NanoFlu™ Vaccine Update: Phase 3 Results

NanoFlu™ Vaccine Update: Phase 3 Results

Vivek Shinde, MD, MPH, VP Clinical Development

October 1st, 2020

Gaps in current seasonal influenza vaccine technologies

NanoFlu was developed to addresses major gaps in vaccine performance

Vaccine Type Problem Addressed

Antigenic drift Immunosenescence Egg-adaptive changes

Antibodies T-cells

Standard Inactivated1,2,3,4,5

High-dose inactivated1,3,4,5

MF-59 Adjuvanted4,5,6,7

Cell-derived inactivated5,8

Recombinant2,3,4

NanoFlu9,10

[recombinant + adjuvanted]

• Only NanoFlu induces BOTH broadly cross-reactive antibodies AND potent polyfunctional CD4+ T-cell

responses; AND avoids egg-adaptive antigenic changes

??

1. Diaz-Granados, 2015; 2. Dunkle, 2017; 3. Zost, 2017; 4. Cowling 2019; 5. Izurieta, 2019; 6. Frey, 2014; 7. Mannino, 2012; 8. Gouma, 2019. 9. Shinde, 2018;

10. Shinde, 2020

CONFIDENTIAL - NanoFlu Update2

The NanoFlu vaccine:

Recombinant hemagglutinin (HA) nanoparticles• Produced in a Baculovirus/Sf9 insect cell system

• Expressed as recombinant, full-length, wild-type, uncleaved HA0 that assembles into homotrimers

• Purified homotrimers form higher order nanoparticle structures of 20-40 nm with PS-80

• Manufactured in a rapid, high-yield, high purity, process

Potent saponin-based Matrix-M adjuvant• Purified fractions extracted as saponins from the bark of Quillaja saponaria Molina

• Formulated with cholesterol and phospholipid, forming cage-like particles

• Characterized by mechanisms of action that include:

• Enhancement of antigen delivery to draining lymph nodes

• Enhancement of activated T cell, B cell, and APC populations in draining lymph nodes

• Induction of functional, and broadly cross-reactive antibodies (e.g. influenza)

• Enhancement in peak and durability of antibody responses (e.g. RSV)

• Induction of polyfunctional T cells, including CD4+ (e.g. Ebola, influenza), and CD8+ (e.g. Ebola)

• Antigen sparing in the context of novel antigens: pandemic influenza and Ebola antigens

> 4200 adults and children have been exposed to Matrix-M in numerous ongoing and completed clinical trials in the context of various antigens, including external collaborations (e.g. Malaria vaccine)

• Acceptable safety profile

The HA nanoparticle antigen and Matrix-M adjuvant


Phase 1: NanoFlu (tNIV) induced higher wild-type HAI antibody responses (GMFRs) vs. Fluzone-High Dose (IIV3-HD) against 5 generations of antigenically drifted A(H3N2) strains

Improved Titers against Influenza

Drift Variants

with a Nanoparticle Vaccine

V Shinde, et al.

NEJM 378 (2018); 24

Phase 1 design:

330 US adults aged ≥60 years

Randomized 1:1:1• tNIV: 15µg each HA (45µg total) + 50µg Matrix-M, or

• tNIV: 60µg each HA (180µg total) + 50µg Matrix-M, or

• Fluzone High Dose: 60µg each HA (180µg total)

Objectives/endpoints:

• Day 21 wild-type HAI antibody responses

against homologous and drift strains

• Safety profile through 1 year


Phase 1: egg-adapted reagents may give a misleading result

Microneutralization antibody responses (GMFRs) against egg-adapted vs. wild-type A/Singapore A/H3N2 virus

Ratio of Day 21 GMTs

61% ↑ p=0.0002

3.8 3.9

3.5

2.1

0.0

1.0

2.0

3.0

4.0

5.0

180µg NanoFlu IIV3-HD 180µg NanoFlu IIV3-HD

A/Singapore H3N2 (egg-adapted) A/Singapore H3N2 (wild-type)

Ge

om

etr

ic M

ean

Fo

ld T

ite

r R

ise (

95%

CI) Ratio of Day 21 GMTs

61% ↑ p=0.0002

NanoFlu induced

improved neutralization

responses against wild-

type vs. egg-adapted

A/Singapore H3N2

viruses underscoring

the problem of egg-

adaptive mutations

Neutralization antibody

responses against wild-

type circulating viruses

are the most clinically

relevant


Phase 2: High-level summary

Phase 2 Design:

Dose/formulation optimization trial

1375 adults aged ≥65 years

Randomized to 1 of 7 groups• NanoFlu: bedside mix

• NanoFlu: co-formulated

• NanoFlu: increased adjuvant dose (75µg Matrix-M)

• NanoFlu: increased B antigen dose

• NanoFlu: antigen only (no Matrix)

• Fluzone HD

• Flublok

Objectives/endpoints:

• Primary: demonstrate “adjuvant effect”

• Day 28 wild-type HAI antibody responses against

homologous and drift strains

• Safety profile through 1 year

• Exploratory: CD4+ T cell responses

Primary endpoint met:

• Demonstration of an “adjuvant effect”, i.e. Matrix-M adjuvant resulted in

significant enhancement of immune responses when compared to

unadjuvanted formulation.

