A Randomized Trial Comparing CD3/CD19 Depleted or CD3 Depleted/CD56 Selected Haploidentical Donor Natural Killer (NK) Cell Based Therapy for Adults With Acute Myelogenous Leukemia Who Have Failed 1 or 2 Induction Attempts
Phase: NA
Conditions: Acute Myelogenous Leukemia
Sponsor: Miltenyi Biotec B.V. & Co. KG

Clinical Study Protocol 
Protocol Number 1401-NK 
Sponsor Reference Number MT-2014-02 
A Randomized Trial Comparing CD3/CD19 Depleted or CD3 Depleted/CD56 
Selected Haploidentical Donor Natural Killer (NK) Cell Based Therapy for Adults 
With Acute Myelogenous Leukemia Who Have Failed 1 or 2 Induction Attempts 
Sponsor: 
Miltenyi Biotec, Inc. 
120 Beacon Street, Suite 200 
Somerville, MA 02143 
Participating 
Institutions:  
 
 
 
 
 
Biostatistician: 
 
 
Version Date: 
March 9, 2017 
Confidentiality Statement 
This study protocol is the property of Miltenyi Biotec Inc. It may not be reproduced, distributed or 
otherwise used. The rights to the information in this study protocol and the resulting study first 
become the property of the sponsor when the contract stipulations have been fulfilled on the 
side of Miltenyi Biotec Inc. and the investigators. 

CD3-/CD19- or CD3-/CD56+ Haploidentical Donor NK Cell Based Therapy for AML who Have Failed 1 or 2 Inductions 
March 9, 2017 
confidential 
Key Roles and Responsibilities/Contact Information 
Role
Contact Information
IDE Sponsor/Study Funder 
Miltenyi Biotec, Inc. 
 
Study PI, Deputy PI and Lead 
Institution 
  
PI:- 
 
 
 
 
       
  
Deputy PI : - 
 
 
 
 
Other Clinical Sites 
 
 
 
  
 
 
 
 
 
 
 
 
 
Clinical and Regulatory Leads 
at the 
 
Study Coordinator (Clinical): 
 
 
 
 
  
Affiliate Sites Manager: 
 
 
 
ClinicalTrials.gov Registration 
Miltenyi Biotec, Inc. 
Subject Registration 
 via Online Enterprise Research 
Management Environment (OnCoreTM), a web based Oracle® database 
Subject Randomization 
Study Coordinator 
Data Management (eCRFs, 
and product lot release) 
via Online Enterprise Research 
Management Environment (OnCoreTM), a web based Oracle® database  
Adverse Event Reporting 
(FDA) 
Miltenyi Biotec, Inc. 
Study Site Monitoring 
Oversight 
Miltenyi Biotec, Inc. 
Statistical Support 
 
Lab For Research Related 
Sample Processing And 
Analysis 
 
 
 
 
 
 
 
 

CD3-/CD19- or CD3-/CD56+ Haploidentical Donor NK Cell Based Therapy for AML who Have Failed 1 or 2 Inductions 
March 9, 2017 
confidential 
Synopsis 
Study Design: 
This is a Simon’s optimal two-stage phase II trial of related donor HLA-haploidentical NK-cell based 
therapy for the treatment of newly diagnosed acute myelogenous leukemia (AML) (except acute 
promyelocytic leukemia) in persons who failed to achieve a complete remission (CR) after one or two 
standard induction attempts. Failure is defined as ≥ 30% bone marrow blasts in a bone marrow of at least 
20% cellularity at the mid-cycle (~day 14) bone marrow biopsy or residual AML on ~day 28 bone marrow 
biopsy by morphology, flow, PCR or FISH.  
In this study, after a preparative regimen of cyclophosphamide and fludarabine, patients receive a single 
infusion of CD3-/CD19- NK cells or CD3-/CD56+ NK cells followed by a short course of Interleukin-2 (IL-2) 
to facilitate NK cell survival and expansion.  
The primary endpoint of the study is complete remission (CR and CRi and CRp) by day +42 (±3 days)  
post NK cell infusion with  

absolute neutrophil count > 1,000/mm3, platelet count > 100,000/ mm3, no leukemic blast in the
peripheral blood and < 5% blast in the marrow, (CR)

a morphologic complete remission with incomplete blood count recovery (CRi)

