CliniMACS® CD34 Reagent System FDA Advisory Panel Miltenyi Biotec CliniMACS® CD34 Reagent System September 23, 2011 With Sincere Thanks and Appreciation To:

Director, Regulatory Affairs Miltenyi Biotec Incorporated The CliniMACS® CD34 Reagent System FDA Advisory Panel September 23, 2011 Nancy Johansen Director, Regulatory Affairs Miltenyi Biotec Incorporated

Agenda and Presenters Company Overview, Product Introduction and Registration of the CliniMACS® CD34 Reagent System as a Humanitarian Use Device Overview of the Clinical Indication and Unmet Medical Need and Summary of the BMT CTN 0303 Clinical Trial Summary of the CliniMACS® CD34 Reagent System Performance Summary of the BMT CTN 0303 versus BMT CTN 0101 Data Analysis Protocol Findings Summary of Safety and Probable Benefit of the CliniMACS® CD34 Reagent System Nancy Johansen Director, Regulatory Affairs Miltenyi Biotec Incorporated Steven Devine, M.D. Ohio State University Arthur G. James Cancer Center Carolyn Keever-Taylor, PhD Medical College of Wisconsin Marcelo Pasquini, M.D., M.S. Center for International Blood and Marrow Transplant Research (CIBMTR) Medical College of Wisconsin Kai Pinkernell, M.D. Head of Clinical Development Miltenyi Biotec GmbH Maybe summary instead of Review in both slides?

Miltenyi Biotec Corporate Overview Founded in 1989 in Bergisch Gladbach, Germany Roughly 1100 employees worldwide Subsidiaries in 10 countries; N.A. Headquarters in Auburn, CA Miltenyi Biotec sells products and services worldwide. Employees: D: >800 EU: 80 US: 100 Asia-Pacific: 60

CliniMACS® CD34 Reagent System The overall function of the CliniMACS® CD34 Reagent System is to select CD34+ cells from heterogeneous hematologic cell populations Thereby passively depleting T-cells

CliniMACS® CD34 Reagent System CliniMACS® Tubing Sets (Standard and Large Scale) Standard: 0.6 x 109 CD34+ Cells from 60 x 109 Cells Large Scale: 0.6-1.2 x 109 CD34+ Cells from 60-120 x 109 Cells CliniMACS®plus Instrument CliniMACS® PBS/EDTA Buffer

The Principle of the CliniMACS® CD34 Reagent System Elution of the labeled cell fraction Magnetic labeling Magnetic Separation (elution of the non-labeled cell fraction)

Location of CliniMACS®plus Instruments in the USA 162 instruments within 97 institutions in the U.S.

CliniMACS® CD34 Reagent System Protocols (as of July 2011) 84 IDE protocols utilize the CliniMACS® CD34 Reagent System Strict distribution procedures are in place to ensure that investigational products are provided only for FDA/IRB approved protocols

Installation, Training and Customer Support CliniMACS® plus Instruments are installed by qualified Miltenyi personnel Installation and Operational Qualification (IQ/OQ) is performed at time of installation All subsequent servicing and Preventative Maintenance are performed by qualified Miltenyi personnel Customer Training Performed by qualified Miltenyi personnel Training validated by written test Emergency Hotline Support Monday through Friday (9 a.m. – 9 p.m. EST) If pre-arranged, outside of normal business hours

CliniMACS® CD34 Reagent System Registration Pathway 2004-2008 BMT CTN Study Conducted 2011 HDE Submitted CliniMACS® CD34 Reagent System Registration Pathway 1998 US MF Submitted 2010 DAP Finalized 2004 Pre-IDE Meeting 1997 CE marked 2005-HUD Designation Dec 2009 FDA pre-HDE meeting

Humanitarian Use Device The device is designed to treat or diagnose a disease or condition that affects fewer than 4,000 individuals per year in the U.S. The device is not available otherwise, and there is no comparable device available to treat or diagnose the disease or condition; and The device will not expose patients to unreasonable or significant risk, and the benefits to health from the use outweigh the risks Clinical data must support “safety” and “probable benefit” argument Exempt from “effectiveness requirement” consistent with the HDE requirements

Clinical Trial Supporting the HUD Registration of the CliniMACS® CD34 Reagent System* Miltenyi supported a Phase II multi-center clinical trial sponsored by the BMT CTN (BB-IDE 11965) which enrolled 47 AML patients from October 2005-December 2008 Evaluated the use of the CliniMACS® CD34 Reagent System for selecting CD34+ cells from HLA-matched related donors for allogeneic stem cell transplantation after myeloablative therapy in patients with Acute Myeloid Leukemia (AML) in 1st or 2nd CR, without additional GVHD prophylaxis * Miltenyi provided material goods only in the support of this study. MBI has negotiated rights through the National Marrow Donor Program (NMDP) to cross reference the BMT CTN 0303 IDE submission for purposes of product registration