Higher wild-type HAI antibody responses against homologous

A/H3N2 and drifted A/H3N2 strains as compared to Fluzone HD.

Similar wild-type HAI antibody responses against homologous and

drifted strains as compared to Flublok.

Strong induction of polyfunctional CD4+ T cell responses, which

were higher than both Fluzone HD and Flublok.

NanoFlu was well-tolerated.

Induction of Cross-reactive

Hemagglutination Inhibiting Antibody

and Polyfunctional CD4+ T-cell

Responses by a Recombinant

Matrix-M-Adjuvanted Hemagglutinin

Nanoparticle Influenza Vaccine

V Shinde, et al. 2020. medRxiv.

doi: 10.1101/2020.05.11.20098574


NanoFlu granted accelerated approval and fast track designation

FDA accelerated approval pathway granted for NanoFlu

• “These regulations allowed vaccines for serious conditions that filled an unmet medical need to be approved based on a surrogate endpoint.” - FDA

• For seasonal influenza vaccines, the HAI antibody response may be an acceptable surrogatemarker of activity that is reasonably likely to predict clinical benefit.

FDA fast track designation granted for NanoFlu

• “Designed to facilitate the development, and expedite the review of vaccines to treat serious conditions and fill an unmet medical need.“ - FDA

Phase 3 trial design

• Non-inferiority immunogenicity (based on HAI using egg-adapted reagents) against any licensed comparator to obtain initial licensure. We chose Fluzone Quadrivalent for several reasons.

• Commitment to conduct a post-licensure efficacy trial.

An opportunity for NanoFlu to advance rapidly to US licensure


Phase 3: a non-inferiority immunogenicity trial

Goals

• Demonstrate immunologic non-inferiority to licensed influenza vaccine (Fluzone Quadrivalent [SD]) on 4 homologous strains

• Establish pivotal clinical trial dataset to support filing of BLA via accelerated approval pathway

Design

• 2650 adults ≥65 years of age, across 19 US sites

• Randomized to 1:1 to either NanoFlu or Fluzone Quadrivalent

• Stratified by receipt of prior year seasonal influenza vaccine

• Single dose of test vaccine on Day 0

Goals and design

Day 0 Trial Treatment Injection

Subjects Per

GroupTreatment

GroupVaccine

HA Dose per Strain,

μg

(H1N1/H3N2/BV/BY)

Matrix-M1

Adjuvant

Dose, µg

Formulation

A NanoFlu (qNIV) 60, 60, 60, 60 75 Co-form 1325

BFluzone Quad

[standard dose]15, 15, 15, 15 N/A N/A 1325

Total Trial Subjects 2650

Primary contrast for

Non-inferior immunogenicity


Phase 3 design

Primary

• Demonstrate non-inferior immunogenicity of NanoFlu vs. Fluzone Quadrivalent:

• Day 0 and 28 egg-propagated virus HAI titers against the 4 vaccine homologous strains

• Describe the safety profile of both vaccines

Secondary

• Describe immunogenicity of NanoFlu vs. Fluzone Quadrivalent

• Wild-type HAI titers against homologous and drift strains

Exploratory

• Evaluate superior immunogenicity of qNIV vs. Fluzone SD (wild-type and egg-propagated HAI)

• Describe polyfunctional CD4+ T cell response (CMI)

• Describe immunogenicity of NanoFlu vs. Fluzone-SD in terms of microneutralization titers (MN)

[available in 2H 2020]

Objectives


Phase 3 HAI testing plan

Substantial A/H3N2 strain diversity and scope of HAI testing

2018 2019

A/Kansas/14/17

A/Swiz/8060/17

A/Singapore/16

3c3a

A2

A3

135N

131K

135N

A1b

A4

A/Tokyo/EH1801/18

A/Neth/1268/19

A/ID/13/18

A/CA/194/19

A/Cardiff/508/19

xA/South Australia/34/19

A1b

Purple – vaccine strains

Purple with box – current N and S hemisphere strains

A2

A1

Red dot – drifted strains to be tested in Phase 3


Phase 3 demographic dataTreatment groups similar at baseline

N 1333 1319

Mean age (SD) 72.5 (5.7) 72.5 (5.7)

