Patent Evaluation

Nootropic agents
stimulate neurogenesis
Brain Cells, Inc.: WO2007104035

1. Introduction Philippe Taupin

2. Chemistry Dublin City University, School of Biotechnology, Glasnevin, Dublin 9, Ireland

3. Biology and action

The application is in the field of adult neurogenesis, neural stem cells and
4. Expert opinion cellular therapy. It aims to characterize the activity of nootropic agents on
adult neurogenesis in  vitro. Nootropic agents are substances improving
cognitive and mental abilities. AMPA (α-amino-3-hydroxyl-5-methyl-4-isoxazole-
Expert Opin. Ther. Patents Downloaded from informahealthcare.com by McMaster University on 01/08/15

propionate) and nootropic agents were assessed for the potential to

differentiate human neural progenitor and stem cells into neuronal cells
in  vitro. They were also tested for their behavioural activity on the novel
object recognition task. AMPA, piracetam, FK-960 and SGS-111 induce and
stimulate neuronal differentiation of human-derived neural progenitor and
stem cells. SGS-111 increases the number of visits to the novel object. The
neurogenic activity of piracetam and SGS-111 is mediated through AMPA
receptor. The neurogenic activity of SGS-111 may contribute and play a role
in its nootropic activity. These results suggest that nootropic agents may
elicit some of their effects through their neurogenic activity. The application
For personal use only.

claims the use of nootropic agents for their neurogenic activity and for the
treatment of neurological diseases, disorders and injuries, by stimulating or
increasing the generation of neuronal cells in the adult brain.

Keywords: AMPA receptor, differentiation, neural stem cells, neuron, piracetam, proliferation

Expert Opin. Ther. Patents (2009) 19(5):727-730

1.  Introduction

Neurogenesis occurs throughout adulthood in mammals  [1,2]. It occurs primarily

in two regions of the adult brain, the hippocampus and subventricular zone,
in various species including humans  [3,4]. Newly generated neuronal cells in the
dentate gyrus and olfactory bulb establish functional connections with neighbouring
and target cells and survive for extended period of time  [5-7]. Adult neurogenesis
is modulated by a broad range of environmental stimuli, physiological and patho-
logical processes, trophic factors/cytokines and drugs. Adrenalectomy, enriched
environment, voluntary exercise, hippocampal dependent learning and antidepressants
stimulate neurogenesis in the adult hippocampus, whereas adrenal hormones,
stress, age and drugs of abuse negatively influence neurogenesis  [8]. Newly generated
neuronal cells of the adult hippocampus are involved in a broad range of
physiological and pathological processes, particularly learning and memory,
Alzheimer’s disease, depression and schizophrenia  [9-12].
It is hypothesized that new neuronal cells in the adult brain originate from a
population of residual stem cells. Neural stem cells (NSCs) are the self-renewing
multipotent cells that generate the main phenotypes of the nervous system. In
support of this contention, neural progenitor and stem cells have been isolated
and characterized from various regions of the adult CNS, including the striatal
area containing the subventricular zone, the hippocampus and the spinal
cord  [13-15]. The existence of NSCs in the adult brain suggest that it may
be amenable to repair  [16,17]. To this aim, drugs and compounds that promote

