NZ622653B2 - Vaccine therapy for the treatment of memory and/or cognitive impairments in subjects with down’s syndrome - Google Patents
Vaccine therapy for the treatment of memory and/or cognitive impairments in subjects with down’s syndrome Download PDFInfo
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- NZ622653B2 NZ622653B2 NZ622653A NZ62265312A NZ622653B2 NZ 622653 B2 NZ622653 B2 NZ 622653B2 NZ 622653 A NZ622653 A NZ 622653A NZ 62265312 A NZ62265312 A NZ 62265312A NZ 622653 B2 NZ622653 B2 NZ 622653B2
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Classifications
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- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
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- C—CHEMISTRY; METALLURGY
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Abstract
Disclosed is a use of an of an antigenic peptide fragment derived from amyloid protein or amyloid-like protein selected from the group consisting of prion protein, tau protein, alpha-synuclein, huntingtin and amyloid beta for the preparation of a medicament for use in the treatment, alleviation or prevention of memory and/or cognitive impairments or abnormalities in subjects with Down’s syndrome; wherein the subjects with Down’s syndrome have not yet developed amyloid protein-associated or amyloid-like protein-associated, plaques in the brain. revention of memory and/or cognitive impairments or abnormalities in subjects with Down’s syndrome; wherein the subjects with Down’s syndrome have not yet developed amyloid protein-associated or amyloid-like protein-associated, plaques in the brain.
Description
VACCINE THERAPY FOR THE TREATMENT OF MEMORY AND/OR COGNITIVE
IMPAIRMENTS IN SUBJECTS WITH DOWN’S SYNDROME
The present invention provides means for treating amyloid-related
pathology in young to middle aged subjects with Down’s syndrome (DS). In
particular, the present invention provides antigenic peptide fragments derived from
amyloid protein or amyloid-like protein for use in the treatment of Alzheimer’s
disease (AD)-like dementia in young to middle aged subjects with Down’s syndrome
(DS).
The genetic disorder Down’s syndrome (DS) is the most common trisomy
and it occurs in one out of 700-1000 newborns. Estimates suggest that 25% or more
of individuals with Down syndrome over age 35 show the signs and symptoms of
Alzheimer's-like dementia (Stanton L.R and Coetzee R.H , 2004). The percentage
increases with age. In DS, the entire or at least a part of the chromosome 21 is
present in triplicate (Antonarakis et al., 2004; Moncaster et al., 2010). Consequently,
the three copies of the gene of the amyloid precursor protein (APP) lead to the
generation of an excess of Amyloid-β (Aβ), one of the main abnormal proteins well
known to be responsible for Alzheimer’s disease (AD) (Ballard et al., 2011).
Therefore, Aβ is suggested to have a possible role for the AD-like dementia in most
DS people (Lee et al., 2005;Liu et al., 2005;Gilman et al., 2005).
In people with Down syndrome, AD-like memory defects can be related to
several pathological proteins, such as Aβ, Tau, Dyrk1A, Apo E etc. Aß increase
begins already in the embryonic stage, progresses at birth and continues to build
up with increasing age (Stoltzner et al., 2000). In addition, Cerebrospinal fluid (CSF)
levels of Aβ42 were lower and tau levels were higher in older (>40 years) than in
younger DS people (Tapiola et al., 2001). Once Aβ is deposited in plaques, Apo E
can be detected in many plaques at age 12 and augments steadily with age
(Lemere et al., 1996). Neurofibrillary tangles (NFTs) are detected in the brain of DS
during the fourth decade of life. These NFT are believed to result from tau
accumulation. It has been shown that hyperphosphorylation might be caused by
overexpression of a kinase named Dyrk1A (dual-specificity tyrosine-phosphorylated
and regulated kinase 1A) (Lemere et al., 1996;Liu et al., 2008). Therefore, people
with DS can develop amyloid-related pathology by age 40 and most have the clinical
symptoms similar to Alzheimer’s, such as cognitive decline and memory impairment,
by age 60 (Stanton 2004).
