Microneedle Patches for Vaccine Stabilization and Enhanced ...
Microneedle Patches for Vaccine Stabilization and Enhanced Immunization Anne Moore University College Cork, Ireland [email protected]
Issues with Current Vaccine Distribution Vaccine Potency
Cost €€
Current Immunization Programme Issues: Summary • HIGH COST: Vaccines and cold packs stored in cold chain • LOW VOLUME: Deployment of new vaccines constrained by lack of cold chain volume; e.g., rotavirus. • Heat/cold sensitivity of different vaccines: correct storage in the right place is critical • The last mile; potential for controlled temperature conditions.
• High rates of vaccine wastage – Discard vaccine vials open at the end of a session. – Open a 10 dose vial even if only 1 child comes to the session to avoid missed opportunities
Immunization Costs: Unsustainable
Gandhi et al., Vaccine 31S (2013) B137–B148 Lydon et al., Vaccine (2008) 26, 6706
ImmuPatch: Solutions to Vaccination Obstacles For Healthcare systems: – Stability; eliminate cold chain & permit stockpiling – Eliminate training costs (easy to administer) – Eliminate reconstitution – Eliminate needlesticks – Single dose – Reduced logistics cost Courtesy of WHO
For the manufacturer: – Integrated into manufacturing process; alternative fill finish – Reduced COGs (must be low cost fabrication method) – Potential for dose-sparing For the User: – Self-administered? Minimal pain.
Development History 1st Generation: Silicon MN (solid & hollow)
2nd Generation: Coated MN Film-coated (top) Spray-coated (bottom)
3rd Generation: Dissolvable MN Laminate Layered MN (method 1; top) Horizontal MN (method 2: bottom)
FITC loaded trehalose CMC + Glyceri CMC + Glycerin
Drug 1 Drug 2
Formulation Considerations Characteristic • Safe for skin administration • Known & uniform distribution • Standard processing conditions
-
Previously included in an injected product or implant in body.
– Uses off the shelf equipment – Can be used in a GMP environment – Can be scaled up to a commerical scale
• Formulation and equipment can work together. • Maintain stability of vaccine - Long term 40oC in hermetically sealed vials under storage conditions - Short term 1 hour after removal from • Retain ability to penetrate skin packaging (40C/75% RH) after storage • Dissolution profile under storage - Mechanical testing conditions
Coated microneedles Screen polymer and process parameters in combination on flat silicon disks Taguchi design of experiments (6 variables 2 levels, 8 runs per polymer, polymer 2 levels )
McGrath, M.G. et al (2011) Int. J. Pharm. 415(1-2)
Coated microneedles; Film coating
McGrath, M.G. et al (2011) Int. J. Pharm. 415(1-2)
Coated microneedles Dip coating; doesn’t work for these MN
Correct parameters: vaccine localised around MN, not spread on base Vrdoljak. et al (2011) J Control Release. 2012 159(1):34-42
Coated microneedles; Delivers live vaccines No vaccine
AdV5 expressing b-gal
MVA expressing b-gal
Vrdoljak. et al (2011) J Control Release. 2012 159(1):34-42
%CD8+
Microneedles delivered vaccine; Skewed T cell responses
0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0
IFN+TNF+ TNF+ IFN+
i.d.
