Increased Frequency of AlloStim(TM) Dosing in Combination With Cryoablation in Metastatic Breast Cancer Patients

In-Situ Cancer Vaccine: Phase I/IIb, Open-Label Study to Assess Safety of AllostimTM in Combination With Cryoablation in Metastatic Breast Cancer Previously Treated With an Anthracycline, a Taxane and Capecitabine

This phase I/II study is designed to compare different treatment schedules of a personalized anti-cancer vaccine protocol which combines the cryoablation of a selected metastatic lesion with intra-tumor immunotherapy. The cryoablation causes the tumor to release tumor-specific antigens into the surrounding environment. The injection of bioengineered allogeneic immune cells, AlloStim(TM), into the lesion is designed to modulate the immune response and educate the immune system to kill other tumor cells.

The study will assess three different dosing schedules. A standard 3 plus 3 study design will be used. The starting dose for each dosing schedule will be escalated in subsequent groups of patients. The study will evaluate safety of increased frequency of AlloStim (TM) dosing and anti-tumor effect of the new proposed dose and frequency schedule.

Inclusion Criteria:

1. Women w/ histologically/cytologically confirmed breast carcinoma

2. Documented progressive metastatic disease not amenable to curative surgery/radiotherapy

3. Age ≥18 and ≤70 years

4. Prior treatments that included capecitabine and both an anthracycline and a taxane drug and resistant to taxane therapy

   1. ER+ patients: minimum cumulative dose of anthracycline (≥ 180 mg/m² of doxorubicin or ≥ 300 mg/m² of epirubicin) or resistance to anthracycline, capecitabine and anti-hormonal therapy
   2. Resistance is defined as tumor progression while receiving treatment or progression within 4 months of the last dose in the metastatic setting, or recurrence within 12 months in the neoadjuvant/adjuvant setting

5. Post-menopausal ER+ and/or PR+ must have received at least 2 lines of prior anti-estrogen therapy, which includes an aromatase inhibitor

6. Her2+ patients: at least 1 Her2+ targeted regimen containing trastuzumab alone or with pertuzumab/lapatinib. Trastuzumab/pertuzumab must have been discontinued at least 4 weeks before treatment

7. Prior radiation therapy completed >4 weeks before treatment

8. Measurable disease according to revised RECIST v.1.1 guidelines with at least 1 lesion deemed to be safely accessible for serial biopsy

9. ECOG <2

10. Adequate hematological function

    1. Absolute granulocyte count ≥ 1,500/mm3
    2. Platelet count ≥ 100,000/mm3
    3. PT/INR ≤ 1.5
    4. INR correctable to ≤ 1.5 or a PT/PTT correctable to normal limits. Patients receiving anti-coagulation treatment with agent such as warfarin/heparin may participate. For patients on warfarin, INR should be monitored weekly prior to any intervention to assure INR is stable. Heparin/warfarin must be withheld before biopsy
    5. Hemoglobin ≥ 9 g/dL (may be corrected by transfusion)

11. Adequate organ function

    1. Creatinine ≤ 1.5 mg/dL
    2. Total bilirubin ≤ 1.5 times ULN
    3. Alkaline phosphatase≤2.5 times ULN (≤5 times normal if liver involvement)
    4. Aspartate aminotransferase (AST/SGOT) ≤ 5.0 times ULN
    5. Alanine aminotransferase (ALT/SGPT) ≤ 5.0 times ULN

12. EKG without clinically relevant abnormalities

13. Pre-menopausal with child bearing potential subjects must use adequate contraception

14. Informed consent in the native language of the subject

Exclusion Criteria:

1. Peritoneal carcinomatosis
2. Moderate-large ascites accumulation requiring/likely to require paracentesis
3. Clinical/radiological evidence of brain metastasis/leptomeningeal involvement
4. Pulmonary lymphangitis/symptomatic pleural effusion (grade ≥ 2) that results in pulmonary dysfunction requiring active treatment
5. History of 2nd primary malignancy, except: bilateral breast carcinoma, in situ carcinoma of the cervix, adequately treated non-melanomatous carcinoma of the skin, and other malignancy treated at least 5 years with no evidence of recurrence
6. >3 prior chemotherapy regimens for metastatic disease
7. History of severe hypersensitivity to monoclonal antibody drugs/any contraindication to study drugs
8. Pregnant or breast feeding
9. Any serious, concurrent uncontrolled medical disorder
10. Prior hepatectomy, liver chemoembolization, liver cryoablation/ radiofrequency ablated
11. Symptomatic pulmonary disease
12. Bevacizumab (Avastin®) within 3 weeks of accrual
13. Prior allogeneic bone marrow/stem cell or solid organ transplant
14. Chronic use (> 2 weeks) of greater than physiologic doses of corticosteroid agent (dose equivalent to > 10 mg/day of prednisone) within 30 days of the first day of study treatment. Topical and inhaled corticosteroids are permitted
15. Concomitant active autoimmune disease
16. Prior experimental therapy/cancer vaccine treatment
17. Current immunosuppressive therapy, including: cyclosporine, antithymocyte globulin, or tacrolimus within 1 month of study entry
18. History of blood transfusion reactions
19. Known allergy to bovine products
20. Know allergy to murine products
21. Progressive viral/bacterial infection. All infections must be resolved and the patient must remain afebrile for 7days without antibiotics prior to enrollment
22. Cardiac disease of symptomatic nature or cardiac ejection fraction < 45%
23. History of HIV positivity or AIDS
24. Psychiatric/addictive disorders or other condition that, in the opinion of the investigator, would preclude study participation
25. Concurrent medication known to interfere with platelet function or coagulation (e.g., aspirin, ibuprofen, clopidogrel, or warfarin) unless can be discontinued for an appropriate time period based on the drug half-life and known activity (e.g., aspirin for 7 days) prior to cryoablation procedure
26. Use of low molecular weight heparin preparations unless can be discontinued 8 hours prior to cryoablation