253 replies to this topic

[niner]

Below, I have added the PDFs for the HNMR, Chiral HPLC and HPLC results. One step closer

HNMR: https://drive.google...dit?usp=sharing
Chiral HPLC: https://drive.google...dit?usp=sharing
HPLC: https://drive.google...dit?usp=sharing

Hi guys, Dazzcat asked me to take a look at these. I tracked down the synthesis patent for EVP-6124 and related compounds, and was able to get a listing of NMR peaks. (Example 69) The spectrum that they sent us matches reasonably well, although the 3 peaks at the high end are somewhat displaced from where they were in the patent. The NMR solvent was deutero-DMSO. You can tell that by the solvent peak at 2.5 ppm. The peak at 0.0 is a standard. The peak integration shows 18 protons, so they must have run it with the quinuclidine nitrogen protonated. The most downfield peak, at 10.49 is probably that. It comes in at 10.03 in the patent, which is the most different of all the peaks. Do we know what counterion they used in the analysis? It was probably hydrochloride but they don't say. That might be the cause of the peak shifts, if it was different. (It was HCl in the patent)

The HPLC tells us that the material is 99.5% pure, and the Chiral HPLC shows a big fat single peak. This means that it's all one stereoisomer, or at least close enough that any stereoisomer impurities aren't resolved. The CHPLC doesn't show any impurities at all, which suggests that it's not that great at separating similar compounds unless they are chiral.

From this data, it looks to be of reasonable purity, and it probably is what they claim it to be.

[socialpiranha]

is this data being provided by the synth lab itself or a third party?

[megatron]

is this data being provided by the synth lab itself or a third party?

By the lab itself. That's why we decided to go with a lab that others have had success with in the past. The amount of drug required for a third party analysis, would be a significant amount and pretty expensive. Although, of course it would have been optimal to have it tested by a third party.

The lab just told me the product has been shipped

[Dazzcat]

What a brilliant response Niner, thank you greatly for going out of your way to do this for us. The input of an experienced chemist is highly valuable on these forums, especially in this case where safety is a concern and incorrectly synthesized compounds can lead to all kinds of unpredictable outcomes.

[Dazzcat]

EVP-6124 being a partial agonist should indeed it would appear be a superior therapy for such conditions and may yield enhanced cognitive functions in those not prior impaired

Varenicline is a full agonist; I wonder if there is any negative consequence to this degree of modulation/stimulation. I do not know the selectivity offhand as relates to its primary therapeutic pathway of partial agonist activity at alpha2/beta4 relative to the full agonist activity at alpha7.

Interestingly EVP-6124 has it greatest effects on evoking inward current through receptors at doses lower than expected to cause any significant channel opening. The primary mechanism of action was proposed to work through co-agonistic behavior with ACh, at higher doses where evp- is expected to cause channel opening by occupying both binding sites leads to receptor desensitization, which I believe is detrimental to improving memory. I would suspect it's similar for any agonist at nAChRs regardless if it's partial or full, if the compound has high affinity, then low doses is where it's at.

In case you missed it earlier in the thread, here is the main study again, explains it far better than I can and gives some good insights into this pathway.

Also, regarding memory improvement with scopolamine imparied rats, there is evidence that evp- also improves memory of healthy rats, both demonstrated using the object recognition test.

Edited by Dazzcat, 25 March 2014 - 12:33 AM.

[Dazzcat]

Also, regarding memory improvement with scopolamine imparied rats, there is evidence that evp- also improves memory of healthy rats, both demonstrated using the object recognition test.

Figure 4A and Figure 5A&B in the paper linked above.

[VERITAS INCORRUPTUS]

EVP perhaps is best classified as an allosteric agonist (alo-agonist). Within this it indeed appears optimal for maintaining superior enhanced levels of alpha7 activity, minimizing the potential for overstimulation and desensitization possible with full agonists and potentially even partial agonists.

[Dazzcat]

EVP perhaps is best classified as an allosteric agonist (alo-agonist). Within this it indeed appears optimal for maintaining superior enhanced levels of alpha7 activity, minimizing the potential for overstimulation and desensitization possible with full agonists and potentially even partial agonists.

I don't believe evp-'s action is through allosteric activity, which would mean evp would be binding else where other than an agonist binding site increasing the affinity of ACh. From what I recall allosteric wasn't mentioned in that paper either as a mechanism of action. However they do seem very similar in that both mechanisms would potentiate the activity of normal concentrations of ACh at the receptor and work at sub agonistic doses.

I could be wrong here, perhaps they are the same thing?

Edited by Dazzcat, 25 March 2014 - 08:58 PM.

[VERITAS INCORRUPTUS]

EVP perhaps is best classified as an allosteric agonist (alo-agonist). Within this it indeed appears optimal for maintaining superior enhanced levels of alpha7 activity, minimizing the potential for overstimulation and desensitization possible with full agonists and potentially even partial agonists.

