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 FLOW OF PRESENTATION

01 02 03 04
INTRODUCTION AND STUDY OBJECTIVE TRIAL DESIGN SELECTION AND
BACKGROUND WITHDRAWL OF
INFORMATION SUBJECTS

05 06 07 08
TREATMENT OF ASSESSMENT OF STATISTICS Strength &
IMMUNOGENECITY Weakness
SUBJECTS
AND SAFETY
 ABBREVIATIONS:
ca: cold adapted
wt: wild type

ts : temperature sensitive
att : attenuated
HI: Hemagglutination Inhibition
EID50: Embryo Infection Dose 50
ELISA: Enzyme Linked Immunosorbent Assay

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 INTRODUCTION AND BACKGROUND
INFORMATION
Influenza-A

INFLUENZA Influenza-B

Influenza-C

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BASED ON SEQUENCE AND ANTIGENIC
ANALYSIS: H(1-16) AND N(1-9)
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ANTIGENIC DRIFT VS ANTIGENIC SHIFT:
 INFLUENZA OUTBREAKS
During seasonal: highest in
young children (30%)
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In younger adults and Complications: secondary
elderly : less than 10% bacterial pneumonia, primary
viral pneumonia, otitis media,
encephalopathy and
encephalitis, sepsis like
syndrome and febrile seizures
in young children.
Worsening leads to COPD,
diabetes, CHF
7
 INFLUENZA
OUTBREAKS:
 In U.S.A: Highest rate of influenza related
hospitalization occurred among those <2 year of age and
>64 years of age and highest death in>64 years of age.
 Serological evaluation
 Currently seasonal influenza vaccine from Solvey,
Novartis and Sanofi are available in Indian market
 Influenza –A subtypes: H3N2 and H1N1
 Influenza –B : two distinct lineages i .e B/Yamagata-like
and B/Victoria-like viruses.

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VACCINES AGAINST INFLUENZA:

Live Attenuated
Influenza Intranasally
Vaccine(LAIV)
Vaccines against
Influenza
Inactivated Influenza
Intramuscularly
Vaccine(IIV)

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 CLASSICAL
REASSORTMENT:
 Reassortant vaccine virus strains containing
HA and NA genes from wt virus and six
internal protein genes of MDV are
generated by classical reassorment method
 First they replicate efficiently at 25-33◦C
but not at temperatures above 39 ◦C.
 Phenotypic traits: ca , ts and in vivo att
which are transferable to 6:2 reassortant
viruses.

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 US VACCINE: Flumist
(2003)
 Trivalent
 2 Influenza-A+ 1 Influenza-B
 Influenza A/Ann Arbor(AA)/6/60(H2N2)
and influenza B/Ann Arbor/1/66 virus
assorted annually with HA and LAIV for
Pandemic Influenza 111NA gene segments
from predicted circulating wild-type(wt)
influenza a(H3N2 and H1N1) and
influenza B viruses.
 Confirmed efficacy levels of 87-93% for
H3N2 and Type-B in young children.

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 RUSSIAN VACCINES:
 Live attenuated influenza vaccines derived from ca MDVs
 Licensed for use in Russia
 Influenza A vaccines were based on the influenza
A/Leningrad/134/57 (H2N2) virus which was passaged in
embryonated hens’ eggs at least 20 times, leading to attenuation; this
strain subsequently underwent 17 or 47 further passages in eggs at
reduced temperatures (primarily at 25–26°C), resulting in two ca
donor viruses: the influenza A/Leningrad/137/17/57 ca (H2N2)
(A/Len/17 ca) virus and the influenza A/Leningrad/137/47/57 ca
(H2N2) (A/Len/47 ca) virus, respectively.
 The live attenuated vaccines based on these viruses and the
influenza B donor virus, B/USSR/60/69, have been widely used in
Russia for annual influenza vaccination in both children and adults.

