Skin immunization against rotavirus using microneedles

Dr. Baoming Jiang

Centers for Disease Control & Prevention

Atlanta, USA

bxj4@cdc.gov

San Antonio, TX

8 October 2014

Rotavirus

• Most common cause of severe diarrhea in children

• All children infected by age 5

• First infections symptomatic• Natural immunity • Various strains/serotypes in circulation• Improvements in sanitation

don ’t prevent infection

Rotavirus Mortality by Country~453,000 deaths, 2008

100 to 1000 deaths per 100,000 50 to 100 deaths per 100,00010 to 50 deaths per 100,000

< 10 deaths per 100,000

Rotavirus Structure

RNA Segment Protein

1

2

34

5

6

7

8

9

10

11

VP1

VP2

VP3VP4

NSP1

VP6

NSP2

NSP3

VP7

NSP4

NSP5

VP2

VP4 Neutralization

antigen

VP6Subgroupantigen

VP7Neutralization

antigen

Subcore

P[8]G47.5%

P[8]G152.2%

P[6]G9 P[8]G92.3%

other18.2%

P[8]G32.8%

P[4]G211.5%

Summary of P & G Types of Rotavirus Childhood Diarrhea From 66 Published Studies

(1993-2003)

N=21,256Rare or regionally common strains (23 strainstotal): P[4]G1 (1.3%), P[6]G2 (0.8%), P[6]G1 (0.6%), P[6]G8 (0.6%), P[4], G3 (0.5%)

5.5%

Gentsch JID 2005

Two Live Oral Rotavirus VaccinesRotaTeq is Pentavalent & Rotarix is Monovalent

6

G1 G3

G2 G4

P[8]

Five bovine-humanreassortant rotavirus

strains

G1P[8]

Single human rotavirus strain

RotarixRotaTeq

Rotavirus Vaccines in US

Feb 2006 – RotaTeq licensed by FDA

Feb 2006 – RotaTeq recommended by ACIP

April 2008 – Rotarix licensed by FDA

June 2008 – Rotarix recommended by ACIP

Vaccine Introduced 2006

9

GAVI Co-Sponsored Trials in Africa and Asia

Kenya

South Africa

Malawi

Ghana

Mali

Rotarix

RotaTeq

Slide: K Neuzil, PATH

Current Status of Rotavirus Vaccine Introduction

Rotavirus Vaccine Experience to Date

11Borrowed from: http://dannybrown.me/wp-content/uploads/2011/01/success_baby.jpg (Courtsey of U Parashar)

12

Issues of safety ORVs- (real or implied) remain! Post-Licensure Data

Source: www.cdc.gov/vaccines/acip/meetings

RV1 RV5

Australia 5.0 5.9

Mexico 1.2

USA 5.3 0.5-1.5

Intussusception cases /100,000 vaccinees

Other safety issues: PCV, antigenemia, AGE

RV Vaccines Need a Separate Cold ChainCost and Volume Comparison with Existing Vaccines

US$4,687.50 *US$4,687.50 *

625 dosesof rotavirus vaccine.

District vaccine store, Brazil.

625 dosesof rotavirus vaccine.

District vaccine store, Brazil.

4,100 dosesof polio and measles

vaccines. Rural hospital storage,

Mozambique.

4,100 dosesof polio and measles

vaccines. Rural hospital storage,

Mozambique.

US$635.50US$635.50

*Older photo—improvements in packaging volume have been achieved. Courtesy: Darin Zehrung

Rotarix* & RotaTeq† Efficacy vs Social Economic Status

Nelson & Glass, Lancet 2010

▲Bangladesh*

▲China*

Risk Factors for Lower Performance Of Live Oral Rotavirus Vaccine

Factors that lower viral titer

• Breast milk

• Maternal antibodies

• Stomach acid

• Prior exposure

Factors that impair immune

response• Malnutrition - Zn, Vit A

• Interfering microbes- viruses and bacteria

• Other infections- HIV, malaria, TBC

Others: Novel & diverse strains

Host genetic diversity

Studies to improve efficacy of ORVs

• Transplacentally transferred maternal serum Ab - Change schedule: time & no. of doses - S. Africa, Pakistan, Bangladesh, India

• High levels of Ab in breast milk- Withhold breast feeding at time of vaccination- S. Africa, India, Pakistan, Bangladesh, Nicaragua

● Add Nutritional supplements - zinc, vitamin A, probiotics, etc.- India, Pakistan

• Increase Rotarix titer ?

• Search for RV vaccine -- OPV interference - South Africa, Bangladesh

None has led to major improvement !

Inactivated Rotavirus vaccine (IRV)

Efficacy ▪ Not subject to interference/gut enteropathy seen with oral vaccines▪ more effective for infants in low income countries

Safety▪ No stigma of ORVs - intussusception, PCVs▪ No vaccine-acquired disease or new reassortant virulent strain

Combination vaccines – ease delivery, lower administration cost, increase vaccine coverage

Technology well established/tested (e.g., IPV, HAV)

Would represent insurance against problems with ORVs

IRV could be a Game Changer !

CDC-9 as a monovalent IRV candidate

G1P8G1P8 G2P4G2P4

G1P8 G1P8

● 107 ~ 108 titer in Vero cells

● Predominant (>90%) triple-layered

● Stable during USP & DSP

ffu

/ml

IRV: Strain & dose matter !