While individuals with DS receive medical care mainly for their various
health problems (such as heart defect, infections and hypothyroidism), specific-
treatment for neuropathological traits, i.e. mental disability and memory deficiency,
is rarely considered. Currently, only few clinical trials are ongoing, all aimed to
enhance mental abilities or to reduce nerve damage. The drugs for treating DS are
also used in AD and all act on the cholinergic system or the glutamatergic
neurotransmission such as Rivastigmine or Donepezil, a cholinesterase inhibitors
and Memantine, a NMDA receptor antagonist (Prasher, 1993; Prasher, 2004).
However, evidence of efficacy is lacking for people with DS. Currently, there are
many amyloid-modifying treatments under review, including targeting-Aβ-
immunotherapy (Rafii, 2010). Several vaccines have recently reached clinical
phases (Weiner and Frenkel, 2006) after having shown efficient reduction of
cerebral Aβ burden and reversing cognitive decline in mouse AD models. In contrast
to AD, immunotherapies targeting Aβ are not being addressed in DS.
Of the several approaches for treating cognitive impairment in adults with
DS, existing studies using acetylcholinesterase inhibitors (AChEIs) or NMDA
receptor antagonists have shown little or no effect (Fernandez et al., 2007, Hanney
et al., 2012). In contrast, there is significant support for the hypothesis that amyloid
plays a role in the cognitive deterioration of people with DS; Netzer and colleagues
showed that altering APP cleavage, by a γ-secretase inhibitor, has a beneficial
effect on the memory of a DS mouse model (Netzer et al., 2010). These
observations build on studies in mouse models showing that increased gene dose
for APP, the parent protein of the Abeta peptide responsible for amyloid deposition,
is necessary for the age-linked degeneration of neurons that characterizes DS and
AD (Salehi et al 2006, 2009). Therefore, targeting amyloid can be a promising
therapeutic strategy for the prevention of AD progression. The development of an
immunotherapy against Aβ is based on the hypothesis that if a molecule targets and
sequesters Aβ (in soluble form and as oligomers) in situ, the molecule can enhance
removal of Abeta from the brain and bring clinical benefits to people with DS. It is
anticipated that the antibodies generated by the anti- Aβ vaccine will bind to fibrillar
Aβ deposits to eventually solubilize or inhibit the growth of the pre-plaques. The Aβ
oligomers are today considered as the most toxic Aβ species, which impairs most
cognitive functions in DS people (Teller et al., 1996).
There is therefore a need for a prevention therapy which aims at
preventing or slowing down the development of clinical symptoms associated with
amyloid-related pathology in subjects with Down’s syndrome (DS), such as,
particularly, memory and/or cognitive impairments or abnormalities.
In particular, there is a need for a treatment of clinical symptoms
associated with amyloid-related pathology in subjects with Down’s syndrome (DS), ,
which leads to an improvement in learning capabilities in subjects with Down’s
syndrome (DS) and/or improvement or restoration of memory and/or cognitive
capacities.
It is an object of the present invention to provide the measures and means
which help to meet this need; and/or to provide the public with a useful choice. In
particular, the present invention is directed to antigenic peptides for use in
prevention therapy and in the treatment of AD-like cognitive impairments in children
and in young to middle-aged subjects with Down’s syndrome.
In a first embodiment, the present invention provides a use of an antigenic
peptide fragment derived from amyloid protein or amyloid-like protein selected from
the group consisting of prion protein, tau protein, alpha-synuclein, huntingtin and
amyloid beta, for the preparation of a medicament for use in the treatment,
alleviation or prevention of memory and/or cognitive impairments or abnormalities in
subjects with Down’s syndrome; wherein said subjects with Down’s syndrome have
not yet developed amyloid protein-associated or amyloid-like protein-associated,
plaques in the brain.