Median age 72.0 71.0

Male (%) / Female (%) 40.6% / 59.4% 35.9% / 64.1%

Race/Ethnicity

White (%) 90.6% 91%

Black/African American (%) 7.9% 7.8%

All other (%) 1.5% 1.2%

Flu vaccine in 2018-19 (%) 83.8% 83.5%

Any flu vaccine w/i 3 yrs (%) 90.8% 91.0%

Fluzone Quad (SD)NanoFluVariable


Phase 3 top-line safety dataNanoFlu well tolerated

N 1333 1319

Counts (%) of Subjects with Events

Any treatment emergent adverse

event (TEAE)659 (49.4) 551 (41.8)

Any Solicited TEAE 551 (41.3) 420 (31.8)

Local solicited 372 (27.9) 243 (18.4)

Severe local solicited 8 (0.6) 2 (0.2)

Systemic Solicited 369 (27.7) 292 (22.1)

Severe systemic solicited 15 (1.1) 11 (0.8)

Unsolicited TEAE 248 (18.6) 241 (18.3)

Severe unsolicited 23 (1.7) 12 (0.9)

Severe & related unsolicited 10 (0.8) 2 (0.2)

Medically-attended unsolicited 99 (7.4) 104 (7.9)

Serious adverse events (SAEs)* 11 (0.8) 5 (0.4)

NanoFlu Fluzone Quad (SD)Safety events (thru Day 28)


*No SAEs in either treatment group were assessed by study investigators as related to vaccine

Phase 3 immunogenicity: homologous strainsDay 28 egg-based or wild-type HAI GMTs and GMT ratios (NanoFlu / Fluzone)

NanoFlu Fluzone Quad D28 GMT Ratio

Assay Strain D28 GMT D28 GMT (NanoFlu / Fluzone) 95% CI

HAI: EGG A/Brisbane/02/2018 (H1N1) pdm09 (Homologous) 49.3 45.0 1.09 ( 1.03, 1.15)

A/Kansas/14/2017 (H3N2) (Homologous) 151.5 126.8 1.19 ( 1.11, 1.27)

B/Maryland/15/2016 (Vic) (Homologous) 110.7 106.3 1.03 ( 0.99, 1.07)

B/Phuket/3073/2013 (Yam) (Homologous) 168.5 133.9 1.23 ( 1.16, 1.29)

HAI: WT A/Brisbane/02/2018 (H1N1) pdm09 (Homologous) 76.6 62.7 1.24 ( 1.17, 1.32)


B/Maryland/15/2016 (Vic) (Homologous) 62.8 47.2 1.32 ( 1.26, 1.39)

B/Phuket/3073/2013 (Yam) (Homologous) 118.3 78.4 1.47 ( 1.40, 1.55)

• GMT ratio success criteria met for non-inferiority

• NanoFlu: 3—23% better using egg-based HAI

• NanoFlu: 24—66% better using wild-type HAI

• “Superiority” criteria met for homologous A/H3N2 (66% better)

Success:

All 95% CI

lower

bounds are

≥ 0.67


Phase 3 immunogenicity: drifted A/H3N2 and B-Victoria strainsDay 28 wild-type HAI GMTs and GMT ratios (NanoFlu / Fluzone)

NanoFlu Fluzone Quad D28 GMT Ratio

Assay Strain D28 GMT D28 GMT (NanoFlu / Fluzone) 95% CI

HAI: WT A/Brisbane/02/2018 (H1N1) pdm09 (Homologous) 76.6 62.7 1.24 ( 1.17, 1.32)


B/Maryland/15/2016 (Homologous) 62.8 47.2 1.32 ( 1.26, 1.39)

B/Phuket/3073/2013 (Homologous) 118.3 78.4 1.47 ( 1.40, 1.55)

A/California (“Drifted” H3N2; Clade 3C2a1b-131K) 115.0 80.6 1.41 ( 1.33, 1.50)

A/Cardiff (“Drifted” H3N2; Clade 3C2a1b-135N) 63.9 45.4 1.34 ( 1.27, 1.43)

A/Netherlands (“Drifted” H3N2; Clade 3C3a) 102.3 74.7 1.38 ( 1.30, 1.46)

A/So. Aus. (“Drifted” H3N2; Clade 3C2a1b-131K) 98.1 70.4 1.36 ( 1.28, 1.44)