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 ADVANTAGES OF LAIV:

 No of doses produced per egg are  Significant potential for indirect

much higher than inactivated vaccines. protection or “herd immunity”

 Administered in nose through a simple  More protective than IIV

device

 Induces more rapid and broader

immune response

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  Developed a Pandemic influenza vaccine
(human, live attenuated) A(H1N1) 2009

 Live monovalent, intranasal spray

SIIL’S EXPERIENCE ON H1N1

PANDEMIC INFLUENZA VACCINE  Virus strain antigenically similar to
A/California/7/2009 (ca, ts ,att)

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 SIIL H1N1 PANDEMIC INFLUENZA VACCINE

Preclinical
Studies: Phase-1 Study: Large Phase 2/3 Study Additional animal
 Tested single dose and repeated  Found to be safe and tolerable study on Ferrets
 No SAE
dose toxicity studies in 2 rodent  No SAE reported nor any
species.  Few solicited reactions.  Ferrets are
unsolicitated event reported ,no
 No vaccine related mortality or effect on vitals.  Immunogenicity exquisitely
abnormal clinical signs, susceptible to
 Few solicitated reactions like evaluation by
abnormal lab values and gross or infection with
nasal discomfort, stuffy nose, “significant
histopathological changes were human influenza
sneezing, runny nose, uptake”(HI, MN,
observed in any animal viruses and are
headache, chills, fatigue, sore serum IgG, nasal
widely believed to
throat, cough, myalgia, IgA,etc.) and
 IV at doses 20,30 and 60 be the ideal small
arthralgia, irritability, loss of seroconversion.
microlitres per rat was not animal model for
teratogenic in Witsar rat. appetite, nausea and diarrhoea influenza research
were observed.
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10/01/2022 16
RESULTS:
Placebo: Vaccinated animals group:
 No detectable HI  Decreased body weight loss.
titers to the new  Low relative lung weight.
H1N1 virus and  Absence of fever.
following
challenge  Little or no affected lung parenchyma
infection when compared to animals of placebo
 All animals group.
showed fever  Viral loads in these animals in the
and weight loss.
 Increased upper (nose and throat) as well as
relative lung lower (lungs) respiratory tract were
weights also decreased or even absent in these
 60%macroscopic animals in comparison with placebo
ally affected animals
lung parenchyma
 DCGI approved use on July 3,2010
 Post marketing surveillance generated in 2000 subjects

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  Human, Live attenuated, Freeze dried (Type A & B)
 Seasonal, Trivalent (SII LAIV)
 Dose of not less than 106.5 EID50 of
A/17/California/09/38 (H1N1) and A/17/Perth/09/87
(H3N2); and dose not less than 106 EID50 of
B/56/Brisbane/60/08, as per the recommendation of
the World Health Organization (WHO) for the
Southern hemisphere 2011 and Northern hemisphere
2011-12 influenza season.
 Same as that of H1N1 as the same MDV is used;
SIIL’S SEASONAL INFLUENZA except addition of two more strains.
VACCINE  WHO Guidelines
 Phase 1 in adults and Phase2/3 in adults ,elderly and
pediatric.

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 STUDY OBJECTIVES

Primary Objective Secondary Objective

To assess safety To assess

immune response

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 End Points
Primary end point
‣ Incidence of solicited local and systemic reactions within 7 days.
‣ Incidence of unsolicited adverse events, serious adverse events and new onset chronic
medical conditions throughout the entire study period of 42 days.

Secondary end point

‣ Proportion of subjects with seroconversion for HI and IgG antibodies on Day 21
‣ Proportion of subjects with seroconversion for IgA antibodies on Day 7
‣ Geometric mean titre of HI and IgG on Day 0 and 21 and IgA on Day 0 and 7.