In a second embodiment, the invention provides a use of a composition
comprising (i) an antigenic peptide fragment derived from amyloid protein or amyloid-like
protein selected from the group consisting of prion protein, tau protein, alpha-synuclein,
huntingtin and amyloid beta described herein and (ii) a pharmacologically acceptable
carrier or excipient and/or an adjuvant, for the preparation of a medicament for use in
the treatment, alleviation or prevention of memory and/or cognitive impairments or
abnormalities in subjects with Down’s syndrome; wherein said subjects with Down’s
syndrome have not yet developed amyloid protein-associated or amyloid-like protein-
associated, plaques in the brain.
In a third embodiment, the invention provides a use of an anti-amyloid antibody for the
preparation of a medicament for use in the treatment, alleviation or prevention of
memory and/or cognitive impairments or abnormalities in subjects with Down’s
syndrome; wherein said anti-amyloid antibody binds an antigenic peptide, and wherein
said antigenic peptide is described herein.
Also described is a method for producing a vaccine comprising formulating the antigenic
peptide described herein with a pharmacologically acceptable carrier or excipient and/or
an adjuvant.
Also described is a vaccine produced by the method of the invention.
Also described is an antigenic peptide fragment derived from amyloid
protein or amyloid-like protein, particularly an antigenic peptide fragment derived
from amyloid-beta protein, or a composition comprising said antigenic peptide, for
use in the treatment and/or alleviation and/or prevention of memory and/or cognitive
impairments or abnormalities, particularly of AD-like memory and/or cognitive
impairments or abnormalities, particularly impairments and abnormalities originating
in the hippocampus and/or the prefrontal cortex and/or the entorhinal cortex of the
brain, in young to middle-aged subjects with Down’s syndrome, in particular in
young subjects with Down’s syndrome, who have not yet developed amyloid
protein- or amyloid-like protein-associated plaques, particularly Aβ-associated
plaques, in the brain.
Also described is an antigenic peptide fragment derived from amyloid
protein or amyloid-like protein, particularly an antigenic peptide fragment derived
from amyloid beta protein, or a composition comprising said antigenic peptide, for
use in the treatment, and/or alleviation and/or prevention of memory and/or
cognitive impairments or abnormalities, particularly of AD-like memory and/or
cognitive impairments or abnormalities, particularly impairments and abnormalities
originating in the hippocampus and/or the prefrontal cortex and/or the entorhinal
cortex of the brain, in middle-aged subjects with Down’s syndrome, in particular in
middle-aged subjects with Down’s syndrome, who have not yet developed amyloid
protein- or amyloid-like protein-associated, particularly Aβ-associated, plaques in
the brain.
Also described is an antigenic peptide fragment derived from amyloid
protein or amyloid-like protein, particularly derived from amyloid beta protein, or a
composition comprising said antigenic peptide, for use in the treatment, and/or
alleviation and/or prevention of memory and/or cognitive impairments or
abnormalities, particularly of AD-like memory and/or cognitive impairments or
abnormalities, particularly impairments and abnormalities originating in the
hippocampus and/or the prefrontal cortex and/or the entorhinal cortex of the brain, in
young subjects with Down’s syndrome, in particular in young subjects with Down’s
syndrome, who have not yet developed amyloid protein- or amyloid-like protein-
associated, particularly Aβ-associated-, plaques in the brain.
Also described is an antigenic peptide fragment derived from amyloid
protein or amyloid-like protein, particularly derived from amyloid beta protein, or a
composition comprising said antigenic peptide, for use in the treatment, and/or
alleviation and/or prevention of memory and/or cognitive impairments or
abnormalities, particularly of AD-like memory and/or cognitive impairments or
abnormalities, particularly impairments and abnormalities originating in the
hippocampus and/or the prefrontal cortex and/or the entorhinal cortex of the brain, in
childrens with Down’s syndrome, in particular in children subjects with Down’s
syndrome, who have not yet developed amyloid protein- or amyloid-like protein-
associated, particularly Aβ-associated, plaques in the brain.
