The "LNPs Go Throughout the Body"

In which I rant. And why just about everyone is missing what the industry is admitting to with saRNA and mNEXSpike (from Moderna)

I’m sorry everyone. If I hear that LNPs go throughout the body one more time I’m going to lose it. I usually stop watching videos or read further if I find this statement. So please stop doing that. For my sanity, at least.

Why?

In Which I Rant

This statement obscures what the LNP biodistribution is all about. Drugs go throughout the body as well. Every therapeutic product that I have studied, reviewed and assessed. That’s what happens to all small organic molecules, peptides, proteins, fats etc.

But drugs are designed and even nutraceuticals to be specific, or semi-specific for a receptor or ligand on the cell membrane. This recognizes the drug or nutraceutical as being very very similar (or close enough) to the natural ligand. Or sometimes it is the “anti-ligand” so to speak.

Lets compare Transfection vs Drug-Ligand binding

Overall

• transfection affects the WHOLE cell, drugs affect specific biomolecules,

• transfection is not immediately reversible, drugs tend to be less permanent and are often rapidly reversible (due to half-life or antidotes), because most act via non-covalent binding to one specific protein (which then gets regenerated…..well most of the time, even in older people like me). Many drugs are designed that way.

• transfection uses stealth LNPs which has a biological identity in vivo which cannot be predicted in vitro and which disrupt then entire cell membrane, whereas drugs bind via specific hydrogen bonds, van der waal forces and are usually predictable from in vitro studies to in vivo activity (because of known stucture activity relationships).

• transfection is non-efficient (ie lots of cells gets transfected to get the one you want), drugs are much more specific. So drugs are delivered “throughout the body” but because the binding tends to be specific for a certain biomolecule, if the cell doesn’t have lots of them, the drug doesn’t affect that cell (usually, there are always exceptions, ok?)

Do you see how specific this drug molecule is? Each specific area has a specific role and is required. And you need to think in 3D. This is what we call STRUCTURE ACTIVITY RELATIONSHIPS in medicinal chemistry. BTW, after 30 years, we have little idea of the structure activity relationships for the LNPs and the cell membrane. ¹

A significant challenge for LNP development is insufficient understanding of structure–activity relationship (SAR). Small changes in chemical composition and process parameters can affect LNP structure, significantly impacting performance in vitro and in vivo.

Change one little thing (ie the PEG concentration from 1% to 1.5%) you change the whole LNP’s other actions, not just what you need the PEG lipid for (ie stabilization, stealth effect, size). The other actions mean the ability to transfect certain cells, endosomal trafficking etc etc.

So it doesn’t matter that LNPs “travel throughout the body” but that transfection with LNPs

• affect the whole cell

• do not target a specific protein, ligand or receptor

• have long lasting effects on the cell’s bi-lipid membrane with one dose compared to drugs which usually do not affect the same

• expose cells to extra LNPs but do not provide any benefit (if we had a drug like that it would never get to market unless it had NO or little pharmacological action in the non-targeted cells. Even then. This would depend on the severity of the disease that is being treated of course. For vaccines it would be as close to zero as possible).

The Targeting Problem with LNPs

Because the LNPs affect the WHOLE cell and are not specific for a protein, enzyme, ligand etc, a considerable amount of money, sweat and tears have been spent to help target these LNPs to the specific cells that are to be transfected.

This includes

1. ionizable lipid optimization

   1. libraries of ionizable lipids have been synthesized. Thousands. Weird stuff like bend lipids², and squaramide lipids by Moderna for example.³ The money spent on this must be insane. So far, little progress

   2. fooling around with the molar ratio or PEG ratio

   3. lipids with specific pKa values

   4. pH sensitive LNPs

2. Active targeting (ligand based) or trying to make the LNPs act more like drugs

   1. antibodies or antibody fragments⁴ (the most common approach)

   2. peptides⁵ which target integrins on tumor cells or epithelial cells

   3. aptamers⁶ (probably the best approach imho) BTW aptamers are nucleic acid ligands which bind specific cell surface markers with high affinity, like drugs do. They don’t enter the cells on their own.

   4. small molecule drugs⁷ (I talk about it here)

   5. metalloproteinase-sensitive linkers for tumor targeting⁸ among others

3. Surface charge modulation, aka the Zeta potential

   1. doesn’t work, and is inefficient

   2. non-specific in general

   3. affected (mostly negated) by the biocorona which the LNPs get as soon as they contact human plasma, blood or extracellular liquid.

4. Cell specific promoters (for mRNA) THIS IS THE SCARIEST IMHO

   1. genetic targeting in the mRNA contrust⁹

      1. basically, they gene silence everything else so you only get mRNA production is specific targeted cells (that does not seem like an advance imho)

b. immune cell-specific promoters

1. include coding for a miRNA for example¹⁰

2. hey, in built adjuvant and cell specific! what’s not to like?

