IFNα(attenuated) is more efficacious than standard MM treatments (lenalidomide, bortezomib, dexamethasone) and exhibits strong synergy with lenalidomide and with bortezomib in xenograft models. Our findings suggest that tumor-targeted attenuated cytokines such as IFNα can promote robust tumor killing while minimizing systemic toxicity.

Abstract A13: Anti-CD38-attenukine: A myeloma-targeting immunocytokine containing an engineered IFNα that provides >10,000-fold enhanced tumor-specific activity compared to native IFNα.

Multiple myeloma (MM) remains an incurable disease despite recent approvals of novel therapies. Treatments which utilize alkylating agents, corticosteroids, proteasome inhibitors and immunomodulatory drugs have provided significant survival benefits; however, tumor relapse eventually occurs with the use of these agents. The cytokine, interferon-α (IFNα), has been used clinically to treat MM for over 30 years. IFNα exerts good anti-myeloma tumor activity but durable responses are rarely achieved due to dose limiting toxicities. Tumor specificity can be modestly increased by attaching IFNα to an anti-CD38 antibody, thus targeting the cytokine to MM cells. However, by introducing an attenuating mutation to the IFNα portion of this fusion protein, the activity of the cytokine is dramatically reduced on CD38 negative cells while exhibiting a similar potency to native IFNα on CD38+ MM cells. As a result, the anti-CD38-attenuated IFNα, referred to as anti-CD38-attenukine™, has over 10,000-fold greater myeloma-specificity than native IFNα in vitro. In vivo, even though the off-target activity is decreased, anti-CD38-attenukine™ has profound anti-tumor effects in murine MM and NHL xenograft models. Treatment with this fusion protein can cure mice with large (>700mm3) tumors—an effect not observed with any other MM agents tested. Here we report that in a Velcade (bortezomib) refractory, IFNα-insensitive primary MM xenograft model, the combination of Velcade with anti-CD38-attenukine™ has potent anti-tumor activity, resulting in tumour elimination and curing of mice. Our findings indicate that anti-CD38-attenukine™ may be a well tolerated, potent anti-MM treatment that may be utilized alone or in combination with Velcade. Citation Format: Sarah Pogue, Tetsuya Taura, Mingying Bi, Glen Mikesell, Yong Yun, Angela Sho, Eric Sanchez, Haiming Chen, James Berenson, Collette Behrens, Maxwell Stevens, Teresa Domagala, Maya Sokolovsky, Hussein Hallak, Moti Rosenstock, Anthony Doyle, David Wilson. Anti-CD38-attenukine: A myeloma-targeting immunocytokine containing an engineered IFNα that provides >10,000-fold enhanced tumor-specific activity compared to native IFNα. [abstract]. In: Proceedings of the AACR Special Conference on Hematologic Malignancies: Translating Discoveries to Novel Therapies; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(17 Suppl):Abstract nr A13.

Abstract 2660: Targeting an attenuated cytokine to tumor cells via antibody fusion results in enhanced tumor killing with significantly reduced off-target activity

Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA Many cytokines have potential therapeutic applications and several, including interferon (IFN)α and IFNβ, have been approved for the treatment of cancer and/or autoimmune disease. While these agents are effective, they are associated with significant dose-limiting toxicities that prevent their use at levels sufficient to promote optimal therapeutic benefit. The toxicity of type I interferons, like other cytokines, is likely mediated via cognate receptor activation on cells other than those that mediate their therapeutic effects. For example, IFNα2b, which is indicated for the treatment of multiple myeloma (MM), has a narrow therapeutic index due to significant systemic toxicity. Approaches that seek to minimize dose-limiting systemic toxicity while maximizing tumor-specific cytokine exposure could greatly enhance the therapeutic index and effectiveness of type I IFNs and other cytokines. Other groups have previously shown that a moderate degree of tumor-specificity can be achieved by attaching a cytokine such as IFNα to a tumor-targeting antibody; such first generation antibody-targeted cytokines are highly active but show only moderate tumor-specificity since the cytokine can still potently stimulate its receptor on antigen-negative cells. We sought to improve the therapeutic index of antibody-targeted cytokines by mutating the cytokine portion to significantly reduce affinity for its receptor, thereby making it more dependent on antibody-based cell-targeting. Here we demonstrate that such fusion proteins, consisting of mutant or attenuated cytokines (Attenukines™) attached to tumor-targeting antibodies, are 1,000 to 100,000-fold more potent on target-positive cells compared to target-negative (normal) cells. This is shown for antibody-Attenukine™ fusion proteins based on multiple tumor antigens (CD20, CD38, CD138, HMW-MAA, HLA) and multiple attenuated mutants of IFNα, IFNβ, IL-4 and IL-6. Furthermore, we have evaluated an anti-CD38-attenuated IFNα molecule (anti-CD38-Attenukine™) in various CD38+ MM xenograft models and found that this molecule retains potent tumor-targeting activity and anti-tumor efficacy. Moreover, in non-human primates, we have confirmed that the attenuating mutation in IFNα indeed decreases non-targeted IFNα biomarker responses by greater than 100-fold. Taken together, our findings suggest that the administration of antibody-attenuated cytokine fusion proteins to cancer patients may promote robust cytokine-dependent tumor-killing while minimizing systemic toxicity. Citation Format: Sarah L. Pogue, Tetsuya Taura, Mingying Bi, Glen Mikesell, Yong Yun, Angela Sho, Collette Behrens, Maxwell Stevens, Teresa Domagala, Maya Sokolovsky, Hussein Hallak, Moti Rosenstock, Anthony Doyle, David S. Wilson. Targeting an attenuated cytokine to tumor cells via antibody fusion results in enhanced tumor killing with significantly reduced off-target activity. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2660. doi:10.1158/1538-7445.AM2014-2660