microneedle patches for vaccine stabilization and … patches for vaccine stabilization and enhanced...

Microneedle Patches for Vaccine

Stabilization and Enhanced

Immunization

Anne Moore

University College Cork, [email protected]

mailto:[email protected]

Cost €€

Vaccine

Potency

Issues with Current Vaccine Distribution

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

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.

Courtesy of WHO

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)

Drug 1

Drug 2

CMC + Glyceri CMC + Glycerin

FITC loaded

trehalose

Characteristic

• Safe for skin administration

• Known & uniform distribution

• Standard processing conditions

• Formulation and equipment can

work together.

• Maintain stability of vaccine

under storage conditions

• Retain ability to penetrate skin

after storage

• Dissolution profile under storage

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

- Long term 40oC in hermetically sealed vials

- Short term 1 hour after removal from

packaging (40C/75% RH)

- Mechanical testing

Formulation Considerations

Coated microneedles

McGrath, M.G. et al (2011) Int. J. Pharm. 415(1-2)

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 )

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

Microneedles delivered vaccine; Skewed T

cell responses


i.d. Microneedle Patch Flat Patch0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

IFN+

IFN+TNF+TNF+

%C

D8

+

MN-mediated delivery of MVA-PbCSP induced a higher proportion of

TNF+CD8+ T cells compared to id delivery

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)

Dermis (1-3mm)

Epidermis (50-100mm)

Stratum Corneum (≈10µm)

Dissolvable Microneedle Fabrication

1

Figure 1

Figure 1

Atomised Spray Technique

McGrath, M.G., et al. (2014) Eur J Pharm Biopharm.;86(2):200-11

CMC

CMC +

Glycerol

CMC

CMC +

Glycerol

Silicon

Comparison of Dissolvable Microneedle Fabrication Methods

Vacuum/centrifuge

Conventional Dissolvable Microneedle Fabrication

Methods

mould

Apply excess formulated vaccine to mould

Remove excess, add to original formulation

Dry the formulation in mould

Remove microneedles from mould

mould

Formulated vaccine

Dispenser

Dispense into pores on (pre-treated) mould


Remove microneedles from mould with tape

Vrdoljak, J Control Release. 2016 Mar 10;225:192-204

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, cost-

effective manufacture

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

0.25 mm

1cm

Drug 1

Drug 2

Dissolvable ImmuPatch

Dissolvable microneedles composed of live virus expressing b-

galactosidase 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

TT (high dose) SC-

TT (low dose) SCTT-alum SC

TT (low dose) DMNTT-alum DMN

2.0

2.5

3.0

3.5

4.0

4.5

5.0

En

dp

oin

t titr

e (L

og

10)

day 34

day 56, 3wks post boost

*

* *

Prime

Boost

Olivia Flynn

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.

H1N1 HA

H3N2 HA

D a y

H1

HA

( m

g)

0 1 5 3 0 4 5 6 0 7 5 9 0

0

3

6

9

1 2

1 5

1 8

D M N (3 m g /d o s e )

L iq u id (1 5 m g /d o s e )

M o n th

H1

HA

( m

g)

0 3 6 9 1 2

0

1

2

3

4

* *

D a y

H3

HA

( m

g)

0 1 5 3 0 4 5 6 0 7 5 9 0

0

3

6

9

1 2

1 5

1 8


TIV: Trivalent inactivated Influenza

Vaccine:, 2011/12

A/California/7/2009 (H1N1)

A/ A/Perth/16/2009 (H3N2)

B/Brisbane/60/2008

Dissolvable ImmuPatch: Immunogenicity in mice: TIV

H1N1

Day post prime

An

ti H

1 H

A E

nd

po

int (L

og

10)

0 28 56 84 1122.0

2.5

3.0

3.5

4.0

4.5

5.0

2.5%DMN

2.5% IM

20% DMN

20% IM

*

** *

H3N2

Day post primeA

nti-H

3 H

A E

nd

po

int (L

og

10)

0 28 56 84 1122.0

2.5

3.0

3.5

4.0

4.5

5.0

2.5%DMN

2.5% IM

20% DMN

20% IM

*****

*

Dose sparing (3μg or 0.375μg)