10.1517/13543770902721303 © 2009 Informa UK Ltd ISSN 1354-3776 727

All rights reserved: reproduction in whole or in part not permitted
 Nootropic agents stimulate neurogenesis
and stimulate adult neurogenesis in  vitro and in  vivo are
candidates for regenerative medicine, for the treatment of
neurological diseases and injuries.
The hippocampus is a region of the brain located in
the temporal lobe. It belongs to the limbic system. The
hippocampus play a central role in learning and memory,
particularly in episodic and spatial memory, general
declarative memory, forming new memories, anterograde
and retrograde amnesia, storing and processing spatial
information. It is also the site of particular forms of plasticity
such as long-term potentiation, a model of memory  [18,19].
Nootropic agents are substances thought to enhance
cognitive function and/or mental activity, among them
Expert Opin. Ther. Patents Downloaded from informahealthcare.com by McMaster University on 01/08/15
piracetam, FK-960 [N-(4-acetyl-1-piperazinyl)-p-fluorobenz-
amide monohydrate] and SGS-111. Piracetam is a pyrrolidone
derivative. It improves concentration and enhances memory.
FK-960 is an antidementia piperazine derivative. SGS-111 is
an analogue of piracetam  [20-22].
The application aims to characterize the activity of the
glutamate agonist, AMPA (α-amino-3-hydroxyl-5-methyl-4-
isoxazole-propionate), and nootropic agents, like piracetam
(Nootropil or 2-oxo-l-pyrrolidineacetamide), FK-960 and
SGS-111 (Figure 1), on adult neurogenesis in vitro. It claims
the use of nootropic agents to stimulate or increase adult
For personal use only.
neurogenesis and for the treatment of neurological diseases
and injuries, by stimulating or increasing neurogenesis in
the adult brain.
2.  Chemistry
2.1  Cell culture
Human neural progenitor and stem cells (hNSCs) were
isolated and cultured as a monolayer, in defined medium in
presence of trophic factors.
2.2  Differentiation
Various concentrations of drugs and compounds were
tested for their differentiating potential on human-derived
neural progenitor and stem cells, in the absence of trophic
factors. Among them, AMPA, an agonist of the AMPA
ionotropic glutamate receptor, PEPA an allosteric potentiator
of AMPA receptor, the nootropic agents piracetam,
FK-960 and SGS-111, NBQX (2,3-dihydroxy-6-nitro-7-
sulfamoyl-benzo[f ]quinoxaline-2,3-dione) an antagonist of
the AMPA receptor. Differentiation was assessed and quantified
by immunocytochemistry.
2.3  Immunocytochemistry
Immunocytochemistry was performed according to standard
protocols. The cells were stained with primary antibodies against
class III β-tubulin isotype, a marker of immature neurons  [23].
2.4  Novel object recognition task
Male F344 rats were administered intraperitoneally various
drugs and compounds, once daily for 7  days. Animals
728 Expert Opin. Ther. Patents (2009) 19(5)
were tested after 7 days of drug administration on the novel
object recognition task, as previously described  [24].
3.  Biology and action
Results show that AMPA induces and stimulates, in a
dose-dependent fashion, the differentiation of human-derived
neural progenitor and stem cells into immature neuronal
cells, immuno-positive for class III β-tubulin isotype. In the
presence of AMPA, PEPA promotes the differentiation of
human-derived neural progenitor and stem cells into
immature neuronal cells that are immunopositive for class III
β-tubulin isotype. Piracetam, FK-960 and SGS-111
induce and promote the differentiation of human-derived
neural progenitor and stem cells into immature neuronal
cells, which are immunopositive for class III β-tubulin
isotype. AMPA promotes the neuronal differentiation of
human-derived neural progenitor and stem cells, in
presence of SGS-111. NBQX inhibits the stimulation of
neuronal differentiation by the neurogenic agents AMPA
and piracetam. Administration of SGS-111 results in a
significant increase in the number of visits to the novel
object when compared to the familiar object, in the novel
object-recognition task.
This shows that AMPA-induced neuronal differen-
tiation in  vitro is mediated through the AMPA receptors.
Piracetam and SGS-111 exert some of their neurogenic
effects in vitro through AMPA receptor activation. SGS-111
acts as cognitive enhancer. The neurogenic activity
of SGS-111 may contribute and play a role in its
nootropic activity.
4.  Expert opinion
AMPA, piracetam, FK-960 and SGS-111 stimulate the
differentiation of human neural progenitor and stem cells
into the neuronal pathway in  vitro. Piracetam and
SGS-111 exert their neurogenic activity in  vitro through
AMPA receptor activation. These agents may be used
to promote the generation of neuronal cells in the adult
brain, to treat and cure neurological diseases, disorders
and injuries.
Nootropic agents are memory enhancer. Hippocampal-
dependent learning enhances neurogenesis in the adult
hippocampus, increases neurogenesis in the adult hippocampus
and stimulates hippocampal dependent-learning and memory
performance in rodents  [25-27]. The neurogenic activity of
nootropic agents on human neural progenitor and stem cells
in  vitro suggest that nootropic agents may exert some of
their activity by stimulating adult neurogenesis in the adult
hippocampus. However, this remains to be evaluated in
experimental studies. Neurogenesis decreases with ageing in
adult rodents  [28]. Nootropic agents, like piracetam, are known
to enhance memory in ageing adults. It would be of interest
to assess the activity of nootropic agents in  vivo in ageing
 Taupin

A. O B.

O N H O
H
N N
N N O
H
O
F O

Figure  1. Structure of the nootropic agents FK-960 and SGS-111. The application claims the nootropic agents, FK960 (A) and
SGS-111 (B), to stimulate or increase adult neurogenesis and for the treatment of neurological diseases and injuries, by stimulating or
increasing neurogenesis in the adult brain.
Expert Opin. Ther. Patents Downloaded from informahealthcare.com by McMaster University on 01/08/15

rodents, in adult neurogenesis and in hippocampal-dependent in the adult brain. Further experimental studies are required
learning and memory performance, like the Morris water to characterize the neurogenic potential of nootropic
maze task  [29]. agents in  vivo.
This application claims the use of nootropic agents
to stimulate or increase adult neurogenesis and the use Declaration of interest
of nootropic agents, alone or in combination with other
substances, for the treatment of neurological diseases The author states no conflict of interest and has received no
and injuries, by stimulating or increasing neurogenesis payment for this manuscript.
For personal use only.

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Patent details
Title: Modulation of neurogenesis by nootropic
agents
Assignee: Brain Cells, Inc.
Inventors: Barlow C, Carter TA, Morse A,
Treuner K, Lorrain KI, Gitnick D, Pires JC.
Priority data: 21/06/2006
Filing date: 08/03/2007
Publication date: 13/09/2007
Publication no.: WO2007104035
Affiliation
Philippe Taupin
Dublin City University,
School of Biotechnology,
Glasnevin, Dublin 9, Ireland
Tel: 01 700 7058;
E-mail: philippe.taupin@dcu.ie