US Patent #10702600: clues to vaccine composition
Here’s the MRNA patent on their experimental injectable composition. This is a link directly to the claims:
When reading a patent, I go to claim #1 first. It contains the broadest possible description:
A composition, comprising: a messenger ribonucleic acid (mRNA) comprising an open reading frame encoding a betacoronavirus (BetaCoV) S protein or S protein subunit formulated in a lipid nanoparticle.
So: the injectible is an Open Reading Frame, formulated in a Lipid Nanoparticle.
The rest of the claims in that section (2-16) help us to fill in the blanks. The experimental injectible is – possibly – claim #16. Assembling these together, the definition looks like this:
Open Reading Frame: BetaCoV S protein (S1 subunit + S2 subunit), a 5′ untranslated region (UTR) and a 3′ UTR, a poly(A) tail, a 5′ cap analog ( 7mG(5′)ppp(5′)NlmpNp), plus a Chemical Modification (a 1-methylpseudouridine modification or a 1-ethylpseudouridine modification), where at least 80% of the uracil in the open reading frame has a chemical modification.
Lipid Nanoparticle: an ionizable cationic lipid (Compound 25 *), a neutral lipid (DSPC), a sterol (cholesterol), and a PEG-modified lipid (PEG-DMG) or PEG-cDMA).
1. A composition, comprising: a messenger ribonucleic acid (mRNA) comprising an open reading frame encoding a betacoronavirus (BetaCoV) S protein or S protein subunit formulated in a lipid nanoparticle.
2. The composition of claim 1, wherein the open reading frame encodes a BetaCoV S protein.
3. The composition of claim 1, wherein the open reading frame encodes an S protein subunit selected from an S1 subunit and an S2 subunit.
4. The composition of claim 1, wherein the mRNA further comprising a 5′ untranslated region (UTR) and a 3′ UTR.
5. The composition of claim 4, wherein the mRNA further comprises a poly(A) tail.
6. The composition of claim 4, wherein the mRNA further comprises a 5′ cap analog.
7. The composition of claim 6, wherein the 5′ cap analog is 7mG(5′)ppp(5′)NlmpNp.
8. The composition of claim 1, wherein the mRNA comprises a chemical modification.
9. The composition of claim 8, wherein the chemical modification is a 1-methylpseudouridine modification or a 1-ethylpseudouridine modification.
10. The composition of claim 8, wherein at least 80% of the uracil in the open reading frame has a chemical modification.
11. The composition of claim 1, wherein the lipid nanoparticle comprises an ionizable cationic lipid, a neutral lipid, a sterol, and a PEG-modified lipid.
12. The composition of claim 11, wherein the lipid nanoparticle comprises 20-60% ionizable cationic lipid, 5-25% neutral lipid, 25-55% cholesterol, and 0.5-15% PEG-modified lipid.
13. The composition of claim 12, wherein the lipid nanoparticle comprises 50% ionizable cationic lipid, 10% neutral lipid, 38.5% sterol, and 1.5% PEG-modified lipid.
14. The composition of claim 11, wherein the ionizable cationic lipid is Compound 25.
15. The composition of claim 11, wherein the neutral lipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), the sterol is cholesterol, and the PEG-modified lipid is 1,2-dimyristoyl-racalycero-3-methoxypolyethylene glycol-2000 (PEG-DMG) or PEG-cDMA.
From the patent:
A “5′ untranslated region” (5′UTR) refers to a region of an mRNA that is directly upstream (i.e., 5′) from the start codon (i.e., the first codon of an mRNA transcript translated by a ribosome) that does not encode a polypeptide.
A “3′ untranslated region” (3′UTR) refers to a region of an mRNA that is directly downstream (i.e., 3′) from the stop codon (i.e., the codon of an mRNA transcript that signals a termination of translation) that does not encode a polypeptide.
An “open reading frame” is a continuous stretch of DNA beginning with a start codon (e.g., methionine (ATG)), and ending with a stop codon (e.g., TAA, TAG or TGA) and encodes a polypeptide.