Microneedle Patch
Flat Patch
MN-mediated delivery of MVA-PbCSP induced a higher proportion of TNF+CD8+ T cells compared to id delivery
Vrdoljak. et al (2011) J Control Release. 2012 159(1):34-42
Dissolvable Microneedles targeted to epidermis or upper dermis Dissolvable Microneedles (DMN): Apply patch to skin, microneedles penetrate past the stratum corneum, then dissolve Typically 300-600µm height for DMN (epidermis & dermis)
Stratum Corneum (≈10µm)
Epidermis (50-100mm)
Dermis (1-3mm)
Dissolvable Microneedle Fabrication Figure 1
Atomised Spray Technique
CMC
CMC
Silicon
CMC + Glycerol
McGrath, M.G., et al. (2014) Eur J Pharm Biopharm.;86(2):200-11
CMC + Glycerol
Comparison of Dissolvable Microneedle Fabrication Methods Conventional Dissolvable Microneedle Fabrication Methods Apply excess formulated vaccine to mould
Dispenser
mould Formulated vaccine
Dispense into pores on (pre-treated) mould Vacuum/centrifuge
Remove excess, add to original formulation
Dry the formulation in mould
Remove microneedles from mould
Vrdoljak, J Control Release. 2016 Mar 10;225:192-204
Dry the formulation in mould
Remove microneedles from mould with tape
mould
Dissolvable ImmuPatch Unique Production process: Simple
Water is the only waste material No vaccine is lost or re-used: improved GMP transfer. Scalable Vaccine formulation can be concentrated to needle tip or to different microneedles Continuous, automated fabrication system for fast, costeffective manufacture
0.25 mm
1cm
Excipients: Safe, Stabilise and Strengthen
Stabilises drug/vaccine in dry form outside of cold chain. Fast dissolution of microneedle for vaccines Sufficient mechanical strength to penetrate into skin Already approved for injectable products
Result: Stable, potent vaccine-loaded microneedles
Adhesive tape
Drug 1
Drug 2
Dissolvable ImmuPatch Dissolvable microneedles composed of live virus expressing bgalactosidase was administered to ex vivo porcine skin. All microneedles penetrated and delivered virus (blue areas) Fabrication process does not destroy sensitive material (live virus successfully infects the skin & expresses b-gal)
Intradermal injection of live virus expressing b-galactosidase using needle-and-syringe
WHO’s 26 vaccine preventable diseases Bacteria: Common Paediatric – Diphtheria – Group A, C meningococcal infections – Hib – Pertussis – Pneumococcal infections – Tetanus – Tuberculosis Others: – Anthrax – Cholera – Typhoid fever (S. typhi)
Virus Common Paediatric – Measles – Mumps – Rubella – Poliomyelitis – Varicella: chickenpox & shingles – Rotavirus Seasonal: – Influenza Adolescent/adult/Travellers’ – HPV: Cervical cancer – Yellow fever – Japanese encephalitis – Rabies – Hepatitis A, Hep B – Smallpox
Dissolvable ImmuPatch: Alum-based subunit vaccines Preliminary DMN (dissolvable microneedle) patch is equivalent to SC (subcutaneous) administration
* *
*
Endpoint titre (Log10)
5.0
Prime Boost
Olivia Flynn
4.5
day 34 day 56, 3wks post boost
4.0 3.5 3.0 2.5 2.0
TT (high dose) SC TT (low dose) SC TT (low dose) DMN TT-alum SC TT-alum DMN
Dissolvable ImmuPatch: Enhanced Vaccine Stability Influenza vaccine Antigen (2011/12) in ImmuPatch is stable at 45C and 75% RH for at least 6 months and is significantly more stable than current, liquid formulation. 4
15
L iq u id ( 1 5 m g /d o s e )
12
D M N ( 3 m g /d o s e )
9 6
H 1 H A (m g )
H1N1 HA
H 1 H A (m g )
18
**
3
2
1 3 0
0 0
15
30
45
60
75
90
0
D ay
3
6
9
M o n th
18
H3N2 HA
H 3 H A (m g )
15
TIV: Trivalent inactivated Influenza Vaccine:, 2011/12 A/California/7/2009 (H1N1) A/ A/Perth/16/2009 (H3N2) B/Brisbane/60/2008
12 9 6 3 0 0
15
30
45
D ay
Vrdoljak, J Control Release. 2016 Mar 10;225:192-204
60
75
90
12
Dissolvable ImmuPatch: Immunogenicity in mice: TIV
H1N1
5.0 4.5
20% DMN
4.0
2.5%DMN
3.5
20% IM
3.0
** *
2.5 2.0
*
0
28
56
84
2.5% IM
112
Day post prime
TIV: Trivalent inactivated Influenza Vaccine:, 2011/12 A/California/7/2009 (H1N1) A/ A/Perth/16/2009 (H3N2) B/Brisbane/60/2008 Vrdoljak, J Control Release. 2016 Mar 10;225:192-204
Anti-H3 HA Endpoint (Log 10)
Anti H1 HA Endpoint (Log 10)
Dose sparing (3μg or 0.375μg) H3N2 5.0
20% DMN 4.5
2.5%DMN
4.0
20% IM
3.5
**
*
2.5% IM
***
3.0 2.5 2.0
0
28
56
84
Day post prime
112
Dissolvable ImmuPatch: Broader Immunity: TIV Chimeric Pseudotype Particle Assay H 5 V ie tn a m /1 1 9 4 H 1 S th C a r o lin a /1 9 1 8 H 1 1 h e a d / H 1 s ta lk
1200
H 11
*
1000
IC 5 0
800 600 400
*
200 0
From: Rino Rappuoli F1000 Med Reports 2011,
Vrdoljak, J Control Release. 2016 Mar 10;225:192-204
0 .3 m g
0 .3 m g
0 .3 m g
0 .3 m g
d28 DMN
d28
d56
d56
IM
DMN
IM
Nigel Temperton, U. Kent
Recombinant Adenovirus Vectors as Vaccine Delivery Platforms Prophylactic & Therapeutic Attenuated Recombinant Vaccines for Malaria, HIV, TB Influenza, HBV, HCV Tumours
Replication incompetent; in vivo Ag “factory”
Ebola virus
Approx 100nm in size. Very homogenous particles Highly immunogenic; Ab & T cell Stability AdHu5; pre-existing immunity may reduce efficacy; overcome with alternative human or chimp serotypes prevents repeated use •
Insert size limitations.