CliniMACS® CD34 Reagent System Proposed Humanitarian Use Indication “Humanitarian Use Device: Authorized by U.S. Federal law for processing allogeneic HLA-matched hematopoietic progenitor cells-apheresis (HPC-A) from a related donor to obtain a CD34+ cell enriched population intended for hematopoietic reconstitution following a myeloablative preparative regimen without the need for additional graft-vs-host disease (GVHD) prophylaxis in patients with acute myeloid leukemia (AML) in first or second morphologic complete remission”

Steven Devine, M.D. Professor of Internal Medicine Director, Blood and Marrow Transplant Program The Ohio State University Arthur G. James Cancer Center BMT CTN 0303 Co-Study Chair

Acute Myeloid Leukemia (AML) Most common leukemia diagnosis in adults1 U.S. incidence: ~ 12,330 new cases diagnosed annually Mortality is roughly 8,950 cases per year However, less than 2,500 patients progress to transplant Allogeneic stem cell transplantation (SCT) is the single most effective therapy currently available for the prevention of relapse and shows significant survival benefit in AML patients with intermediate and poor risk cytogenetics in first complete remission (CR1)2 Acute myeloid leukemia (AML), also known as acute myelogenous leukemia or acute non-lymphocytic leukemia, is a clonal malignant disease characterized by an over-proliferation in the myeloid stem cell compartment. This results in an accumulation of myeloid precursor cells in the blood and the bone marrow, arrested in an early stage of maturation. These immature and dysfunctional cells in the marrow interfere with normal hematopoiesis by inhibition of normal differentiation, which leads to suppression of the erythroid, myeloid and megakaryocytic series, resulting in bone marrow failure. Leukemic cells may also spread outside the blood and bone marrow, and invade other parts of the body, including the central nervous system, skin, gums, and testes. annual incidence rising from 3.8 cases per 100,000 to 17.9 cases per 100,000 adults aged 65 and older High mortality (also for younger adults, when untreated) / poor survival (5 year relative survival is 21% overall and decreasing with higher age) 1 http://www.Cancer.org; 8/26/10 2 Koreth, J et al. JAMA 2009 16

Graft Versus Host Disease (GVHD) Complicates Allogeneic Transplantation From Matched Related Donors (MRD)1-6 Reduced incidence of leukemic relapse and overall survival often not realized due to complications caused by GVHD Acute GVHD (aGVHD) risk is 35-45% 33-81% of these patients will develop chronic GVHD (cGVHD), resulting in post-transplant morbidity, mortality and reduced quality of life Immunosuppressive agents used to prevent and treat aGVHD do not affect incidence of cGVHD Previous studies demonstrate that T cell depletion reduces the risk of severe acute and chronic GVHD 46 Ferrara JLM, et al: Graft-versus-host disease. Lancet 373: 1550-61, 2009. 47 Chao NJ, et al: Cyclosporine, methotrexate, and prednisone compared with cyclosporine and prednisone for prophylaxis of acute graft-versus-host disease. N Eng J Med 329: 1225-30, 1993. 48 Devine SM, et al: Recent advances in allogeneic hematopoietic stem-cell transplantation. J Lab Clin Med 141:7-32, 2003. 49 Clift RA, et al: Allogeneic marrow transplantation in patients with acute myeloid leukemia in first remission: a randomized trial of two irradiation regimens. Blood 77: 1660-5, 1991. 50 Nash RA, et al: Phase 3 study comparing methotrexate and tacrolimus with methotrexate and cyclosporine for prophylaxis of acute graft-versus-host disease after marrow transplantation from unrelated donors. Blood 96: 2062-8, 2000. 51 Ratanatharathorn V, et al: Phase III study comparing methotrexate and tacrolimus (prograf, FK506) with methotrexate and cyclosporine for graft-versus-host disease prophylaxis after HLA-identical sibling bone marrow transplantation. Blood 92: 2303-14, 1998. 1 Ferrara J, et al. Lancet 2009 2 Chao N, et al. NEJM 1993 3 Devine S, et al. J Lab Clin Med 2003 4 Clift R, et al. Blood 1991 5 Nash R, et al. Blood,2000 6 Ratanatharathorn V, et al. Blood 1998

Chronic GVHD of the Skin Walker, I; 2011 BMT Tandem Meetings CBMTG Section; Clinical Trials Network Meeting. www.cbmt.org

Chronic GVHD of the Oral Mucosa Walker, I; 2011 BMT Tandem Meetings CBMTG Section; Clinical Trials Network Meeting. www.cbmt.org

