decide™ - based dc manufacturing - pact...

PACT Meeting, Houston Spencer

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DeCIDe™ - based DC manufacturing

October 22, 2012

David M Spencer, CSO

How to destroy a target without collateral damage?

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Solution: Smart Bomb

• Active guidance + self‐destruct = No risk of collateral damage

• Payload as powerful as it needs to be

• Enabling technology: remote guidance and control

B lli i l i thi• Bellicum is applying this approach to the field of medicine, for the first time

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How Does a Therapeutic Vaccine Work?

CONFIDENTIAL 4


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Problem: CD40, the Master Thermostat

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Solution: Replace with iCD40 CID Switch

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How Does a DeCIDe™ Vaccine Work?

• Initial product: BPX‐101

• Targets PSMA antigen

– Expressed by prostate p y pcancer and tumor blood vessels (i.e., endothelium)

• Lead indication: metastatic castrate resistant prostate cancer (mCRPC)

– Potential application in other solid tumorsother solid tumors

• Extensive published preclinical data, including tumor eradication

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BPX‐101 Manufacturing Process Flow, Day 0

Starting Material Manufacturing Steps Comments

Step 0: Start with good Leukapheresis!

Cobe Spectra™ 6-12 L, NMT 1.0 mL/min

Li C l 1 3% H t

PBMCsMonocytes > 1E9

PMN < 3%

Testing

Leukapheresis!Line Color 1- 3% Hct

Step 1: Receiving & Testing

Sterility/Cell Counts/Viability Step 2: Dilution ApheresisDilute 1:1 w HBSS/1%HSA

Tf to cGMP Facility, Sample

Step 3: Isolation MonoctyesGambro Elutra™Cell SeparationCounts/Phenotypeay 0

PMN 3%RBC < 7.5 mL

Step 3: Isolation MonoctyesGambro Elutra Cell SeparationCounts/PhenotypeD

Step 4: Generation imDCs VueLife gas-permeable bags

w CellGenix DC media (+ hGM-CSF & IL-4)1 cm, 6.8E6 cells/mL


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BPX‐101 Manufacturing Process Flow, Days 5‐6

Testing Manufacturing Steps Comments

Harvest DCs w Cobe 2991 (closed), adjust 4E6/mL in bag,

+ PSMA 2h5

Step 5: Harvest imDC, Td, PSMA, LPS, AP1903

+ PSMA 2h, + Ad5f35-iCD40 4h,

+ Dilute 4X w CellGenix DC,+ PSMA, AP1903, LPS

Incubate 16-20 h @ 37˚C

Step 6: Harvest mDC WashHarvest Cobe 2991, Extensive

Counts/Viability/Myco

Day

5

IL-12p70 (bkgd)

IL-12p70, Cell Step 6: Harvest mDC, WashHarvest Cobe 2991, Extensive

wash w PlasmaLyte 3% HSA

Step 7: Formulation, Vialing, Cryo

DP: BPX-101

Adjust dose to 4, 12.5, 25E6 (per 1-1.6 mL) in 7.5% DMSO, 12.5% HSA,

PlasmaLyte 2 mL bags CR Freeze LN2

count/Viability/Pheno(CD83+), Endo, Myco

DP Release: Sterility

Day

6

Planned BPX‐201 Manufacturing Equipment

Plasma Press

Elutra COBE 2991 Bag CentrifugeControlled-

Rate Freezer


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BP‐PC‐001 Phase I/II Clinical Trial Synopsis

• Phase I/IIa 3+3, BPX‐101 dose escalation in 18 patients with documented mCRPC

– 12 patients in Cohorts 1‐3 (EOW x 6 dose escalation)

– 6 patients enrolled in Cohort 4 (Q4W x 3, high dose)

• Trial conducted at Memorial Hermann/UT Health Science Center, P.I. Dr Guru Sonpavde

Leukapheresis

Week 12 assessmentBaseline scans Week 0

Maintenance (Q 8 weeks until progression)Induction (EOW x 6)

BPX‐101

AP1903

• BPX‐101 made at MD Anderson (6‐day process following leukapheresis)

• Average Halabi‐predicted survival 13.8 months

– Dogma is that patients with < 18 months to live don’t benefit from vaccine therapy

