ΑΠΟΚΑΛΥΠΤΟΥΜΕ πως παρασκευάζεται το εμβόλιο με τεχνητή γενετική τροποποίηση του γνησίου φυσικού ιού στο εργαστήριο, δια της τεχνολογίας recombinant technology.
Τα εμβόλια αυτά που είναι "γενετικά τροποποιημένοι οργανισμοί" για να πάρουν άδεια κυκλοφορίας στην Ευρώπη από την Κομισιόν μόνο, απαιτούν να έχουν γίνει πρώτα εκτεταμένες και πολύχρονες περιβαλλοντολογικές μελέτες για τους κινδύνους που προκύπτουν από την χρήση των για το περιβάλλον και τον άνθρωπο, σύμφωνα με το αρ.6 του Κανονισμού 726/2004 και τις Οδηγίες 2001/18/ΕΚ και 2001/83/ΕΚ. Τέτοιες μελέτες δεν έχουν φυσικά γίνει και γι αυτό το εμβόλιο αυτό, όσο κι αν φαίνεται ΑΠΙΣΤΕΥΤΟ, ΔΕΝ ΕΧΕΙ ΑΔΕΙΑ ΚΥΚΛΦΟΡΙΑΣ ΚΑΙ ΧΡΗΣΗ, διότι τέτοια άδεια ΔΕΝ έχει δημοσιευτεί στην Εφημερίδα της ΕΕ (αντίστοιχο ΦΕΚ), όπως προβλέπεται στο αρ.13.2 Κανονισμού 726/2004.
Κι όχι μόνο αυτό: Στην παρ. 2.3.4 (σελ.51) της έκθεσης της ΕΜΑ, αναφέρεται ότι περιβαλλοντολογική αξιολόγηση δεν απαιτείται!! « Ecotoxicity/environmental risk assessment As the active substance is a vaccine product (which additionally is based on naturally degradable mRNA and lipids), no ERA is considered necessary» (σελ.51), ενώ στην σελ.50 αναφέρεται ότι δεν έχει γίνει ούτε μελέτη για τις επιπτώσεις που έχει το εμβόλιο στα αναπαραγωγικά κύτταρα (ωάρια και σπερματοζωάρια): Genotoxicity No genotoxicity studies have been provided. This is acceptable as the components of the vaccine formulation are lipids and RNA that are not expected to have genotoxic potential (σελ.50).
Σας χρησιμοποιούν ως πειραματόζωο..
Δημ. Αντωνίου
ΣΗΜΑΝΤΙΚΟ:
Η παρακάτω περιγραφή κατασκευής του εμβολίου, που περιγράφει την μέθοδο
κατασκευής του και τα άλλα στοιχεία που προαναφέρω, δημοσιεύεται στην
επίσημη έκθεση της ΕΜΑ (Ευρωπαικός ιός) ΑΛΛΑ δεν δημοσιεύεται στην Ελληνική του μετάφραση !
Δείτε εδώ ολόκληρη την έκθεση της ΕΜΑ...
ΕΞΑΙΡΕΤΙΚΑ ΕΠΕΙΓΟΥΣΑ ΑΝΑΦΟΡΑ
Του Δρ. Δημητρίου Αντωνίου PhD, FRCS, Χειρουργού
Δ/νση: Οδός Δημ. Βώκου 6, Χαλκίδα ΤΚ 34132- ΕΛΛΑΔΑ-GREECEΑΔΤ ΑΙ 986650/ΤΑ Χαλκίδος, ΑΦΜ 059325986 ΔΟΥ Χαλκίδος
τηλ.22210-62743, e-mail: dr.dim.antoniou@gmail.com
ΘΕΜΑ:
- Έκθεση σε άμεσο κίνδυνο της Υγείας και Ζωής των εμβολιασθέντων με εμβόλιο που είναι «γενετικά τροποποιημένος οργανισμός» χωρίς ΑΔΕΙΑ από τις αρμόδιες Ευρωπαικές (κι Ελληνικές) Αρχές.