I don't believe evp-'s action is through allosteric activity, which would mean evp would be binding else where other than an agonist binding site increasing the affinity of ACh. From what I recall allosteric wasn't mentioned in that paper either as a mechanism of action. However they do seem very similar in that both mechanisms would potentiate the activity of normal concentrations of ACh at the receptor and work at sub agonistic doses.

I could be wrong here, perhaps they are the same thing?

EVP-6124 undeniably has allosteric properties.

They term it a co-agonist as I assume it must producing partial agonist and PAM via binding to the same site as the endogenous agonist ACh, the endogenous agonist binding site (orthosteric binding site). A classical allosteric agonist binds both to a site other than the orthosteric binding site (allosteric binding site) inducing a conformational change that promotes the capacity to modulate (allosteric modulation) the effect of the endogenous agonist (in a positive or negative manner; PAM or NAM).

EVP-6124 produces both agonistic and allosteric modulatory effects I assume (as I have not reviewed the full texts) within only binding the orthosteric binding site and thus has been termed a co-agonist, something not as yet documented prior within the scientific literature. I personally do not like the term co-agonist for a substrate producing both agonist and allosteric receptor modulation.

The terminology orthoallosteric-agonist seems a bit bulky so I went with the simpler conjoined 'allo-agonist' denoting the dual effect upon the same binding site (implicitly orthosteric to engender any direct agonist modulation), distinct of course from the classical allosteric agonist that acts upon both the orthosteric bindng site and an allosteric binding site. It was my own terminology for this type and manner of activity (though I typoed 'alo-agonist' instead of what I meant to write as allo-agonist).

I should have been clear I was designating a new terminology for this activity that prior had never been documented before. I was curious if anyone would pick up on it
On that note, such a substrate with a positive allosteric modulation (PAM) at the orthosteric biding site as well as direct agonist activity could then be termed a PAMagonist, lol

I assume though since they were first to discover and document such a 'phenomena' the term co-agonist will stick. Maybe we can look to form a consensus here and petition for a change as co-agonist just does not cut it in my book, LOL.

BTW, wiki gives an excellent detailed explanation of allosteric modulators and allosteric agonists:
http://en.wikipedia....teric_modulator

Edited by VERITAS INCORRUPTUS, 27 March 2014 - 01:44 AM.

[YOLF]

Update:
Not sure when this is due to arrive, but I've got an infection at present. If I still have it when it arrives I'll wait until I'm done with the abx before I open it and split it up. So there may be a delay. Got the infection tested and I'm going to need an IV/IM abx (refusing to take aminoglicosides when safe IV/IM options exist), so that may add some delay as well. The treatment regimen will be 7-14 days, 14 if I get my wishes, I don't want to give it any opportunity to build resistance. Called my doc earlier today, waiting for a return phone call.

[Jbac]

Update:
Not sure when this is due to arrive, but I've got an infection at present. If I still have it when it arrives I'll wait until I'm done with the abx before I open it and split it up. So there may be a delay. Got the infection tested and I'm going to need an IV/IM abx (refusing to take aminoglicosides when safe IV/IM options exist), so that may add some delay as well. The treatment regimen will be 7-14 days, 14 if I get my wishes, I don't want to give it any opportunity to build resistance. Called my doc earlier today, waiting for a return phone call.

what the heck?

[YOLF]

Would you rather I take a risk on spreading an abx resistant pathogen that I had to go so far as to get tested to get rid of? Or were you commenting on the difficultly of getting a safe and effective abx?

[YOLF]

Package arrived today.

[Dazzcat]

Package arrived today.

That's awesome, I'm glad we have you on board cryonicsculture, take as much time as you need to get well first.

Edited by Dazzcat, 28 March 2014 - 07:23 PM.

[MasterHerb]

Has anyone tried it?

[megatron]

Package arrived today.

Jeez, that was quicker than I had expected . I see the other package has arrived customs in Hungary, so Bonee should also have it shortly.

Bonee thought maybe it would be better to put the drug in gelatin capsules, so that less of the drug goes to waste compared to directly putting it in plastic bags. I think it would be easier scooping the drug out of the capsules compared to the plastic bags. Also, to let nothing go to waste one can also dissolve the entire capsule in DMSO. What do you guys think?

Edited by Megatrone, 29 March 2014 - 10:47 PM.

[YOLF]

I always recommend against DMSO for just about anything... Caps sound like a good idea though. You could also rip the baggie and submerge it into the mixing bottle if you're adding it to water when you receive it if you use a mason jar.

[bkaz]

Caps would be nice.

[YOLF]

A concern came up during a pm discussion with Dazzcat:

I had initially suggested using caps so each dose could be accurately measured as not everyone has a scale at their immediate disposal. However, the more it is handled the more loss there may be to surfaces and spillage and given the 30-90mg amounts I'll be shipping the loss could be substantial. I'd like to change my recommendation to the following should it be possible given the chemistry concerns:

What's needed:
Clean mason jars, graduated flasks, or some kind of bottles (vessels)
A bottle of distilled water ($1 for a gallon at your local supermarket) or ethanol from sources such as grain or corn.