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Patient Demographics

Subject # 1001 1002 1003 1004 1005 1006

Age 73 72 81 80 66 73

KPS (at screening) 90% 90% 80% 80% 100% 90%

Gleason Score 7 7 9 10 8 8

Prior Chemotherapy None None Taxotere Taxotere None None

Clinical Subtype 4 5 3 4 4 5

Baseline PSA (ng/mL) 5.8 11.1 312.8 46.5 69.0 30.9

Pre‐Treatment PSADT (Months) 4.9 7.3 5.0 1.7 1.4 1.6

Subject # 1007 1008 1009 1010 1011 1012

Age 67 80 69 85 79 70

KPS (at screening) 100% 90% 90% 80% 90% 100%

l b

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Gleason Score 8 10 8 9 8 LN biopsy

Prior Chemotherapy (Abiraterone) Taxotere None Taxotere Taxotere Taxotere

Clinical Subtype 4 5 4 4 4 3

Baseline PSA (ng/mL) 2.4 55.8 26.0 1070.0 818.9 3.2

Pre‐Treatment PSADT (Months) N/A 7.7 9.1 1.8 0.25 6.0


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AEs: Injection Site Reactions

CONFIDENTIAL

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Taxotere

500

600

Subject 1003 – Alive with KPS 90 at 21 months

100

200

300

400

PSA

, ng/m

L

0

‐40 ‐32 ‐24 ‐16 ‐8 0 8 16 24 32 40 48 56 64

Weeks Relative to First Dose

CONFIDENTIAL 14


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Subject 1003: Tumor Shrinkage & Antivascular Effect

Baseline (Week –7) Week 12

1 of 8

Week 52

Exam

ple

2 of 8

36 x 29 mm 31 x 24 mm 25 x 21 mm

Exam

ple

25 x 23 mm 19 x 16 mm

CONFIDENTIAL 15

13 x 10 mm

Subject 1003: Soft Tissue Partial Response

25

30

m

• 8 measurable lymph node lesions at baseline

• Steady decrease in all 8 LNs

Normal LN size range

SD

LD

5

10

15

20

Average Size, m

m over >1 year

• Partial Response (PR) per RECIST criteria at 1 year time point

• Greatest rate of decrease during induction treatment

0

‐8 0 8 16 24 32 40 48 56


gphase– Tumor growth between baseline

and first dose (7 weeks) likely

CONFIDENTIAL 16


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Cytokine Spikes – Subject 1003

100

13,086

1,680

3,760118

268

20

40

60

80

old In

crease @

1 week

I

G

M

M

MI

R

T

I

0

20

1 2 3 4 5 6 7 8 9 10 11

Fo

Dose #

CONFIDENTIAL 17

2,500

3,000

Subject 1003: Restoration of Taxotere Sensitivity?

Taxotere Taxotere Taxotere Taxotere

500

1,000

1,500

2,000

PSA

, ng/m

L

0

‐84 ‐72 ‐60 ‐48 ‐36 ‐24 ‐12 0 12 24 36 48 60 72 84 96


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Subject 1003 – Gone Fishin’!

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Shown with permission

Subject 1006: Durable Complete Response

60

70

0.3“Undetectable” PSA

Taxotere‐based chemo

10

20

30

40

50

PSA

, ng/m

L

0.0

0.1

0.2

20 24 28 32 36 40

Standard

Ultrasensitive

Liver function normalized Off hormones

Scan Scan Scan

0

‐16 ‐8 0 8 16 24 32 40 48 56 64



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Subject 1006: First Complete Response

• Large biopsy‐proven prostate cancer metastasis in

Baseline (Week ‐8) Week 12 Week 34

Benign renal cyst confirms patient identity!

metastasis in the liver at baseline

• Liver function returned to normal by 15 weeks

• No detectable viable tumor (i l di

12.3 x 11.5 cm8.1 x 7.1 cm

(including lung, LN and bone lesions at baseline, not shown) at 34 weeks

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Subject 1007: Severe Tissue Inflammatory Response

1. Tumor cells express PSMA 3. Tumor destruction

CONFIDENTIAL 22

Source: Subject 1007 prostate biopsy, after 5 doses of vaccine

2. Inflammatory infiltration


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BPX‐101, evidence of efficacy, but a bit cumbersome

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“It’s a cell phone, man!”

iMyD88 Replaces LPS Activation of TLR Signaling

Lipid Rafts

TLR4

TLR4 TLR4

ER

TLR3

TLR4TLR7

TLR8

TLR4Myr, MFyn

PIP2 Binding Motif

Amino acidlinker

RT2

Tamoxifen

NOD2MyD88

Downstream

Adaptors

CONFIDENTIAL 24

IRF3,7 NF‐BSEAP Reporter Assay Screen

RIG‐I

NOD2

TRIFMyD88


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iMyD88/CD40 (iMC) Vector replaces iCD40+LPS

35,000

40,000

45,000

nits)

