TrumpLyftAlles | 6 points
Ivermectin Docks to the SARS-CoV-2 Spike Receptor-binding Domain Attached to ACE2 (US 2020-06-19)tada! this is the proof we all knew for months now
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7261036/figure/Fig2/
ivermectin may not have been engineered to fight covid specifically but it most definitely has well documented antiviral action and is a practical alternative to unaccessable $xxxx per dose hospital-only IV drugs
the question is how many more months into this will the USA finally say "oh yeah, okay that's useful" after the HCQ nightmare, maybe never because we're stupid that way
[-] TrumpLyftAlles | 2 points
this is the proof we all knew for months now
It's just a docking study, not the kind of evidence the FDA bases drug policy on.
Did you see my depressing essay suggesting that it may be more than two years before BorodyMectin(tm) is available? I'm sorry to say that it's probably going to require a US trial to get the FDA to budge on ivermectin, and then after that there's the sizable tasks of setting up manufacture and distribution -- and I suppose, sending out the pharmaceutical reps to educate the MDs. It's discouraging.
well at least it's still available off-label for educated doctors but so many are missing out on easy help and that's sad, it's either an ER visit or suffer at home with no medication
also the price in the USA is very very wrong, more than $1 a pill, should be like $10 a box when it's $30
[-] TrumpLyftAlles | 2 points
it's still available off-label for educated doctors
If the CDC or FDA would make a mild endorsement of ivermectin:
Ivermectin is safe and may be therapeutically beneficial, although that has yet to be sufficiently proven.
That would give docs enough cover for them to feel comfortable prescribing ivermectin.
the price in the USA is very very wrong
The first use of horse paste ivermectin I came across was a guy using it to treat scabies. He turned to the horse paste because prescriptions for human ivermectin were too costly. I wonder why it costs so much in the US?
[-] TrumpLyftAlles | 1 points
This is a docking study. What does that mean?
Concept and definition of docking. Molecular docking is the study of how two or more molecular structures (e.g., drug and enzyme or protein) fit together [50]. In a simple definition, docking is a molecular modeling technique that is used to predict how a protein (enzyme) interacts with small molecules (ligands).
So I guess docking studies look at how well molecules bind together -- a strong binding being more probably and stable.
The linked page only has a few images. Here are some excerpts from the PDF found on the linked page.
This one caught my attention as yet another retort to the favorite attack on ivermectin, CONCENTRATION TOO HIGH!!! which is always based on the April 3 Monash 48 hours study by Caly et al:
The Vero-hSLAM cell assays performed by Caly et al. might not be entirely relevant to human SARS-Cov2 infection. Vero-hSLAM is an African green monkey kidney epithelial cell line that does not express human ACE2 (13). Therefore, Caly et al. had to use high ivermectin concentrations.
Green monkey cells don't have ACE2 receptors. Wow.
Why is ACE2 important?
SARS-CoV-2 viral spike protein binds to the membranebound form of angiotensin-converting enzyme 2 (ACE2) to infect the host cell.
Monash 48 hours tested ivermectin against cells that don't have ACE2, which is how the virus infects the cell. Presumably lower concentrations of ivermectin would work against human cells, which were what this study used.
Monash was published early, perhaps before the key role of ACE2 was known?
This is a good summary of ivermectin's action against parasites:
One drug that has attracted interest is the antiparasitic compound ivermectin, a macrocyclic lactone derived from the bacterium Streptomyces avermitilis. Ivermectin kills parasites by interfering with nervous system and muscle function and enhancing inhibitory neurotransmission. Ivermectin binds to glutamate-gated chloride channels in the membranes of invertebrate nerve and muscle cells, causing increased permeability to chloride ions, resulting in cellular hyper-polarization, followed by paralysis and death
The researchers used human cells that have ACE2 receptors.
Materials and Methods
We used the program AutoDock Vina Extended to perform the docking study (7). The ivermectin molecule is from PubChem CID: 6321424 (Figure 1). Crystal structure of SARS-CoV-2 spike receptor-binding domain bound with ACE2 was deposited in the Protein Data Bank 2020-02-21, released: 2020-03-18 (8). We analyzed the human ACE2 receptor because this enzyme apparently differs among species, and the affinity of the virus for the human form may explain its particular infectivity for humans (9).
The software turned up one strong binding site between ivermectin and ACE2.
