Janssen Johnson and Johnson ad26 Adenovirus Vector

1. Single-shot Ad26 vaccine protects against SARS-CoV-2 in rhesus macaques - 30 July,2020 Here we show the immunogenicity and protective efficacy of a single dose of adenovirus serotype 26 (Ad26) vector-based vaccines expressing the SARS-CoV-2 spike (S) protein in non-human primates. Fifty-two rhesus macaques (Macacamulatta) aged 6–12 years old were immunized with Ad26 vectors that encoded S variants or sham control, and then challenged with SARS-CoV-2 by the intranasaland intratracheal routes9,10. Macaques received a single immunization of 1 × 1011viral particles of Ad26 vectors by the intramuscular route without adjuvant at week 0. We observed receptor-binding-domain (RBD)-specific binding antibodies by enzyme-linked immunosorbent assay (ELISA) in 31 out of 32vaccinatedmacaquesby week 2, and in all vaccinated macaques by week 4 Median titres of neutralizing antibodies in the macaquesvaccinatedwithAd26-S.PPwerefourfold higher than median titresof neutralizing antibodies in previously reported cohorts of 9 convalescent macaques9and 27 convalescent humans after recoveryfrom SARS-CoV-2 infection10 The Ad26-S.PP vaccine also induceddetectable S-specific IgG and IgA responses in bronchoalveolar lavage (BAL) samples. Ad26-S.PP is a wild-type leader sequence with full-length S and mutation of the furin cleavage siteand proline-stabilizing mutations Analysis of a cohort of 10 similarly immunized macaquesdemonstrated that a single immunization of 1 × 1011viral particles of Ad26-S.PP elicited consistent IFN-γ ELISPOT responses but minimal or no IL-4ELISPOT responses which suggestsinduction of type 1 T helper (TH1)-biased responses. At week 6, all macaques were challenged with 1.0 × 105 50% tissue culture infectious dose (TCID50) of SARS-CoV-2 by the intranasaland intratrachealroutes9,10. Consistent with previous observations, clinical disease was minimal in all macaques after challenge9,10. Viral loads in BAL and nasal swabs were assessed by reverse transcription PCR (RT–PCR) specific for subgenomic mRNA (sgRNA), which is thoughtto measure replicating virus9,20. By contrast, macaques that were treated with the Ad26-S.PP vaccine immunogendid not show anamnestic neutralizing antibody responses and only had lowT cell responses against spike proteins (S1 and S2), which was the vaccine antigen, after challenge.

2. *Our data extend recent preclinical studies of inactivated virus vaccines and DNA vaccines for SARS-CoV-2 in non-human primates10,21. Whereas inactivated virus vaccines and nucleic acid vaccines typically require two or moreimmunizations, some adenovirus vectors can induce robust and durable neutralizing antibody responses after a single immunization22–24. A single-shotSARS-CoV-2 vaccine would have important logistical and practical advantages compared with a two-dose vaccinefor mass vaccination campaigns and control of the pandemic. However, we would expect that a two-dosevaccine with Ad26-S.PP would be more immunogenic. *Our previous data demonstrate that a homologous boost with Ad26-HIV vectors augmented antibody titres by more than tenfoldin both non-human primates and humans25–27 A limitation of our study is that we did not evaluate the durability of neutralizing antibody responses elicited by these vaccines, andfuture studiesare planned to investigate this question. Additional studies could also evaluatemucosal delivery of this vaccine. Our studies were also not specifically designed to assess safety or the possibility of vaccine-associated enhanced respiratory disease or antibody-dependent enhancement of infection29. However, it is worth noting that the Ad26-S.PP vaccine elicitedTH1-biased rather than TH2-biasedT cell responses The optimalAd26-S.PP vaccine from this study, termed Ad26.COV2.S, is currently being evaluated in clinical trials. 1INTERVENTION GROUPS AND DURATION Participants will be vaccinated at the study site according to the schedules detailed above: Ad26.COV2.Ssupplied at a concentration of 1×1011vp/mL in single-use vials, with an extractable volume of 0.5 mL, and dosed at 5×1010vp(virus particles) Placebo: 0.9% sodium chloride (NaCl) solution. For blinding purposes, all participants will receive Ad26.COV2.S or placebo at Day 1, using the same volume (ie, 0.5 mL). The occurrence of COVID-19-related hospitalization and COVID-19-related complications (such as but not limited to hyperinflammatorysyndrome, pneumonia, neurological or vascular complications, severe neurological or vascular events, acute respiratory distress syndrome, renal complications, sepsis, septic shock, death)awill be monitored throughout the study. 1Janssen Vaccines & Prevention*B.V. Clinical Protocol – ENSEMBLE Protocol VAC31518COV3001; Phase 3 VAC31518 (JNJ-78436735) - *Johnson & Johnson