Chronic GVHD and Quality of Life Quality of life (QoL) at 6 or 12 months is significantly worse in patients with acute or cGVHD after transplant1 QoL at 12 months improves unless cGVHD develops1 Significantly less patients return to work if they develop cGVHD after allogeneic transplantation2 Only 41% of patients with cGVHD return to work at 3 years, compared to 95% of patients without cGVHD2 1 Lee et al. BMT 2006; (38) 305-10 2 Wong et al. Blood. 2010; 115(12)2508-19

Chronic GVHD Treatment The treatment of cGVHD with long-term corticosteroids increases the risk of cataract formation, avascular necrosis, and osteoporosis1 The 10-year survival is less than 5% for patients affected by severe cGVHD There are no effective options for the prevention or treatment of chronic GVHD 1 Horowitz, ME., and Sullivan, KM. Blood Reviews (2006); 20: 15–27

Unmet Medical Need Served by The CliniMACS® CD34 Reagent System The incidence and severity of GVHD are most effectively reduced by ex vivo T cell depletion (TCD) of the allograft There is currently no approved method for ex vivo TCD for allogeneic SCT in the United States If approved, the CliniMACS® CD34 Reagent System will be the only FDA approved method of CD34+ enrichment and passive TCD available

Clinical Study Rationale Use of ex vivo T cell depletion (TCD) has been limited by logistical difficulties and variability in TCD methods lack of an FDA-approved method concerns regarding potential risk of graft rejection and leukemic relapse A multi-center trial of TCD in AML patients in complete remission (CR1/CR2) using standard eligibility criteria and a uniform method of TCD was warranted

HLA-Identical Sibling-Matched, CD34+ Selected, T cell Depleted Peripheral Blood Stem Cells Following Myeloablative Conditioning For First or Second Remission Acute Myeloid Leukemia (AML): Results of Blood and Marrow Transplant Clinical Trials Network* (BMT CTN) Protocol 0303 S Devine, S Carter, R Soiffer, M Pasquini, P Hari, A Stein, H Lazarus, C Linker, E Stadtmauer, E Alyea, C Keever-Taylor, and R O'Reilly * Devine, S et al; BBMT, 2011

Blood and Marrow Transplant Clinical Trials Network (BMT CTN) Established: Sept. 2001; renewed July 2011 Funded by NHLBI/NCI 20 Core Center cooperative agreements 1 DCC cooperative agreement >80 affiliate centers access trials through DCC Goal of the Program: Provide the infrastructure needed to allow promising HCT therapies to be developed/ evaluated in high quality multicenter studies. 25

BMT CTN Centers, 2011 >100 centers have enrolled >3900 patients since 2003 = PBMTC Centers = Core Centers = Affiliate Centers 26 Mmh06_16.ppt

BMT CTN 0303 Historical References Single center studies show reduced GVHD without increased relapse rates in AML patients receiving T cell depleted (TCD) allografts in CR1-3 One randomized, prospective, multi-center TCD trial showed no increase in relapse in AML patients receiving TCD allografts4 1 Aversa et al. Blood Cells Mol and Disease 2008 2 Pappadopoulos et al, Blood, 1998 3 Soiffer et al; Blood 1997 4 Wagner, J. et al. Lancet 2005 27

Randomized, Prospective, Multi-center T Cell Depletion (TCD) Vs Randomized, Prospective, Multi-center T Cell Depletion (TCD) Vs. Methotrexate and Cyoclosporine (M/C) Trial Unrelated Bone Marrow Wagner et al., Lancet 366:733, 2005

BMT CTN 0303 Statistical Sampling and Time Points The sample size was 45 patients, wherein 47 patients were enrolled and 44 completed treatment There were no blinding or randomization aspects to this trial The median follow up of the patients was 34 months (Range: 11.5- 51.5 months)

BMT CTN 0303 Study Eligibility AML in CR1 or CR2 Age 18-65 HLA-identical sibling available No more than 2 induction cycles of chemotherapy required to induce remission No more than six months from CR to transplant (three months for CR2) Other standard organ function criteria No uncontrolled bacterial/fungal/viral infections Karnofsky performance status > 70% 30

BMT CTN 0303 Study Endpoints Primary Endpoint: Disease-Free Survival (DFS) at 6 months >75% Secondary Endpoints: Acute and chronic GVHD Overall Survival (OS) Disease-Free Survival at 2 years Transplant-Related Mortality (TRM) Relapse Engraftment/graft failure Infusional Toxicities Incidence of EBV reactivation and PTLD Proportion of grafts containing > 5 x 106 CD34+ cells/kg and < 1 x 105 CD3+ cells /kg

Eight Centers Enrolled Patients onto BMT CTN 0303 Site Patients Accrued Dana Farber/Partners Cancer Center 18 Ohio State University 8 Memorial Sloan Kettering Cancer Center 7 Medical College of Wisconsin City of Hope National Medical Center 3 University Hospitals of Cleveland 2 University of CA, San Francisco 1 University of Pennsylvania TOTAL 47 44 patients were evaluable on study 32