Results
The root-mean-square deviations of atomic positions (RMSD in Angstroms) are tabulated in Table I. Lower values of RMSD indicate that docking is validated with higher accuracy. RMSD values of 3 or more indicate that no docking has occurred. Only one docking position with RMSD=0 is highly valid, and this docking position is shown in Figures 2 and 3. The next 9 docking calculations were greater than 22 Angstroms RMSD, indicating that no docking was detected (Figure 4 and Table I).
Ivermectin docked in the region of leucine 91 of the spike and histidine 378 of the SARS Cov2-ACE2 receptor complex, between the SARS-Cov2 protein and the ACE2 protein. The binding energy of ivermectin to the spike-ACE2 complex was –18 kcal/mol and binding constant was 5.8×10–8.
I wish I knew whether these numbers are remarkable, -18 etc. Anyone?
The Vero-hSLAM cell assays performed by Caly et al. might not be entirely relevant to human SARS-Cov2 infection. Vero-hSLAM is an African green monkey kidney epithelial cell line that does not express human ACE2 (13). Therefore, Caly et al. had to use high ivermectin concentrations.
Conclusion
The ivermectin docking site we identified, between the viral spike and the ACE2 receptor, may interfere with the attachment of the spike to the human cell membrane. Our observation is consistent with the findings of Caly et al. and Patel et al. Clinical trials now underway should determine whether ivermectin is an effective treatment for SARS-Cov2 infection.
It's interesting that the authors cite Patel et al's mid-April ivermectin study.
Patel et al. found that the administration of ivermectin during COVID-19 illness in hospitalized patients is associated with a lower mortality and hospital length of stay, although these findings have been called into question.
That study is based on the same Surgesphere data was used for Lancet and BMJ papers that were withdrawn when the data was attacked and Patel (who owned the data) refused to allow an audit. IMO the Patel study is valid: lower mortality and shorter length of stay is consistent with the clinical trials and reports.
[-] movethroughit | 1 points
My question is can we get a sample from a positive patient that's been treated with Ivermectin to verify this in a lab? This study is "in Silico" which as I understand has a high degree of error.
[-] TrumpLyftAlles | 1 points
I don't understand what you're suggesting. Do you want a blood sample? Do you want to somehow verify the proposal "Ivermectin docked in the region of leucine 91 of the spike and histidine 378 of the SARS Cov2-ACE2 receptor complex"?
[-] movethroughit | 2 points
I don't even really know what kind of useful sample could be harvested, perhaps nasopharyngeal? And yes, verify it docked as described in the study, potentially activating the FDA's STUFU receptors.
[-] TrumpLyftAlles | 1 points
verify it docked as described in the study
Is that possible? I have no idea if the technology exists to do that. Do you want to bring the study to /r/science or another suitable sub and ask there?
potentially activating the FDA's STUFU receptors.
Please don't mean that you're concerned about an FDA conspiracy to suppress ivermectin. I really don't want to go there.
[-] movethroughit | 3 points
No, I'm just kind of irked with them.
[-] TrumpLyftAlles | 2 points
I got discouraged when someone on twitter alleged that the FDA will probably only pay attention to US trials, it's biased that way. Bummer: there are only 2 underway, AFAIK, and one won't be done for 17 months.
[-] movethroughit | 2 points
Yeah, seems like we could be doing a lot more than telling people to keep from dying horribly until a vaccine gets shoved through or the virus mutates itself out of business. At least NIH is saying 'not outside a clinical trial' but wtf happened to ICON?
[-] TrumpLyftAlles | 1 points
wtf happened to ICON
Not sure what you mean. Are you asking "How did the ICON MDs get away with using ivermectin?" or "Why didn't the FDA pay attention when ICON came out?"
[-] movethroughit | 1 points
AFAIK, it's still in preprint and hasn't been published yet.
[-] TrumpLyftAlles | 1 points
Rumor is, no journal wants to accept the paper. The "design" was absent, sorry to say, not just terrible.
[-] movethroughit | 2 points
"Is that possible? I have no idea if the technology exists to do that. Do you want to bring the study to /r/science or another suitable sub and ask there?"
An excellent question and a very good suggestion.
[-] TrumpLyftAlles | 5 points | Sep 01 2020 22:19:13
I sent this to Dr. Wagstaff, co-author of Monash 48 hours, via LinkedIn:
HI, Dr. Wagstaff.