TIV: Trivalent inactivated Influenza Vaccine:, 2011/12

A/California/7/2009 (H1N1)

A/ A/Perth/16/2009 (H3N2)

B/Brisbane/60/2008


Dissolvable ImmuPatch: Broader Immunity: TIV

Chimeric Pseudotype Particle Assay

Nigel Temperton, U. Kent

0 .3 m g

d 2 8

D M N

0 .3 m g

d 2 8

IM

0 .3 m g

d 5 6

D M N

0 .3 m g

d 5 6

IM

0

2 0 0

4 0 0

6 0 0

8 0 0

1 0 0 0

1 2 0 0

IC5

0

H 1 1

H 1 1 h e a d / H 1 s ta lk

H 1 S th C a ro lin a /1 9 1 8

H 5 V ie tn a m /1 1 9 4

*

*

From: Rino Rappuoli F1000 Med Reports 2011,


Recombinant Adenovirus Vectors as Vaccine Delivery

Platforms

Replication incompetent; in vivo Ag “factory”

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.

Malaria, HIV, TB

Influenza,

HBV,

HCV

Tumours

Ebola virus

Prophylactic & Therapeutic Attenuated Recombinant Vaccines for

Liquid vaccine stored at -80C

DMN: Ad stability: Accelerated Conditions

0 4 8 1 2

1 0 3

1 0 4

1 0 5

1 0 6

1 0 7

1 0 8

1 0 9

1 0 1 0

K in e t ic 0 3 0 4 1 5

W e e k

Ad

tit

re

(if

u/m

l)

D M N 4 0 C

L iq u id 4 0 C

D M N 2 5 C

Ad in DMN are stable up to at least 12 weeks.

Ad in liquid has degraded by week 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 |

ID

-

ID

ID

ID

M N

M N

ID

M N

M N

2 .0

2 .5

3 .0

3 .5

4 .0

4 .5

5 .0

5 .5

d 5 4

d 7 4

d 3 5

* * * *

* *

An

ti-M

SP

En

dp

oin

t (l

og

10

)

P r im e

B o o s t

A

Ab responses to malaria antigen

ID

-

ID

ID

ID

M N

M N

ID

M N

M N

1 .5

2 .0

2 .5

3 .0

3 .5

4 .0

4 .5

5 .0

5 .5

d 5 4

d 7 4

d 3 5* * *

* *

* *

An

ti-V

ec

tor E

nd

po

int

(lo

g 1

0)

P r im e

B o o s t

B

Ab responses to adenovirus vector

A d /M ID /ID 5 /6

0 1 0 2 0 3 0

0

2 0

4 0

6 0

8 0

1 0 01

2

3

4

5

6

% p

ara

sit

em

ia

†

A d /A d ID /ID : 3 /6

0 1 0 2 0 3 0

0

2 0

4 0

6 0

8 0

1 0 0

7

8

9

1 0

1 1

1 2

D a y

%p

ara

sit

em

ia

†

†

†

A d /A d M N / ID : 6 /6

0 1 0 2 0 3 0

0

2 0

4 0

6 0

8 0

1 0 0

1 9

2 0

2 1

2 2

2 3

2 4

D a y

%p

ara

sit

em

ia

A d /A d M N / M N : 3 /6

0 1 0 2 0 3 0

0

2 0

4 0

6 0

8 0

1 0 0

2 5

2 6

2 7

2 8

2 9

3 0

D a y

%p

ara

sit

em

ia

†

†

†

N a iv e : 0 /5

D a y

%p

ara

sit

em

ia

0 1 0 2 0 3 0

0

2 0

4 0

6 0

8 0

1 0 0

3 7

3 9

4 0

4 1

4 2

†

† † †

†

Si-MN overcome anti-vector immunity & enhance

vaccine efficacy

Carey et al., Scientific Reports, 2014

Efficacy

ID ImmuPatch

MVA-RFP, ex vivo porcine skin

Why is ImmuPatch different to ID?

