Ya bana ne Ali\u2019den, Veli\u2019den, Ay\u015fe\u2019den \u2013 Fatma\u2019dan\u2026
\nHans\u2019dan \u2013 Kurt\u2019dan\u2026
\nNamusum ve \u015ferefim \u00fczerine yemin ederim, bakm\u0131yorum, ilgilenmiyorum\u2026
\nSadece dikkatimi \u00e7ekenler, tepemi att\u0131ranlar\u2026
\n>>> Namusum ve \u015ferefim \u00fczerine yemin ederim <<<
\nBen\u2026
\nSeviniyorum okuyor insanlar diye\u2026
\n\u00d6zellikle gen\u00e7ler\u2026
\nBelki\u2026
\nBir faydam olur diye, ONLAR\u2026
\nGelece\u011fimiz.<\/p>\n
*<\/p>\n
Ve KORONA ve profesyonel sporcular. Gittik\u00e7e onlarda bu konudan muzdarip\u2026 Onlar bu durumdaysa\u2026 *<\/p>\n „The EMA covid-19 data leak, and what it tells us about mRNA instability Leaked documents show that some early commercial batches of Pfizer-BioNTech\u2019s covid-19 vaccine had lower than expected levels of intact mRNA, prompting wider questions about how to assess this novel vaccine platform, writes Serena Tinari<\/p>\n As it conducted its analysis of the Pfizer-BioNTech covid-19 vaccine in December, the European Medicines Agency (EMA) was the victim of a cyberattack.1 More than 40 megabytes of classified information from the agency\u2019s review were published on the dark web, and several journalists\u2014including from The BMJ\u2014and academics worldwide were sent copies of the leaks. They came from anonymous email accounts and most efforts to interact with the senders were unsuccessful. None of the senders revealed their identity, and the EMA says it is pursuing a criminal investigation.<\/p>\n The BMJ has reviewed the documents, which show that regulators had major concerns over unexpectedly low quantities of intact mRNA in batches of the vaccine developed for commercial production.<\/p>\n EMA scientists tasked with ensuring manufacturing quality\u2014the chemistry, manufacturing, and control aspects of Pfizer\u2019s submission to the EMA\u2014worried about \u201ctruncated and modified mRNA species present in the finished product.\u201d Among the many files leaked to The BMJ, an email dated 23 November by a high ranking EMA official outlined a raft of issues. In short, commercial manufacturing was not producing vaccines to the specifications expected, and regulators were unsure of the implications. EMA responded by filing two \u201cmajor objections\u201d with Pfizer, along with a host of other questions it wanted addressed.<\/p>\n The email identified \u201ca significant difference in % RNA integrity\/truncated species\u201d between the clinical batches and proposed commercial batches\u2014from around 78% to 55%. The root cause was unknown and the impact of this loss of RNA integrity on safety and efficacy of the vaccine was \u201cyet to be defined,\u201d the email said.<\/p>\n Ultimately, on 21 December, EMA authorised Pfizer-BioNTech\u2019s vaccine. The agency\u2019s public assessment report, a technical document published on its website, noted, \u201cthe quality of this medicinal product, submitted in the emergency context of the current (covid-19) pandemic, is considered to be sufficiently consistent and acceptable.\u201d2<\/p>\n It\u2019s unclear how the agency\u2019s concerns were satisfied. According to one of the leaked emails dated 25 November, positive news had come from an undisclosed source in the US: \u201cThe latest lots indicate that % intact RNA are back at around 70-75%, which leaves us cautiously optimistic that additional data could address the issue,\u201d the email said.<\/p>\n A near miss? EMA says the leaked information was partially doctored, explaining in a statement that \u201cwhilst individual emails are authentic, data from different users were selected and aggregated, screenshots from multiple folders and mailboxes have been created, and additional titles were added by the perpetrators.