A/Idaho (“Drifted” H3N2– Clade 3C3a) 202.5 136.8 1.46 (1.37, 1.56)

A/Tokyo (“Drifted” H3N2– Clade 3C2a2) 78.0 54.5 1.39 (1.31, 1.48)

B/Washington (“Drifted B-Victoria) 88.2 71.4 1.23 (1.18, 1.28)

• NanoFlu: 34—46% better on drifted H3N2s using wild-type HAI and 23% better on drifted

B-Vic strain


Phase 3 immunogenicity: homologous strainsDay 28 egg-based or wild-type HAI SCR and SCR difference (NanoFlu - Fluzone)

NanoFlu Fluzone Quad Absolute SCR Difference

Assay Strain SCR SCR

NanoFlu -

Fluzone Quad 95% CI

HAI:EGG A/Brisbane/02/2018 (H1N1) pdm09 (Homologous) 22.0% (282/1280) 17.0% (219/1286) 5.0 ( 1.9, 8.1)

A/Kansas/14/2017 (H3N2) (Homologous) 41.8% (535/1280) 34.4% (443/1286) 7.3 ( 3.6, 11.1)

B/Maryland/15/2016 (Vic) (Homologous) 11.2% (143/1280) 10.7% (137/1286) 0.5 ( -1.9, 2.9)

B/Phuket/3073/2013 (Yam) (Homologous) 31.3% (401/1280) 22.9% (294/1286) 8.5 ( 5.0, 11.9)

HAI:WT A/Brisbane/02/2018 (H1N1) pdm09 (Homologous) 32.7% (419/1280) 21.4% (275/1286) 11.4 ( 7.9, 14.7)


B/Maryland/15/2016 (Vic) (Homologous) 25.1% (321/1280) 13.5% (173/1286) 11.6 ( 8.6, 14.6)


• Seroconversion (SCR) difference success criteria met

• NanoFlu: 0.5—8.5% increased SCR using egg-based HAI

• NanoFlu: 11.4—20.4% increased SCR using wild-type HAI

• “Superiority” criteria met for homologous A/H3N2

Success:

All 95% CI

lower bounds

are ≥ -10


Phase 3 immunogenicity: drifted A/H3N2 and B-Victoria strainsDay 28 wild-type HAI SCR and SCR difference (NanoFlu - Fluzone)

NanoFlu Fluzone Quad Absolute SCR Difference

Assay Strain SCR SCR

NanoFlu -

Fluzone Quad 95% CI

HAI:WT A/Brisbane/02/2018 (H1N1) pdm09 (Homologous) 32.7% (419/1280) 21.4% (275/1286) 11.4 ( 7.9, 14.7)


B/Maryland/15/2016 (Vic) (Homologous) 25.1% (321/1280) 13.5% (173/1286) 11.6 ( 8.6, 14.6)


A/California (“Drifted” H3N2; Clade 3C2a1b-131K) 37.1% (475/1280) 20.5% (264/1286) 16.6 ( 13.1, 20.0)

A/Cardiff (“Drifted” H3N2; Clade 3C2a1b-135N) 32.7% (419/1280) 18.6% (239/1286) 14.1 ( 10.8, 17.5)

A/Netherlands (“Drifted” H3N2; Clade 3C3a) 38.4% (492/1280) 21.7% (278/1284) 16.8 ( 13.3, 20.2)

A/S. Aus (“Drifted” H3N2; Clade 3C2a1b-131K) 34.4% (440/1280) 19.6% (252/1284) 14.7 ( 11.3, 18.1)

A/Idaho (“Drifted” H3N2– Clade 3C3a) 57.5% (735/1278) 38.7% (497/1283) 18.8 (14.9, 22.5)

A/Tokyo (“Drifted” H3N2– Clade 3C2a2) 34.7% (444/1280) 18.0% (231/1284) 16.7 (13.3, 20.0)

B/Washington (“Drifted B-Victoria) 17.7% (226/1278) 9.7% (124/1283) 8.0 (5.4, 10.7)

NanoFlu: 14.1—18.8% increased SCR against drifted A/H3N2 strains, and 8% increased

against drifted B-Vic strain


Phase 3 CMI:

• Subset of 140 subjects total: 70/group

• Day 0 and 7 available now

• Parameters assessed (all descriptive)

• Effector CD4+ T cells (including effector memory - Tem) [gated on CCR7-CD45RA-] in terms of various

polyfunctional phenotypes:

• Double cytokine positive *At least 2 of 4 of: IFN-γ, TNF-α, IL2, or CD40L+

• Triple cytokine positive *At least 3 of 4 of: IFN-γ, TNF-α, IL2, or CD40L+

• Quadruple cytokine positive *All 4 of: IFN-γ, TNF-α, IL2, and CD40L+

• Total CD4+ T cells, in terms of various polyfunctional phenotypes:

• Same definitions as above

• Reported as:

• Medians, Geometric Mean Counts (GMC), GMFR (Post/Pre), baseline adjusted ratio of GMCs (NanoFlu/Fluzone)

• Box plots

• RCD plots

Assessments


Phase 3 CMI: RCD of Day 0 and 7 counts of double cytokine+ effector CD4+ T cells against A/Kansas (H3N2)

• NanoFlu substantially right shifted distribution (“cleans out basement”)

• Similar pattern of CMI responses seen against B/Maryland


Phase 3 CMI:RCD of Day 0 and 7 counts of IFN-γ/double/triple/quadruple cytokine+ effector CD4+ T cells against A/Kansas (H3N2)

NanoFlu substantially right shifted distribution (“cleans out basement”)

IFN-γ+

Double+ Quadruple+

Triple+


Phase 3 CMI:Day 0 and 7 counts of IFN-γ/double/triple/quadruple cytokine+ effector CD4+ T cells against A/Kansas (H3N2)

• NanoFlu induced markedly higher responses than Fluzone Quad

• Similar pattern of CMI responses seen against B/Maryland

IFN-γ+

Double+ Quadruple+

Triple+


Phase 3 CMI: comparison to recent Univ of HK study [Cowling, 2019]Day 7 / 0 geometric mean fold rise (GMFRs) of IFN-γ cytokine+ total CD4+ T cells against A/H3N2 or B-Victoria strain

GM

FR

at

Day 7

• NanoFlu induced substantially higher fold-rises of IFN-γ+ CD4+ T cells as compared to Fluzone HD,

Flublok, or FLUAD based on comparable literature estimates [Cowling, 2019]


5.1

1.6 1.22.0 2.6

1.8

7.9

2.51.7 2.2

1.42.6

0.0

2.0

4.0

6.0

8.0

10.0

12.0

NanoFlu FluzoneQuad

FluQuadri FluzoneHD

Flublok Fluad NanoFlu FluzoneQuad

FluQuadri FluzoneHD

Flublok Fluad

qNIV-E-301 Cowling et al qNIV-E-301 Cowling et al

A/Kansas (E301) or A/Switzerland (Cowling) (H3N2) B/Maryland (E201) or B/Brisbane (Cowling) (B-Vic)

Primary endpoint met: • Demonstrated immunologic non-inferiority to Fluzone Quad in terms of hemagglutination inhibition (HAI) antibody responses

(assayed with egg-derived virus reagents) against all four vaccine homologous strains (per CBER criteria).

Statistically significant higher HAI antibody responses (assayed with wild-type reagents) compared to Fluzone Quad:

• 24—66% improved Day 28 GMTs against homologous strains

• 34—46% improved Day 28 GMTs against drifted A/H3N2 strains

• 11.4—20.4% increased Day 28 seroconversion rate against homologous strains

• 14.1—18.8% increased Day 28 seroconversion rate against drifted H3N2 strains

Substantially and statistically significantly improved polyfunctional CD4+ T-cell responses across both Total and Effector CD4+ T cells subsets

• Unlike Fluzone Quad, virtually all NanoFlu subjects became “CMI responders”, including, most notably, those with low baseline CMI

• Substantially greater induction of CMI than what has been reported for other “enhanced” influenza vaccines in older adults

NanoFlu was well-tolerated

Phase 3: High-level summary and conclusions


NanoFlu developed with the intent to meet an urgent public health need for an improved seasonal influenza vaccine in older adults by overcoming the challenges of:

• Antigenic drift

• Egg-adaptative changes

• Immunosenescence

• Poor induction of T cell immunity

The full-length recombinant nanoparticle structure and Matrix-M adjuvant are both critical for: • Induction of clinically relevant immune responses against circulating wild-type viruses (thereby avoiding problems

of egg-adaptive antigenic changes)

• Induction of broadly cross-reactive antibody responses through recognition of conserved broadly neutralizing head-and stem-based epitopes

• Potent (best in class) induction of polyfunctional and cross-reactive CD4+ T cells with a “cleaning out of the basement” (elimination of the non-responder phenotype; and potentially mitigating effects of immunosenescence)

Next steps• Conduct lot-to-lot consistency trial

• Clinical efficacy trial

• BLA filing

The NanoFlu vaccine: summary and next steps


Documents

NanoFlu™ Vaccine Update: Phase 3 Results