Carey et al., Scientific Reports, 2014

Skin TNF

0 20 40 600.000

0.002

0.004

0.006

0.008

0.010ID

MN

Naive

**

*

**

Time

2d

elta

CT

Skin IL-1

0 20 40 600.000

0.001

0.002

0.003

0.004

0.005

*

ID

MN

Naive

Time

2d

elta

CT

Skin IL-10

0 20 40 600.000

0.002

0.004

0.006

0.008

0.010

*

* ID

MN

Naive

**

Time

2d

elta

CT

Skin type I IFN

0 20 40 600.000

0.001

0.002

0.003

0.004

0.005

Naive

MN

ID *

Time

2d

elta

CT

ImmuPatch: Non-inflammatory AdV Priming


ImmuPatch: Non-inflammatory AdV Priming

LN TNF

0 20 40 600.00

0.01

0.02

0.03

0.04 *ID

MN

Naive**

Time

2d

elta

CT

LN IL-1

0 20 40 600.0

0.1

0.2

0.3

0.4

0.5ID

MN

Naive

**

**

Time

2d

elt

aC

T

LN IL-10

0 20 40 600.000

0.001

0.002

0.003

0.004

$

ID

MN

**

Naive

Time

2d

elta

CT

LN Type I IFNb

0 20 40 600.000

0.002

0.004

0.006

0.008

0.010ID

MN**

Naive

**

Time

2d

elta

CT


Translation of MN manufacturing to large scale; HOW?

Vacuum/centrifuge

Conventional Dissolvable Microneedle Fabrication

Methods

mould

Apply excess formulated vaccine to mould

Remove excess, add to original formulation


Remove microneedles from mould

mould

Formulated vaccine

Dispenser

Dispense into pores on (pre-treated) mould


Remove microneedles from mould with tape



Formulation A:

Physical Parameters 1

Unstable droplet formation

Formulation B: Physical

Parameters 2

Stable droplet formation

Formulation A

Physical Parameters 1

Stable droplet formation

Manipulation of meniscus

formation

Allen et al., Int J Pharm. 2016 Mar 16;500(1-2):1-10


Sugar/polymer + Congo red Sugar +congo red

Control of formulation volume dispensing and drug/vaccine position



Effect of dispensing parameters on Hemagglutinin H1 A/California/7/2009 integrity

C o n tr o l 3 0 V /1 0 0 0 H z 5 0 V /1 0 0 0 H z 8 0 V /1 0 0 0 H z 3 0 V /5 0 H z 3 0 V /1 6 0 0 0 H z

0

2 0

4 0

6 0

HA

A/C

ali

forn

ia C

on

c.

(ug

/ml)

1 2 3 4 5


Dissolvable Microneedle Physical Stability

Dissolvable microneedles are composed of hydrophilic materials with large surface

area in contact with atmosphere following unpacking before application

Need to balance fast dissolution in vivo with no dissolution in

environment.

0

10

20

30

40

50

60

70

80

90

99.6

99.8

100.0

100.2

100.4

100.6

100.8

101.0

101.2

0 30 60 90 120 150 180 210 240

Ta

rge

t R

H (

%)

Ch

an

ge

In

Ma

ss

(%

)

Time/mins

DVS Trehalose DMN Method 1

dm

© Particular

Sciences Ltd.

DVS - The Sorption

Solution

60%

70%

80%

50%

0

10

20

30

40

50

60

70

80

90

99.6

99.8

100.0

100.2

100.4

100.6

100.8

101.0

101.2

-50 50 150 250

Ta

rge

t R

H (

%)

Ch

an

ge In

Ma

ss

(%

)

Time/mins

DVS Trehalose DMN Method 2

© Particular

Sciences Ltd.

DVS - The

Sorption Solution

60%

70%

80%

Method 1 Recrystallisation event detected 60% RH

Method 2 Recrystallisation event detected 80% RH

Dissolvable Microneedle Physical Stability

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.

• 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.

Formulation Considerations

• 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 needle-

and-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

Vaccine-loaded MN; New Frontiers

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

CollaboratorsSimon Draper, Jenner Institute, University of Oxford

Nigel TempertonU. Kent

National Immunisation OfficeBrenda Corcoran

Michael McAuliffeCork institute of Technology

Abina Crean, Conor O’Mahony

microneedle patches for vaccine stabilization and … patches for vaccine stabilization and enhanced...

Documents