Liquid vaccine stored at -80C
DMN: Ad stability: Accelerated Conditions K in e t ic 0 3 0 4 1 5 1 0 10
A d t i t r e ( i f u / m l)
10 9 10
DM N 40C
8
DM N 25C
10 7
L iq u id 4 0 C
10 6 10 5 10 4 10 3 0
4
8
W eek
Ad in DMN are stable up to at least 12 weeks. Ad in liquid has degraded by week 12
12
Silicon MN: Skewed Antibody Response to the Ad-encoded Antigen
Ratio of Ab titre to Antigen versus AdHu5 at day 50 post-prime
Carey et al., Sci Reports 2014 | 4 : 6154 |
Si-MN overcome anti-vector immunity & enhance vaccine efficacy Ab responses to malaria antigen
Efficacy
A
**
A d /A d ID /ID : 3 /6
*
†
7 8
5 .0
d35
4 .5
d54 4 .0
% p a r a s it e m ia
80
5 .5
†
100
†
9
60
10 11
40
12
20
% p a r a s it e m i a
A n ti-M S P E n d p o in t (lo g 1 0 )
***
A d /M ID /ID 5 /6
†
100
1 2
80
3 4
60
5 40
6
20
d74 0
3 .5
0 0
10
20
30
0
10
20
30
D ay
3 .0 2 .5 2 .0
N a iv e : 0 /5
ID
ID
M N
M N
P r im e
-
ID
M N
ID
M N
Boost
† 100
A d /A d M N / M N : 3 /6
†
100
25
†
26
% p a r a s it e m ia
80
Ab responses to adenovirus vector
†
60
28 29
40
30
20
B
†
† 37 39
60
40 40
41 42
20
0 0
**
10
20
0 0
10
**
20
30
D ay
D ay
***
d35
5 .5
A d /A d M N / ID : 6 /6
d54 5 .0
100
19
d74
4 .5
20
80 % p a r a s it e m ia
A n t i- V e c t o r E n d p o in t ( lo g 1 0 )
27
††
80 % p a r a s it e m ia
ID
4 .0 3 .5 3 .0
21 60
22 23
40
24
20
2 .5 0
2 .0
0
10
20
30
D ay
1 .5 ID
ID
ID
MN
MN
P r im e
-
ID
MN
ID
MN
Boost
Carey et al., Scientific Reports, 2014
30
Why is ImmuPatch different to ID? ID
MVA-RFP, ex vivo porcine skin
ImmuPatch
Carey et al., Scientific Reports, 2014
ImmuPatch: Non-inflammatory AdV Priming Skin TNF
Skin IL-1 0.005
0.003 0.002
*
0.001 0.000
0
20
40
**
0.006
*
0.004
0.000
60
40
Skin IL-10
Skin type I IFN
0.002
40
0.003 0.002 0.001
* 20
ID MN Naive
0.004
2deltaCT
**
60
*
0.005
ID MN Naive
0.006
0
20
Time
*
0.004
0
Time
0.008
2deltaCT
ID MN Naive
0.002
0.010
0.000
**
0.008
2deltaCT
0.004
2deltaCT
0.010
ID MN Naive
60
Time
Carey et al., Sci Reports 2014 | 4 : 6154 |
0.000
0
20
40
Time
60
ImmuPatch: Non-inflammatory AdV Priming LN TNF
LN IL-1
0.3 0.2
** 0.02 0.01
0.1 0.0
ID MN Naive
0.03
2deltaCT
2deltaCT
ID MN Naive
**
0.4
*
0.04
0.5
** 0
0.00
20
40
0
20
60
40
60
Time
Time
LN IL-10
LN Type I IFNb
**
0.004
0.002 0.001
$
0
20
40
ID MN Naive
** 0.008
2deltaCT
2deltaCT
0.003
0.000
0.010
ID MN Naive
0.006
**
0.004 0.002
60
Time
Carey et al., Sci Reports 2014 | 4 : 6154 |
0.000
0
20
40
Time
60
Translation of MN manufacturing to large scale; HOW? Conventional Dissolvable Microneedle Fabrication Methods Apply excess formulated vaccine to mould
Dispenser
mould Formulated vaccine
Dispense into pores on (pre-treated) mould Vacuum/centrifuge
Remove excess, add to original formulation
Dry the formulation in mould
Remove microneedles from mould
Vrdoljak, J Control Release. 2016 Mar 10;225:192-204
Dry the formulation in mould
Remove microneedles from mould with tape
mould
Dissolvable Microneedle Fabrication
Formulation A: Physical Parameters 1 Unstable droplet formation
Formulation B: Physical Parameters 2 Stable droplet formation
