Trial Information

AML08: A Phase II Randomized Trial of Clofarabine Plus Cytarabine Versus Conventional Induction Therapy And A Phase II Study Of Natural Killer Cell Transplantation In Patients With Newly Diagnosed Acute Myeloid Leukemia

The overall objective of this protocol is to improve the cure rate of acute myeloid leukemia
(AML).

We will compare the immunologic complete response rate after one course of therapy in
patients who receive cytarabine + daunorubicin + etoposide (ADE) with that in patients who
receive clofarabine + cytarabine (Clo/AraC)

Secondary objectives include

- To estimate the event-free survival (EFS) of standard risk (SR) patients who receive
chemotherapy alone and the EFS of SR patients who receive chemotherapy followed by
natural killer (NK) cell transplantation.

Exploratory Objectives:

- To genotype natural killer (NK) cell receptors and measure their expressions at
diagnosis and after induction therapy, and to explore the associations of these
features with treatment outcome

- To assess the prognostic value of levels of minimal residual disease in peripheral
blood at day 8 of induction I

- To validate new markers and methods for minimal residual disease (MRD) detection

- To identify new prognostic factors by applying new technologies to study patient
material

- To identify pharmacogenetic, pharmacokinetic and pharmacodynamic predictors for
treatment-related outcomes in the context of the systemic therapy used in the protocol

- To describe the impact of antibiotic and antifungal prophylaxis on invasive bacterial
and fungal infections, febrile neutropenia, hospitalization, and antibiotic resistance.

- To determine the performance characteristics of broad-range, molecular diagnostic
methods for detection of bacterial, fungal, and viral agents, in comparison to methods
currently in routine clinical use

Treatment will be based on cytogenetic and molecular characteristics, morphology, and
response to therapy as assessed by flow cytometry. Risk groups are defined below. The
general treatment plan will consist of chemotherapy for LR patients, chemotherapy ± NK cell
therapy for SR patients, and chemotherapy + stem cell transplant (SCT) for HR patients. HR
patients who do not have a suitable stem cell donor or who decline SCT will be eligible for
NK cell therapy.

Low-risk (LR) criteria (not eligible for SCT or NK cell therapy)

- Core binding factor (CBF) leukemia [t(8;21)/AML1-ETO or inv(16)/t(16;16)/CBF-MYH11,]
and MRD < 0.1% at day 22,regardless of other genetic features.

- Patients with CBF leukemia who have MRD ≥ to 0.1% at day 22 or who have increasing
levels of fusion transcript will be considered SR and thus eligible for NK cell
therapy.

Standard-risk (SR) criteria (eligible for NK cell therapy)

- Absence of low-risk or high-risk features.

- CBF leukemia with MRD ≥ 0.1% at day 22 or increasing levels of fusion transcript

- FLT3-ITD and MRD < 0.1% at day 22

High-risk (HR) criteria (candidates for SCT; eligible for NK cell therapy)

Presence of one of the following features:

- t(6;9), t(8;16), t(16;21), -7, -5, or 5q-

- FAB M0 or M6

- FAB M7 without t(1;22)

- Treatment-related (secondary) AML

- RAEB-2 or AML arising from prior MDS

- FLT3-ITD and MRD ≥ 0.1% at day 22

- All other patients with poor response to therapy (must have one of the following
features) MRD ≥ to 5% at day 22 MRD ≥ to 0.1% after Induction II

Induction therapy (2 courses)

All patients will receive two courses of induction therapy that will include one course of
either high dose cytarabine, daunorubicin, and etoposide (HD-ADE) or one course of
clofarabine and cytarabine (Clo/AraC), followed by one course of low dose cytarabine,
daunorubicin, and etoposide (LD-ADE). Patients will be randomly assigned to receive one of
the following induction regimens.

Induction I: HD-ADE

Cytarabine: 3 g/m2 IV over 3 hours q12 hours x 6 doses (days 1, 3, 5) Daunorubicin: 50 mg/m2
(1.67 mg/kg for patients less than 10 kg) IV over 6 hours on days 2, 4, 6 (3 doses)
Etoposide: 100 mg/m2 IV over 4 hours on days 2-6 (5 doses)

Induction I: Clo/AraC

Clofarabine: 52 mg/m2 IV over 2 hours on days 1-5 (5 doses) Cytarabine: 1 gram/m2 IV over 2
hours on days 1-5 (5 doses; each dose to start 4 hours after the start of clofarabine)

Induction II: LD-ADE

Cytarabine: 100 mg/m2 IV over 30 minutes q12 hours on days 1-8 (16 doses), Daunorubicin: 50
mg/m2 (1.67 mg/kg for patients less than 10 kg) IV over 6 hours on days 2, 4, 6 (3 doses)
Etoposide: 100 mg/m2 IV over 4 hours on days 1-5 (5 doses)

Induction II for patients with FLT3-ITD: LD-ADE + Sorafenib

Patients with FLT3-ITD will take Sorafenib, 400 mg/m2 per day, orally in two divided doses
(200 mg/m2/dose BID) starting one day after the completion of Induction II and continuing
for 21 days Patients with FLT3-ITD who do not experience toxicity related to Sorafenib will
also receive a 21-day course of Sorafenib after subsequent courses of chemotherapy.