Also described is an antigenic peptide fragment derived from amyloid
protein or amyloid-like protein, particularly derived from amyloid beta protein, or a
composition comprising said antigenic peptide, for use in prevention of memory
and/or cognitive impairments or abnormalities, particularly of AD-like memory and/or
cognitive impairments or abnormalities, particularly impairments and abnormalities
originating in the hippocampus and/or the prefrontal cortex and/or the entorhinal
cortex of the brain, in children and in young to middle-aged subjects with Down’s
syndrome, in particular in young subjects with Down’s syndrome, who have not yet
developed amyloid protein- or amyloid-like protein-associated, particularly Aβ-
associated plaques in the brain.
In still another embodiment of the disclosure the antigenic peptide is
administered to a subject with Down’s syndrome, particularly a subject with Down´s
syndrome suffering from memory and/or cognitive impairments or abnormalities,
particularly of AD-like memory and/or cognitive impairments or abnormalities,
particularly impairments and abnormalities originating in the hippocampus and/or
the prefrontal cortex and/or the entorhinal cortex of the brain, wherein said subject is
below age 60, particularly below age 55, particularly below age 50, particularly
below age 45, particularly below age 40, particularly below age 35, particularly
below age 30, particularly below 25, particularly below age 20, particularly below
age 15, particularly below age 10, particularly below age 5, particularly below age 3.
In a specific embodiment of the disclosure the antigenic peptide is
administered to a subject with Down’s syndrome, particularly a subject with Down´s
syndrome suffering from memory and/or cognitive impairments or abnormalities,
particularly of AD-like memory and/or cognitive impairments or abnormalities,
particularly impairments and abnormalities originating in the hippocampus and/or
the prefrontal cortex and/or the entorhinal cortex of the brain, wherein said subject is
below age 35, particularly below age 30, particularly below 25, particularly below
age 20, particularly below age 15.
In a specific embodiment of the disclosure the antigenic peptide is
administered to a subject with Down’s syndrome, particularly a subject with Down´s
syndrome suffering from memory and/or cognitive impairments or abnormalities,
particularly of AD-like memory and/or cognitive impairments or abnormalities,
particularly impairments and abnormalities originating in the hippocampus and/or
the prefrontal cortex and/or the entorhinal cortex of the brain, wherein said subject
is below age 15, particularly below age 10, particularly below age 5, particularly
below age 3.
In a specific embodiment of the disclosure the antigenic peptide is
administered to a subject with Down’s syndrome, particularly a subject with Down´s
syndrome suffering from memory and/or cognitive impairments or abnormalities,
particularly of AD-like memory and/or cognitive impairments or abnormalities,
particularly impairments and abnormalities originating in the hippocampus and/or
the prefrontal cortex and/or the entorhinal cortex of the brain, wherein said subject
is below age 10, particularly below age 5, particularly between 0 and 10, particularly
between 1 and 10, particularly between 2 and 10, particularly between 3 and 10,
particularly between 4 and 10, particularly between 5 and 10.
In one embodiment, treatment with the antigenic peptide as described
herein prevents the development of Aβ-associated plaques in the brain, particularly
in the hippocampus and/or the prefrontal cortex and/or the entorhinal cortex of the
brain.
In certain embodiments, contemplated herein is using the antigenic
peptide fragment derived from amyloid protein or amyloid-like protein, or a
composition comprising said antigenic peptide as described herein in the treatment,
and/or alleviation and/or prevention of memory and/or cognitive impairments or
abnormalities, particularly of AD-like memory and/or cognitive impairments or
abnormalities, particularly impairments and abnormalities originating in the
hippocampus and/or the prefrontal cortex and/or the entorhinal cortex of the brain, in
children and in young to middle-aged subjects with Down’s syndrome who have
already developed Aβ-associated plaques in the brain.
In another embodiment of the disclosure treatment with the antigenic
peptide as described herein reduces the amount of Aβ-associated plaques in the
brain, particularly in the hippocampus and/or the prefrontal cortex and/or the
entorhinal cortex of the brain.
In one embodiment, the patient group to be treated with the antigenic
peptide or the composition described herein comprises young to middle-aged
subjects with Down’s Syndrome, particularly subjects, which are below the age of
65, particularly below the age of 60, 55 or 50. In one embodiment, subjects with an
age of between 0 and 65, particularly of between 5 and 55, particularly of between
and 50, particularly of between 15 and 45, particularly of between 20 and 40,
particularly of between 25 and 35, may be treated with the antigenic peptide or the
composition described herein.