Allen et al., Int J Pharm. 2016 Mar 16;500(1-2):1-10
Formulation A Physical Parameters 1 Stable droplet formation Manipulation of meniscus formation
Dissolvable Microneedle Fabrication Control of formulation volume dispensing and drug/vaccine position
Sugar/polymer + Congo red
Allen et al., Int J Pharm. 2016 Mar 16;500(1-2):1-10
Sugar +congo red
Dissolvable Microneedle Fabrication
H A A /C a lif o r n ia C o n c . ( u g /m l)
Effect of dispensing parameters on Hemagglutinin H1 A/California/7/2009 integrity
60
40
20
0 C o n tro l
3 0 V /1 0 0 0 H z
1
5 0 V /1 0 0 0 H z
2
Allen et al., Int J Pharm. 2016 Mar 16;500(1-2):1-10
8 0 V /1 0 0 0 H z
3
3 0 V /5 0 H z
4
3 0 V /1 6 0 0 0 H z
5
Dissolvable Microneedle Physical Stability Need to balance fast dissolution in vivo with no dissolution in environment. Dissolvable microneedles are composed of hydrophilic materials with large surface area in contact with atmosphere following unpacking before application
Dissolvable Microneedle Physical Stability DVS Trehalose DMN Method 2
DVS Trehalose DMN Method 1 dm
90
101.2
80
101.0
70
100.8
90
80%
101.0
80%
70%
100.6
60
50%
50
100.4
40
100.2
30
100.0
20
99.8
Change In Mass (%)
60%
70%
Target RH (%)
Change In Mass (%)
100.8
99.6
0 0
30
DVS - The Sorption Solution
60
90 120 150 180 210 240 Time/mins © Particular Sciences Ltd.
70
60%
60
100.6
50 100.4
40 100.2
30
100.0
20
99.8
10
99.6 -50 DVS - The Sorption Solution
Method 1 Recrystallisation event detected 60% RH Method 2 Recrystallisation event detected 80% RH
80
Target RH (%)
101.2
10 0 50 150 250 Time/mins © Particular Sciences Ltd.
Solid state form and location of drug: Drug Stability in Individual Microneedles
Drug- loaded DMN dried method 1
Drug-loaded DMN dried method 2
No difference was observed between the drug in the DMN (irrespective of the drying conditions) and the drug raw material.
Formulation Considerations •
Be aware of regulatory status of your formulation.
•
Need experimental design to select formulation in conjunction with processing parameters.
•
Formulation can be optimised but also processing parameters; important to consider both!
•
Process parameters should be considered in relation to what is sensible.
•
Need analytical methodology to deal with 3D micron devices.
•
Fabrication process is critical to final structure, distribution, form need to study actual products.
•
Need to consider interaction with backing layers, dissolution methodology and in vivo delivery.
•
Need to address challenge of fast dissolution and environmental physical stability.
Vaccine-loaded MN; New Frontiers • Manufacturing Translation: – Nano-fluidics and biologics that will be used clinically, not as IVD. – Assaying solid state biological in a 3D format.
• Clinical translation; will they be better than current needleand-syringe? • Utility of their use? – Complexity introduced by adjuvants. Alum, o/w etc? – Limit to live and inactivated whole vaccines?
• Push and Pull; – Adopted by manufacturer; practically all vaccines administered with a needle and syringe. – Desired for by healthcare providers and vaccinees.