Only today did I learn that your breakthrough April 3 ivermectin study was performed on monkey cells that don't express ACE2 receptors. As I'm sure you know, the main line of attack on ivermectin for covid-19 is the "concentration too high!!!" argument based on the concentrations in your study. Since ACE2 is the mechanism for covid-19 to infect cells, presumably your current study will show that lower concentrations are effective, presuming that you're using human cells this time.
Can you share the ETA of your current study, please? Can't wait!
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[-] TrumpLyftAlles | 3 points | Sep 02 2020 03:39:39
Dr. Wagstaff replied!
Takeways:
Key quote:
I am happy to tell you that we have absolutely conducted these studies in human lung cells since then (and many other conditions too) and I am definitely not concerned about the IC50 issue based on the results we have seen.
The green monkey cells do "express ACE2 to some degree" (but less than human cells, is implied)
Dr. Wagstaff (any many others) feel that the failure of lower concentrations to affect covid-19 is probably due to the use of the monkey cells.
Dr. Wagstaff's "current hypothesis is that SARS-Cov-2 inhibits interferon production by blocking imp a/b and ivermectin alleviates this"
She "still strongly of the opinion that ivermectin works in concert with the immune system, which cannot be replicated in a cell monolayer and is likely to mean that doses lower than the IC50 are required."
Her current study uses human lung cells! :)
We need to wait for the RCTs
Personal take-away: Dr. Wagstaff is an excellent scientist and outstandingly nice!
Hi Hoyt,
Thanks for reaching out about our study. Our Vero line is permissible to SARS-CoV-2 infection so it does express ACE2 to some degree, however it is not an ideal cell line for examining ivermectin as it does not express interferon alpha or beta, which is fundamentally the process by which I think ivermectin works in this instance (my current hypothesis is that SARS-Cov-2 inhibits interferon production by blocking imp a/b and ivermectin alleviates this). We utilised these cells as these experiments were performed very early (1st week of Feb) using the first batch of the first virus to be grown outside China and we didn't know enough about the virus back then to use anything but the cells that had been successfully used to grow it.
This is a very typical route for virology to take in the lab.
One final point I should make is that I often see that people are concerned that we used too much ivermectin in our study, which is largely from people misunderstanding the data (or only reading the abstract possibly). The panels C-F in our figure show that we used a range of ivermectin concentrations from very high (5uM0 all the way down to low nano molar, but we didn't see effect at the lower concentrations in these cells. As I and many others have pointed out this is largely probably due to the cell line choice (explained above) and I am still strongly of the opinion that ivermectin works in concert with the immune system, which cannot be replicated in a cell monolayer and is likely to mean that doses lower than the IC50 are required.
I can't announce any specific results yet (and I believe strongly in peer review so any new study will go through that process rather than appear directly on a pre-print server) but I am happy to tell you that we have absolutely conducted these studies in human lung cells since then (and many other conditions too) and I am definitely not concerned about the IC50 issue based on the results we have seen.
As for ivermectin being a treatment for COVID-19 we still unfortunately do need to wait for the results of properly powered RCTs that are ideally double blinded before we can conclude anything definitively. Until then Ivermectin use should be only under consultation with you own personal health care provider.
Hopefully these comments help you understand the science better
Stay safe and well
Kind regards
Kylie
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[-] jpdowlin | 4 points | Sep 02 2020 13:21:14
I am definitely not concerned about the IC50 issue based on the results we have seen
This is as big a give-away as you will get. She has mechanism-of-action evidence for Ivermectin working in-vitro at lower concentrations. This is one more "objection" that medical researchers will have to ditch when arguing against Ivermectin.
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[-] TrumpLyftAlles | 1 points | Sep 02 2020 16:07:37
Thanks for pointing that out, I missed it.
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[-] TrumpLyftAlles | 2 points | Sep 02 2020 04:14:03
I sent Dr. Wagstaff a follow-up question.
Having taken the time to write-up your answer (my method of making my brain understand it), I have a new question, if you would be so kind (again).
Will your current study be able to examine your hypothesis that ivermectin's action against covid-19 is blocking imp a/b?
That theory is offered in several places. You and your colleagues published "Ivermectin is a specific inhibitor of importin α/β-mediated nuclear import able to inhibit replication of HIV-1 and dengue virus" -- but that paper didn't address covid-19.
Will your next paper speak to that action with respect to covid-19? (Hope so!)
Thanks again, very much! - Hoyt
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[-] stereomatch | 2 points | Sep 04 2020 04:27:34
Great work.