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 Study Design

Phase
Parallel
II/III Randomized
Group

Observer Active- Multi-

Blind Controlled center

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Study Groups Vaccine
Group I (1st dose Influenza vaccine (Human, Live attenuated) Freeze dried (Type A &
B) (Seasonal, Trivalent) (SII LAIV) (Formulation A):
group)
Dose of not less than 107 EID50 of A/17/California/09/38 (H1N1) and
A/17/Perth/09/87 (H3N2); and dose of not less than 106.5 EID50 of
B/56/Brisbane/60/08.

Group II (2nd dose Influenza vaccine (Human, Live attenuated) Freeze dried (Type A &
B) (Seasonal, Trivalent) (SII LAIV) (Formulation B):
group)
Dose of not less than 106.5 EID50 of A/17/California/09/38 (H1N1) and
A/17/Perth/09/87 (H3N2); and dose not less than 106 EID50 of
B/56/Brisbane/60/08.

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 Study Groups

Population Age Groups Subject/

Group I Group II Population

Paediatric 2-17 Years 55 55 110

Adult 18-49Years 35 35 70

Elderly ≥ 50 Years 55 55 110

No. of Subject per group 145 145 290

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 Ingredients Concentration
A/17/California/09/38 (H1N1) Not less than 107 EID50/dose
A/17/Perth/09/87 (H3N2) Not less than 107 EID50/dose
B/56/Brisbane/60/08 Not less than 106.5 EID50/dose
Partially hydrolysed gelatin 2.5%
Sorbitol 5.0%
L-Alanine 0.1%
L-Histidine 0.21%
Tricine 0.3%
L-Arginine hydrocloride 1.6%
Lactalbumin hydrolysate 0.35%
Phosphate buffer saline Base

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 • LAIV (1 dose vaccine)
• Diluent vial (2.5ml)
• Syringe (1ml/5ml)
Packing • Syringe (1ml) Luerlock
• Intra nasal spray device
• Vial adapter
• Protocol number
• Name of study vaccine
• Expiry date
Labelling • Storage conditions
• Cautionary Statement
• 6 digit number

Storage • Safe, Locked, Secure place

• Refrigerator (+2°C to +8°C)

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 Preparation

reconstituted
vial contains one
using 0.5ml (A)/
Lyophilized dose (A)/ three Administered 0.25
1.6ml (B) - sterile
vaccine doses (B) of ml per nostril
water for
0.5ml vaccine
inhalation

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 Withdraw 0.25 ml vaccine in 1 ml syringe;
Procedure
Apply spray device

Subject - upright position

Insert tip of sprayer inside a nostril;

push the plunger to spray

Fine spray - deposits the vaccine in the nose and nasopharynx

Remove spray device;

Repeat

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Study Day Day 0 Day 7 Day 21 Day 42
Visit Visit 1 Visit 2 Visit 3 Visit 4
Study Day D0 D0+1 D0+3 D0+7
Interval (days) (+7;+8) (+21;+24) (+42;+49)
Informed consent and informed assent x
Randomization x
Medical history x x x x
Vitals x x x x
Physical examination x x x x
Vaccine administration x
Blood collection x x
Nasal wash samples x x
Dispensing of diary x
Diary card review x
Solicited reaction x x
Adverse events x x x x
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SAE x x x x
 Precautions
• Anaphylactic shock
• Asthma
• Urticaria and Other allergic reaction
• Axillary temperature (≥38°C)
• Medical Treatment, Staff, Supervision
• Rescue Medications: Adrenaline, Anti- Histaminics, corticosteroids,
Resuscitation measure

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 Selection and Withdrawal of Subjects

Inclusion Criteria
 Normal healthy subjects of appropriate age
 Parents/legal guardians (2-17 years) wiling to give informed consent.
 The subjects (>-18 years) willing to give informed consent
 The subjects (13-17 years) willing to give informed assent
 Free of obvious health problems
 The participants should be the resident of the study areas
 A women of child bearing potential must agree not to become
pregnant and man must not to conceive a child.