A “polyA tail” is a region of mRNA that is downstream, e.g., directly downstream (i.e., 3′), from the 3′ UTR that contains multiple, consecutive adenosine monophosphates …. In a relevant biological setting (e.g., in cells, in vivo) the poly(A) tail functions to protect mRNA from enzymatic degradation, e.g., in the cytoplasm, and aids in transcription termination, export of the mRNA from the nucleus and translation.
And, of course, Compound 25:
There was mention of that flagellin adjuvant; not part of the claims, but definitely part of the description. It may have been a claim that was disallowed by the PTO as being an obvious combination. Is this in there? My guess – just a guess – is yes, because:
Surprisingly, in some aspects, it has also been shown that efficacy of mRNA vaccines can be significantly enhanced when combined with a flagellin adjuvant, in particular, when one or more antigen-encoding mRNAs is combined with an mRNA encoding flagellin.
Who doesn’t want their experimental injectible to be significantly enhanced?
So what’s a flagellin?
Flagellin is an approximately 500 amino acid monomeric protein that polymerizes to form the flagella associated with bacterial motion. Flagellin is expressed by a variety of flagellated bacteria (Salmonella typhimurium for example) as well as non-flagellated bacteria (such as Escherichia coli). Sensing of flagellin by cells of the innate immune system (dendritic cells, macrophages, etc.) is mediated by the Toll-like receptor 5 (TLR5) as well as by Nod-like receptors (NLRs) Ipaf and Naip5. TLRs and NLRs have been identified as playing a role in the activation of innate immune response and adaptive immune response. As such, flagellin provides an adjuvant effect in a vaccine.
Examples further include:
Salmonella typhi (UniPro Entry number: Q56086), Salmonella typhimurium (A0A0C9DG09), Salmonella enteritidis (AOAOC9BAB7), and Salmonella choleraesuis.
So sticking a flagellin subcomponent onto this Open Reading Frame ends up (my guess) being the the equivalent of throwing a piece of meat into the immune system shark tank. “Holy shit guys, its salmonella!” And it responds. In some way.
If you’ve had salmonella before, might you have a different reaction from someone who didn’t? Who knows!
And there’s also the matter of “PEG” allergy (from the “lipid nanoparticle”), which many people in the US have. Just one example:
Injected into tens of millions, around the world. At Warp Speed.
I’ve been reading about Polyethylene Glycol and Potassium Chloride
as ingredients in the various Covid19 vaccines.
Ethylene Glycol being antifreeze … toxic to humans, mammals, birds, etc.
I do wonder about the wisdom of letting oneself be injected with small quantities of Polyethylene Glycol.
Potassium Chloride being the stuff they give during Death Penalty proceedings, official reason “to stop the patient’s heart” … I also wonder about taking even small quantities of it.
I understand, there’s a big difference between a Zima and Everclear (2% vs. 100% ethanol alcohol.)
Would like to hear the life sciences PP STEM’ies weigh in. About the PEG and the KCl.
*Disclaimer: I am not a lab biologist. All of this developed after the completion of my medical school and is outside my specialty reading. So I come at this topic with the background of a lay person.
What is flagelin and what does role does it play?
Role of flagellin in the pathogenesis of shock and acute respiratory distress syndrome: therapeutic opportunities (Abstract)
Author: Csaba Szabó
Objective: To provide an overview of the role of flagellin as an immunostimulatory and proinflammatory factor.
Main results: Flagellin, a protein of 40-60 kD, is the principal constituent of the flagellum, a prominent surface structure found in motile bacteria. Recent work reveals that monomeric flagellin, a protein component of flagellated bacteria, can act as a soluble immunostimulatory and proinflammatory factor, activating the immune/inflammatory axis via the toll-like receptor 5-nuclear factor-kappaB axis. Monocytes, macrophages, and intestinal and pulmonary epithelial cells respond to monomeric flagellin at low concentrations. Monomeric flagellin can induce prominent local and systemic immune/inflammatory responses.
Conclusions: Recognition of the flagellin-toll-like receptor 5 pathway offers novel opportunities for the experimental therapy of various forms of shock, sepsis, and acute respiratory distress syndrome.
So this is like the boy scout building his first campfire who adds a cup of gasoline to the kindling to “help.” Just like you said DaveF.
Several other links at the bottom of the above article explore the mechanism further.