3. 1045 participants Other Names: JNJ-78436735 Ad26COVS1

4. Our innovative vaccine technology platform – AdVac – offers real hope for the accelerated development of vaccines and is designed to help the global health community better prepare for and combat life-threatening infectious disease outbreaks. This technology was used in the development of our Ebola vaccine regimen, which was approved by the European Commission in July 2020 and has been deployed in the Democratic Republic of the Congo (DRC) and Rwanda. The AdVac technology was also used to construct our HIV, RSVand Zika vaccine candidates. During the Ebola epidemic in West Africa, Janssenresponded by accelerating its Ebola vaccine development effort and scale up, resulting in the manufacturing of two million vaccine regimens in less than one year. This two-dose regimen, Zabdeno (Ad26.ZEBOV) and Mvabea (MVA-BN-Filo), is now approved ADVAC® VIRAL VECTOR TECHNOLOGY Adenoviruses are a group of viruses that cause the common cold - so they're good for transporting things into humans. Janssen’s AdVac vectors are based on a specific type of adenovirus, which has been genetically modified so that it can no longer replicate in humans and cause disease. The AdVac technology works by using an adenovirus as a vector(a carrier) of an antigen’s genetic code, to mimic components of a pathogen (a bacterium, virus, or other disease-causing organism). Antigens(components of a pathogen) are produced to mimic the pathogen, without causing severe disease. On March 30, 2020, Johnson & Johnson announced a lead vaccine candidate for COVID-19, Ad26.COV2.S.

5. 60,000 participants

6. 44,325 participants

7. The ENSEMBLE 2 (NCT04614948) study, which is being conducted in collaboration with the UK National Institute for Health Research (NIHR), will enrol up to 30,000 participants globally. This trial will run in parallel with the Phase III ENSEMBLE study of the single-doseregimen of JNJ-78436735 - also known as Ad26.COV2.S - which is proceeding to enrol up to 60,000 participants across the globe The randomised, double-blind, placebo-controlledENSEMBLE 2 trial will analyse the safety and efficacy of a two-dosevaccine regimen versus placebo in adults with and without comorbidities associated with an increased risk for severe Covid-19. It will analyse efficacy of the vaccine candidate after the firstand seconddoseto evaluate the protection providedagainst the virus and possible incremental benefits for duration of protection with a second dose The initiation of the Phase III ENSEMBLE and ENSEMBLE 2 trials follow positive interim data from Janssen’s on-going Phase I/IIa trial analysing the safety pro In a separate development, J&J expanded partnership between its Janssen and the US Biomedical Advanced Research and Development Authority(BARDA) for the on-going development of Janssen’s Covid-19 vaccine candidate. As per the deal, Janssenwill commit around $604m while BARDAwill offer $454m to support the Phase III ENSEMBLE trial. Last month, J&J temporarily paused dosing in all its vaccine candidate trials for Covid-19, including Phase III ENSEMBLE trial, after a study participant reported an unexplained illness.