BMT CTN 0303 Patient Characteristics Patient Age Mean (range) 46.3 (21-59 ) Donor age 46.2 (16-63) Gender Male 16 (36%) Female 28 (64%) Leukemia stage CR1 37 (84%) CR2 7 (16%) Cytogenetic Risk (CR1/CR2) Favorable 0 / 2 Intermediate 25 / 3 Unfavorable 10 / 1 Unknown* 2 / 1 * Unknown cytogenetic risk due to lack of metaphase during testing 33

Patient Conditioning Regimen BMT CTN 0303 Patient Conditioning Regimen Day of Tx -9 -8 -7 -6 -5 -4 -3 -2 -1 0 TBI 1375 cGy* 11 total doses; administered on days -9 through -6 Thiotepa @ 5mg/kg Thymoglobulin @ 2.5 mg/kg Cyclophosphamide @ 60mg/kg CliniMACS® CD34-enriched cells

BMT CTN 0303 Donor Mobilization & Leukapheresis Received daily G-CSF for mobilization following screening and enrollment Leukapheresis performed according to institutional standards Daily leukapheresis with subsequent CD34+ cell selection using the CliniMACS® CD34 Reagent System continued until a post-selection target dose of > 5.0 x 106 CD34+ cells/kg and < 1 x 105 CD3+ cells/kg was met

BMT CTN 0303 Steering Committee Laboratory Representative Carolyn Keever-Taylor, PhD Professor of Medicine Director BMT Laboratories Division of Hematology and Oncology Medical College of Wisconsin BMT CTN 0303 Steering Committee Laboratory Representative

The Manuscript Entitled: “Characteristics of CliniMACS® System CD34-Enriched T Cell-Depleted Grafts in a Multi-Center Trial for Acute Myeloid Leukemia-Blood and Marrow Transplant Clinical Trials Network (BMT CTN) Protocol 0303” has been submitted and accepted for publication by the peer reviewed journal, “Biology of Blood and Marrow Transplantation”    

Secondary Endpoint: CliniMACS® CD34 Reagent System Performance Leukapheresis collections from a Matched Related Donor were performed in order to obtain a minimum of ≥ 2.0 x 106 CD34+ cells/kg Target of > 5.0 x 106 CD34+ cells/kg and < 1 x 105 CD3+ cells/kg Up to three collections were allowed to achieve the minimum CD34+ cell dose The CliniMACS® Tubing Sets (Standard or Large Scale) were used based on starting nucleated cell counts

Cellular Testing Requirements Cell Viability (7-AAD), TNC and CD34+ cell content At product receipt After platelet and antibody wash On CliniMACS® CD34-enriched fraction CD3+ T cell content Other cellular testing on CliniMACS® CD34-enriched fraction only CD14+ monocytes CD19+ or CD20+ B cells CD56+ NK Cells Similar point is to identify testing requirements. Not on slide but will indicate that Flow was expected to have been validated and labs were to have participated in proficiency programs and be CLIA certified.

Donors and Products 47 patients enrolled, 44 proceeded to transplant 86 products collected Total lots (cells from one tubing set) assessed=84 Collections pooled for 2 patients 4 sites processed from 9 to 34 lots 4 sites processed ≤ 4 lots May not be needed, but sets stage for data

CliniMACS® CD34 Reagent System Post-Processing Performance N = 84 % CD34+ Recovery % CD34+ Purity Log10 TCD % Viability Mean 66.06 93.0 4.78 96.57 SD ±20.25 ±8.3 ±0.55 ±3.84 Min 29.9 61.5 3.2 74.0 Max 125.6 99.8 5.9 100.0 Overall outcome for all sites. Could be dropped.

Center to Center Statistical Analysis Sites processing ≥ 9 lots compared individually (N=4) Sites processing ≤ 4 lots pooled (N=4) Multivariate analysis used a linear mixed effect model to account for repeated measures (≥ 2 lots for most patients) Pairwise center comparisons were performed with Tukey-Kramer adjustment for multiple comparisons Not sure what Adam will present. This as much as I would be willing to say about the stats

Pre-Processing Cell Counts We first looked to see if the starting products were similar for each of the groups and found that there were some differences in each of the parameters. The median TNC, CD34+ cells, and CD3+ cells were highest for center two at which the first two donors were mobilized using 16 mg/kg G-CSF initially required by the protocol. Subsequent donors were mobilized at 10 mg/kg. The figure shows the products that split. Viability by 7-AAD also differed between center 4 and most of the other centers by having more products with a lower viability. However, this center performed it’s analysis using a flow method that does not include wash steps, which is known to results in somewhat lower viability using 7-AAD, since dead cells are not washed away.