\u201d3<\/p>\n But the documents offer the broader medical community a chance to reflect on the complexities of quality assurance for novel mRNA vaccines, which include everything from the quantification and integrity of mRNA and carrier lipids to measuring the distribution of particle sizes and encapsulation efficiency. Of particular concern is RNA instability, one of the most important variables relevant to all mRNA vaccines that has thus far received scant attention in the clinical community. It is an issue relevant not just to Pfizer-BioNTech\u2019s vaccine but also to those produced by Moderna, CureVac, and others,4 as well as a \u201csecond generation\u201d mRNA vaccine being pursued by Imperial College London.5<\/p>\n RNA instability is one of the biggest hurdles for researchers developing nucleic acid based vaccines. It is the primary reason for the technology\u2019s stringent cold chain requirements and has been addressed by encapsulating the mRNA in lipid nanoparticles (box).<\/p>\n \u201cThe complete, intact mRNA molecule is essential to its potency as a vaccine,\u201d professor of biopharmaceutics Daan J.A. Crommelin and colleagues wrote in a review article in The Journal of Pharmaceutical Sciences late last year. \u201cEven a minor degradation reaction, anywhere along a mRNA strand, can severely slow or stop proper translation performance of that strand and thus result in the incomplete expression of the target antigen.\u201d6<\/p>\n Crommelin and colleagues note that specific regulatory guidance for mRNA based vaccines has yet to be developed, and The BMJ\u2019s attempts to clarify current standards were unsuccessful.<\/p>\n Transparency and confidentiality The Medicines and Healthcare products Regulatory Agency, the UK\u2019s medicines regulator, acknowledged the lack of a specified percentage RNA integrity, but declined to provide further detail. \u201cThe specification limit acceptance criteria are commercially confidential,\u201d the agency said in an email.<\/p>\n The US Food and Drug Administration (FDA) directed The BMJ to read its guidance documents78 and its review of Pfizer\u2019s vaccine,9 but none of these specify the percentage RNA the agency is requiring. Asked to comment, the regulator pointed to Pfizer: \u201cinformation that you seek that is not addressed in the FDA Review Memorandum should be directed to Pfizer.\u201d<\/p>\n In subsequent correspondence, FDA, EMA, and Canadian government department Health Canada all stated that specific information related to the acceptability criteria is confidential.<\/p>\n EMA did acknowledge, however, that vaccine efficacy depends on the presence of suitable amounts of intact mRNA. In the case of the commercial batches that first raised alarm bells, the agency told The BMJ that the levels of truncated mRNA \u201cand the amounts of a potential protein produced by the truncated mRNA would be too low to constitute a safety risk.\u201d EMA did not comment on how truncated mRNA might affect efficacy. The issue was satisfactorily addressed, the agency underlined, when further information was supplied by the manufacturer.<\/p>\n Health Canada told The BMJ that Pfizer had conducted investigations into the root cause of reduced integrity in the commercial vaccine batches, and \u201cchanges were made in their processes to ensure that the integrity was improved and brought in line with what was seen for clinical trial batches.\u201d Health Canada said the three agencies subsequently determined that \u201cthere was no concern with the RNA integrity or any other product specifications.\u201d<\/p>\n Correspondence in the leaked documents suggests that FDA, Health Canada, and EMA were aligned on clinically qualified specifications of percentage mRNA integrity. Health Canada has confirmed to The BMJ that regulators \u201chave worked together to align those requirements,\u201d but all agencies declined to share with The BMJ any specifics on grounds that such information was commercially sensitive.