8. ENSEMBLE is being initiatedin collaboration with the Biomedical Advanced Research and Development Authority(BARDA), part of the Office of the Assistant Secretary for Preparedness and Response at the U.S. Department of Health and Human Services (HHS) under Other Transaction Agreement HHSO100201700018C, and the National Institute of Allergy and Infectious Diseases (NIAID), part of the National Institutes of Health (NIH) at HHSand the COVID-19 Prevention Trials Network (COVPN) and in collaboration with Operation Warp Speed (OWS),. The Janssen COVID-19 vaccine candidateleverages the Company’s AdVac technology platform, which was also used to develop and manufacture Janssen’s European Commission approved Ebola vaccineand construct its Zika, RSV, and HIV vaccine candidates. Janssen’s AdVac technology platform has been used to vaccinate more than 100,000 people to date across Janssen’s investigational vaccine programs. With Janssen’s AdVac technology, the vaccine, if successful, is estimated at launch to remain stable for two years at -20 °Cand at least three months at 2-8° C. This makes the vaccine candidate compatible with standard vaccine distribution channels and would not require new infrastructure to get it to the people who need it. Ad26.COV2.S (previously known as Ad26COVS1) is a monovalent vaccine composed of a recombinant, replication-incompetent adenovirus type 26(Ad26) vector, constructed to encode the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) spike (S) protein “We used a highly scientific and evidence-basedapproach to select this vaccine candidate.”

9. Adenoviral-vectored Vaccines Recombinant, replication-incompetent adenoviral vectorsare attractive candidates for expression of foreign genes for a number of reasons. The adenoviral genome is well characterized and comparatively easy to manipulate. Adenoviruses exhibit broad tropism, infecting a variety of dividing and non-dividing cells. The adenoviral vaccine (AdVac®) vector platform, developed by CrucellHolland B.V. (now Janssen Vaccines & Prevention B.V.) allows for high-yield production of replication-incompetent adenovirus vectors, eg, Ad26, with desired inserts. The adenovirus E1 region is deleted to render the vector replication-incompetent and create space for transgenes (GMOs), with viral replication taking place in cells that complement for the E1 deletion in the virus genome. Ad26has been selected as a potential vaccine vector because there is substantial nonclinical and clinical experience with Ad26-based vaccines that demonstrate their capacity to elicit strong humoral and cellular immune responses and their acceptable safety profile, irrespective of the antigen transgene(see also Section 2.3.1). Furthermore, these data support an immunogenicity profile with emphasis on T-helper cell type 1 (Th1) responsesand demonstrate predominantly interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) production in CD4+ and CD8+ T cells.3,35,45 The parental S protein sequence was derivedfrom a SARS-CoV-2 clinical isolate(Wuhan, 2019; whole genome sequence NC_045512). Genomic sequencing was performed on bronchoalveolarlavage fluid samples collected from patients with viral pneumoniaadmitted to hospitals in Wuhan, which identified a novel RNA virus from the family Coronaviridae.42,66 Phylogenetic analysis of the complete viral genome revealed that the virus, SARS-CoV-2, is part of the subgenus Sarbecovirus of the genus Betacoronavirus Vaccine-associated enhanced disease has been described in some animal models for SARS and MERS in which candidate vaccines induced a Th2 biased immune response,1,7,22,31,32but proof of human SARS- or MERS-vaccine-associated enhanced disease does not exist as these candidate vaccines were never tested for efficacy nor used in outbreak situations. Nonclinical Pharmacology Nonclinical studies were performed to test the immunogenicity of different vaccine candidates, leading to the selection of the current vaccine for this development program. In addition, VE of Ad26.COV2-S has been shown in Syrian hamsters and NHP. Details are provided in the IB.33,34 1Janssen Vaccines & Prevention*B.V. Clinical Protocol – ENSEMBLE Protocol, December 14, 2020 VAC31518COV3001; Phase 3 VAC31518 (JNJ-78436735) - *Johnson & Johnson