Steps:
1. Measure out a TBD amount of distilled water or ethanol into vessel A.
2. Pour the plastic pouch into vessel B.
3. Take some liquid from vessel A and put it into the plastic pouch and swish it around.
4. Pout liquid from plastic pouch into vessel A.
5. Repeat steps 3 and 4 as necessary or until there is no more liquid in vessel A
6. Put the remaining liquid from vessel A into vessel B.

After EVP-6124 is dissolved divide the amount of liquid used into the number of doses you would like to have from your 30-90mg and you will have accurate doses.

As long as EVP-6124 is orally available and will be stable in water or ethanol, this is the best option. I'm not currently aware of the stability of EVP-6124 in various substances, so this will have to be determined before this decision is made.

Depending on the necessary amount of liquid, I could also skip the bags and for about $.86ea (+ shipping costs) put it into 1oz amber boston round dropper bottles from specialty bottle and the recipient could just fill it with the liquid we decide on if the budget permits.

[megatron]

It could possibly be dissolved in alcohol, but solubility in water is very poor.

[Dazzcat]

I always recommend against DMSO for just about anything... Caps sound like a good idea though. You could also rip the baggie and submerge it into the mixing bottle if you're adding it to water when you receive it if you use a mason jar.

Given that we only need 1-2mg per dose, can it be acceptably safe to only use a small volume of DMSO per dose?

Edited by Dazzcat, 01 April 2014 - 09:19 PM.

[Dazzcat]

This is interesting, first time I seen evp considered to treat cognitive decline from tobacco abstinence.

Dr. A. Eden Evins:
Proof-of-Concept Trial of an Alpha-7 Nicotinic Agonist for Nicotine Dependence (Evins PI) NIDA R01DA030992 (04/01/2011-01/31/2016)
When people quit smoking, a major symptom of nicotine withdrawal is cognitive impairment. This type of impairment can make individuals more likely to start smoking again in order to get rid of that side effect. The Alpha 7 acetylcholine receptor agonist, EVP 6124, has been shown to improve mental performance, such as ability to pay attention and focus for extended periods of time. This study aims to determine whether by improving cognitive performance with EVP 6124, individuals who are trying to quit smoking are more likely to sustain abstinence. This study is currently enrolling 450 healthy male and female smokers aged 18-65 who have smoked an average of at least 15 cigarettes per day during the past year.

http://www.massgener...ion_trials.aspx

Edited by Dazzcat, 01 April 2014 - 10:35 PM.

[YOLF]

I always recommend against DMSO for just about anything... Caps sound like a good idea though. You could also rip the baggie and submerge it into the mixing bottle if you're adding it to water when you receive it if you use a mason jar.

Given that we only need 1-2mg per dose, can it be acceptably safe to only use a small volume of DMSO per dose?

If alcohol will work, why bother/risk using DMSO?

[Dazzcat]

I always recommend against DMSO for just about anything... Caps sound like a good idea though. You could also rip the baggie and submerge it into the mixing bottle if you're adding it to water when you receive it if you use a mason jar.

Given that we only need 1-2mg per dose, can it be acceptably safe to only use a small volume of DMSO per dose?

If alcohol will work, why bother/risk using DMSO?

Good point, though the trouble is there is no alcohol solubility figures for evp as far as I'm aware, so this may require some guess work, it was speculated to work by someone at the start of this thread. How would one go about determining solubility in ethanol, add drop wise until substance dissolves? I'm thinking 1ml/1mg would be convenient, so hopefully it's safe to add 30ml of ethanol to 30mg evp and see if it dissolves, if it fails then adding water until it dissolves would be fine?

Edited by Dazzcat, 02 April 2014 - 01:34 AM.

[megatron]

I really prefer not receiving it pre-dissolved, because I have not idea how long it will be stable for.

[YOLF]

The idea is to just put the powder in the bottle and let the recipient dissolve it. They could always split it up into additional bottles or bags for later reconstitution. Is there anyone here that can figure out what the stability in alcohol would be?

[VERITAS INCORRUPTUS]

This molecule should be highly stable in EtOH
Solubility in EtOH should likely even be superior to that of within DMSO as well.
I understand the concerns, but I speculate they are wholly unwarranted.
Pure EtOH is the way to go...

[Dazzcat]

I just have one more question, how easy is it to have access to pure EtOH?

[YOLF]

Are you 21?

[Dazzcat]

Are you 21?

Well over. The drinking age is 18 in NZ, which is crazy when you consider that many late teens can legally drink while their brain is still developing.

My question was referring to pure ethanol, I thought that stuff was reserved as a lab solvent, therefore not something easy to get. I think the highest alcohol concentration sold in liquor shops here is around 60%.

Edited by Dazzcat, 02 April 2014 - 08:33 PM.