Adjuvant Therapy With an Alpha-lactalbumin Vaccine in Triple-Negative Breast Cancer

Phase I Trial of Adjuvant Therapy With an Alpha-lactalbumin Vaccine in Patients With Non-Metastatic Triple-Negative Breast Cancer at High Risk of Recurrence

The purpose of this study is to determine the safety as well as the most effective dose of the alpha-lactalbumin vaccine (aLA breast cancer vaccine) to treat patients with non-metastatic triple negative breast cancer as well as a cohort of cancer-free women at risk for triple-negative breast cancer.

This is an open-label, phase I dose-escalation trial that will be performed on two successive cohorts. The first cohort is comprised of participants with high-risk triple-negative breast cancer. The second is made up of participants scheduled to undergo bilateral prophylactic mastectomy due to their genetic risk of triple-negative breast cancer. Both cohorts will be treated with successively higher doses of α-lactalbumin and zymosan This aLA breast cancer vaccine is an investigational (experimental) drug that the study team believes will work by stimulating the immune system to fight the participant's cancer, in a way similar to the way the immune system fights off an infection after a vaccination for that infection. α-lactalbumin Vaccine is experimental because it is not approved by the Food and Drug Administration (FDA). A traditional "3+3" Phase I trial design will be employed for both cohorts to determined the Maximum Tolerated Dose (MTD). After identification of the MTD, if at least 1 participant has an immunologic response (correlative measurement), successively lower dose levels will be expanded to a total of 6 participants and immunologic response assessed. Enrollment will stop if a dose level is reached for which no responses are observed. Dose-Limiting toxicities (DLTs) in 2 or more of 6 participants, the next lower dose will be considered the new MTD. Objectives are to determine MTD, DLT incidence, and Lowest Immunologic Dose (LID) for both cohorts. Toxicity will be assessed every 2 weeks until day 56 and at day 84 or at off-study. Participants will be offered participation in long-term follow-up involving contact or in-person follow-up for late toxicity and survival every 3 months for 2 years, every 6 months for an additional 3 years, and then annually for 10 years.

Pathologic Stage IIA-IIIC Triple-Negative Breast Cancer, TNBC - Triple-Negative Breast Cancer, Residual Disease

Participants diagnosed with triple negative breast cancer (TNBC) will be treated with successively higher doses of α-lactalbumin and zymosan in a traditional 3 + 3 phase I trial design. Treatment will involve a course of 3 vaccinations given every 2 weeks. Participants will be enrolled sequentially into 1 of 5 different dose levels each comprised of cohorts of 1-6 participants until the MTD has been identified (intra-patient dose escalation not permitted), after which the MTD will be expanded to 6 participants. Successively lower doses will be expanded to 6 participants until the lowest DL associated with immune response has been expanded. Dose Level (DL) 1: 10 microgram (mcg) a-lactalbumin + 10 mcg Zymosan DL2: 100 mcg a-lactalbumin + 10 mcg Zymosan DL3: 500 mcg a-lactalbumin + 10 mcg Zymosan DL4: 500 mcg a-lactalbumin + 30 mcg Zymosan DL5: 500 mcg a-lactalbumin + 60 mcg Zymosan

Participants with a genetic risk for developing TNBC who plan to undergo prophylactic mastectomy will be treated with α-lactalbumin and zymosan at doses based on the TNBC cohort. Treatment will involve a course of 3 vaccinations given every 2 weeks. Participants enrolled in the prevention cohort we will be enrolled at the dose level being used in the TNBC cohort if no DLTs above Grade 1 have been observed. If the TNBC cohort proceeds to the next dose level before another prevention cohort patient is enrolled, the next prevention patient will be enrolled on the next dose level along with the TNBC cohort. Dose Level (DL) 1: 10 microgram (mcg) a-lactalbumin + 10 mcg Zymosan DL2: 100 mcg a-lactalbumin + 10 mcg Zymosan DL3: 500 mcg a-lactalbumin + 10 mcg Zymosan DL4: 500 mcg a-lactalbumin + 30 mcg Zymosan DL5: 500 mcg a-lactalbumin + 60 mcg Zymosan

α-lactalbumin vaccine will be administered subcutaneously in rotating sites (vaccine will not be administered in the arms of any participant, due to likelihood of prior bilateral mastectomy). DL1: 10 mcg DL2: 100 mcg DL3-5: 500 mcg

MTD of an a-lactalbumin vaccine in participants at risk for TNBC who are scheduled for prophylactic double mastectomy.

LID of α-lactalbumin vaccine in participants with operable triple-negative breast cancer, based on ELISPOT assays to assess the ability to induce a pro-inflammatory T cell response consistent with tumor protection. This assessment will be determined using the ELISPOT assay to determine peripheral blood frequencies of T cells that produce interferon-gamma (IFNγ; type-1) and IL-17 (type-17) in response to recombinant human α-lactalbumin

LID of α-lactalbumin vaccine in participants at risk for TNBC who are scheduled for prophylactic double mastectomy, based on ELISPOT assays to assess the ability to induce a pro-inflammatory T cell response consistent with tumor protection. This assessment will be determined using the ELISPOT assay to determine peripheral blood frequencies of T cells that produce interferon-gamma (IFNγ; type-1) and IL-17 (type-17) in response to recombinant human α-lactalbumin

