Experimental trials of
new nootropics
Illia Dorosh,
Biotechnology
student

Definition
• Nootropics – neurometabolic stimulators
• Cognitive function (Memory, learning, creativity,
motivation, attention)
• Resilience to hypoxia and cognitive loads

N06BX Other psychostimulants and nootropics
• N06BX01 Meclofenoxate
• N06BX02 Pyritinol
• N06BX03 Piracetam
• N06BX04 Deanol
• N06BX05 Fipexide
• N06BX06 Citicoline
• N06BX07 Oxiracetam
• N06BX08 Pirisudanol
• N06BX09 Linopirdine
• N06BX10 Nizofenone
• N06BX11 Aniracetam
• N06BX12 Acetylcarnitine
• N06BX13 Idebenone
• N06BX14 Prolintane
• N06BX15 Pipradrol
• N06BX16 Pramiracetam
• N06BX17 Adrafinil
• N06BX18 Vinpocetine
• N06BX21 Tetramethylglycoluril
• N06BX22 Phenibut
Anatomical Therapeutic Chemical (ATC)
Classification System

N06BX Other psychostimulants and
nootropics
• N06BX01 Meclofenoxate
• N06BX02 Pyritinol
• N06BX03 Piracetam
• N06BX04 Deanol
• N06BX05 Fipexide
• N06BX06 Citicoline
• N06BX07 Oxiracetam
• N06BX08 Pirisudanol
• N06BX09 Linopirdine
• N06BX10 Nizofenone
• N06BX11 Aniracetam
• N06BX12 Acetylcarnitine
• N06BX13 Idebenone
• N06BX14 Prolintane
• N06BX15 Pipradrol
• N06BX16 Pramiracetam
• N06BX17 Adrafinil
• N06BX18 Vinpocetine
• N06BX21 Tetramethylglycoluril
• N06BX22 Phenibut
Anatomical Therapeutic Chemical (ATC)
Classification System

History
• Piracetam 1964 by Corneliu E. Giurgea

Prescribed for:
• Post Hypoxia recovery (stroke)
• Alzheimer’s disease patients
• Developmental disorders
Downregulated expression of nicotinic receptor is observed in

Models
• Aplysia sp. (sea slug)
10,000 cells

Time-lapse movie of cultured Xenopus retinal
ganglion cell axons. Supplementary Video 2
from Koser, et al. 2016. Nature Neuroscience

Chronic toxicity trials for repeated use
Carcinogenic toxicity
Mutagenic toxicity
LD50 =
5.6 g/kg щурів
20 g/kg мишей
Piracetam

Abuse potential evaluation
Profile of the receptor binding in vivo
Sales rules (Prescription or over the counter)
Local recommendations FDA, EMA
New CNS actions mechanisms
 additional precilinical trials

ICH guideline M3
1. drug discrimination
2. self-administration of the compound
3. assessment of withdrawal
15. Nonclinical abuse liability

• If a substance shows the signals, similar to
known models of abuse or a new mechanism
of addiction, additional nonclinial trials are
needed in parallel (for examphle, phase III).

Drug
wheel

Антидепресанти

operant conditioning methods
• innovative studies of Barry
(1968) and Morrison and Stephenson (1969),
two-bar operant conditioning techniques
• relative response rates, rather than upon
absolute response rates
• one of the major problems in behavioural
pharmacology, the distinction between
specific drug effects and general excitatory or
sedative effects, is very largely avoided.

Drug discrimination typically involves training an
animal to produce a particular response in a given
drug state for a food reinforcer and to produce a
different response for the same food reinforcer in
the placebo or nondrug state. The interoceptive cue
state that is produced by the drug controls behavior
as a discriminative stimulus or cue that informs the
animal to make the appropriate response to gain
reinforcement. The choice of the response that
follows the administration of an unknown test
compound can provide valuable information about
the similarity of that drug's interoceptive cue
properties to those of the training drug.