Post Processing Outcomes Here the post processing outcomes are shown by center, and although some differences are seen these are mostly limited to a one or at most two centers. CD34 recovery was significantly lower for center 4 as compared to center 3 and 1, and center 1 had significantly higher recovery than center two, though this was clearly affected by the four lots that exceeded 100% recovery. Purity and log TCD were less variable, with very high purities achieved for most products with a few outliers at center 2, 4, and the combined 5-8. Log TCD significantly differed only between center 3 and 4, and again viability was generally high, with the only significant difference seen for center 3 with a uniformly high viability near 100%. All centers were able to process grafts that met the study criteria

CD34+ Cells x 106/kg of Patient Weight Infused CD34+/kg CD34+ Target Dose CD34+ Minimum Dose CD34 infused. Nicely shows that not only were the targets met, but they were mostly exceeded. I have not presented anything regarding the number of collections for each patient to achieve the doses, Figure that would be part of the clinical presentation, but let me know. All patients received the minimum CD34+ dose (> 2.0x106 cells/kg) 84.1% of patients received > 5 x 106 CD34+ cells/kg 45

Patient Weight Infused CD3+ Cells x 105/kg of Patient Weight Infused CD3+/kg Upper limit of CD3+ dose This represent the cells infused. All final graft products were under the upper limit. No patients received more than 1.0x105 CD3+ cells/kg 46

Final Cellular Product Summary Parameter Result Median CD34+ dose 7.92 x 106/kg Range 2.4 - 30.3 x 106/kg Median CD3+ dose 0.7 x 104/kg 0.1 – 8.3 x 104/kg Median Log10 TCD 4.9 logs 3.2 – 5.9 logs The grafts contained a median of nearly 8e6 CD34+ cells/kg, with only 0.7e4 T cells. The products were sterile, as tested by gram stain and culture, with endotoxin below the detectable limits in all cases. No significant infusional related toxicities were observed. All gram stains/14 day cultures were negative All endotoxin < 5.0 EU/kg No significant infusion related toxicities observed

Conclusions - Secondary Endpoint CliniMACS® CD34 Reagent System Performance All sites, and all products met and most exceeded study goals for: CD34+ cell infusion dose > 2 x 106/kg 84% met the goal of > 5 x 106/kg CD3+ cell infusion dose < 1 x 105/kg The performance of the CliniMACS® CD34 Reagent System was stable and reproducible, resulting in a consistently high degree of CD34+ cell enrichment, T cell depletion and sterility in a multi-center setting

Steven Devine, M.D. Professor of Internal Medicine Director, Blood and Marrow Transplant Program The Ohio State University Arthur G. James Cancer Center BMT CTN 0303 Co-Study Chair

Primary Endpoint 6 Month Disease-Free Survival of >75% 81.8% @ 6 months (95% CI 66.9-90.5)

Secondary Endpoints Neutrophil/Platelet Engraftment Analysis N Median Days to Engraftment Day 30 Estimate Day 100 Estimate Platelet Engraftment >20K/µL 44 16 days 93.2% (95% CI: 85.2-100) 97.7 % 92-100) Cumulative Incidence of Neutrophil Engraftment >500/µL 12 days 100% 85.5-100) No primary graft failures One secondary graft failure at Day +54 after initially engrafting on Day +12 51

Stopping guideline of <30% TRM at 1 year was not exceeded Secondary Endpoint Cumulative Incidence of Transplant-Related Mortality (1yr) All Patients By CR1/CR2 13.6% @ 1yr (95% CI: 3.4-23.8) Stopping guideline of <30% TRM at 1 year was not exceeded TRM at 2 years was 19.9% (95% CI: 7.1-32.7)

Secondary Endpoint Cumulative Incidence of EBV Reactivation All Patients By CR1/CR2 18.2% @ 2 yrs 13.6% (95% CI: 3.4-23.8) 18.9% @ 2 yrs 14.3% @ 2 yrs 8 patients treated for EBV DNA levels >1000 copies/mL One case of PTLD with subsequent death EBV monitoring performed weekly

Secondary Endpoint Cumulative Rate of Relapse All Patients By CR1/CR2 57.1% @ 2 yrs 23.7% @ 2 yrs 20.6% @ 1 yr 17.4%* @ 2 yrs * N=7 patients treated in CR2; 4 patients relapsed (95% CI: 14.6-99.6%)

Secondary Endpoint - Cumulative Incidence of Acute GVHD Grades II-IV All Patients By CR1/CR2 22.7% @ 100 days (95% CI: 10.2-35.2) No Grade IV acute GVHD observed Published acute GVHD risk 35-45% 55

Secondary Endpoint - Cumulative Incidence of Acute GVHD Grades III-IV All Patients By CR1/CR2 4.5% @ 100 days (95% CI: 0 – 10.8%) No Grade IV acute GVHD observed