For the purpose of the present disclosure, children refer to subjects who
are below the age of 18 particularly subjects with an age of between 0 and 18
particularly of between 1 and 10, particularly of between 2 and 9, particularly
between 3 and 9, particularly of between 4 and 9, particularly between 5 and 18.
For the purpose of the present disclosure, young subjects refer to
subjects who are below the age of 35, particularly subjects with an age of between 0
and 35 particularly of between 1 and 30, particularly of between 5 and 25.
For the purpose of the present disclosure, middle-aged subjects refer to
subjects with an age of between 36 and 65 particularly of between 40 and 60,
particularly of between 45 and 55.
In one embodiment, treatment of a children or young to middle-aged
subject with Down’s syndrome and suffering from memory and/or cognitive
impairments or abnormalities, particularly from AD-like memory and/or cognitive
impairments or abnormalities, particularly impairments and abnormalities originating
in the hippocampus and/or the prefrontal cortex and/or the entorhinal cortex of the
brain, using the antigenic peptide or the composition as described herein, leads to
an amelioration or restoration of said impairments or abnormalities, particularly to
memory amelioration, particularly amelioration or restoration of the recognition
memory and/or the contextual associative memory.
In one embodiment, treatment of a children or young to middle-aged
subject with Down’s syndrome and suffering from cognitive impairments or
abnormalities, particularly from AD-like memory and/or cognitive impairments or
abnormalities, using the antigenic peptide or the composition as described herein,
leads to an increase in the retention or a complete restoration of cognitive memory
capacity in the treated subject.
The treatment of AD-like cognitive impairments or abnormalities in
children and young to middle-aged subjects with Down’s syndrome with the
antigenic peptide and as described herein shows the therapeutic effects as
disclosed herein without inducing unwanted side effects such as
meningoencephalitis and microhemorrhage.
In certain embodiments of the invention, the subject is an children-aged
subject, particularly a young-aged subject, particularly a middle-aged subject as
defined herein, with Down’s syndrome and suffering from memory and/or cognitive
impairments or abnormalities, particularly from AD-like memory and/or cognitive
impairments or abnormalities, particularly impairments and abnormalities originating
in the hippocampus and/or the prefrontal cortex and/or the entorhinal cortex of the
brain.
In a further embodiment, the treatment with the antigenic peptide or the
composition as described herein, prevents the development of Aβ-associated
plaques in the brain, particularly in the hippocampus and/or the prefrontal cortex
and/or the entorhinal cortex of the brain, in subjects with Down’s syndrome,
particularly in the brain of young- to middle-aged, particularly in the brain of children-
aged, particularly in the brain of young-aged, particularly in the brain of middle-aged
subjects, with Down’s syndrome.
In one embodiment, the antigenic peptide as described herein is derived
from an amyloid protein or amyloid-like protein selected from the group consisting of
prion protein, tau protein, alpha-synuclein, huntingtin and amyloid-β or a
combination of one or more of the above peptides.
Said Aβ antigenic peptide fragment corresponds in one embodiment of
the disclosure to the N-terminal part of the Aβ peptide, particularly to the N-terminal
part comprising at least 5, particularly at least 6, particularly at least 7, particularly at
least 8, particularly at least 9, particularly at least 10, particularly at least 11,
particularly at least 12, particularly at least 13, particularly at least 14, particularly all,
amino acid residues from the Aβ1-15 fragment.
In one embodiment, the Aβ antigenic peptide fragment corresponds to the
N-terminal part of the Aβ peptide comprising at least 5, particularly at least 6,
particularly at least 7, particularly at least 8, particularly at least 9, particularly at
least 10, particularly at least 11, particularly at least 12, particularly at least 13,
particularly at least 14, particularly at least 15, particularly all, amino acid residues
from the Aβ1-16 fragment, the Aβ1-17 fragment, the Aβ1-18 fragment, the Aβ1-19
fragment, the Aβ1-20 fragment, the Aβ1-22 fragment, the Aβ1-23 fragment, the
Aβ1-24 fragment, the Aβ1-25 fragment or, the Aβ1-26 fragment, or the 3-15 Aβ
fragment.