Effectiveness trials
• step down
• step through
• two compartment test
• up-hill avoidance
• scopolamine induced test
• ischemia induced amnesia memory
impairments in basal forebrain
• Electroencephalography

Morris water navigation test

Bees

• Радіолігандні дослідження краще виконувати, маючи
мічену речовину, що досліджується [24]. Однак її синтез
стає економічно доцільним на етапі клінічних
досліджень. На доклінічному етапі вивчають вплив
неміченої сполуки на зв'язування з синаптосомами
мозку специфічних мічених лігандів: [3Н]-
флунітразепаму (ліганд бензодіазепінових рецепторів),
[3Н]- спіперону (ліганд D2 дофамінових і 5-НТ2
рецепторів), [3Н]-іпсапірону (ліганд 5-НТІА рецепторів).
Вивчають вплив ФЗ на зв'язування цих мічених лігандів
мембранами різних структур мозку - стріатума,
гіпокампа, фронтальної кори. У дослідах з міченим
спіпероном стріатум розглядають як джерело D2, а кору
- як джерело 5-НТ2 рецепторів.

Requirements
• Pass through the blood brain barrier
biopharmaceuticals
• B. Subtilis SCK6 масляну кислоту

biopharmaceuticals

• Immediately after decapitation, the necessary brain structures are isolated
on ice from the brains of several rats, weighed and homogenized in a 32-
fold volume of 50 mM TRIS buffer (pH = 7.4) in a glass-Teflon homogenizer
on ice. The homogenate is centrifuged at 30,000 g for 20 minutes. The
resulting precipitate is resuspended in the same buffer and centrifuged
again. The final sediment is dissolved in the same buffer so that the
protein content in the sample is 0.15 mg. The binding of [ 3H]-
flunitrazepam (0.125 nM) is studied with a total incubation volume of 500
μl. Nonspecific binding is determined in the presence of 10 μM
flunitrazepam. The substance under investigation is used in several
concentrations (from 1010 to 10"4 M). After 60 minutes of incubation (on
ice), the reaction in all test tubes is stopped by rapid filtration through
glass fiber filters GF/B (Whatman). The filters are washed three times
cooled buffer. A day before measuring radioactivity, filters are placed in 5
ml of scintillation liquid (ACS). Radioactivity is measured using a liquid
scintillation counter and IC50 is determined - the concentration of a
substance that inhibits the specific binding of [3H]-flunitrazepam by 50%
[25] Under similar conditions, the membrane suspension is incubated with
labeled spiperone (final concentration 0.1-0.25 nM) or [3H]-ipsapyrone (2
nM) in the absence and presence of various concentrations of the
substance under investigation. that is being studied are measured in
nanomolar concentrations, the conclusion about the high affinity of the
compound to the receptors corresponding to the labeled ligand is correct

References
• Evaluation of the specific pharmacological activity of medicinal
products
• Komissarov I.V., Abramets I.I. New look at the molecular
mechanisms of action of psychopharmacological agents//Arch.
wedge, and experiment. Medicine .- 1993.- Т.2, №1.- С.6-12.
• ICH guideline M3(R2) on non-clinical safety studies for the conduct
of human clinical trials and marketing authorisation for
pharmaceuticals
• Pardridge WM. Re-engineering biopharmaceuticals for delivery to
brain with molecular Trojan horses. Bioconjug Chem. 2008
Jul;19(7):1327-38. doi: 10.1021/bc800148t. Epub 2008 Jun 12.
PMID: 18547095.
• Hesp, Casper & Smith, Ryan & Parr, Thomas & Allen, Micah &
Friston, Karl & Ramstead, Maxwell. (2019). Deeply Felt Affect: The
Emergence of Valence in Deep Active Inference.
10.31234/osf.io/62pfd.
• Arish, M. R., Tong Michelle, T. T. Making “Good” Choices: Social
Isolation in Mice Exacerbates the Effects of Chronic Stress on
Decision Making. Frontiers in Behavioral Neuroscience, 14(81)

References II
• Establishing Natural Nootropics: Recent Molecular
Enhancement Influenced by Natural Nootropic
https://doi.org/10.1155/2016/4391375
Evidence-Based Complementary and Alternative
Medicine
Hindawi Publishing Corporation
• Вивчення здатності до навчання та пам'яті:
формування умовних рефлексів як з позитивним,
так і з негативним підкріпленням та збереження
отриманих навичок (Я. Бурені та співавт., 1991).
• (Вікторов О.П., 2011)

Thanks for your attention and memory