   Controlling the site of transcript expression could also limit systemic inflammation. miRNAs are short RNA sequences found across mammalian species that, when bound to complementary mRNA sequences, form double-stranded RNAs that inhibit translation and induce transcript recycling [11], effectively silencing protein translation. Organs have different miRNA signatures, enabling the design and incorporation of multiple sequences in the untranslated region of an mRNA construct

Pieter Cullis who basically invented these LNPs says he out-lasted 5 post-docs trying to solve the targeting problem. I believe this is pretty well unsolvable, for mRNA/LNP vaccines.

The whole video is worthwhile watching but especially the Q&A at the end. Time 47:17 onwards. He talks about structure activity relationships, targeting etc. He doesn’t say the LNPs go everywhere because, in truth, they don’t. First you need to target the specific tissues, then you need to protein production off and on….maybe with external magnetic or other similar techniques???!!!!. Holy Toledo.

The Inefficiency Problem with the LNPs

OK, the SECOND major problem with LNPs is the INEFFICIENCY PROBLEM. What do I mean by that?

ONLY 3-5% OF THE LNPs DELIVERED, ACTUALLY PRODUCE SPIKE PROTEIN

For some strange reason I cannot get this through to people. I keep saying this and it goes in one ear and out the other. NO ONE really mentions it. This happens because of the poor endosomal escape which is the rate-limiting step.

Bitounis talked about this in his seminal paper published in Nature Reviews and Drug Toxicity.¹¹

THE PATHOGENIC POTENTIAL OF THE REMAINING 98% IS UNSTUDIED.

PARTICULATE TOXICITY WHICH I BELIEVE I AM THE FIRST ONE TO BRING THIS ISSUE AS ONE OF THE CAUSES OF THE VACCINE TOXICITY ESPECIALLY THE EARLY BATCHES. See here¹² , here¹³ and here¹⁴

Ok if you are a manufacturer and you’ve kind of come to the conclusion targeting LNPs is a huge task, what do you do? What is happening to 95% of the LNPs? Well, we are pretty sure they are not passive. Here are empty LNPs doing its thing.¹⁵

HOLY TOLEDO. The LNPs activate the monocytes and innate immunity all on their own. I’m sure Moderna (especially) and BioNTech knew this. And they INSISTED it was just an excipient and should not be assessed as an adjuvant. This regulatory mess needs to be straightened out before we use LNPs for ANY indication.

MAYBE UP TO HALF OF THE LNPs ARE EMPTY!

Does everyone know this? It depends on how you measure them because LNPs are extremely tricky to measure. You need several methods.

At the present time they are using encapsulation efficiency using RiboGreen to figure out how many LNPs have the RNA inside. Results are usually 85% or better.

BUT, as explained in this paper¹⁶, it is how you measure them, or the analytical method which determines the final result.

They conclude:

So yes, half of the LNPs are empty. Imagine that. Analytical methods are an ongoing problem with mRNA/LNP formulations and why it has taken forever to get compendial standards because things change everytime you measure the LNPs (not so much for mRNA or the DNA). I talk about that here. Which is why I call them “shape-shifting balls of evil”, lol.

WHAT TO DO WHAT TO DO?

What can the manufacturers do without tipping everyone off on how bad the LNPs are? How about decreasing the AMOUNT of LNPs you need to give somebody. Well, lets develop saRNA and a more reactive mRNA vaccine

Self-amplifying mRNA vaccines¹⁷

An alternative to mRNA vaccines is self-amplifying RNA (saRNA)-based vaccines. saRNA are based on alphaviruses, with a replicase complex that amplifies both the transgene and the vaccine construct. A major benefit of the saRNA platform compared with non-replicating mRNA is a reduction in the dose needed to induce a protective immune response.

In preclinical studies, 1–10 µg of saRNA often matches or exceeds the immunogenicity of 30–100 µg of conventional mRNA as shown by our own Canadian Anna Blakeny.¹⁸ And an Arcturus (ARCT-154) press releases¹⁹

“Low-dose self-replicating RNA vaccine (5ug) demonstrates immunogenicity comparable to 100 µg mRNA vaccine”

So less LNPs are needed. SURPRISE!!! saRNA vaccines use less LNP per dose simply because they use less RNA, and the LNP:RNA ratio remains constant. This is one of the key advantages of saRNA platforms: high protein yield at low RNA AND LNP dose. And the adverse reaction response is about the same, and maybe even LESS myocarditis? (no cases reported in 8,000 patients and they were looking for it but who knows. We will need more data).

Also, saRNA doesn’t need codon N-1-methylpseudouridine, though they are codon optimized. So less or no frameshifting. WIN WIN WIN!!!