In one embodiment, the Aβ antigenic peptide fragment corresponds to the
C-terminal part of the Aβ peptide comprising at least 5, particularly at least 6,
particularly at least 7, particularly at least 8, particularly at least 9, particularly at
least 10, particularly at least 11, particularly at least 12, particularly at least 13,
particularly at least 14, particularly at least 15, particularly all amino acid residues
from the Aβ20-40 or Aβ20-42 fragment, the Aβ21-40 or Aβ21-42 fragment, the
Aβ22-40 or Aβ22-42 fragment, the Aβ23-40 or Aβ23-42 fragment, the Aβ24-40 or
Aβ24-42 fragment, the Aβ25-40 or Aβ25-42 fragment, the Aβ26-46 or Aβ27-42
fragment, or the Aβ27-40 or Aβ27-42 fragment, or the Aβ29-40 .
In one embodiment, the Aβ antigenic peptide fragment corresponds to the
middle part of the Aβ peptide comprising at least 5, particularly at least 6,
particularly at least 7, particularly at least 8, particularly at least 9, particularly at
least 10, particularly at least 11, particularly at least 12, particularly at least 13,
particularly at least 14, particularly at least 15, particularly all amino acid residues
from the Aβ15-35, particularly the Aβ20-35 fragment.
In another embodiment, the Aβ antigenic peptide fragment corresponds to
the central part of the Aβ peptide, particularly the Aβ14-29 fragment.
In a specific embodiment the Aβ antigenic peptide fragment corresponds
to the C-terminal part of the Aβ peptide, particularly the C-terminal Aβ22-35
fragment.
In another embodiment, the full length Aβ1-39, Aβ1-40, or Aβ1-42
fragment may be used.
In certain embodiments, the Aβ antigenic peptide fragment as disclosed
herein may contain one or more modified or non-natural amino acid residues.
In certain embodiments, the use of Aβ antigenic peptide fragments is
contemplated, which are not fragments consisting of a single or repetitive stretch of
between 13 and 15 contiguous amino acid residues from the N-terminal part of the
Aβ peptide, particularly not fragments, wherein said contiguous stretch of 13 to 15
amino acid residues is obtained from the N-terminal fragment 1-16 or 1-17 of the Aβ
peptide, particularly from the N-terminal part of the Aβ peptide selected from the
group consisting of residues 1-15, 1-14, and 1-13, particularly consisting of Aβ
1-15
peptide antigen as given in SEQ ID NO: 1 and Aβ as given in SEQ ID NO: 3
1-16(∆14)
disclosed in . In one embodiment, the antigenic peptide as
described herein is presented reconstituted in a carrier such as, for example, a
vesicle, a particulate body or molecule, but particularly reconstituted in a liposome.
In one embodiment, the antigenic peptide as described herein is
presented in a single or repetitive array on the surface of the carrier or liposome.
In one embodiment, said highly repetitive array on the surface of the
carrier or liposome comprises at least 10 repetitive antigenic units/carrier molecule,
particularly at least 50 repetitive antigenic units/carrier molecule, particularly at least
100 repetitive antigenic units/carrier molecule, particularly at least 200 repetitive
antigenic units/carrier molecule, particularly at least 300 repetitive antigenic
units/carrier molecule; particularly at least 400 repetitive antigenic units/carrier
molecule, particularly at least 500 repetitive antigenic units/carrier molecule.
The antigenic composition as described herein may comprise a
conformational antigen, particularly an Aβ antigen as described herein, wherein
more than 30%, particularly more than 40%, particularly more than 50%, particularly
more than 60%, particularly more than 70%, particularly more than 80%, particularly
more than 90%, particularly more than 95% and up to 100% is in a beta-sheet
conformation.