Interesting to see the Monash paper authors comment on the IC50 issue - which was a factual result in their paper.
However that paper was hugely influential in bringing ivermectin to the attention of the public - and which led to all the current front line researchers who have reported results on patients.
Even though they say this time they wont release a pre-print - it should and will become public if it has the results expected ie ivermectin works even better on humans than the green monkey cell line.
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[-] stereomatch | 1 points | Sep 04 2020 04:48:38
Our Vero line is permissible to SARS-CoV-2 infection so it does express ACE2 to some degree, however it is not an ideal cell line for examining ivermectin as it does not express interferon alpha or beta, which is fundamentally the process by which I think ivermectin works in this instance (my current hypothesis is that SARS-Cov-2 inhibits interferon production by blocking imp a/b and ivermectin alleviates this). We utilised these cells as these experiments were performed very early (1st week of Feb) using the first batch of the first virus to be grown outside China and we didn't know enough about the virus back then to use anything but the cells that had been successfully used to grow it.
The panels C-F in our figure show that we used a range of ivermectin concentrations from very high (5uM0 all the way down to low nano molar, but we didn't see effect at the lower concentrations in these cells. As I and many others have pointed out this is largely probably due to the cell line choice (explained above) and I am still strongly of the opinion that ivermectin works in concert with the immune system, which cannot be replicated in a cell monolayer and is likely to mean that doses lower than the IC50 are required.
You will note her emphasis on ivermectin acting on another pathway ("SARS-Cov-2 inhibits interferon production by blocking imp a/b and ivermectin alleviates this") is in line with what Adam Gaertner has been emphasizing - ie ivermectin counteracts the disruption covid19 is doing to interferon production. Notably in the Dr Yo interview he mentions this and Dr Yo is surprised by that possible mechanism or pathway and so there is some discussion on that in that video:
https://youtu.be/qfEKz5sS2iM Dr Yo interviews Adam Gaertner
Basically the Monash researcher is saying that the earliest Monash study correctly showed a lower response to ivermectin (lower doses of ivermectin had lesser response). This result may not hold for humans - but first Monash study was very early and they worked with what they had - ie the green monkey cells.
However she points out that not only is the lower ACE2 a factor, but the ivermectin salvaging interferon route (that is hard to replicate in a petri dish) as it does not demonstrate the interferon pathway as well.
So for humans it may be the ACE2 being more but that in real patients the interferon pathway may be a part of the recovery demonstrated by patients on ivermectin.
So very interesting to see she is inclined to believe ivermectin works at much smaller doses in humans (real patients) than green monkeys (and in petri dish ie in vitro).
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[-] TrumpLyftAlles | 2 points | Sep 04 2020 05:04:44
So very interesting to see she is inclined to believe ivermectin works at much smaller doses in humans (real patients) than green monkeys (and in petri dish ie in vitro).
Agree. I think that's foreshadowing her next paper. :) I like your other inferences.
Except for what you seem to be implying here:
You will note her emphasis on ivermectin acting on another pathway ("SARS-Cov-2 inhibits interferon production by blocking imp a/b and ivermectin alleviates this") is in line with what Adam Gaertner has been emphasizing - ie ivermectin counteracts the disruption covid19 is doing to interferon production.
Dr. Wagstaff and her team have been have been doing Monash 48 hours-style research for years, going back to 2012 at least. The imp a/b1 inhibition was in their April 3 paper. It has been the prevailing theory about ivermectin's action since then -- really the only theory until recently. Wagstaff et al published another article about ivermectin and imp a/b1 on May 12, less than 6 weeks after Monash 48 hours.
Ivermectin is a specific inhibitor of importin α/β-mediated nuclear import able to inhibit replication of HIV-1 and dengue virus
I start my write-up of that article with "More verification of ivermectin's importin α/β mechanism", referring to the prior suggestion about that from Monash 48 hours. (I should probably re-read Monash; I'll recognize more words than I did when it came out. ;)
Gaertner is an amateur who reads papers and makes himself out to be an expert. He's got a pretty big brain and bigger balls. He's not a scientist. He has never come up with an original research result. He doesn't know anything he hasn't read in someone else's paper. Gaertner got it from Wagstaff.