10. Clinical Studies At the time of initial protocol writing, no clinical datawith the Ad26.COV2.S vaccine were available. As of 10 September 2020, a single injection of Ad26.COV2.S has been administered to 805 adult participants, aged 18 and older. The FIH(First-in-Human) studyVAC31518COV1001will be on-going at the time of initiation of study VAC31518COV3001. Study VAC31518COV1001 is a randomized, double-blind, placebo-controlled, Phase 1/2a multicenter study in adults aged ≥18 to ≤55 years and aged ≥65 years. The safety, reactogenicity, and immunogenicity of Ad26.COV2.S will be evaluated at 2 dose levels(5×1010vp and 1×1011vp), administered IM as a single-dose or 2-dose schedule, with a single booster vaccination administered in 1 cohort. Overall, a target of 1,045 adult participants in these 2 age groups will be randomly assigned in this study. Th1/Th2 Profile of Ad26-based Vaccines in Clinical Studies In the 1960s, a formalin-inactivated RSV vaccine was associated with enhanced respiratory disease(ERD) in young children, characterized by an increased rate of RSV-mediated, severe lower respiratory tract infection in the vaccinated individuals compared with the control group.18,28,37,38 Although the mechanisms for ERD are not fully understood, it is thoughtthat FI-RSV may have: 1) failed to induce adequate neutralizing antibody titers; 2) led to an overproduction of binding antibodies promoting immune complex deposition and hypersensitivity reactions; 3) failed to induce adequate numbers of memory CD8+ T cellsimportant for viral clearance; and 4) induced a Th2-skewed type T-cell response.47Vaccine-induced ERD has also been described for SARS-CoV and MERS-CoV in animal models,34but proof of human SARS-CoV or MERS-CoV vaccine-associated enhanced disease does not exist as these candidate vaccines were never tested for efficacy nor used in outbreak situations. For SARSand MERS, the mechanism of enhanced disease observed in mice has been associated with aTh2-mediated eosinophilicinfiltration in the lung, which is reminiscent of ERD effects observed after RSV infection of mice immunized with FI RSV. Similar to RSV vaccines, enhanced disease has been shown for whole-inactivated SARS-CoV vaccines, as well as subunit vaccines inducing aTh2-type immune response, which can be rescuedby formulating vaccines in Th1-skewingadjuvants. Participants of childbearing potential will be required to agree to practicing an acceptable effective method of contraception and agree to remain on such a method of contraception from providing consent until 3 monthsafter receiving study vaccine

11. 8.1.1. Pre-specified Criteria for Suspected COVID-19 The criteria for suspectedCOVID-19(ie, the triggers to proceed with home-collection of the nasal swabs on COVID-19Day 1-2 and to proceed with the COVID-19 Day 3-5 visit) are pre-specified as follows:  A positive RT-PCR result for SARS-CoV-2, through a private or public laboratory independent of the study, whether symptomaticor asymptomatic OR  New onset or worsening of any 1 of the symptoms, which lasts for at least 24 hours, not otherwise explained: Headache, Malaise (appetite loss, generally unwell, fatigue, physical weakness), Myalgia (muscle pain), Chest congestion, Cough, Runny nose, Shortness of breath or difficulty breathing (resting or on exertion), Sore throat, Wheezing, Eye irritation or discharge, Chills, Fever (≥38.0°C or ≥100.4°F), Pulse oximetry value ≤95%, which is a decreasefrom baseline, Heart rate ≥90 beats/minute at rest, which is an increasefrom baseline. Gastrointestinal symptoms(diarrhea, vomiting, nausea, abdominal pain), Neurologicsymptoms (numbness, difficulty forming or understanding speech), Red or bruised looking toes, Skin rash, Taste loss or new/changing sense of smell, Symptoms of blood clots: pain/cramping, swelling or redness in your legs/calves, Confusion, Bluish lips or face As several of the pre-specified criteria for suspected COVID-19 overlap with vaccine-related reactogenicity, investigators' clinical judgement is required to exclude vaccine-related eventswhen assessing suspected COVID-19.

12. 10.3.3. Informed Consent Process Consentof each participant must be obtained according to local requirements after the nature of the study has been fully explained. The informed consent(s)must be obtained before performance of any study-related procedure. The informed consent should be in accordance with principles that originated in the Declaration of Helsinki, 2. It is the duty of the physicianto promoteand safeguard the health of the people. The physician’s knowledge and conscienceare dedicated to the fulfilment of this duty. 22. The subject should be informed of the right to abstain from participation in the study or to withdraw consent to participate at any time without reprisal. After ensuring that the subject has understood the information, the physician should then obtain the subject’s freely-given informed consent, preferably in writing.

13. People of any age with the following conditions are at increased risk of severe illness from COVID-19: Cancer  Chronic kidney disease  COPD (chronic obstructive pulmonary disease)  Immunocompromised state (weakened immune system) from solid organ transplant  Obesity(body mass index [BMI] of 30 or higher)  Serious heart conditions, such as heart failure, coronary artery disease, or cardiomyopathies  Sickle cell disease  Type 2 diabetes mellitus