In one embodiment the composition comprises the antigenic peptide as
described herein in the various embodiments in a therapeutically effective amount
together with a pharmaceutically acceptable carrier and/or excipient for use in the
treatment, and/or alleviation and/or prevention of memory impairments or
abnormalities in subjects with Down’s syndrome.
In another embodiment, the composition comprises the antigenic peptide
as described herein in the various embodiments in a therapeutically effective
amount together with a pharmaceutically acceptable carrier and/or excipient, for use
in the treatment, and/or alleviation and/or prevention of cognitive impairments or
abnormalities in subjects with Down’s syndrome.
In particular said memory and/or cognitive impairments originate in the
hippocampus and/or the prefrontal cortex and/or the entorhinal cortex of the brain.
In various specific embodiments the composition comprises the antigenic
peptide as described herein in the various embodiments in a therapeutically
effective amount together with a pharmaceutically acceptable carrier and/or
excipient for use in the treatment and/or alleviation and/or prevention of impairments
of recognition memory and/or impairments of the contextual associative memory
and/or impairments of the associative learning and/or impairments of the declarative
memory for facts and events and/or episodic memory impairments and/or language
dysfunction such as aphasia and/or visuospatial impairments, such as
misplacement of items and difficulty navigating in unfamiliar and familiar terrain
and/or decreased executive functions, such as apathy, disinhibition, social isolation,
poor judgment, difficulties with planning and/or poor abstract reasoning and/or
personality changes and/or emotional changes such as apathy, agitation and
psychosis and/or apraxia and/or impairments of performing learned motor tasks
and/or other neurological signs including pyramidal and extrapyramidal findings as
well as myoclonus or seizures.
In particular said impairment or abnormality concerns the recognition
memory.
In a specific embodiment, said antigenic peptide for use in enhancing or
restoring recognition memory in subjects with Down’s syndrome is derived from the
N-terminal part of the Aβ peptide, particularly said antigenic peptide consists of all or
part of amino acid residues Aβ1-15, Aβ1-16, Aβ1-17, Aβ1-18, Aβ1-19, Aβ1-20, Aβ1-
22, or Aβ1-23.
In particular said antigenic peptide consists of amino acid residues Aβ1-
In another specific embodiment, said antigenic peptide for use in
enhancing or restoring recognition memory in subjects with Down’s syndrome is
derived from the N-terminal part of the Aβ peptide, particularly said antigenic peptide
consists of all or part of amino acid residues
In particular said impairment or abnormality concerns the contextual
associative memory.
In a specific embodiment, said antigenic peptide for use in enhancing or
restoring contextual associative memory in subjects with Down’s syndrome is
derived from the N-terminal part of the Aβ peptide, particularly said antigenic peptide
consists of all or part of amino acid residues Aβ1-15, Aβ1-16, Aβ1-17, Aβ1-18,
Aβ1-19, Aβ1-20, Aβ1-22, or Aβ1-23.
In particular said antigenic peptide consists of amino acid residues Aβ1-
In particular said impairment or abnormality concerns the associative
learning.
In a specific embodiment, said antigenic peptide for use in enhancing or
restoring associative learning in subjects with Down’s syndrome is derived from the
N-terminal part of the Aβ peptide, particularly said antigenic peptide consists of all or
part of amino acid residues Aβ1-15, Aβ1-16, Aβ1-17, Aβ1-18, Aβ1-19, Aβ1-20, Aβ1-
22, or Aβ1-23.
In particular said antigenic peptide consists of amino acid residues Aβ1-
In another specific embodiment, said antigenic peptide for use in
enhancing or restoring associative learning in subjects with Down’s syndrome is
derived from the C-terminal part of the Aβ peptide, particularly said antigenic peptide
consists of all or part of amino acid residues Aβ20-36, Aβ20-40, Aβ20-42, Aβ21-36,
Aβ21-40, Aβ21-42, Aβ22-36, Aβ22-40 or Aβ22-42.
