Synthesis of Psilocin and Development of Psilocin Transdermal Patch, followed by an In-Vitro Permeation Study with Fresh Human Skin (with photos)

Agilent1100DAD

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Alright guys! Time to share some knowledge with you. I will describe a synthesis I did for psilocin, complete with photos. I used the psilocin produced and developed a simple adhesive patch. In order to test the permeation of the psilocin in the patch, we utilized a diffusion apparatus I pieced together. It involves several membranes, with a 1 cm^2 patch put on top of a 1 cm^2 piece of fresh human skin @ 400 um thickness (donated from a tummy tuck surgical center). Below the skin, there is a constant flow of water that is at pH 7.4 and at body temperature. The diffusion apparatus is able to collect a 1mL fraction per hour, which is then analyzed via LC-MS. The amount of psilocin in the 1mL vial will give the flux (mg/hr/cm^2). This dictates how much psilocin passes per hour with a patch size of cm^2.

This gets deeper, as psilocin is highly unstable.The following publication, Structure Elucidation and Spectroscopic Analysis of Chromophores Produced by Oxidative Psilocin Dimerization, goes over the issue. When the dimer forms, it is undetectable via HPLC analysis and ATR-FTIR which is strange. This gives research groups false positives for their psilocin purity. I have found a way to stabilize psilocin in the patch.

Anyways, lets begin! I did every step myself, with the assistance of my partner who is a chemical engineer. This was back when I had a schedule1 and 3 DEA license not only for research, but for Manufacturing as well.


Part I (Psilocin Synthesis)

step 1: synthesis of 3-[2-(Dimethylamino)-2-oxoacetyl]-1H-indol-4-yl Acetate


1a. A 2000 mL, four-necked, round-bottomed flask was equipped with an overhead stirrer, J-Kem temperature controller, a 250 mL dropping funnel, and rubber septum through which a positive pressure of dryN2 was inserted. The septum was removed and the flask was chargeds equentially with 1H-indol-4-yl acetate (5; 50.1 g, 285 mmol, 1 equiv)
nd anhyd Et2O (700 mL). The flask was resealed with the septum and is flushed with N2. The suspension was stirred for 10 min and then
cooled to 0 °C in an ice-water bath for 30 min.


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1b. The dropping funnel was charged with a solution of oxalyl chloride (37.1 mL, 428 mmol, 1.5 equiv) in Et2O (60 mL). The oxalyl chloride solution was added dropwise at a rate sufficient to keep the temperature at or below 5 °C, to minimize formation of dimer and other possible by-product. As the addition progressed, a yellow slurry of 10 was formed and when the addition was completed, the mixture was stirred for 4 h.


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1c. After this time, heptane (400 mL) was addedand the mixture stirred for 30 min at 0 °C. The yellow solid obtained was quickly filtered and rinsed successively with heptane (2 × 300 mL), which was quickly used in the next step. . A 2.0 M solution of dimethylamine in THF (175 mL) was added dropwise at a rate sufficient to maintaintemperature below 5 °C in order to minimize side reactions. After the
addition was complete, pyridine (46 mL) in of THF (100 mL) was add-ed dropwise and the mixture was stirred well for 60 min. Heptane (600 mL) was added and the flask contents were suction filtered via aBüchner funnel. The filtered residue was transferred into a round-bottomed flask and deionized H2O (1000 mL) was added, stirred for 30 min, and filtered via Büchner funnel. The off-white solid was tritu-rated sequentially for 40 min in EtOAc (600 mL) and heptane (400mL). The slurry was filtered via Büchner funnel and the solid wasdried in an oven at 40 °C overnight to afford 6 as a light-yellow solid;yield: 66.1 g (81%); mp 205–207 °C.


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ATR-FTIR confirmed the synthesis of synthesis of 3-[2-(Dimethylamino)-2-oxoacetyl]-1H-indol-4-yl Acetate. This compound can be stored at 4C for several months!