0

5,000

10,000

15,000

20,000

25,000

30,000

,

SEAP Activity (Un

Control (M.Fv'Fvls)

iCD40

CD4/TLR4

iMyD88

iMyD88/CD400

0.001 0.01 0.1 1 10 100

Dimerizer Concentration (nM)

25

+ +

- iC iC iML iMCiMC

CID

Adv

S

+++

-

-

-

-

-

- +-1 5

2.0

2.5

3.0PBSCD40L+LPSAd-LuciMCol

ume

(cm

3 )

iMyD88/CD40 (iMC) Vector replaces iCD40+LPS

p-IKK

p38

p-ERK

p-p38IKK/

JNKp-JNK

+ ---+ +- - - ---

LPS

CD40L

- - - -

-

0 10 20 30 40 50 600.0

0.5

1.0

1.5

Tum

or v

o

60

80

100

surv

ival

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-Actin

p-ERKERK

Aktp-Akt

0 10 20 30 40 50 60 700

20

40

60

Per

cent

s


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iMC‐PSMA Induces Higher Levels of IL‐12 than iCD40

30 000

35,000

‐ AP1903 + AP1903 • Data compares:

– iCD40 @ 10,000 VP/cell vs.

– iMC‐P @ 1 000 VP/cell

10,000

15,000

20,000

25,000

30,000

IL‐12, p

g/m

L

iMC P @ 1,000 VP/cell

• Higher background with iMC‐P

– May be sufficient to stimulate migration without AP1903 pre‐activation during cell processing

0

5,000

iCD40 + LPS iMC‐P

CONFIDENTIAL

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BPX‐201: Optimization of vp/cell and [AP1903]

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Optimization of vp/cell and [AP1903]

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iMC‐DCs produce copious AP1903‐dependent IL‐12p70

Adherent CD14+ Elutra

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Planned BPX‐201 Manufacturing Process

Testing Manufacturing Steps Comments

St 1 P t

1) 1:1 dilution fresh leukopak in HBSS/MgCl2/Pulmozyme@30’

2) El t i ti t > diy 0

Count/viability/Flow/Sterility

Step 1: Prepare monocytes2) Elutriation monocytes-> media3) Culture 2E6/mL (GM-CSF, IL4),

80% volume

Step 2: Re-feed cells1) Re-feed cells w added 20% CellGro

DC media w GM-CSF/IL4

Day

IL-12p70, Cell Day

3

Count/viability/Flow

Step 3: Formulation,Vialing, Cryo

DP: BPX-101

1) Harvest by COBE29912) Td w Ad5f35-iMC-RP-FL (2-3h)3) Harvest, Wash, Resuspend in

CryoStor CS54) Adjust to 1.6 mL dose (w

overage), cryopreserve CRF

count/Viability/Pheno(CD83+), Endo, Myco

DP Release: SterilityDay

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BPX‐101 vs. BPX‐201 Product Improvement Elements

Component BPX‐101 Product Element BPX‐201 Product Element

Adenoviral Vector Used Ad5f35‐iCD40 Ad5f35‐iMC‐RP‐FL

Recombinant Protein added Vector encoded under RSVPSMA Source

Recombinant Protein added to DC Culture

Vector‐encoded under RSV Promoter Control

DC Maturation/Activation

Stimulant SourceLPS added to DC Culture

Vector‐encoded using AP1903‐inducible MyD88/CD40 (iMC)

fusion protein

iCD40 transgene induced in manufacturing process iMC transgene induced in

Induction Methodology

manufacturing process through AP1903 addition ex vivo followed by additional

patient in vivo dosage

iMC transgene induced in patient only by in vivo AP1903

dosage

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BPX‐101 vs. BPX‐201 Product Improvement Elements

Component BPX‐101 Product Element BPX‐201 Product Element

Incubator Racks Standard incubator racksPerforated racks to elevate bags and maximize gas

exchange area

Mid‐Process Media Feed None (Day 5 infection) Day 3 (Day 6 infection)

Timing of infection4h infection, ~20h

incubation w LPS/AP19032‐3h infection

WashingExtensive Cobe 2991 and bag washes to remove LPS

Use 3x PlasmaLyte/1% HSAprior to CS5 formulation

Freezing Media7.5% DMSO/12.5% HSA/PlasmaLyte A

CryoStor CS5 (5% DMSO) removes variability (GMP)

Release Testing Sterility, IL‐12, CD11c/CD83Sterility, MycoTooL (PCR), IL‐

12, CD11c/CCR7

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Acknowledgements

• Dazzling gratitude to: (TBA)

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