• Regulatory definitions
Abina Crean, Core Researchers Anto Vrdoljak Marie McGrath John Carey Timmy Doody Caroline O’Sullivan
Kate Dillane Sonja Vucen Evin Allen Olivia Flynn Joanne McCaffrey Cristina Walter Agnese Donadei
Conor O’Mahony
Collaborators Simon Draper, Jenner Institute, University of Oxford Nigel Temperton U. Kent
National Immunisation Office Brenda Corcoran Michael McAuliffe Cork institute of Technology
Issues with Current Vaccine Distribution Vaccine Potency
Cost €€
Current Immunization Programme Issues: Summary • HIGH COST: Vaccines and cold packs stored in cold chain • LOW VOLUME: Deployment of new vaccines constrained by lack of cold chain volume; e.g., rotavirus. • Heat/cold sensitivity of different vaccines: correct storage in the right place is critical • The last mile; potential for controlled temperature conditions.
• High rates of vaccine wastage – Discard vaccine vials open at the end of a session. – Open a 10 dose vial even if only 1 child comes to the session to avoid missed opportunities
Immunization Costs: Unsustainable
Gandhi et al., Vaccine 31S (2013) B137–B148 Lydon et al., Vaccine (2008) 26, 6706
ImmuPatch: Solutions to Vaccination Obstacles For Healthcare systems: – Stability; eliminate cold chain & permit stockpiling – Eliminate training costs (easy to administer) – Eliminate reconstitution – Eliminate needlesticks – Single dose – Reduced logistics cost Courtesy of WHO
For the manufacturer: – Integrated into manufacturing process; alternative fill finish – Reduced COGs (must be low cost fabrication method) – Potential for dose-sparing For the User: – Self-administered? Minimal pain.
Development History 1st Generation: Silicon MN (solid & hollow)
2nd Generation: Coated MN Film-coated (top) Spray-coated (bottom)
3rd Generation: Dissolvable MN Laminate Layered MN (method 1; top) Horizontal MN (method 2: bottom)
FITC loaded trehalose CMC + Glyceri CMC + Glycerin
Drug 1 Drug 2
Formulation Considerations Characteristic • Safe for skin administration • Known & uniform distribution • Standard processing conditions
-
Previously included in an injected product or implant in body.
– Uses off the shelf equipment – Can be used in a GMP environment – Can be scaled up to a commerical scale
• Formulation and equipment can work together. • Maintain stability of vaccine - Long term 40oC in hermetically sealed vials under storage conditions - Short term 1 hour after removal from • Retain ability to penetrate skin packaging (40C/75% RH) after storage • Dissolution profile under storage - Mechanical testing conditions
Coated microneedles Screen polymer and process parameters in combination on flat silicon disks Taguchi design of experiments (6 variables 2 levels, 8 runs per polymer, polymer 2 levels )
McGrath, M.G. et al (2011) Int. J. Pharm. 415(1-2)
Coated microneedles; Film coating
McGrath, M.G. et al (2011) Int. J. Pharm. 415(1-2)
Coated microneedles Dip coating; doesn’t work for these MN
Correct parameters: vaccine localised around MN, not spread on base Vrdoljak. et al (2011) J Control Release. 2012 159(1):34-42
Coated microneedles; Delivers live vaccines No vaccine
AdV5 expressing b-gal
MVA expressing b-gal
Vrdoljak. et al (2011) J Control Release. 2012 159(1):34-42
%CD8+
Microneedles delivered vaccine; Skewed T cell responses
0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0
IFN+TNF+ TNF+ IFN+
i.d.
Microneedle Patch
Flat Patch
MN-mediated delivery of MVA-PbCSP induced a higher proportion of TNF+CD8+ T cells compared to id delivery
Vrdoljak. et al (2011) J Control Release. 2012 159(1):34-42
Dissolvable Microneedles targeted to epidermis or upper dermis Dissolvable Microneedles (DMN): Apply patch to skin, microneedles penetrate past the stratum corneum, then dissolve Typically 300-600µm height for DMN (epidermis & dermis)
Stratum Corneum (≈10µm)
Epidermis (50-100mm)
Dermis (1-3mm)
Dissolvable Microneedle Fabrication Figure 1
Atomised Spray Technique
CMC
CMC
Silicon
CMC + Glycerol
McGrath, M.G., et al. (2014) Eur J Pharm Biopharm.;86(2):200-11
CMC + Glycerol
Comparison of Dissolvable Microneedle Fabrication Methods Conventional Dissolvable Microneedle Fabrication Methods Apply excess formulated vaccine to mould
Dispenser