- Εισαγγελική έρευνα για τις αιτίες εισαγωγής στην ΜΕΘ του Νοσ/μείου Νίκαιας Πειραιά, Υποδ/του Νοσ/μείου Αττικής, εμβολιασθέντος πριν λίγες μέρες με το παραπάνω παράνομο εμβόλιο
ΠΡΟΣ:
Αστυνομική Δ/νση Πειραιά και Δυτ. Αττικής
Για τον αρμόδιο Εισαγγελέαadpeiraias@hellenicpolice.gr dapeiraia.public@astynomia.gr dadattikis@astynomia.gr
Αξιότιμε Κε Διοικητά, Κε Εισαγγελέα
Η Ευρωπαική Επιτροπή, με επικεφαλής την Πρόεδρό της της Ursula von der Leyen αγόρασε από την Pfizer, διένειμε σε όλη την Ευρώπη και παρότρυνε την χορήγησή του εμβολίου BNT162b2, το οποίο είναι «γενετικά τροποποιημένος ορφανισμός», χωρίς να πληρούνται οι προϋποθέσεις προστασίας του ανθρωπο-περιβάλλοντος του Κανομισμού 726/2004.
Έτσι, άρχισαν ήδη στην Ελλάδα, με την άμεση συνέργεια του Υπουργού Υγείας και Γ.Γ. Πολιτικής προστασίας και θα επεκταθούν τις επόμενες ημέρες, όπως ανακοινώθηκε από τον αρμόδιο υπουργό Υγείας, οι εμβολιασμοί σε διάφορα νοσοκομεία της χώρας με το εμβόλιο BNT162b2.
Ο εμβολιασμός όμως αυτός στην Ελλάδα με το εμβόλιο BNT162b2, διεξάγεται χωρίς να τηρούνται οι προϋποθέσεις κυκλοφορίας και χρήσης φαρμάκων-εμβολίων που είναι «γενετικά τροποποιημένοι-μεταλλαγμένοι οργανισμοί», επιπλέον δε –και το σημαντικότερο- διενεργείται και ΧΩΡΙΣ ΑΔΕΙΑ από τις αποκλειστικά αρμόδιες αρχές της ΕΕ, ήτοι την Επιτροπής (Commission), όπως απαιτούν οι παρακάτω Ευρωπαϊκές Οδηγίες και Κανονισμοί, που συνιστούν αντίστοιχα και στοιχεία του ελληνικού δικαίου, και ισχύουσες πάσης αντίθετης εθνικής διάταξης:
- αρ. 1,1 παρ. 4,5 παρ.2,3,6 της Οδηγίας 2001/83/ΕΚ/06-11-2001), Παράρτημα Ι Μέρος
ΙV παρ. 3.2.1.1.
- αρ.1,2 παρ.4,12-19-24 της Οδηγίας 2001/18/ΕΚ/12-03-2011, Παράρτημα ΙΑ
και ΙΙ,ΙΙΙ
- αρ.2,3,4,6 παρ.2α,γ,3 και αρ. 10,12, 14 παρ.7,8,9 Κανονισμού 726/31-03-2004, που διαβάζεται σε συνδυασμό με τις δύο προαναφερόμενες Οδηγίες και του ν.1316/1983.
Η άδεια εμβολίου για την Ευρώπη δημοσιεύεται στην επίσημη εφημερίδα της ΕΕ (αρ. 13.2 Κανονισμού 726/2002) και τέτοια δημοσίευση ΔΕΝ υπάρχει.
Κι ο λόγος έλλειψης άδειας είναι ακριβώς η γνώση των κρατικών υπευθύνων για την επικινδυνότητα του και προς αποφυγή των οποιονδήποτε συνεπειών εις βάρος των από τις συνέπειες της διεξαγωγής του.
Για τους λόγους αυτούς, ο εμβολιασμός ΟΛΩΝ εγκυμονεί άμεσους και θανατηφόρους κινδύνους για την ζωή των κι ανυπολόγιστων συνεπειών στο περιβάλλον, οργανικό μέρος του οποίου είναι κι ο άνθρωπος.