Secondary Endpoint Cumulative Incidence of Chronic GVHD (2 yrs) Includes Limited and Extensive All Patients By CR1/CR2 19% @ 2 years (95% CI: 6.8-31.1) Published chronic GVHD risk 33-81%

Secondary Endpoint Cumulative Incidence of Extensive Chronic GVHD (2 Years) All Patients By CR1/CR2 6.8% @ 2 years (95% CI: 0-14.4) 58

Secondary Endpoint Disease-Free Survival at 2 Years All Patients By CR1/CR2 By Stage 65.7% @ 1 yr 56.4% @ 2 yrs 61.9% @ 2 yrs 28.6% @ 2 yrs Check graph #2 by CR1/CR2 the line is partially missing Historical data estimates 2 year DFS < 60% in CR11-3 2º endpoints were > 70% for CR1 and > 60% for CR2 1 Suciu Blood 2003 2 Brunet et al, Hematologica 2004 3 Cornelissen et al, Blood 2007

Secondary Endpoint Overall Survival at 2 Years 77.3% @ 1 yr 59.4% @ 2 yrs 60

Adverse Events (AEs) Toxicities were reported as AEs No Unexpected Grade 3-5 AEs were reported AEs were due to regimen related events and toxicities common to allo HCT and could not be attributed to the CliniMACS® CD34 Reagent System The most common toxicities within 1 year post-transplant: Regimen related Grade 3 or 4 GI toxicity (40.9%) Abnormal liver function (36.4%) Graded using the NCI’s CTCAE version 3.0

Causes of Death by 2 Years Post-Transplant Recurrence 6 Infection 4 Idiopathic Pneumonia Organ failure Other* 2 1 PTLD Total 16 * Possibly cardiac related 62

BMT CTN 0303 Conclusions HCT following myeloablative preparative regimen for patients with AML in CR1 or CR2 can be performed in a multicenter setting using the CliniMACS® CD34 Reagent System without additional post-transplant pharmacologic GVHD prophylaxis All 1º and most 2º endpoints were met, demonstrating: 81.8% Disease-Free Survival 6 months post TX No primary graft failure; consistent neutrophil and platelet engraftment Acute GVHD grades II-III <23%. No Grade IV aGVHD Chronic GVHD at 2 years 19%; extensive 6.8% TRM <20% at 2 years Overall risk of relapse was low at 23.7% at 2 years The CliniMACS® CD34 Reagent System consistently produced a graft with > 2 x 106 CD34+ cells/kg and < 1 x 105 CD3+ cells/kg with no reported device related toxicities

Principal Investigator Data Analysis Protocol Marcelo Pasquini, M.D., M.S. Assistant Professor of Medicine Medical College of Wisconsin Assistant Scientific Director Center for International Blood and Marrow Transplant Research (CIBMTR) Principal Investigator BMT CTN 0101 versus BMT CTN 0303 Data Analysis Protocol

A Single Arm, Multi-center Phase II Trial of Transplants of HLA-Matched, CD34+ Enriched T cell Depleted Peripheral Blood Stem Cells Isolated by the CliniMACS® System in the Treatment of Patients with AML in First or Second Morphologic Complete Remission (BMT CTN) Protocol 0303 A Randomized Double-blind Trial of Fluconazole versus Voriconazole for the Prevention of Invasive Fungal Infections in Allogeneic Blood and Marrow Transplant Patients (BMT CTN) Protocol 0101

The Data Analysis Protocol (DAP) Performed according to FDA approved Data Analysis Protocol #1001-34 which was a prospective statistical comparison of retrospective data This comparison of the 0101 and 0303 data has been submitted for publication. The manuscript is currently under review.

Enrollment Period BMT CTN 0101 versus BMT CTN 0303 2003 2006 2005 2008 Enrolling Centers by Protocol, and Their Contribution to Each Study 15 3 5 23% of 0101 47.5% of 0303

Data Analysis Protocol (DAP) Objective Test the hypothesis that T cell depletion using the CliniMACS® CD34 Reagent System as the sole form of immune suppression (BMT CTN 0303) demonstrates a comparable safety profile as compared to conventional GVHD prophylaxis post-transplant (BMT CTN 0101) in AML patients in CR1/CR2 receiving a Peripheral Blood Stem Cell (PBSC) allograft from a matched related donor

The Following One Year* Endpoints Were Statistically Compared Disease-Free Survival (DFS) Overall Survival (OS) Engraftment Graft Failure Transplant-Related Mortality (TRM) Relapse Incidence and Severity of Acute GVHD (aGVHD) Incidence and Severity of Chronic GVHD (cGVHD) * BMT CTN 0101 follow-up was limited to 1 year