mould Formulated vaccine
Dispense into pores on (pre-treated) mould Vacuum/centrifuge
Remove excess, add to original formulation
Dry the formulation in mould
Remove microneedles from mould
Vrdoljak, J Control Release. 2016 Mar 10;225:192-204
Dry the formulation in mould
Remove microneedles from mould with tape
mould
Dissolvable ImmuPatch Unique Production process: Simple
Water is the only waste material No vaccine is lost or re-used: improved GMP transfer. Scalable Vaccine formulation can be concentrated to needle tip or to different microneedles Continuous, automated fabrication system for fast, costeffective manufacture
0.25 mm
1cm
Excipients: Safe, Stabilise and Strengthen
Stabilises drug/vaccine in dry form outside of cold chain. Fast dissolution of microneedle for vaccines Sufficient mechanical strength to penetrate into skin Already approved for injectable products
Result: Stable, potent vaccine-loaded microneedles
Adhesive tape
Drug 1
Drug 2
Dissolvable ImmuPatch Dissolvable microneedles composed of live virus expressing bgalactosidase was administered to ex vivo porcine skin. All microneedles penetrated and delivered virus (blue areas) Fabrication process does not destroy sensitive material (live virus successfully infects the skin & expresses b-gal)
Intradermal injection of live virus expressing b-galactosidase using needle-and-syringe
WHO’s 26 vaccine preventable diseases Bacteria: Common Paediatric – Diphtheria – Group A, C meningococcal infections – Hib – Pertussis – Pneumococcal infections – Tetanus – Tuberculosis Others: – Anthrax – Cholera – Typhoid fever (S. typhi)
Virus Common Paediatric – Measles – Mumps – Rubella – Poliomyelitis – Varicella: chickenpox & shingles – Rotavirus Seasonal: – Influenza Adolescent/adult/Travellers’ – HPV: Cervical cancer – Yellow fever – Japanese encephalitis – Rabies – Hepatitis A, Hep B – Smallpox
Dissolvable ImmuPatch: Alum-based subunit vaccines Preliminary DMN (dissolvable microneedle) patch is equivalent to SC (subcutaneous) administration
* *
*
Endpoint titre (Log10)
5.0
Prime Boost
Olivia Flynn
4.5
day 34 day 56, 3wks post boost
4.0 3.5 3.0 2.5 2.0
TT (high dose) SC TT (low dose) SC TT (low dose) DMN TT-alum SC TT-alum DMN
Dissolvable ImmuPatch: Enhanced Vaccine Stability Influenza vaccine Antigen (2011/12) in ImmuPatch is stable at 45C and 75% RH for at least 6 months and is significantly more stable than current, liquid formulation. 4
15
L iq u id ( 1 5 m g /d o s e )
12
D M N ( 3 m g /d o s e )
9 6
H 1 H A (m g )
H1N1 HA
H 1 H A (m g )
18
**
3
2
1 3 0
0 0
15
30
45
60
75
90
0
D ay
3
6
9
M o n th
18
H3N2 HA
H 3 H A (m g )
15
TIV: Trivalent inactivated Influenza Vaccine:, 2011/12 A/California/7/2009 (H1N1) A/ A/Perth/16/2009 (H3N2) B/Brisbane/60/2008
12 9 6 3 0 0
15
30
45
D ay
Vrdoljak, J Control Release. 2016 Mar 10;225:192-204
60
75
90
12
Dissolvable ImmuPatch: Immunogenicity in mice: TIV
H1N1
5.0 4.5
20% DMN
4.0
2.5%DMN
3.5
20% IM
3.0
** *
2.5 2.0
*
0
28
56
84
2.5% IM
112
Day post prime
TIV: Trivalent inactivated Influenza Vaccine:, 2011/12 A/California/7/2009 (H1N1) A/ A/Perth/16/2009 (H3N2) B/Brisbane/60/2008 Vrdoljak, J Control Release. 2016 Mar 10;225:192-204
Anti-H3 HA Endpoint (Log 10)
Anti H1 HA Endpoint (Log 10)
Dose sparing (3μg or 0.375μg) H3N2 5.0
20% DMN 4.5
2.5%DMN
4.0
20% IM
3.5
**
*
2.5% IM
***
3.0 2.5 2.0
0
28
56
84
Day post prime
112
Dissolvable ImmuPatch: Broader Immunity: TIV Chimeric Pseudotype Particle Assay H 5 V ie tn a m /1 1 9 4 H 1 S th C a r o lin a /1 9 1 8 H 1 1 h e a d / H 1 s ta lk
1200
H 11
*
1000
IC 5 0
800 600 400
*
200 0
From: Rino Rappuoli F1000 Med Reports 2011,
Vrdoljak, J Control Release. 2016 Mar 10;225:192-204
0 .3 m g
0 .3 m g
0 .3 m g
0 .3 m g
d28 DMN
d28
d56
d56
IM
DMN
IM
Nigel Temperton, U. Kent
Recombinant Adenovirus Vectors as Vaccine Delivery Platforms Prophylactic & Therapeutic Attenuated Recombinant Vaccines for Malaria, HIV, TB Influenza, HBV, HCV Tumours
Replication incompetent; in vivo Ag “factory”
Ebola virus
Approx 100nm in size. Very homogenous particles Highly immunogenic; Ab & T cell Stability AdHu5; pre-existing immunity may reduce efficacy; overcome with alternative human or chimp serotypes prevents repeated use •
Insert size limitations.