ΑΝΑΖΗΤΕΙΣΤΕ ΤΗΝ ΑΔΕΙΑ ΚΥΚΛΟΦΟΡΙΑΣ ΤΟΥ ΕΜΒΟΛΙΟΥ ΚΙ ΕΜΒΟΛΙΑΣΜΟΥ…κι επειδή ΔΕΝ υπάρχει, λάβετε όλα τα αναγκαία μέτρα διακοπής του και τιμωρίας των υπαιτίων για την απόπειρα αυτή γενοκτονίας που επιχειρούν, από αύριο Δευτέρα 04-01-2021 στο Παν/κό Νοσ/μείο Ηρακλείου, Ρεθύμνου και Χανίων, με την συνέργεια των Δ/των των νοσοκομείων αυτών και των υπαιτίων ιατρών.
ΕΙΣΩΓΩΓΗ ΣΤΗΝ ΜΕΘ Εμβολιασθέντος Υποδ/του Νοσοκομείου
Όλο το πανελλήνιο κατέστη μάρτυρας του δημόσιου εμβολιασμού την 27-12-2020 της ΠτΔ, Πρωθ/ργού κι άλλων πολιτικών και συνεχίστηκαν τις επόμενες ημέρες με τον εμβολιασμού ιατρών και νοσηλευτών σε νοσ/μεία της Αθήνας.
Σήμερα 03-01-2021 ανακοινώθηκε ότι εμβολιασθής Υποδ/της «μεγάλου Νοσ/μείου της Αττικής» (τα δημοσιεύματα δεν αναφέρουν το όνομά του ή το νοσ/μείο που υπηρετεί) 55 μόλις ετών, κατέρρευσε ξαφνικά και νοσηλεύεται στην ΜΕΘ του νοσοκομείου Νίκαιας του Πειραιά, ΠΙΘΑΝΩΣ από παρενέργειες του παράνομου αυτού εμβολίου.
Για τον λόγο αυτό και λόγω της σοβαρότητας του θέματος για την δημόσια υγεία όλων των Ελλήνων, ζητώ από τον εισαγγελέα να ερευνήσει εάν προκύπτουν ποινικές ευθύνες για πρόκληση επικίνδυνης σωματικής βλάβης με ενδεχόμενο δόλο στον ιατρό αυτό δια της διενέργειας του εμβολισμού του με το παραπάνω παράνομο και χωρίς άδεια εμβόλιο, κατά των υπαιτίων, ήτοι του Δ/του του Νος/μείου που έγινε το εμβόλιο, του επιβλέποντος ιατρού (-ών) του εμβολιασμού και των υπευθύνων πολιτικών προσώπων προμήθειας και «διαταγής» για την διενέργεια του εμβολιασμού.
Ταυτόχρονα ζητώ να ληφθούν άμεσα όλα τα μέτρα αποφυγής παρόμοιων συμβάντων.
Μάρτυρας: Σωτήρης Τσιόδρας, καθηγητής Θριάσιου Νοσ/μείου, που επελήφθη του περιστατικού αυτού, όπως δημοσιεύτηκε.
Χαλκίδα 03-01-2021 Δημ. Αντωνίου
Περιγραφή κατασκευής του εμβολίου, που περιγράφει την μέθοδο κατασκευής του Δείτε εδώ ολόκληρη την έκθεση της ΕΜΑ...