leukemia clearance (< 5% marrow blast and no circulating peripheral blasts) and neutrophil
recovery but with incomplete platelet recovery.
However, a secondary endpoint is the expansion and persistence of NK cells. This endpoint will be used 
in a randomized sub-study in stage 1 to choose one of two T cell depleted NK cells products (CD3-/CD19- 
versus CD3-/CD56+) to be used for the remainder of the study. 
The first 24 patients enrolled in the study (stage I) will be randomized to either CD3-CD19- or CD3-CD56+ 
in a parallel comparison of the two products. An additional 17 patients (stage 2) will receive the optimal NK 
cell product based on success of NK cell expansion from the initial randomized sub-study. Between the 2 
stages, 29 patients will receive the chosen product and be evaluable for the primary endpoint of CR, CRi 
and CRp by day +42.  
Patients achieving a remission may proceed to standard of care consolidation or hematopoietic stem cell 
transplantation at the discretion of the treating physician and independent of this study.  
Patients (including those who proceed to transplant or consolidation therapy) will be followed for disease 
free survival until 1) disease progression, 2) start of a new anti-leukemia therapy, 3) discharge to hospice  
and then for overall survival for a maximum of 2 years from the NK cell infusion,  In addition all patients 
will be followed through day 100 to rule-out for treatment related morality (early study stopping rule 
events) and GvHD 
Primary 
Objective: 
To determine the rate of complete remission defined as CR (leukemia clearance with blood count 
recovery) CRi (morphologic complete remission with incomplete blood count recovery) and CRp 
(leukemia clearance with incomplete platelet recovery) by day +42 post NK cell infusion. 
Secondary 
Objectives: 
Expansion and persistence of NK cells - this endpoint will be used  in a randomized sub-study in stage
1 to identify the preferred cell product processing method
Incidence and severity of adverse events
Incidence of infusional toxicities
Incidence of treatment related mortality
Incidence of “proceeding” to hematopoietic cell transplantation
Incidence of acute graft versus host disease (GvHD) by day +42
Overall survival at 12 and 24 months
Disease-free survival at 12 and 24 months
Correlative 
Objectives: 
Incidence of successful in vivo expansion of adoptively transferred haploidentical NK cells as measured
by the percentage of patients achieving 40% donor DNA and 40% of lymphocytes are NK cells at day 7
OR 20% donor DNA and 20% of lymphocytes are NK cells at day 14
Feasibility of using the appropriate CliniMACS® Reagent System to deplete CD3+ cell and deplete
CD19+ cells (CD3-/CD19-), or to deplete CD3+ cells and enrich CD56+ cells(CD3-/CD56+) to consistently
obtain the product specific targeted NK cell dose
Incidence of minimal residual disease
 

CD3-/CD19- or CD3-/CD56+ Haploidentical Donor NK Cell Based Therapy for AML who Have Failed 1 or 2 Inductions 
March 9, 2017 
confidential 
Disease 
Related 
Criteria: 
Newly diagnosed acute myelogenous leukemia (except acute promyelocytic leukemia) and has failed to 
achieve remission after 1 or 2 standard induction attempts with failure defined as  

≥ 30% bone marrow blasts in a bone marrow of at least 20% cellularity  on the mid-course (~day 14)
bone marrow biopsy or

residual AML at ~day 28 bone marrow biopsy  by morphology, flow, PCR or FISH
AML that has progressed from MDS is eligible provided no prior MDS directed treatment.
Patients enrolling after only 1 failed induction attempt must meet at least one of the following additional
eligibility criteria of high risk: 1) ≥ 60 years of age  2) adverse cytogenetics or molecular characteristics
as defined in section 4.1
Other Key 
Inclusion  
Criteria: 

Available related HLA haploidentical NK cell donor aged 12-70 years
•
≥ 18, but < 75 years of age, Karnofsky performance status ≥ 60%

Adequate organ function within 14 days of study registration (30 days for pulmonary and
cardiac) as defined in section 4.5

Ability to be off prednisone and other immunosuppressive drugs for at least 3 days prior to the
NK cell infusion (excluding preparative regimen pre-meds)

No prior hematopoietic transplant

Not pregnant or lactating

Sexually active females of childbearing potential and males with partners of child bearing
potential must agree to use birth control
Statistical  
Considerations: 
This trial uses a Simon’s two-stage design to estimate the complete remission (CR, includes CRi, 
CRp) rate at day +42 post NK cell infusion. In addition, the trial includes an initial randomized sub- 
study of 24 patients during stage 1 to choose which of the enriched NK cell products (CD3-/CD19- 
versus CD3-/CD56+) should be used to complete the trial. Twelve patients will be randomized to 
each product. As resources are limited, the selection of the best product will partly be a clinical 
decision, based in part on which meets the correlative endpoint of successful in vivo NK cell 
expansion. This is defined as 40% donor DNA and 40% of lymphocytes are NK cells at day 7 post 
infusion OR 20% donor DNA and 20% of lymphocytes are NK cells at day 14 post infusion. As the 
CD56+ is a more pure product, CD3-/CD19- processing only will be used if there is dramatically 
better NK cell expansion and if the safety profile is acceptable. If neither product achieves success 
at the end of stage 1, the study will stop and the platform redesigned. If the trial continues after 
stage 1, an additional 17 patients will be enrolled using the optimal NK cell product to complete the 
trial. 
The null hypothesis is that the CR (including CRi, CRp) rate is ≤ 38% and the alternative 
hypothesis is that CR rate is ≥ 63% at day +42. If 16 or more patients (of 29 using the optimal 
product) have achieved CR, the regimen will be deemed worthy of further study.  
Sample Size: 
A maximum of 29 evaluable (defined as alive at day +14 and receiving all study therapy with at 
least 3 doses of IL-2) patients receiving treatment with the optimal product will be sufficient to 
maintain an overall type I error of 6.5% while providing 80% statistical power. Including Part I 
where the two products are compared side by side, the maximum number of patients will be 41 
and the minimum number of patients will be 24. 
Enrollment Plan: 
Based on enrolling 8 to 10 patients per year, accrual should be completed within 3 to 4 years