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-GREECE ΑΔΤ
ΑΙ 986650/ΤΑ Χαλκίδος, ΑΦΜ 059325986 ΔΟΥ
Χαλκίδος τηλ.22210-62743, e-mail: dr.dim.antoniou@gmail.com
www.zoidosia.blogspot.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 Εισαγγελέα
adpeiraias@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
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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