EMA ASSESSMENT
About the product BNT162b is a mRNA vaccine for prevention of COVID-19. The vaccine is made of a mRNA encoding for the full-length SARS-CoV-2 spike glycoprotein (S) encapsulated in lipid nanoparticles (LNPs). The sequence of the S protein was chosen based on the sequence for the “SARS-CoV-2 isolate Wuhan-Hu1”, which was available when the program was initiated: GenBank: MN908947.3 (complete genome) and GenBank: QHD43416.1 (spike surface glycoprotein). The active substance consists of a single-stranded, 5'-capped mRNA that is translated into a codonoptimized sequence encoding the spike antigen of SARS-CoV-2. The RNA contains common structural elements optimized for mediating high RNA stability and translational efficiency (see section 2.2). The LNPs protect the RNA from degradation by RNAses and enable transfection of host cells after intramuscular (IM) delivery. The mRNA is translated into the SARS-CoV-2 S protein in the host cell cytosol. The S protein is then expressed on the cell surface where it induces an adaptive immune response. The S protein is identified as a target for neutralising antibodies against the virus and is therefore considered a relevant vaccine component. The vaccine, BNT162b2 (30 µg), is administered intramuscularly (IM) in two 30 µg doses of the diluted vaccine solution given 21 days apart. Intended indication: ‘Active immunisation to prevent COVID-19 disease caused by SARS-CoV-2 virus, in individuals 16 years of age and older’ (σελ.13)
Control of materials
An adequate overview of the raw materials and solutions used in the active substance manufacturing process is provided. Representative certificates of analysis have been provided. The submitted information supports the appropriate quality of raw materials. It is recommended that the applicant should implement relevant testing strategies to ensure an adequate microbiological control for the starting materials (REC1) and should implement a relevant testing strategy to ensure that HEPES (Pfizer) raw material, included in the formulation buffer of FP, is free from contaminating RNases (REC2). The description of synthesis of 5’cap and its related impurities were requested during the procedure. Appropriate information was given. The applicant should implement in-house functional activity analytical methods for release testing of enzymes used in the manufacturing process at all relevant manufacturing sites, by Q1 2021 (REC3). The BNT162b2 active substance is manufactured by in vitro transcription using a linear DNA template, produced via plasmid DNA from transformed Escherichia coli cells. The linear DNA template is not part of the final product but defines the sequence of the mRNA product and therefore it is fundamental to ensure the adequate control of the active substance. Changes to the manufacturing process of the linear DNA template (e.g. change to plasmid host cell) may result in a different impurity profile in the active substance. Additional details on the manufacturing process and the control strategy for this starting material, initially only shortly described, have been provided and the dossier will be updated accordingly. The cell banks involved in the plasmid manufacturing process are described. Master cell bank (MCB) and working cell bank (WCB) qualification tests are listed. Relevant specifications are set and data from the current MCB and WCB are provided. The plasmid MCBs and WCBs are enrolled in a cell bank stability program. The strategy is considered adequate, noting that the dossier will be updated as appropriate. A protocol for establishment of future WCBs is provided. Following fermentation, the cells are harvested and chemically lysed to recover the plasmid DNA. After this lysis step, the circular plasmid DNA is purified. The circular plasmid DNA is filtered and stored frozen. The strategy for establishing the initial shelf-life is endorsed and data provided support the proposed shelf life. A list of the raw materials as well as other materials used in the manufacture of the Assessment report EMA/707383/2020 Page 17/140 linear DNA template is provided. All materials used are animal origin free and sourced from approved suppliers. Specifications for the circular plasmid DNA as well as for the DNA linear template are provided. Process- and product-related impurities including host cell genomic DNA, RNA, proteins, endotoxins, bioburden and plasmid isoforms, for the plasmid DNA, are routinely quantified. The reference material is described. Implementation of any changes in the manufacture of the linear DNA template should be applied for in a variation application. (ΣΕΛ.15-17)