Step 2: Psilocin Synthesis

Step 2A: A 2000 mL, four-necked, round-bottomed flask was equipped with an overhead stirrer, J-Kem temperature controller, a
250 mL dropping funnel, and rubber septum through which a positive pressure of dry N2 was inserted. The septum was removed and the
flask charged sequentially with 3-[2-(dimethylamino)-2-oxoacetyl]-1H-indol-4-yl acetate (6; 31.5 g, 115 mmol) and 2-CH3-THF (1000
mL). The flask was immersed in an ice-bath at 0 °C and a solution of 2.3 M LiAlH4 in 2-CH3-THF (140 mL, 322 mmol) was added through
the 250 mL dropping funnel. The dropping funnel was rinsed with additional 2-CH3-THF (20 mL). The LiAlH4 solution was added dropwise
at a rate to maintain a temperature below 20 °C. After the addition, the ice-water bath was removed and the mixture stirred for 30 min.
.The light-yellow solution was heated to reflux (80 °C) with aheating mantle and became ivory-colored after 3 h. Accumulation of
yellow solids was observed on the sides of the round-bottomed flask

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Step 2C. The heating mantle was removed, and the flask was allowed to cool to 50 °C. The flask was again
chilled to 20 °C. The reaction was quenched by sequential addition of 3 drops of aq 1 M NaOH and 3 drops of deionized H2O. The mixture
was diluted with THF (500 mL) and stirred for 20 min. The mixture was filtered via Büchner funnel and the filtrate was kept under N2.
The filter cake was quickly reslurried with 200 mL of [10% solution of (7% ammonia in MeOH) in CH2Cl2] and THF (500 mL). The filtrates
were then combined and concentrated to give a green solid. The solid was triturated with 1:1 EtOAc/heptane (50 mL), then filtered via
Büchner funnel. The dark green solid was dried in an oven at 40 °C overnight to provide dry psilocin (7) as a dark green solid; yield: 20.7
g (91%); mp 167–169 °C.


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Psilocin has been synthesized.

Part II: Development of Transdermal Psilocin Patch and In-Vitro Study (CAUTION: YOU WILL SEE FRESH HUMAN FLESH, ITS GROSS
I then dissolve the psilocin into a proprietary blend of acrylate polymer (in heptane), I then cast the solution into a release line and use a casting knife to get the adhesive layer to 100 um. Once dried, I apply the backing layer and punch out a 1cm^2 patch. This patch is used for the in vitro study.

The following photo is a snapshot of the diffusion apparatus (patch on top of skin with water running below)

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Hope you enjoyed!
 

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HerrHaber

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beautiful synthesis, congratulations and kindest regards!
 

Agilent1100DAD

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Here is some bonus photos and also the data for one run with the diffusion apparatus. We can run 16 samples at once.Just to add, all of this done was done under a DEA license, incase the alphabet team gets any ideas. At the time, we may have been one of the few that performed an in-vitro study using a psilocin patch. To purchase psilocin from a private company would cost about $5,000 a gram. Many people think that psilocybin is a better candidate for transdermal or sublingual, but that big phosphate group greatly reduces its permeation.

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(Photo of our skin-permeation lab)


(Our analytical room, which includes an HPLC w/DAD and fluorometer detector, LC-MS/MS and an ATR-FTIR.)
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(Example of the patch, silver layer is the "front of the patch, facing outwards", the middle layer is the drug in adhesive and the clear layer is the release liner.)

BEWARE, THE VIDEO BELOW IS NOT FOR THE FAINT OF HEART (SKIN DERMATOMING)


d

In summary, a small 1 cm^2 would deliver about 1.8mg over the course of 24 hrs.
This would be ideal for an indication, such as reducing the inflammation in children suffering from fragile x syndrome.

Also...ketamine freebase is HIGHLY permeable. Convert 100 mg of ketamine HCl to ketamine freebase (dissolve ketamine HCl in water, add 2M NaOH until the pH reaches 9, filter, dry). Just rub some of the freebase on your skin and it will absorb within seconds.