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 Exclusion criteria
 History of confirmed/suspected H1N1
 Subject with asthma with active wheezing
 Known allergy to eggs or components of vaccine
 Intranasal medication within 2 weeks
 Positive UPT or breastfeeding women
 Subjects (2-17 yrs) with history of receiving aspirin
 Subject participation in other clinical trail
 History of previous severe allergic reactions
 Altered immune status or chronic medication of
 Major congenital defects or serious chronic illness
immunosuppressants within 6 months
 History of Guillain barre syndrome
 Acute febrile illness or acute infectious disease
 Acute or chronic medical history which in opinion
 Subject with any nasal pathology
of investigator might interfere with study
 History of ongoing allergic rhinitis.

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 Subjects develops
hypersensitivity or any
SAE causally related
to vaccine
Subject Withdrawal Criteria
Development of
immunosuppressive
condition or disease,
pregnancy, asthma
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Administration of study
Subjects can withdraw
vaccine within 48 hrs of
consent at any time.
cessation of H1N1 antiviral
therapy
Any immune suppressants or
Any condition that affect the
immune modifying drugs (>3
safety or rights of volunteer
days) or any drug/vaccine
other than study vaccine
participating in the study
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 Treatment of Subjects
1. Study vaccine and dosing

0.5 ml of
Subjects formulation A or 0.25 ml per nostril
B

2. Concomitant therapy and rescue medication

The use of the prophylactic medications for potential adverse reactions and receipt of any other
investigational or non registered drug or vaccine during the study period will not be allowed. Use of
aspirin therapy or aspirin containing therapy will not be allowed during the trial in children and
adolescents (2-17 yrs).

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 Monitoring Subject Compliance
 Each subject will be instruct to visit the study site on specified study visit dates and contact investigator at any time
to report AE
 If the subject misses a visit he/she will be contacted by telephone or home visit can also be done if required.

Assessment of Immunogenicity
Immunogenicity
parameters

Hemagglutination inhibition (HI)

Blood sample on
day 0 and 21
ELISA (IgG)

Mucosal IgA Nasal samples on

day 0 and 7

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 Methods for Assessing, Recording and Analyzing Immunogenicity Parameters

 Blood will be obtained from each subject prior to first dose and on day 21.
 Blood will be allowed to clot and serum obtained by centrifugation.
 Serum stored at -20°C to -80°C in three aliquots
 One aliquot will be sent to NIV, second to SIIL and third will be kept as a back up

HI assay: Serum samples ELISA IgG assay: This is

will be tested at an initial based on detection of virus
dilution of 1:10, and those specific antibodies by
that are negative for the ELISA. Type specific
hemagglutinin antibody will (against influenza A and B
be assigned a titer of 1:5. type) IgG will be measured.

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 Nasal Wash/Swab Samples

 Nasal wash will be carried out in adult and elderly subjects

 Nasal swab will be performed in children.
 Tested by ELISA method for mucosal IgA at SIIL against all three vaccine strains

For each subject the seroconversion for HI, IgG and IgA will be determined as
 HI: subject is considered seroconverted if there is ≥2-folds rise in post-vaccination
HI titer.
 IgG: subject is considered seroconverted if post-vaccination titer is > baseline (pre-
vaccination)
 IgA: Subject is considered seroconverted if post-vaccination titer is > baseline (pre-
vaccination)

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 ANALYSIS

Subjects with seroconversion in HI and IgG

antibodies on day 21

Proportion of subjects with

seroconversion IgA antibodies on day 7

Geometric mean of HI and IgG on day 0

and 21 and IgG on day 7

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 Assessment of safety
Safety parameters include-
 Occurrence of Solicited local or systemic
reactions within 7 days of vaccine
administration.
 Unsolicited adverse events and serious adverse
events(SAE).
 New onset Chronic medical condition
throughout the entire study of 42 days.