Active substance Overall, the information provided is satisfactory. However, certain information is still pending due to the very short time frame of product development and will either be updated in the dossier imminently or further addressed in specific obligations and other post-approval measures. Information on the manufacturing process and process controls for the manufacturing sites Andover and BNT Mainz & Rentschler have been provided and are considered satisfactory. Two active substance processes have been used during the development; Process 1 and 2. The major changes between AS Process 1 and 2 are: increased process scale, DNA template changed from a PCR template to linearised plasmid DNA, magnetic bead purification replaced with proteinase K digestion and UFDF steps. Based on the differences observed between batches manufactured by active substance Process 1 and 2 for the CQA mRNA integrity and lack of characterisation data, a MO was raised regarding comparability, characterisation and clinical qualification of the one proposed acceptance criteria. Biological characterisation of the active substance was limited, and additional data and discussion were requested to address functionality. Additional characterisation data for the active substance are to be provided to confirm the identities of the observed Western Blot (WB) bands obtained by the in vitro expression assay (SO1). Truncated RNA species are regarded as product-related impurities and can be expected due to the principle of the in-vitro transcription reaction (i.e. directional polymerase activity) and (theoretical) hydrolysis during manufacturing. Analysis of RNase treated samples showed that all species detected Assessment report EMA/707383/2020 Page 33/140 by the capillary gel electrophoresis (CGE)-based method are composed of RNA. Manufacturing consistency with respect to fragmented species has been sufficiently demonstrated. There were some differences in truncated RNA species level, however further analyses revealed a comparable overall fragmentation profile among Process 1 and Process 2 active substance batches. Additionally, oligonucleotide mapping data demonstrated no significant differences observed between Process 1 and Process 2 active substance batches. The company does not expect truncated transcripts formulated in the finished product to pose a safety or efficacy concern, as in their view no protein expression is expected from truncated transcripts. Further, clinical trials with process 1 material have not revealed major safety concerns to date. Truncated BNT162b2 RNA species lacking either the 5’ cap or the poly(A) tail are expected to be rapidly targeted for degradation in the cytoplasm or would show a decrease or loss of translation efficiency. Preliminary characterization data on isolated fragment species suggest that fragment species predominantly include the 5’-cap but lack the poly(A) tail, supporting the hypothesis that most fragments would arise from premature termination in the IVT reaction. However, as the overall characterisation of the truncated species is still limited, additional analysis of truncated species, additional translated protein characterisation, additional characterisation of lipidrelated impurities and potential lipid-RNA species should be provided to support that they are not impacting clinical performance in terms of safety and/or efficacy. The current specification allows for a certain level of truncated mRNA species to be present however data from recent batches have shown levels of truncated species below that level. No related safety issues have been identified in the clinical studies thus far in subjects who received vaccine containing up to a certain level of truncated species. Therefore, the current specification is considered acceptable subject to the submission of additional data in the frame of a specific obligation (SO1). Based on available data, the proposed specification for active substance is acceptable with respect to the attributes chosen for routine release testing. However, the length of the poly(A) tails in BNT162b2 active substance is critical for RNA stability and translational efficiency and therefore should be introduced in active substance release testing in the frame of a specific obligation (SO2). Moreover, the active substance specification acceptance limits should be re-assessed and revised as appropriate, as further data become available from ongoing clinical trials and in line with manufacturing process capability (SO2). It is noted that the Applicant states that testing is currently in progress on the clinical batches and data for these batches, as well as any new data available for the primary process validation batches, will be provided. Based on the limited stability data presented, a shelf-life is approved for the active substance. Finished product The finished product is a preservative-free, multi-dose concentrate to be diluted for intramuscular injection, intended for 5 doses. The finished product is a sterile dispersion of RNA-containing lipid nanoparticles (LNPs) in aqueous cryoprotectant buffer. The formulation development studies of the RNA containing lipid nanoparticles have been thoroughly described including studies that were performed with available active substance, representative of the mRNA platform and included in the finished product. The development of the manufacturing process is extensively described, and critical process parameters are defined. The manufacturing process includes lipid nanoparticle fabrication and bulk finished product formulation followed by fill and finish, and the process has been acceptably described. Assessment report EMA/707383/2020 Page 34/140 Comparability between the commercial finished product and the clinical finished product has been sufficiently demonstrated for the attributes tested and will be subject to a specific obligation. Limited data on the finished product batches manufactured at the commercial facility (entire manufacturing process at the commercial site Pfizer, Puurs, at commercial scale, active substance from process 2) were presented. A process validation plan for PPQ lots has