Ivt Mrna Therapies: Timelines

* * * TRANSIENT

ivt mRNA ACGU ivt mRNA Cap1-ACGY VACCINATION (TLR agonist) GENE THERAPY

Anti-Cancer Expression of therapeutic protein Anti-Virus e.g. erythropoeitin, antibodies, bispecific Anti-Allergy Cellular reprogramming Genome engineering (Meganuclease, TALEN, CRISPR/CAS9)

VACCINATION (i.m.)

O’Neill et al, 2013. > mRNA IS VERY STABLE (in the abscence of RNases!)

CureVac Wins Two Million EUR from Inaugural European Commission Vaccine Prize TÜBINGEN, Germany/BRUSSELS, Belgium, 10 March 2014 – CureVac, a German clinical stage biopharmaceutical company, today announced that it has won the inaugural European Commission Vaccine Prize. The EU has dedicated the prize of two million Euros in an effort to stimulate innovative solutions for vaccine transportation and storage where cold chain cannot be guaranteed. CureVac’s RNActive® vaccine technology, based on messenger RNA (mRNA), convinced the panel of judges of its potential as a novel vaccine platform with the ability to revolutionize the way vaccines will be developed, manufactured and distributed around the world. As a result of their exceptional stability, RNActive® vaccines eliminate the demand for cold chain logistics.

> RNA-liposome formulations may not be stable (aggregate, change size/form over time or by freeze/thaw)

> RNA in liposome is already an approved drug: Onpattro (Patisiran). Up to 30 mg i.v. every 3 weeks. siRNA. Treatment of polyneuropathy in people with hereditary transthyretin-mediated amyloidosis.

> ivt mRNA vaccines are vegan Mumps (attenuated Virus) Measles (attenuated Virus)

RNA

Lipid Membrane

Rubella (attenuated Virus) ivt mRNA vaccine Production of ivt mRNA

pDNA Fullfill specifications Starting material RNA polymerase Restriction enzyme DNase Nucleotides Buffers Cap analog

First step: Linearisation Restriction enzyme (not needed if PCR) Robust. Two hours

Second step: Transcription T7/SP6 RNA polymerase Robust. Two hours

HPLC Third step: Degradation of DNA DNase Robust. Two hours

Fourth step: Purification of mRNA LiCl precipitation:HPLCHPLC Robust. Two hours

Pascolo. Messenger RNA: The Inexpensive Biopharmaceutical. JMEST. Vol. 4 Issue 3, March - 2017 HPLC HPL C

HPLC

HPLC ivt mRNA therapies: Timelines

Wolff et al describe that direct injection of naked mRNA in mouse muscle gives protein expression

Martinon et al use mRNA in liposomes to vaccinate mice

Conry et al use naked mRNA to vaccinate mice against cancer

Hoerr et al use naked and Protamine formulated mRNA to vaccinate mice

Founding of CureVac

1990 1993 1995 1998 1999 2004 2006 2008 2010

First ivt mRNA inj. in human Herstellungserlaubnis GMP production mRNA

First mRNA vaccine clinical study (naked mRNA) published

Founding of BioNTech

Founding of Moderna Hans-Georg Rammensee Gunther Jung Ingmar Hörr Florian von der Mülbe Steve Pascolo Basic Copyright Notice & Disclaimer

©2020 This presentation is copyright protected. All rights reserved. You may download or print out a hard copy for your private or internal use. You are not permitted to create any modifications or derivatives of this presentation without the prior written permission of the copyright owner.

This presentation is for information purposes only and contains non-binding indications. Any opinions or views expressed are of the author and do not necessarily represent those of Swiss Re. Swiss Re makes no warranties or representations as to the accuracy, comprehensiveness, timeliness or suitability of this presentation for a particular purpose. Anyone shall at its own risk interpret and employ this presentation without relying on it in isolation. In no event will Swiss Re be liable for any loss or damages of any kind, including any direct, indirect or consequential damages, arising out of or in connection with the use of this presentation. ivt mRNA vaccine against SARS-CoV-2 in liposomes, intramuscular

CureVac: Full Length mutated (2 Prolines to keep “pre-fusion conformation”) Spike, non-modified mRNA, 2mg to 12mg; 12mg per dose in Phase III

Moderna: Full Length mutated (2 Prolines to keep “pre-fusion conformation”) Spike, modified mRNA, Injected first volunteer on March 16 > 25 μg, 100 μg, or 250 μg. There were 15 participants in each dose group. An mRNA Vaccine against SARS-CoV-2 - Preliminary Report. N Engl J Med. 2020 Nov Safety and Immunogenicity of SARS-CoV-2 mRNA-1273 Vaccine in Older Adults. N Engl J Med. 2020 Sep 29

BioNTech/Pfizer: 4 mRNA vaccines - Full Length Mutated Spike (modified and unmodified mRNA “pre-fusion conformation”) - RBD (modified mRNA) - Replicating mRNA Injected first volunteer on April 23 > RBD mRNA vaccine at 10μg, 30μg,100μg (35 in US); 1μg 10μg, 30μg, 50μg, 60μg (60 in Germany) Safety and Immunogenicity of Two RNA-Based Covid-19 Vaccine Candidates. N Engl J Med. 2020 Oct 14 Phase I/II study of COVID-19 RNA vaccine BNT162b1 in adults. Nature. 2020 Oct;586(7830):589-593. COVID-19 vaccine BNT162b1 elicits human antibody and TH1 T cell responses. Nature. 2020 Oct Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine. NEJM. 2020 Dec

Safe and well tolerated although there were some side effects up to grade III at higher doses ALL VOLUNTEERS SEROCOVERTED – NEUTRALISING ANTIBODIES AFTER BOOST

Moderna (100mg per dose): 30,000 participants (50% placebo), 196 COVID cases (185 in placebo), 30 severe (30 placebo)

BioNTech (30mg per dose): Side effects: Related adverse events: 21% in vaccine and 5% in placebo Four related serious adverse events were reported among BNT162b2 recipients (shoulder injury related to vaccine administration, right axillary lymphadenopathy, paroxysmal ventricular arrhythmia, and right leg paresthesia). Severe fatigue was observed in approximately 4% of BNT162b2 recipients. Temperature (38.9 to 40°C) in 0.8% (vaccine) and 0.1% (placebo) after the second dose. Efficacy: 9 cases of Covid-19 at least 7 days after the second dose were observed among vaccine recipients and 169 among placebo recipients Basic Copyright Notice & Disclaimer