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38
 Methods for assessing, recording , and analysing
safety parameters
Adverse Events-
 Local solicited reactions:
Nasal discomfort, Sneezing, Stuffy nose, Runny nose, Loss of smell, Red eyes, Lacrimation, Facial
swelling.
 Systemic solicited reactions:
Fever (> 38 0 C), Headache, Chills, Fatigue, Sore throat, Cough, Myalgia, Arthralgia, Irritability,
Wheezing, Loss of appetite.
They are assessed by medical history, and physical Examinations. In addition, for specified time
period (7 days post vaccination), subjects/parents/local guardian are asked to record local reaction and
generalized signs and symptoms in diary cards.
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10/01/2022
Severity grading of
Adverse events

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 Measurement of
Post-vaccination Reactions

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 Adverse event will be
described by-

Description

Duration

Severity of event

Relationship to the investigational

product
Action taken to treat the adverse event

Outcome

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Action taken in response to an adverse event
described on a numerical scale from 0 to 6.
• 0 - No action taken
• 1 - Study vaccine temporarily interrupted
• 2 - Study vaccine permanently discontinued due
to this adverse event
• 3- Concomitant medication taken
• 4- Non-drug therapy given
• 5– Physician visit
• 6- Hospitalization/prolonged hospitalization
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Outcome shall be assessed as at the time of
last observation:
• 1 = Recovered/resolved without sequelae
• 2 = Recovered/resolved with sequelae
• 3 = Ongoing
• 4 = Death
• 5 = Ongoing at time of death but not the cause of
death
• 6 = Unknown. The outcome of the AE is not known
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 Study Contacts for Reporting Serious Adverse Events

Medical monitor (Primary contact)

Dr. Tushar Tewari
GVK Biosciences Pvt. Ltd.
307-309, BPTP Park Centra,
Third floor, Sector-30,
Gurgaon- 122001, Haryana, India
Ph No: +911244324000 Ext -4024 Reporting SAE​​
Mobile No: +919958029977
Adverse events
Reporting Serious

Fax: +911244324001 The investigator must report SAEs within one calendar

Email – tushar.tewari@gvkbio.com day (24 hours) of becoming aware of the event by email
or telefax to the GVK BIO Medical Monitor. If the SAE
Back up Medical monitor is fatal or life-threatening, the Medical Monitor shall be
Dr. Arpit Agarwal informed immediately by telephone.
GVK Biosciences Pvt. Ltd.
The report is then to be followed by submission of a
307-309, BPTP Park Centra,
Third floor, Sector-30, completed SAE Report Form as soon as possible but at

Gurgaon- 122001, Haryana, India latest within 3 calendar days of the telefax detailing
Ph No - +9124432400 Ext -4035 relevant aspects of the SAE in question.
Mob No - +919717992590
Fax - +911244324001
Email – sushant.sehrawat@gvkbio.com
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24/24 hour and 7/7 days availability
 Follow-up of Serious adverse events
All SAEs documented and followed-up until event resolved, stabilized , disappeared or explained. The report
forms with additional/changed information should be forwarded to the GVK BIO Medical Monitor latest within 5
calendar days after receipt of the new information.

Causality assessment of serious adverse events

The degree of certainty with which an SAE can be attributed to administration of the study vaccine(s) (or
alternative causes, e.g. natural history of the underlying diseases, concomitant therapy, etc.) will be determined by
WHO scale of causality assessment . It is made on the basis of the available information at the reporting time point.
Assessment of causality can change according to follow-up information.

Treatment of AE and SAEs

As per current Good Manufacturing Practices and applied measures should be recorded in eCRF.

Management and transmission of SAEs

GVK BIO Medical Monitor will process SAE report, write narrative and send to SIIL for review.
After approval, narrative forwarded to investigator for submission to IRB/IEC within 7 working days.
SIIL will report the SAE narrative to the DCGI within 14 calendar days.
GVK BIO shall forward the same to the other participating sites within 14 calendar days.