Induction II for other HR patients:

LD-ADE + valproic acid Patients with M7 AML without t(1;22) and other HR patients without
FLT3-ITD will be treated with a combination of valproic acid and LD-ADE. Valproic acid will
be given orally for 3 days (Days -2, -1, 0) prior to the initiation of Induction II
chemotherapy and will continue to be given for an additional 6 days during chemotherapy (Day
1 through 6).Valproic acid doses may be increased if necessary to achieve levels > 50
mcg/ml.

Special subgroup HR patients with MRD < 0.1% may proceed directly to SCT after Induction I
if a suitable donor is available and the transplant can be performed without delay.

Consolidation I:

Mitoxantrone: 12 mg/m2 (0.4 mg/kg for patients less than 10 kg) IV over 1 hour on days 3-5
(3 doses) Cytarabine: 1 g/m2 IV over 2 hours every 12 hours on days 1-4 (8 doses)

Consolidation II:

Cytarabine 3 g/m2 IV over 3 hours every 12 hours on days 1, 2, 8, 9 (8 doses). Erwinia
Asparaginase 25,000 Units/m2 (833 Units/kg for infants < 1 month of age, or for infants < 3
months of age who were born significantly prematurely defined as < 36 weeks gestation) IV
over 1 hour, 3 hours after the 4th and 8th doses of cytarabine.

NK cell therapy Standard risk patients who have a KIR-mismatched family member who is
greater than 18 years old will undergo NK cell transplantation. In addition, HR patients who
do not have a suitable stem cell donor or who decline SCT will be eligible for NK cell
therapy if they have a KIR-mismatched family member.

Treatment schema Day -7: Cyclophosphamide 60 mg/kg IV over 1 hour. Mesna 15 mg/kg/dose IV
Days -6 through -2: Fludarabine 25 mg/m2/day IV over 30 minutes (5 doses) Days -1, +1, +3,
+5, +7, +9: IL-2 1 million units/m2 given subcutaneously Day -1: Donor pheresis Day 0: NK
cell infusion

No steroids, including the use of hydrocortisone as pre-medication, may be given to patients
during the 3 days prior to the NK cell infusion or during the first 7 days after the
infusion.

CNS therapy

Triple intrathecal therapy with methotrexate, hydrocortisone, and cytarabine (MHA) will be
used for all CNS therapy at the doses:

< 1 year methotrexate 6 mg, hydrocortisone 12 mg, cytarabine 18 mg, 1-2 years methotrexate 8
mg, hydrocortisone 16 mg, cytarabine 24 mg, 2-3 years methotrexate 10 mg, hydrocortisone 20
mg, cytarabine 30 mg, > 3 years methotrexate 12 mg, hydrocortisone 24 mg, cytarabine 36 mg

Leucovorin rescue (5 mg/m2 per dose; 5 mg maximum per dose) will be given orally or
intravenously at 24 and 30 hours after each IT MHA treatment.

Patients with no evidence of CNS disease \[(i.e., no leukemic blast cells on cerebrospinal
fluid (CSF) cytospin] will receive 4 total doses of intrathecal therapy, given at
approximately one month intervals or at the beginning of each of the first 4 courses of
chemotherapy.IT therapy will not be given before NK cell therapy.

Patients with overt CNS leukemia (less than or equal to 5 leukocytes per l of CSF and the
presence of leukemic blast cells on CSF cytospin) will receive weekly intrathecal therapy
until the CSF is free of blast cells (minimum number of doses, 4). These patients will then
receive 4 additional doses of intrathecal therapy (minimum total number of doses, 8) at
approximately 1-month intervals (generally given with each subsequent course of
chemotherapy).IT therapy will not be given before NK cell therapy.

Patients with < 5 leukocytes per mul of CSF and the presence of leukemic blast cells on CSF
cytospin (CNS2)will receive weekly intrathecal therapy until the CSF is free of blast cells.
These patients will then receive 4 additional doses of intrathecal therapy at approximately
1-month intervals (generally given with each subsequent course of chemotherapy).IT therapy
will not be given before NK cell therapy.

Patients who are unable to undergo lumbar puncture and receive intrathecal therapy prior to
starting induction I should be treated as CNS2 unless they have overt CNS leukemia (CNS3).