Note: There MAY be more protein made overall with saRNA vs regular mRNA vaccines but I am looking at LNPs here.

mNEXSPIKE

The dose is 10ug vs the regular 50ug for Moderna’s SpikeVax

It encodes receptor-binding + N-terminal domains only and seems to work better than Moderna’s Spikevax in the elderly. Similar systemic reaction with LESS local effects. YEAH/sarc

The dose for mNEXSPIKE is 5 times lower so I would expect 5 times less LNPs. Given the LNP:RNA ratio stays relatively constant. But of course, they don’t tell us directly. And no reported cases of myocarditis, but time will tell.²⁰ mNEXSPIKE is codon optimized though and apparently does use N-1-methylpseudouridine.

Did anyone TELL YOU THERE ARE LESS LNPs in these new formulations? FDA? Manufacturers? Others?

Is THAT the reason why toxicity looks like it is less, compared to regular modRNA COVID-19 vaccine despite a more reactogenic RBD (in mNEXSPIKE) for example?

Summary

1. I hope you know now why “LNPs go throughout the body” distracts from the true and significant toxicity of the LNPs. Please refrain, if for nothing else but my own sanity. Thanks. Use “the LNPs are able to non-specifically transfect just about every cell” (except adipose tissue and cartilage BTW and a few others). Doesn’t sound as good I know.

2. The LNPs are quite immunogenic and immunotoxic on their own

3. The LNPs have a targeting issue which as yet looks unsolvable for vaccines imho

4. The LNPs have an efficiency problem as most never make the spike protein

5. Upto half of all LNPs are likely empty raising questions as to efficiency, efficacy etc. This relates to the lack of structure activity relationships that Cullis highlights and the problems with the measurements of critical quality attributes. I discuss some of this regarding the LNPs here.

6. The manufacturers know this and have developed mRNA vaccines with LESS LNPs. The FDA and regulatory authorities likely know this as well.

WHEREIN I RANT ONCE MORE

If you really want to learn how mRNA vaccines work, how the LNPs work, etc then you need to

1. go to manufacturers webinars (they’re FREE!) to see what they know and what the problems are. For biologics, the process is the product so anything they change or upgrade usually means it is important for overall safety and efficacy

2. go to the belly of the beast and attend scientific conferences on RNA therapeutics like yours truly did.

3. Realize the scientific literature doesn’t really tell you a lot (which is why the contract manufacturers tell you what is really going on).

   Spartacus
   had a great meme on what you feel as an outsider going into these conferences. Everyone is siloed and directed and I understood as much as did most everyone else. A surprise.

I’m sure

Genervter Bürger

feels exactly the same way and is a devastating indictment of our scientific class.

4. Read the literature on formulation, pharmacology, pharmaceutics, biophysics, etc and also on nanoparticle technology and NOT only on antibodies schmantibodies

5. Know the analytical methods so finding out the LNPs are mostly empty is NOT a surprise (so not ONLY measurement of DNA but on everything that must be measured, how and why)

Hope this post gives everyone a better idea of the overall issues of the mRNA/LNPs. And why these new types of vaccines are being trialed.

I leave you with this question:

ARE THE LNPs THE MAJOR CAUSE OF mRNA VACCINE TOXICITY

Oh and pray the rosary.

PS hope I don’t have any spelling mistakes

1. https://pubs.acs.org/doi/10.1021/acsnano.3c01186

2. https://www.nature.com/articles/s41467-024-55137-6

3. https://advanced.onlinelibrary.wiley.com/doi/full/10.1002/adfm.202106727

4. https://www.sciencedirect.com/science/article/pii/S2162253125000745

5. https://pubs.acs.org/doi/10.1021/acsnano.4c14625

6. https://pubmed.ncbi.nlm.nih.gov/39067551/

7. https://pubs.acs.org/doi/10.1021/acs.nanolett.3c04415

8. https://pmc.ncbi.nlm.nih.gov/articles/PMC7611664/

9. https://www.sciencedirect.com/science/article/pii/S147149062500078X

10. https://www.sciencedirect.com/science/article/pii/S147149062500078X

11. https://www.nature.com/articles/s41573-023-00859-3

12. A Hypothesis on Why the Early Lots of Pfizer Were So Toxic

13. Moderna's Issues with Particulates

14. Particulates in Pfizer

15. https://www.nature.com/articles/s41541-025-01124-x

16. https://www.nature.com/articles/s41598-024-52685-1

17. https://www.sciencedirect.com/science/article/pii/S216225312200316X

18. https://www.mdpi.com/2076-393X/9/2/97

19. https://ir.arcturusrx.com/news-releases/news-release-details/nature-communications-publishes-pivotal-data-demonstrating

20. file:///C:/Users/mguts/Downloads/TML-article-5111a-2.pdf