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Documentation of Influenza like illness
Subject with at least three Influenza like illness during study participation should be documented . Subject whose
reports having an influenza-like illness will be asked to provide specimens of nasal and throat swabs for virology
testing. Virology testing of nasopharyngeal swab specimens will be performed for three virus strains.

Early Termination Evaluation Visit

Subject wishes to withdraw before study completion, an Early Termination Evaluation Visit will be performed.
possible. All procedures conducted at the Study Termination Visit should be followed including: physical
examination, laboratory investigations (if required), reporting of adverse event and serious adverse events, if any,
and reporting of relevant medication.

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 Discontinuation Criteria

Possible reasons are responsible for

the study premature termination of
the subject:
Making great products
• Serious adverse event
Take risks to lead in product innovation ​
• Death
Listen to customers and learn what they need ​
• Non Serious Adverse Event
Test with customers until you get the product right ​
• Protocol violation
• Subject’s consent withdrawal
• Inappropriate enrollment
• Migrated/moved from the study area
• Lost to follow-up
• Other (specify)
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 Geometric Mean Titre of HI
Statistics
and IgG

Proportion of Proportions of
Subjects with subjects with
Seroconversion in seroconversion in
IgA Ab HI & IgA Ab
Immunogenicity
Analysis
(95% Confidence DAY
0
DAY DAY
7 21
Interval)
Geometric Mean
Titre of IgA

Safety Analysis
(95%confidence
DAY
Interval) DAY
0
DAY
7 42
Incidence of unsolicited
Incidence of adverse events; serious
solicited local and adverse events (SAEs) and
10/01/2022 systemic reactions new onset chronic 4medical
8

conditions
 • Total seroconversion calculated by adding number of
subjects who shows seroconversion in any one of 3 tests
(HI and IgA).
• Statistical significance between two groups will be
tested using Fisher ’s exact test.
• Additional exploratory analysis will be done.(e.g. ≥ 2
fold/4 fold increase in Antibody titre or HI titre
≥ 1:20, ≥ 1:40)

• Mean, standard deviation (SD) and range will be

Vital Signs

presented for continuous parameters and the

frequency and percentage for categorical
parameters.
• The notable vital sign abnormalities will be
presented by study groups using descriptive
statistics.

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Number of subjects planned
Assuming a drop-out rate of at most 15%, 145 subjects should be enrolled in each group to get 290 evaluable
subjects. The subjects in each group are further stratified into adults, elderly and pediatric population.

The level of significance

All confidence intervals for parameters to be estimated will be constructed using two sided test statistic with a
significance level of α=0.05 (i.e., a 95% confidence level).

Interim analysis
An interim analysis will be performed after completion of 21 days study follow up of all the study subjects. It will
include the safety as well as immunogenicity. For performing it, un-blinding is done.

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 Pre-study
Documentation Monitoring

Data Study &

Collection Site
Closure
Quality Control
Data
Management and
and Processing Quality
Data
Management Assurance
Procedures Audits &
Inspection

Data
Verification Analysis of Coding
Procedures Consistency and
Medical Plausibility

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  REGULATORY AND ETHICAL REQUIREMENTS\
o Ethics committee review and communication
o Protocol Amendments
o Subject Information and Informed Consent
o Subject Confidentiality
o Data Protection
o Notification of Primary Care Physician
o Participants information on outcome of the research
o Ethical conduct of the study:

 DATA HANDLING AND RECORD KEEPING

 INSURANCE OF STUDY SUBJECTS
 PUBLICATION POLICY & CONFIDENTIALITY

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STRENGTH
 Protocol includes data about previous non-clinical and clinical studies conducted on monovalent vaccine which
were found to be safe and efficacious.
 Additional information on protective efficacy of SIIL vaccine from animal study also included.
 Population of all age groups enrolled during study.

WEAKNESS
 There is wider range of factors that define the exclusion criteria, so it’s difficult to enrol subjects in the study.
 The Count of number of males and females to be participated in the study is not specified in the protocol.

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THANK YOU!