been provided. A concurrent validation approach will be used due to the urgent need for this product and the pandemic situation. The rationale for this approach has been documented. This concurrent approach requires interim reports to be documented for each individual validation run. An overall report per site will be compiled that summarises all evaluations and contains a comparability assessment of the data of all batches manufactured. Finally, a concluding report linked to this plan will be written that summarises all findings from the different validation reports. Further data was requested in order to conclude on the consistency of finished product manufacturing, to assure comparability between the commercial product with the product used in clinical trials, and to support the claimed finished product shelf-life and storage conditions. A process validation plan for PPQ lots has been provided. Process validation (PPQ) for commercial scale batches were initiated, and a summary report from one PPQ validation batch was provided. In summary, given that an acceptable validation program, also comprising the commercial facility at Puurs, Belgium, has been established, and a summary report from one PPQ validation batch was provided, the information on process validation is considered acceptable subject to the agreed specific obligation that the MAH should provide additional validation data (SO3). The specifications for finished product include a comprehensive panel of relevant tests along with corresponding acceptance criteria. Several issues in relation to the acceptance criteria in the finished product specifications were raised, i.e. the LNP size, polydispersity, RNA encapsulation, in-vitro expression and RNA integrity. Whilst FP specifications were subsequently amended and overall found to be acceptable, the acceptance limits should be re-assessed, and revised as appropriate, as further data becomes available from ongoing clinical trials and in line with manufacturing process capability (SO2). Two novel excipients are included in the LNP. Complete information is not provided for both the cationic lipid ALC-0315 and the PEGylated lipid ALC-0159. In order to assure comprehensive control throughout the lifecycle of the finished product and to ensure batch to batch consistency, further information needs to be submitted regarding the synthetic process and control strategy in line with specific obligations (SO4, SO5). Lipid-related impurities have been observed in some recently manufactured finished product batches. For the batches with lipid-related impurities the existing quality control parameters including RNA integrity remain unchanged. Considering the above and the emergency situation, the characterisation of the active substance and finished product is considered acceptable, and the proposed specifications for RNA Integrity and 5’-Cap are considered to be scientifically justified and acceptable. Nevertheless, additional data to complete the characterisation of the active substance and finished product and additional clinical data from batches currently in use in ongoing clinical studies, are considered important to confirm the clinical qualification of these specifications. These data are requested to be provided as specific obligations to the applied conditional marketing authorisation (SO1, SO2). Efficacy, safety and immunogenicity was demonstrated using clinical batches of vaccine from Process 1. The commercial batches are produced using a different process (Process 2), and the comparability of these processes relies on demonstration of comparable biological, chemical and physical characteristics of the active substance and finished product. Assessment report EMA/707383/2020 Page 35/140 The characterisation and control of active substance and finished product are limited in relation to critical quality attributes and impurities. The suitability of the analytical methods used for control of potency and poly(A) tail have not been fully demonstrated. Data demonstrate the presence of significant amounts of truncated/modified forms of mRNA at somewhat higher levels in the batches manufactured with the commercial process as compared to material used in clinical trials. These forms are poorly characterised, and the limited data provided for protein expression do not fully address the uncertainties relating to the risk of translating proteins/peptides other than the intended spike protein. The control strategy for active substance and finished product is important to guarantee acceptable quality and ensure batch to batch consistency of the finished product. Regarding the proposed control strategy, questions were raised both with regard to the suitability of the test methods used and the acceptance criteria for some tests. Based on the above, the following uncertainties are considered to be of importance for the benefit-risk assessment: • Truncated and modified RNA are present as impurities. Considering the low dose of mRNA (30 µg), the impurities are not considered a safety issue based on general toxicological principles. However, when present in the cell there is a possibility that aberrant proteins will be expressed with possibilities for unwanted immunological events. The risk of this occurring is considered low based on the following observations and considerations: o Such impurities were present in the vaccine used in the Phase 3 