Liquid vaccine stored at -80C
DMN: Ad stability: Accelerated Conditions K in e t ic 0 3 0 4 1 5 1 0 10
A d t i t r e ( i f u / m l)
10 9 10
DM N 40C
8
DM N 25C
10 7
L iq u id 4 0 C
10 6 10 5 10 4 10 3 0
4
8
W eek
Ad in DMN are stable up to at least 12 weeks. Ad in liquid has degraded by week 12
12
Silicon MN: Skewed Antibody Response to the Ad-encoded Antigen
Ratio of Ab titre to Antigen versus AdHu5 at day 50 post-prime
Carey et al., Sci Reports 2014 | 4 : 6154 |
Si-MN overcome anti-vector immunity & enhance vaccine efficacy Ab responses to malaria antigen
Efficacy
A
**
A d /A d ID /ID : 3 /6
*
†
7 8
5 .0
d35
4 .5
d54 4 .0
% p a r a s it e m ia
80
5 .5
†
100
†
9
60
10 11
40
12
20
% p a r a s it e m i a
A n ti-M S P E n d p o in t (lo g 1 0 )
***
A d /M ID /ID 5 /6
†
100
1 2
80
3 4
60
5 40
6
20
d74 0
3 .5
0 0
10
20
30
0
10
20
30
D ay
3 .0 2 .5 2 .0
N a iv e : 0 /5
ID
ID
M N
M N
P r im e
-
ID
M N
ID
M N
Boost
† 100
A d /A d M N / M N : 3 /6
†
100
25
†
26
% p a r a s it e m ia
80
Ab responses to adenovirus vector
†
60
28 29
40
30
20
B
†
† 37 39
60
40 40
41 42
20
0 0
**
10
20
0 0
10
**
20
30
D ay
D ay
***
d35
5 .5
A d /A d M N / ID : 6 /6
d54 5 .0
100
19
d74
4 .5
20
80 % p a r a s it e m ia
A n t i- V e c t o r E n d p o in t ( lo g 1 0 )
27
††
80 % p a r a s it e m ia
ID
4 .0 3 .5 3 .0
21 60
22 23
40
24
20
2 .5 0
2 .0
0
10
20
30
D ay
1 .5 ID
ID
ID
MN
MN
P r im e
-
ID
MN
ID
MN
Boost
Carey et al., Scientific Reports, 2014
30
Why is ImmuPatch different to ID? ID
MVA-RFP, ex vivo porcine skin
ImmuPatch
Carey et al., Scientific Reports, 2014
ImmuPatch: Non-inflammatory AdV Priming Skin TNF
Skin IL-1 0.005
0.003 0.002
*
0.001 0.000
0
20
40
**
0.006
*
0.004
0.000
60
40
Skin IL-10
Skin type I IFN
0.002
40
0.003 0.002 0.001
* 20
ID MN Naive
0.004
2deltaCT
**
60
*
0.005
ID MN Naive
0.006
0
20
Time
*
0.004
0
Time
0.008
2deltaCT
ID MN Naive
0.002
0.010
0.000
**
0.008
2deltaCT
0.004
2deltaCT
0.010
ID MN Naive
60
Time
Carey et al., Sci Reports 2014 | 4 : 6154 |
0.000
0
20
40
Time
60
ImmuPatch: Non-inflammatory AdV Priming LN TNF
LN IL-1
0.3 0.2
** 0.02 0.01
0.1 0.0
ID MN Naive
0.03
2deltaCT
2deltaCT
ID MN Naive
**
0.4
*
0.04
0.5
** 0
0.00
20
40
0
20
60
40
60
Time
Time
LN IL-10
LN Type I IFNb
**
0.004
0.002 0.001
$
0
20
40
ID MN Naive
** 0.008
2deltaCT
2deltaCT
0.003
0.000
0.010
ID MN Naive
0.006
**
0.004 0.002
60
Time
Carey et al., Sci Reports 2014 | 4 : 6154 |
0.000
0
20
40
Time
60
Translation of MN manufacturing to large scale; HOW? Conventional Dissolvable Microneedle Fabrication Methods Apply excess formulated vaccine to mould
Dispenser
mould Formulated vaccine
Dispense into pores on (pre-treated) mould Vacuum/centrifuge
Remove excess, add to original formulation
Dry the formulation in mould
Remove microneedles from mould
Vrdoljak, J Control Release. 2016 Mar 10;225:192-204
Dry the formulation in mould
Remove microneedles from mould with tape
mould
Dissolvable Microneedle Fabrication
Formulation A: Physical Parameters 1 Unstable droplet formation
Formulation B: Physical Parameters 2 Stable droplet formation
Allen et al., Int J Pharm. 2016 Mar 16;500(1-2):1-10
Formulation A Physical Parameters 1 Stable droplet formation Manipulation of meniscus formation
Dissolvable Microneedle Fabrication Control of formulation volume dispensing and drug/vaccine position