Are you aware of anything that Gaertner has written about ivermectin, of greater than tweet-length? I ask because I was struck by his inarticulateness in the Beans interview. Journalists on TV are glib as hell because they have already written about what they're commenting on and they're mostly recapping that writing. Gaertner's hesitation and constant referral to notes on his computer (apparently) indicates to me that he's never written anything. If you're aware of any Gaertner writings, give me link(s) please.
Dr Yo is surprised by that possible mechanism or pathway
Dr Yo must have been familiar with it. It was the theory, and it's still far and away the leading theory. He was prompting Gaertner on a topic he anticipated Gaertner would be able to speak to without too many long pauses looking at his notes. Whoops: I realize now that I was confusing Yo with the Beans guy. I have no idea what Yo knows about ivermectin, maybe very little.
I might be irrationally biased against Gaertner.
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[-] stereomatch | 1 points | Sep 04 2020 05:13:30
Adam Gaertner may be an amateur, but he is putting himself out there - and is the reason or one of the reasons the public is aware of the interferon angle.
I had not noticed it phrased that way - perhaps it had been implied in Medcram, Dr Been videos, but Dr Yo's surprise and subsequent discussion in the Dr Yo interview of Adam Gaertner that it was highlighted.
So despite your low opinion of Adam Gaertner, he is helping visibility into ivermectin and we should appreciate that - unlike many others who would seek to undermine talk of ivermectin he is one of the few figures in the US advocating it - and that counts for something (evidently he has some sort of street cred with the doctors too - a history I am unfamiliar with).
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[-] TrumpLyftAlles | 2 points | Sep 04 2020 05:48:43
he is helping visibility into ivermectin and we should appreciate that
Is he? It's my perception that he makes unfounded claims and doesn't include supporting links. "You can cure everyone in the US with $24 worth of ivermectin." That's not enough money to buy one person two doses of ivermectin at US retail prices. Is that obviously ridiculous "information" helpful to the cause? He can't say ridiculous things like that and retain credibility, IMO. As someone on the sub advised me, everything he says should be taken with a large grain of salt.
He blocked me (again) so I don't know what he's tweeting lately, but during the 2-3 weeks during which I wasn't blocked, he revealed zero new information, IIRC. Maybe that's not his thing, I don't know.
He did pass along at least one paper that I was first to tweet about, without citing me. That's OK, he's under no obligation, and my goal is to get the information out, not rack up likes. His was a lazy tweet, though, just the title and link to the paper. I retweeted his tweet, adding what I had written a few hours later describing the study, to develop interest in the paper. He has four times as many followers as me; I took advantage. :)
I think Dr. Been is probably a much more effective advocate for ivermectin, but I haven't been following him.
If you want to make Gaertner useful (IMO), write down salient "facts" from his videos and let's discuss them. I'm sure he knows stuff we don't. We need to find the wheat among the chaff.
For example, if I understand him, at around 7:35 in the You video you linked, Gaertner says that ivermectin somehow increases interferon production independent of the imp a/b1 thing.
and it also induces the production of interferon alpha and beta inside the cell and I'm still actually trying to work out the mechanism of how it does that.
Have you heard that ivermectin does that? I haven't.
Or I may have simply forgotten. Do you remember anything like that?
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[-] stereomatch | 2 points | Sep 04 2020 18:30:00
For example, if I understand him, at around 7:35 in the You video you linked, Gaertner says that ivermectin somehow increases interferon production independent of the imp a/b1 thing. and it also induces the production of interferon alpha and beta inside the cell and I'm still actually trying to work out the mechanism of how it does that. Have you heard that ivermectin does that? I haven't.
I think he was talking about an as yet less well known pathway, or mechanism.
He did not elaborate, and I am not competent to comment.
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[-] TrumpLyftAlles | 1 points | Sep 04 2020 18:37:39
He did not elaborate, and I am not competent to comment.
That makes two of us, LOL. Maybe three of us (another dig at Gaertner).
It would be great if Adam (to use a friendlier name) would write a paper summarizing his theories, with links. He obviously knows a lot of stuff that I don't know, not that I know much. It would be great to glean learnings from him -- but I can't when all I have is a couple youtube videos.
I take it you see him on twitter? Could you suggest that? IMO, it would give him more credibility, maybe grow his base. Last time I could see his number, he had less than 1200 followers. Maybe he could grow that if he published a paper on Medium or something. Might help him fund his trial, even?
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[-] [deleted] | 2 points | Sep 01 2020 22:19:26
[removed]
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[-] movethroughit | 0 points | Sep 01 2020 22:44:25
Nice!
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