Inclusion Criteria: Triple Negative Cohort: Histologically proven invasive breast cancer. Primary tumor must be ER-negative (ER in <1% of cells), PR-negative (PR in <1% of cells), and HER2-negative (0-1+ by IHC or FISH ratio<2.0 with signal number <6/cell), or consistent with contemporary NCCN guidelines (https://www.nccn.org/). Patients must be high risk, defined as either: Pathologic stage IIA, IIB, IIIA, IIIB, or IIIC by AJCC 8, or Residual invasive cancer in breast or regional nodes following preoperative chemotherapy. Patients must have no convincing evidence of recurrent disease based on one of the following: bone scan and imaging scans of the chest/abdomen/pelvis or FDG PET scan. ≥1 months since last active therapy (chemotherapy, radiation therapy, or surgery) and ≤ 36 months since the initiation of treatment for the current cancer, based on the period of highest risk for patients with Stages I-III triplenegative breast cancer [33, 34]. Treatment prior to enrollment must be consistent with NCCN guidelines extant at the time treatment was given, found at: https://www.nccn.org/. Age greater than or equal to18 years. ECOG Performance Status 0-1. Adequate major organ function, defined as: WBC ≥ 3,000/mcl, hemoglobin ≥ 10.0 gm/dL, platelets ≥ 100,000/mcL, total bilirubin within normal limits, ALT/AST <3 x upper limits of normal (ULN), serum creatinine ≤ 1.5 x ULN. Serum prolactin level must be ≤ upper limits of normal (ULN). Subjects must have the ability to understand and the willingness to sign and provide a written informed consent document. Subjects must have archival tissue available for potential correlative studies (e.g., assays for α-lactalbumin expression or tumor infiltrating lymphocytes), but tumors will not be required to exhibit overexpression of α-lactalbumin for enrollment. Subject agrees not to use alternative therapies from the time of informed consent through 30 days following the last vaccine injection (see section 6.6.1.3). Prevention Cohort: Participant must have a high risk for developing triple-negative breast cancer, defined as: carrying a deleterious mutation in BRCA1 or PALB2 Patients must have no evidence of breast cancer based on both of the following: Negative mammography or breast MRI within 180 days, Negative breast examination by a physician or advanced practice practitioner within 30 days Age ≥ 18 years ECOG Performance Status 0-1. Adequate major organ function, defined as: WBC ≥ 3,000/mcl, hemoglobin ≥ 10.0 gm/dL, platelets >100,000/mcL, total bilirubin within normal limits, ALT/AST <3 x upper limits of normal (ULN), serum creatinine ≤ 1.5 x ULN. Serum prolactin level must be <upper limits of normal (ULN) Subjects must have the ability to understand and the willingness to sign and provide a written informed consent document. Subject agrees not to use alternative therapies from the time of informed consent through 30 days following the last vaccine injection Exclusion Criteria: Triple Negative Cohort: Receipt of cytotoxic chemotherapy within 4 weeks of study entry. Radiation therapy within 4 weeks of study entry. Failure to recover from the toxicity of the previous therapy to CTCAE Grade 0-1, except for alopecia and grade 2 neuropathy. Need for systemic corticosteroid use (except as physiologic replacement, defined as prednisone 10 mg/day or equivalent). Need for immunosuppression (e.g., for a history of organ transplantation). Known HIV infection. Active or planned lactation or pregnancy. Patients taking or planning to take oral contraceptives will be excluded, as there is some evidence that such agents can induce lactational foci. This includes patients using hormone containing IUD's. Refusal to use effective non-hormonal contraception. Acceptable contraception methods include but may not be limited to barrier contraception (diaphragm or condom), non-hormonal intrauterine device, vasectomy of male partner. Subjects receiving any other investigational agents within the last 4 weeks. Subjects with any known recurrence or metastasis. Subjects with a history of another active invasive malignancy within 5 years of study entry. History of allergic reactions to α-lactalbumin, human milk (excluding lactose intolerance), zymosan, or other agents used in this study. Subjects with uncontrolled intercurrent illness including, but not limited to ongoing or active infection, symptomatic congestive heart failure, unstable angina pectoris, cardiac arrhythmia, or psychiatric illness/social situations that would limit compliance with study requirements. Subjects with known hyperprolactinemia. Subjects being treated with drugs known to cause hyperprolactinemia (see appendix 2). Prevention Cohort: Receipt of cytotoxic chemotherapy within 4 weeks of study entry (including for benign indications). Radiation therapy within 4 weeks of study entry (including for benign indications) Need for systemic corticosteroid use (except as physiologic replacement, defined as prednisone 10 mg/day or equivalent). Need for immunosuppression (e.g., for a history of organ transplantation). Known HIV infection. Active or planned lactation or pregnancy. Patients taking or planning to take oral contraceptives will be excluded, as there is some evidence that such agents can induce lactational foci. This includes patients using hormone containing IUD's. Refusal to use effective non-hormonal contraception. Acceptable contraception methods include but may not be limited to barrier contraception (diaphragm or condom), non-hormonal intrauterine device, vasectomy of male partner. Subjects receiving any other investigational agents within the last 4 weeks. Subjects with a history of invasive malignancy within 5 years of study entry. History of allergic reactions to α-lactalbumin, human milk (excluding lactose intolerance), zymosan, or other agents used in this study. Subjects with uncontrolled intercurrent illness including, but not limited to ongoing or active infection, symptomatic congestive heart failure, unstable angina pectoris, cardiac arrhythmia, or psychiatric illness/social situations that would limit compliance with study requirements Subjects with known hyperprolactinemia Subjects being treated with drugs known to cause hyperprolactinemia

Yes, the data will be shared with the FDA, the Department of Defense (DOD), and Anixa Biosciences who has negotiated rights to the drug, and any pharmaceutical partner who may wish to negotiate rights to the drug. All below may be shared, with patient data anonymized

The data will be available to the FDA and DOD. Otherwise, a confidentiality agreement will need to be in place.