clinical trials with an acceptable safety profile. Although the lack of characterisation hinders a full comparability evaluation there is no indication that there would be important qualitative differences in the nature of these impurities. o The high levels of these impurities reflect the instability of RNA resulting in generation of RNA fragments both in the transcription step and thereafter. Based on electrophoretic data it appears that there is a diverse set of fragments. Although not confirmed, it is unlikely that these RNA molecules to a large extent would be mRNA molecules with intact 5’-cap and 3’-polyA. o The level of any individual aberrant mRNA species would in any way be magnitudes lower than the level of the intact mRNA and this would be mirrored by the level of protein expression. The amount of the protein would be expected to be too low to elicit an immune response. The spike protein is a highly immunogenic protein and immunodominance would also ascertain that the immune response to the aberrant protein would be non-significant. • Lipid related impurities were observed in recently produced finished product batches. Based on the low dose (30 µg mRNA) it is considered that the amounts of these impurities are too low to be of toxicological significance. (σελ.32-37)
Mechanism of action SARS-CoV-2 infects the body by the use of the Spike protein (S) to attach to specific cell surface receptors, of which the angiotensin converting enzyme 2 (ACE2) may constitute a major part, as recently suggested. In addition to the initial attachment to a host cell, the S protein is also responsible for viral envelope fusion with the host cell membrane resulting in genome release. Due to its indispensable role, the S protein is a major target of virus neutralizing antibodies and has become a key antigen for vaccine development. By immunisation with the modified RNA (modRNA) product BNT162b2, encoding for the S protein, the intention is to trigger a strong and relatively long-lasting production of high affinity virus neutralizing antibodies, which can act through blocking the S-protein and it’s receptor-binding domain (RBD) interaction with host cell receptors but also by opsonisation mediated virus clearance. In addition, the immunisation with BNT162b2 is also intended to elicit a concomitant T cell response of the Th1 type, supporting the B cells responsible for the production of Sspecific antibodies and cytotoxic T cells that kill virus infected cells. Assessment report EMA/707383/2020 Page 42/140 The S protein is a trimeric class I fusion protein that exists in a metastable prefusion conformation before engaging with a target cell. BNT162b2 encodes a P2 mutant (P2 S) variant of S where two consecutive proline mutations have been introduced in order to lock the RBD in the prefusion conformation. In addition, BNT162b2 is nucleoside-modified by a substitution of 1-methylpseudouridine for uridine and thus its inherent adjuvant activity mediated by binding to innate immune sensors such as toll-like receptors (TLRs) 7 and 8, is dampened, but not abrogated. Furthermore, the structural elements of the vector backbones of the BNT162b2 are optimised for prolonged and strong translation of the antigen-encoding RNA. The potency of the RNA vaccine is further optimised by encapsulation of the RNA into lipid nano particles (LNPs), which protects the RNA from degradation by RNAses and enable transfection of host cells after intramuscular (i.m.) delivery. The functional and ionizable lipid, ALC-0315, is identified as the primary driver of delivery as it allows the LNPs to have a neutral charge in a physiological environment to facilitate internalization; the endosomal environment exhibits a positive charge and therefore triggers the translocation of RNA into the cytosol (Midoux & Pichon, 2015; Hassett et al, 2019; Patel et al, 2019); ALC-0159 is included in the formulation to provide a steric barrier to: 1) facilitate the control of particle size and homogeneity during manufacturing and product storage, and 2) regulate the association of plasma and proteins with the LNP surface. The composition of the LNPs may also affect the distribution of injected BNT162b2. In addition, it cannot be excluded the LNP composition contributes to the overall immunogenicity. Administration of LNP-formulated RNA vaccines IM results in transient local inflammation that drives recruitment of neutrophils and antigen presenting cells (APCs) to the site of delivery. Recruited APCs are capable of LNP uptake and protein expression and can subsequently migrate to the local draining lymph nodes where T cell priming occurs. In general, following endocytosis of LNPs, the mRNA is released from the endosome into the host cell cytosol (Sahay et al, 2010; Maruggi et al, 2019). The process of an RNA vaccine-elicited immune response has been demonstrated in both murine and nonhuman primate models (Pardi et al, 2015; Liang et al, 2017) (σελ.41-42)
Genotoxicity No genotoxicity studies have been provided. This is acceptable as the components of the vaccine formulation are lipids and RNA that are not expected to have genotoxic potential (σελ.50)
2.3.4. Ecotoxicity/environmental risk assessment As the active substance is a vaccine product (which additionally is based on naturally degradable mRNA and lipids), no ERA is considered necessary (σελ.51)
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