Jiang et al, Human Vaccines & Immunotherapeutics 2013

1. Three doses induce cross neut. antibodies to human strains & reassortant G1 strain

2. Little cross reactivity with animal strain or reassortant P8 strain

-------------- ---------------------------------------------------------------------------------Homotypic Heterotypic

Wang et al Vaccine 2010

Vaccine

Placebo

Neutralizing antibody (post dose 3)

Proof of concept for IRV in gnotobiotic piglets

Skin Immunization

● One of the earliest known route of vaccination -- smallpox, TB

● Immunology: rich in APC – Langerhans cells & dermal DC

● No hypodermic needle, no disposal of sharp waste

● No pain

● Combination vaccines, mass vaccination campaigns

● No need for additional cold chain

22 of 53

←Vaccinostyle←Rotary lancet←Surgical needle

Source: Fenner, et al, WHO, 1988

“Multiple Pressure” method

Smallpox Delivery TechniquesEarly Tools for Breaking the Skin

0

1000

2000

3000

4000

5000

6000

7000

IgG

tite

r (G

MT)

MN 5 µg

MN 0.5 µg

IM 5 µg

IM 0.5 µg

IM dis 5 µg

MN mock

0 10 28

Days Post Inoculation

Skin immunization uses fractional doses and enhances antibody response in mice

Moon et al, Vaccine, 2013 (in collaboration with M. Prausnitz)

Dose sparing: 10% MN = 100% IM

Beforecoating

Aftercoating

NanoPass developed MicronJet™, an intradermal (ID) microneedles device

Used for ID delivery of vaccines and large molecules

First true (~0.5mm) microneedle device ever registered with FDA

Microneedles are barely visible to the naked eye

Clinically shown to be almost painless and non-intimidating

Applicable for adults and pediatrics (i.e., IPV)

NanoPass MicronJet™

IntanzaNanoPass

Courtesy: Yotam Levin

Dose sparing effect established

Jiang (unpublished data)

1st Vax 2nd Vax 3rd Vax 1st Vax 2nd Vax 3rd Vax

ID & IM immunization induces comparable IgG & IgA titers in gnotobiotic piglets

ID: 5 ug Ag; IM: 5 ug Ag + 600 ug Al(OH)3; Control: diluent

IRV induces protection against oral challenge in piglets

Jiang (unpublished data)

IRV (ID)Placebo (ID) IRV (IM)

RV shedding in stool was measured by EIA

Day after oral challenge

RV

an

tig

en

in

sto

ol

(OD

va

lue

)

CDC IRV Program – Current status

Established technology (cell substrate, methods, assays, etc.) Strains developed, characterized

- CDC-9 G1P8

Inactivation method - Heat process – simple, robust, maintain structural integrity- Formalin procedure

Process development, formulation, stability Pre-clinical studies

- Proof of concept for serum antibody in macaques- Proof of concept for IRV by IM immunization in mice, guinea pigs & piglets- Proof of concept for IRV by skin immunization in mice & piglets

IP protection (strains & inactivation method) Partnerships (CMO, manufacturers, NGO, etc)

Discovery to Market: IRV Pre-clinical Studies

Clinical trials

▪ cGMP MVB production complete (CMO)

▪ SBIR contracts (CMO)

• Phase I: Optimization of rotavirus vaccine production (complete)

- USP & DSP process development

- Assay development

• Phase II: Preparation of IRV pilot lots for phase I clinical trials

- Validation of processes, scale-up, inactivation & formulation

- Toxicity study in animals

- cGMP production of pilot vaccine lots

▪ 2011 Winner - CDC’s inaugural innovation Fund Challenge▪ 2011 Excellence in Technology Transfer Award - SE Federal Laboratory Consortium▪ 2012 Excellence in Technology Transfer Award - Federal Laboratory Consortium▪ 2013 CDC SBIR Phase I Award▪ 2014 CDC SBIR Phase II Award

Public Support & Public-Private Partnerships

CDC Commercial Partners#

Co

mp

an

y

Pathways Forward

Early Phase (5~7 yrs): Develop stand-alone IRV (IM & ID)

- Demonstrate safety, efficacy, and value

- Determine if IRV is more effective than ORV

Late Phase (3-5 yrs): Combine IRV with multivalent vaccine

(e.g., DTaP, IPV) for IM administration (established vaccine Co.)

- Adds competitive value to multivalent vaccine

- Avoids administration & delivery costs

- Opens up product to global market

Or Combine with target vaccine (e.g., IPV) for ID administration

(Biotech Co.)

In partnerships with several vaccine manufacturers

OPV

IPV then OPV

IPV

Countries Currently Using IPV in National Immunization Programs

Courtesy: Cara Burns

The global move to IPV !

AcknowledgmentsAcknowledgmentsCDC

Yuhuan Wang

Sung-Sil Moon

Daniel Velasquez

Houping Wang

Mathew Esona

Jennifer Hull

Charles Humphrey

Lauren Snipes

Larry Westerman

Umesh Parashar

Jon Gentsch

Roger Glass

Outside CDC

Linda Saif

Anastasia Vlasova

Marli Azevedo

Mark Prausnitz

Chris Edens

Yotam Levin

Penelope Dennehy

Harry Keyserling

Jean-Francois Saluzzo

Stan Cryz

Harry Greenberg

Vic Van Cleave