<\/p>\n Pfizer also declined to comment on what percentage mRNA integrity it is aiming for, nor would it address questions about the cause of the unexpectedly low percentage mRNA integrity in certain batches, leaving open the question of whether it could happen again. Pfizer stressed: \u201cEach batch of vaccines is tested by the official medicinal control laboratory\u2014the Paul Ehrlich Institute in Germany\u2014before final product release. As a result, the quality of all vaccine doses that are placed on the market in Europe has been double tested to ensure compliance with the specifications agreed upon with the regulatory authorities.\u201d<\/p>\n Moderna\u2019s chief corporate affairs officer Ray Jordan declined to respond to any of The BMJ\u2019s questions, stating: \u201cAt this point, Moderna will not be offering additional commentary on these topics.\u201d<\/p>\n CureVac, whose mRNA vaccine was submitted for EMA\u2019s \u201crolling review\u201d in February,10 told The BMJ that \u201cit is too soon to give details.\u201d<\/p>\n The shortage of information may reflect the lack of certainty, even among regulators, about how to assess the evidence fully for this novel technology. Professor Crommelin told The BMJ that, \u201cFor small, low molecular weight products, the active pharmaceutical ingredient integrity is typically close to 100%.\u201d<\/p>\n But for mRNA vaccines? \u201cExperience with mRNA integrity is limited.\u201d<\/p>\n Lipid nanoparticles\u2014where do they go and what do they do? Over the years, researchers attempted to resolve intrinsic instability by encapsulating mRNA in nanocarriers made of polymers, lipids, or inorganic materials. Lipid nanoparticles (LNPs) were chosen by Moderna, Pfizer-BioNTech, CureVac, and Imperial College London for their covid-19 vaccines. This has attracted the attention of specialists in the field of pharmaceutical biotechnology, some of whom have raised concerns about further unknowns.<\/p>\n In a rapid response posted on bmj.com, JW Ulm, a gene therapy specialist who has published on tissue targeting of therapeutic vectors,13 raised concerns about the biodistribution of LNPs: \u201cAt present, relatively little has been reported on the tissue localisation of the LNPs used to encase the SARS-CoV-2 spike protein-encoding messenger RNA, and it is vital to have more specific information on precisely where the liposomal nanoparticles are going after injection.\u201d14<\/p>\n It is an unknown that Ulm worries could have implications for vaccine safety.<\/p>\n Ulm told The BMJ: \u201cPfizer-BioNTech and Moderna did a remarkable job of rapidly scaling up manufacturing of such a novel system in swift fashion, which is genuinely a landmark technological achievement. However, pharmacokinetic studies, with independent laboratory confirmation, are essential to ascertain potential cytotoxicity and macroscopic toxicity, especially given the likelihood of booster injections over months or years, since the tissue trafficking patterns of the mRNA vaccine payload will determine which cells and tissues are killed by cytotoxic T-cells in each round.\u201d Given the variation in LNP formulations, it is unclear how relevant previous animal experiments are to answering this question.<\/p>\n Regulators and manufacturers contacted by The BMJ for this article did not wish to address any of the questions raised by Ulm\u2019s rapid response.<\/p>\n Footnotes Provenance and peer review: commissioned; externally peer reviewed<\/p>\n This article is made freely available for use in accordance with BMJ’s website terms and conditions for the duration of the covid-19 pandemic or until otherwise determined by BMJ. You may use, download and print the article for any lawful, non-commercial purpose (including text and data mining) provided that all copyright notices and trade marks are retained.