So the flux for ketamine free base in a patch is about 120ug/hr/cm^2. The surface area of adults is about 18,000 cm2 (men) or 16,000 cm2 (women). Lets say you were to wrap 10% of a male and a females body in this ketamine patch; for men you would be delivering 216 mg of ketamine per hour and for woman you would be delivering 160 mg of ketamine per hour. Mind you, this could send you into a k-hole for 8-12 hours straight,

I have worked on biodegradable implants that are made up of PLA, PGA, PLGA, etc., and can deliver a consistent amount of drug every hour for either 30, 60, 90, or 180 days. Examples of these drug delivery systems include Subcutaneous Biodegradable Implants for HIV Pre-Exposure Prophylaxis (PrEP), Ozurdex® (dexamethasone intravitreal biodegradable implant), estrogen, etc.

Now imagine you develop an LSD biodegradable implant that delivers 100 ug/hr for 6 months straight...Im sure the feds have thought about it. You apply the polymeric pellet with the LSD API on a dart, you get the point.


Below is the data for the psilocin patch. Thank you for coming to my ted talk.

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G.Patton

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Here is some bonus photos and also the data for one run with the diffusion apparatus. We can run 16 samples at once.Just to add, all of this done was done under a DEA license, incase the alphabet team gets any ideas. At the time, we may have been one of the few that performed an in-vitro study using a psilocin patch. To purchase psilocin from a private company would cost about $5,000 a gram. Many people think that psilocybin is a better candidate for transdermal or sublingual, but that big phosphate group greatly reduces its permeation.

View attachment 9632
(Photo of our skin-permeation lab)


(Our analytical room, which includes an HPLC w/DAD and fluorometer detector, LC-MS/MS and an ATR-FTIR.)
View attachment 9633


View attachment 9634
(Example of the patch, silver layer is the "front of the patch, facing outwards", the middle layer is the drug in adhesive and the clear layer is the release liner.)

BEWARE, THE VIDEO BELOW IS NOT FOR THE FAINT OF HEART (SKIN DERMATOMING)


View attachment 9635d

In summary, a small 1 cm^2 would deliver about 1.8mg over the course of 24 hrs.
This would be ideal for an indication, such as reducing the inflammation in children suffering from fragile x syndrome.

Also...ketamine freebase is HIGHLY permeable. Convert 100 mg of ketamine HCl to ketamine freebase (dissolve ketamine HCl in water, add 2M NaOH until the pH reaches 9, filter, dry). Just rub some of the freebase on your skin and it will absorb within seconds.

So the flux for ketamine free base in a patch is about 120ug/hr/cm^2. The surface area of adults is about 18,000 cm2 (men) or 16,000 cm2 (women). Lets say you were to wrap 10% of a male and a females body in this ketamine patch; for men you would be delivering 216 mg of ketamine per hour and for woman you would be delivering 160 mg of ketamine per hour. Mind you, this could send you into a k-hole for 8-12 hours straight,

I have worked on biodegradable implants that are made up of PLA, PGA, PLGA, etc., and can deliver a consistent amount of drug every hour for either 30, 60, 90, or 180 days. Examples of these drug delivery systems include Subcutaneous Biodegradable Implants for HIV Pre-Exposure Prophylaxis (PrEP), Ozurdex® (dexamethasone intravitreal biodegradable implant), estrogen, etc.

Now imagine you develop an LSD biodegradable implant that delivers 100 ug/hr for 6 months straight...Im sure the feds have thought about it. You apply the polymeric pellet with the LSD API on a dart, you get the point.


Below is the data for the psilocin patch. Thank you for coming to my ted talk.

View attachment 9631
hello1321
Now imagine you develop an LSD biodegradable implant that delivers 100 ug/hr for 6 months straight...Im sure the feds have thought about it. You apply the polymeric pellet with the LSD API on a dart, you get the point.

Below is the data for the psilocin patch. Thank you for coming to my ted talk.
So exciting research! Thank you for sharing indeed.
 

HerrHaber

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Oh I so much miss the days back when I was doing academic research.... there are no words to describe how good I feel when I see science. Keep up the good work!

P.S. Admitting I am actually gelous of your work is said to have psychotherapeutic value so here you go....
 

Agilent1100DAD

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Oh I so much miss the days back when I was doing academic research.... there are no words to describe how good I feel when I see science. Keep up the good work!

P.S. Admitting I am actually gelous of your work is said to have psychotherapeutic value so here you go....
HerrHaberThe memories will forever be within you
 
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