So this theme in vaccine science is recurrent: vaccines are designed with an adjuvant (like a cup of gasoline) added to stimulate the immune system. This is necessary to get a strong immune response from a vaccine. Unfortunately sometimes the stimulation goes too far or in an unexpected direction causing immune attack on tissues of the host.
One example I heard this week from a PP member is Transverse Myelitis, an inflammatory reaction in the spinal cord making a healthy person suddenly unable to walk or control her bladder. Onset 24 hours after her COVID vaccine.
So it seems like some percentage of people are going to have “upstream” or “down stream” preconditions that might mutate or attenuate the intended response into something complete different. Sort of the way injecting stem cells is a little like hoping it catches the right wave (and avoids the sharks). The need for small molecules explains the need for nano compounds.
There’s a Sufi story about a sufi who comes upon a man searching desperately in the dirt for a lost key. The sufi helps search. Eventually the sufi asks “where do you think you lost it?” The man points at the hut, which is dark. The sufi askes then “why are you searching out here?” the man replies; “because it’s too dark in there.”
All I know is that Bill Gates in there and I can’t see what he’s up to but an evil man is most dangerous when he’s “doing well by doing good” (look’n at you Bubba Clinton)
Thanks for the thorough response thread. I’ll dive in when I have more time. So much new technology, so little long-term data.
I had a horrible reaction to shingles vaccine. Discovered today that the adjuvant is QS-21 in a lipid nanoparticle. Apparently, this material, originally derived from tree bark, was so toxic in animal vaccines that it was not approved for humans. But now, they use nanotechnology to enclose it with cholesterol to help prevent it from busting open your cells while you still get a hyper-immune reaction. Emphasis on hyper.
Being prone to allergies, I really don’t need my immune response further enhanced, thanks very much. Will be asking very pointed questions in the future.
We have updated the Covid Data through 1.29.2021. For some reason the interface for VAERS is not updated, but the data is available. So here it is: 172 new deaths last week and 2k more reports. @AlexBerenson https://t.co/OjpVrTpUdB pic.twitter.com/va50nXExtK
— the Arkivist (@the_Arkivist) February 5, 2021
New VAERS data is out. (As of Jan 29th) 172 new deaths added and 2,000 more reports.
Not yet peer reviewed. People who have had COVID have greater reactions to first dose of the vaccine. Naive (unexposed) people display a greater reaction to the second dose.
Raises three questions in my mind:
1) Why would you get the vaccine if you’ve already had COVID? Doesn’t natural immunity count for anything?
2) Is this an example of antibody dependent enhancement? You have natural immunity and react worse when subsequently exposed to spike protein (antigen) that the vaccines causes your body to manufacture?
3) Is the vaccine adjuvant stimulating a greater response upon subsequent exposure to the antigen in those who recovered from COVID?
This vaccine was released under an emergency use authorization (EUA). That means it is experimental.
An EUA cannot be used if there exists an already approved treatment. Both Remdesivir and the vaccines are being made available under EUA, Is that why certain already approved drugs shall not be named?
Direct quote from manuscript
The antibody titers of vaccinees with pre-existing immunity are not only 10-20 times higher than those of naïve vaccinees at the same time points (p <0.0001, two tailed Mann Whitney test), but also exceed the median antibody titers measured in naïve individuals after the second vaccine dose by more than 10-fold.
Common sense suggests that those who have had COVID are already immune, and that they get no doses of the vaccine. Otherwise, we’d be deluged in a wave of reinfections, hospitals would be constantly overrun, and everyone would get COVID once a month.
Is that happening? No.
The “Old WHO” (before June 2020) said previous infection or vaccination leads to herd immunity. I’m going to go with Old WHO on this one.
New WHO appears to be – exclusively – an experimental injectible marketing organization. Just like NIH, and FDA.
[Note that New WHO got their previous, apparently inconvenient description for herd immunity (the one everyone knows) deleted from archive.org. Must be nice to be able to rewrite history like that.]
As I recall, Winston Smith worked at the Ministry of Truth and had the job of rewriting archives to make them mesh with the current official truth.
“Winston works in the Ministry of Truth, where he alters historical records to fit the needs of the Party.”