Eligibility Criteria for 0303 was Matched in 0101 Patient Subsets Patient Age 18-65 Unmodified PBSC Allograft HLA identical sibling donor AML in CR1 or CR2 All cytogenetic risk patients included in 0101 Only CR2 patients in 0303 enrolled favorable cytogenetic risk patients

Patients meeting eligibility Patients meeting eligibility DAP Patient Selection BMT CTN 0101 BMT CTN 0303 Patients Randomized N=600 Patients Enrolled N=47 No Transplant N=1 Received Transplant N=599 Received Transplant N=44 No Transplant N=3 Eligibility Criteria Age 18-65 HLA Match-Sibling PBSC AML in CR1 or CR2 Patients meeting eligibility N=84 Patients meeting eligibility N=44 AML CR1 N=65 AML CR2 N=19 AML CR1 N=37 AML CR2 N=7

Baseline Characteristics % of patients 0303 (N=44) p-Value Significantly More Females in 0303 Female 44% 63.6% 0.04 Male 56% 36.4% Patient Ages Similar in Both Protocols Median Age 45.1 48.5 0.14 % > 50 yrs of Age 32.1% 43.2% 0.22 Karnofsky Performance Status Similar in Both Protocols 100% 22 (26.2%) 17 (38.6%) 0.36 > 90% 46 (54.8%) > 80% 13 (15.5%) 8 (18.2%) > 70% 3 (3.6%) 2 (4.5%)

Cytogenetic Risk Profile By Protocol Control (0101) (N=84) N (%) CD34+ Enriched (0303) (N=44) p Value Favorable 9 (10.7%) 2 (4.5%) 0.34 Intermediate 55 (65.5%) 28 (63.6%) Unfavorable 12 (14.3%) 11 (25.0%) Unknown 8 (9.5%) 3 (6.8%) Slightly more unfavorable cytogenetics in 0303 than in 0101; overall, statistically similar * Slovak M et al. Blood 2001. SWOG/ECOG classification used for profile determination.

Conditioning Regimen BMT CTN 0101 versus BMT CTN 0303 # (%) 0303 (N=44)  Conditioning Regimen TBI based 43 (51) 44 (100) Busulfan based 41 (49) ATG 7 (8) GVHD Prophylaxis T cell depletion Tacrolimus based 48 (57) Cyclosporine A based 37 (43)

Day +30 Neutrophil Engraftment (> 500/µL)* 100% in CD34+ (0303) 96.4% in Control (0101) p=0.002 3 primary graft failures in the Control (0101) 0 primary graft failures in the CD34+ Enriched (0303) 1 secondary graft failure in each trial *Cumulative Incidence

Day +100 Platelet Engraftment (≥ 20,000/µL)* 97.7% in CD34+ (0303) 88.1% in Control (0101) p=0.228 *Cumulative Incidence

Transplant-Related Mortality* All Patients by Protocol CR1/CR2 and Protocol p=0.62 16.7% in Control (0101) 13.6% in CD34+ (0303) *Cumulative Incidence

Incidence of 12 Month Relapse* 20.3% in Control (0101) 20.6% in CD34+ (0303) *Cumulative Incidence

12 Month Relapse* CR1/CR2 By Protocol 57.1% CR2 p=N/A** 31.6% p=0.02 17% CR1 p=0.57 13.7% **Statistical calculations not performed due to low patient numbers 4/7 in 0303 and 6/19 in 0101 relapsed *Cumulative Incidence

12 Month Disease-Free Survival 65.7% in CD34+ (0303) 63% in Control (0101) p=0.59

12 Month DFS in CR1/CR2 By Protocol 72.8% 66.1% p=0.39 CR1 52.6% p=0.7 p=N/A* CR2 28.6% * Statistical comparisons were not performed on CR2 patients due to low patient numbers

DFS Stratification by Age > 50 CR1 and CR2 73.7% 0303 > 50 68.3% 0101 < 50 59.7% 0303 < 50 p=0.88 51.9% 0101 > 50

12 Month Overall Survival All Patients by Protocol CR1/CR2 and Protocol 77.3% in CD34+ (0303) 73.8% in Control (0101) p=0.61 p=0.87

Acute GVHD Grades II-IV* p=0.068 39.3% in Control (0101) 22.7% in CD34+ (0303) *Cumulative Incidence

Acute GVHD Grades II-IV in Patients < 50 Years of Age* 47.4% in Control < 50 (0101) p=0.028 24% in CD34+ < 50 (0303) *Cumulative Incidence

Acute GVHD Grades III-IV * p=0.31 9.5% in Control (0101) 4.5% in CD34+ (0303) *Cumulative Incidence

Limited/Extensive Chronic GVHD* p=0.0004 49.9% in Control (0101) 15.9% in CD34+ (0303) 1 year post-hoc power for the comparison between 0101 and 0303 was 98% *Cumulative Incidence