<\/p>\n https:\/\/bmj.com\/coronavirus\/usage https:\/\/www.bmj.com\/content\/372\/bmj.n627<\/p>\n T\u00fcrkiyeli\u2026 Ya bana ne Ali\u2019den, Veli\u2019den, Ay\u015fe\u2019den \u2013 Fatma\u2019dan\u2026 Hans\u2019dan \u2013 Kurt\u2019dan\u2026 Namusum ve \u015ferefim \u00fczerine yemin ederim, bakm\u0131yorum, ilgilenmiyorum\u2026 Sadece dikkatimi \u00e7ekenler, tepemi att\u0131ranlar\u2026 >>> Namusum ve \u015ferefim \u00fczerine yemin ederim <<< Ben\u2026 Seviniyorum okuyor insanlar diye\u2026 \u00d6zellikle gen\u00e7ler\u2026 Belki\u2026 Bir faydam olur diye, ONLAR\u2026 Gelece\u011fimiz. * Ve KORONA ve profesyonel sporcular. Gittik\u00e7e onlarda bu … <\/p>\n
\nLong CoVid\u2026
\nKALP, kalp, kalp\u2026
\nCi\u011ferler!<\/p>\n
\nSenin vay haline!<\/strong><\/p>\n
\nBMJ 2021; 372 doi: https:\/\/doi.org\/10.1136\/bmj.n627 (Published 10 March 2021)
\nCite this as: BMJ 2021;372:n627<\/p>\n
\nIt\u2019s also unclear whether the events in November constitute a near miss in the commercial manufacturing of mRNA vaccines.<\/p>\n
\nThe BMJ asked Pfizer, Moderna, and CureVac, as well as several regulators, what percentage mRNA integrity they consider acceptable for vaccines against covid-19. None offered any specifics.<\/p>\n
\nConceived three decades ago, RNA based therapeutics11 have long inspired imaginations for their theoretical potential to transform cells of the body into \u201can on-demand drug factory.\u201d12 But despite heavy investment by the biotech industry, bench-to-bedside translation was constantly hindered by the fragility of mRNA.<\/p>\n
\nCompeting interests: I have read and understood the BMJ Group policy on declaration of interests and have no relevant interests to declare.<\/p>\n
\nReferences
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\n\u21b5European Medicines Agency. European public assessment report (EPAR) 2020. https:\/\/www.ema.europa.eu\/en\/documents\/assessment-report\/comirnaty-epar-public-assessment-report_en.pdf
\n\u21b5European Medicines Agency. Cyberattack on EMA\u2014update 6. (press release). 2021. https:\/\/www.ema.europa.eu\/en\/news\/cyberattack-ema-update-6
\n\u21b5World Health Organization. The covid-19 candidate vaccine landscape. 2021. https:\/\/www.who.int\/publications\/m\/item\/draft-landscape-of-covid-19-candidate-vaccines
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\n\u21b5US Food and Drug Administration. Emergency use authorization (EUA) for an unapproved product review memorandum. 2020. https:\/\/www.fda.gov\/media\/144416\/download
\n\u21b5European Medicines Agency. EMA starts rolling review of CureVac\u2019s covid-19 vaccine (CVnCoV) (press release). 2021. https:\/\/www.ema.europa.eu\/en\/news\/ema-starts-rolling-review-curevacs-covid-19-vaccine-cvncov
\n\u21b5Sahin U, Karik\u00f3 K, T\u00fcreci \u00d6. mRNA-based therapeutics–developing a new class of drugs. Nat Rev Drug Discov2014;13:759-80. doi:10.1038\/nrd4278 pmid:25233993CrossRefPubMedGoogle Scholar
\n\u21b5Garde D. The story of mRNA: how a once-dismissed idea became a leading technology in the covid vaccine race. STAT News 2020. https:\/\/www.statnews.com\/2020\/11\/10\/the-story-of-mrna-how-a-once-dismissed-idea-became-a-leading-technology-in-the-covid-vaccine-race\/
\n\u21b5Ulm JW, Perron M, Sodroski J, Mulligan RC. Complex determinants within the Moloney murine leukemia virus capsid modulate susceptibility of the virus to Fv1 and Ref1-mediated restriction. Virology2007;363:245-55.CrossRefPubMedGoogle Scholar
\n\u21b5Ulm JW. Rapid response. Re: Will covid-19 vaccines save lives? Current trials aren\u2019t designed to tell us. BMJ 2020. https:\/\/www.bmj.com\/content\/371\/bmj.m4037\/rr-19“<\/p>\n
\nHadi demi\u015f, hadi\u2026
\nHadi oradan, zibidi!<\/strong><\/p>\n","protected":false},"excerpt":{"rendered":"