Sugar/polymer + Congo red
Allen et al., Int J Pharm. 2016 Mar 16;500(1-2):1-10
Sugar +congo red
Dissolvable Microneedle Fabrication
H A A /C a lif o r n ia C o n c . ( u g /m l)
Effect of dispensing parameters on Hemagglutinin H1 A/California/7/2009 integrity
60
40
20
0 C o n tro l
3 0 V /1 0 0 0 H z
1
5 0 V /1 0 0 0 H z
2
Allen et al., Int J Pharm. 2016 Mar 16;500(1-2):1-10
8 0 V /1 0 0 0 H z
3
3 0 V /5 0 H z
4
3 0 V /1 6 0 0 0 H z
5
Dissolvable Microneedle Physical Stability Need to balance fast dissolution in vivo with no dissolution in environment. Dissolvable microneedles are composed of hydrophilic materials with large surface area in contact with atmosphere following unpacking before application
Dissolvable Microneedle Physical Stability DVS Trehalose DMN Method 2
DVS Trehalose DMN Method 1 dm
90
101.2
80
101.0
70
100.8
90
80%
101.0
80%
70%
100.6
60
50%
50
100.4
40
100.2
30
100.0
20
99.8
Change In Mass (%)
60%
70%
Target RH (%)
Change In Mass (%)
100.8
99.6
0 0
30
DVS - The Sorption Solution
60
90 120 150 180 210 240 Time/mins © Particular Sciences Ltd.
70
60%
60
100.6
50 100.4
40 100.2
30
100.0
20
99.8
10
99.6 -50 DVS - The Sorption Solution
Method 1 Recrystallisation event detected 60% RH Method 2 Recrystallisation event detected 80% RH
80
Target RH (%)
101.2
10 0 50 150 250 Time/mins © Particular Sciences Ltd.
Solid state form and location of drug: Drug Stability in Individual Microneedles
Drug- loaded DMN dried method 1
Drug-loaded DMN dried method 2
No difference was observed between the drug in the DMN (irrespective of the drying conditions) and the drug raw material.
Formulation Considerations •
Be aware of regulatory status of your formulation.
•
Need experimental design to select formulation in conjunction with processing parameters.
•
Formulation can be optimised but also processing parameters; important to consider both!
•
Process parameters should be considered in relation to what is sensible.
•
Need analytical methodology to deal with 3D micron devices.
•
Fabrication process is critical to final structure, distribution, form need to study actual products.
•
Need to consider interaction with backing layers, dissolution methodology and in vivo delivery.
•
Need to address challenge of fast dissolution and environmental physical stability.
Vaccine-loaded MN; New Frontiers • Manufacturing Translation: – Nano-fluidics and biologics that will be used clinically, not as IVD. – Assaying solid state biological in a 3D format.
• Clinical translation; will they be better than current needleand-syringe? • Utility of their use? – Complexity introduced by adjuvants. Alum, o/w etc? – Limit to live and inactivated whole vaccines?
• Push and Pull; – Adopted by manufacturer; practically all vaccines administered with a needle and syringe. – Desired for by healthcare providers and vaccinees.
• Regulatory definitions
Abina Crean, Core Researchers Anto Vrdoljak Marie McGrath John Carey Timmy Doody Caroline O’Sullivan
Kate Dillane Sonja Vucen Evin Allen Olivia Flynn Joanne McCaffrey Cristina Walter Agnese Donadei
Conor O’Mahony
Collaborators Simon Draper, Jenner Institute, University of Oxford Nigel Temperton U. Kent
National Immunisation Office Brenda Corcoran Michael McAuliffe Cork institute of Technology