Patients with Infections Infectious Complications Control (0101) N=84 CD34+ Enriched (0303) N=44 # of Infectious Reports # of Patients 1-3 44 20 4-5 7 6-10 6 3 >10 2 Total # of Patients with Infections (% of Patients) 57 (67.9%) 32 (72.7%)

Types of Infections Seen By Protocol Control (0101) N=84 CD34+ Enriched (0303) N=44 # of Infections and (# of Patients) Bacterial 103 (48) 62 (25) Viral 46 (30) 41 (24)** Fungal 9 (9)* 8 (5) Protozoal 0 (0) 1 (1) Other 4 (3) 0 (0) Total Infection Events 162 112 * Represents confirmed infections only; 10 possible and 3 presumptive infections not included ** 8 of 24 viral infections in CD34+ enriched cohort (0303) were due to EBV reactivation  

Percentage of all deaths due to infection were Infection Events Control (0101) N=84 CD34+ Enriched (0303) N=44 Maximum Severity by Patient # of Patients (% of Patients) None 28 (33.3%) 12 (27.3%) Moderate 30 (35.7%) 15 (34.1%) Severe 23 (27.4%) 13 (29.5%) Life Threatening/Fatal 3 (3.6%) 4 (9.1%) Percentage of all deaths due to infection were 18.2% in 0101 and 20% in 0303

Causes of Death at One Year Post-Transplant Control (0101) N and % of Deaths CD34+ Enriched (0303) Recurrence 9 (40.9%) 3 (30%) Acute GVHD 1 (4.5%) Chronic GVHD Infection 4 (18.2%) 2 (20%) Interstitial Pneumonia Veno-occlusive Disease 1 (4.5%) ARDS Organ Failure 5 (22.7%) PTLD 1 (10%) Total 22 (23.1%) 10 (18.9%)

DAP Statistical Comparison Summary The incidence of chronic GVHD at one year post-transplant was significantly lower for recipients of CD34+ enriched allografts (p= 0.0004) Between patients receiving CD34+ enriched versus unmanipulated grafts, there was no significant difference in: Platelet and Neutrophil Engraftment, Acute GVHD, DFS, OS, TRM, and Relapse Low numbers of CR2 patients in both cohorts make statistical comparisons of relapse inconclusive

The DAP results support a comparable safety profile for CD34+ enriched transplants as compared to patients receiving unmanipulated grafts and conventional GVHD prophylaxis but with a significant reduction in cGVHD DAP Conclusion

Kai Pinkernell, M.D. Head of Clinical Development Miltenyi Biotec GmbH Overall Conclusions of Safety and Probable Benefit of the CliniMACS® CD34 Reagent System

Performance and Safety Conclusions CliniMACS® CD34 Reagent System The CliniMACS® CD34 Reagent System consistently produced a graft with > 2 x 106 CD34+ cells/kg and < 1 x 105 CD3+ cells/kg 84% of the grafts contained > 5 x 106 CD34+ cells/kg There was no significant difference between CD34+ enriched and unmanipulated allografts for: Platelet and Neutrophil Engraftment, Acute GVHD, DFS, OS, TRM, and Relapse While 0303 patients experienced more infectious episodes/patient (112/44 patients in 0303 versus 162/84 patients in 0101), these did not translate to higher infectious death rates (18.2% of deaths in 0101 vs. 20% in 0303) Increased incidence of EBV reactivation reports most likely due to stringent protocol specified monitoring of EBV in 0303

Probable Benefit Conclusions CliniMACS® CD34 Reagent System The CliniMACS® CD34 Reagent System was shown to consistently yield CD34+ enriched, T cell depleted, sterile cellular grafts in a multi-center setting Low rates of both acute and chronic GVHD without the risks associated with traditional pharmacologic GVHD prophylaxis while maintaining consistent engraftment, excellent DFS and OS, low TRM, and a low incidence of relapse Significantly reduced incidence of chronic GVHD without compromising survival

Safety and Probable Benefit Objectives Met The CliniMACS® CD34 Reagent System is safe, and has a probable benefit in significantly reducing the incidence of chronic GVHD while eliminating the need for pharmacologic GVHD prophylaxis for AML patients in complete remission, undergoing PBSC transplantation from a matched related donor

Acknowledgements BMT CTN NHLBI/NCI CIBMTR/EMMES/NMDP Investigators and Contributors Steven Devine Marcelo Pasquini Carolyn Keever-Taylor Richard O’Reilly Robert Soiffer John Wingard Adam Mendizabal, EMMES Shelly Carter, EMMES Nancy DiFronzo, NHLBI Mary Horowitz, CIBMTR Nancy Poland, NMDP Many, Many others The patients and their families who have made these trials possible

Thank You