In particular said antigenic peptide consists of amino acid residues Aβ22-
The formation and stabilization of the desired conformation of the
antigenic peptide may be achieved by presenting the antigenic peptide attached to,
or incorporated or reconstituted, partially or fully, into a carrier such as, for example,
a vesicle, a particulate body or molecule or any other means that can suitably serve
as a carrier/adjuvant for the antigenic peptide. In a specific embodiment of the
invention, the antigenic peptide is attached to, or incorporated or reconstituted in the
carrier through weak interactions such as, for example, van der Waal´s, hydrophobic
or electrostatic interaction, or a combination of two or more of said interactions, such
that the peptide is presented on the carrier surface with a specific conformation,
which is maintained and stabilized by restricting said antigenic peptide in its three
dimensional freedom of movement so that conformational changes are prevented or
severely restricted.
When a vesicle, a particle or a particulate body is used as a
carrier/adjuvant such as, for example, a liposome, the composition of the antigenic
peptide may be chosen such that its overall net charge is identical to that of the
carrier/adjuvant surface to which the peptide is attached. Electrostatic repulsion
forces being effective between the identically charged carrier/adjuvant surface and
the antigenic peptide, but particularly the identically charged carrier surface and the
amino acid residues constituting the antigenic peptide and more particularly the
identically charged carrier surface and the identically charged amino acid residues
comprised in the antigenic peptide, may lead to the antigenic peptide taking on a
defined, highly specific and stabilized conformation which guarantees a high
biological activity. As a result, the antigenic peptide is exposed and presented in a
conformation that is highly biologically active in that it allows the immune system of
the target organism to freely interact with the antigenic determinants contained in
the antigenic construct in the biologically active conformation, which leads to a
strong and conformation-specific immune response, resulting in, for example, a high
antibody titer in the target organism.
The immunogenic response may be further increased by using a liposome
as a carrier, which liposome may function as an adjuvant to increase or stimulate
the immune response within the target animal or human to be treated with the
therapeutic vaccine described herein. Optionally, the liposome may, in addition,
contain a further adjuvant such as, for example, lipid A, alum, calcium phosphate,
interleukin 1, and/or microcapsules of polysaccharides and proteins, but particularly
a detoxified lipid A, such as monophosphoryl or diphosphoryl lipid A, or any other
adjuvant that can be suitably used within the scope of the present disclosure such
as, for example, alum, calcium phosphate, interleukin 1, and/or microcapsules of
polysaccharides and proteins, LPS, CpG ODN, Pam2CSK4, Pam3CSK4, dsRNA,
ssRNA, muramyl dipeptide, Quil Q, QS-21.
Liposomes that can be used in the compositions include those known to
one skilled in the art. Any of the standard lipids useful for making liposomes may be
used. Standard bilayer and multi-layer liposomes may be used to make
compositions described herein. While any method of making liposomes known to
one skilled in the art may be used, the most preferred liposomes are made
according to the method of Alving et al., Infect. Immun. 60:2438-2444, 1992, (Lauer
et al., 1992) hereby incorporated by reference. The liposome may have a dual
function in that it can be used as a carrier comprising the supramolecular construct
as described herein before and, at the same time, function as an adjuvant to
increase or stimulate the immune response within the target animal or human to be
treated with the therapeutic vaccine described herein. Optionally, the liposome may,
in addition, contain a further adjuvant or and immunomodulator or both such as, for
example, lipid A, alum, calcium phosphate, interleukin 1, and/or microcapsules of
polysaccharides and proteins, but particularly a lipid A, more particularly a detoxified
lipid A, such as monophosphoryl or diphosphoryl lipid A, or alum.
The liposome may be composed of constituents selected from the group
consisting of dimyristoyl phosphatidyl choline (DMPC), dimyristoyl phosphatidyl
ethanolamine (DMPEA), dimyristoyl phosphatidyl glycerol (DMPG) and cholesterol.
In one embodiment, contemplated herein is using as a replacement for
cationic lipids in the liposomal membrane, anionic lipids selected from the group
consisting of:
a. diacyl-phospholipids with headgroups phosphatidyl glycerol, phosphatidyl
serine, phosphatidyl inositol, L-α-phosphatidylinositolphosphate or
phosphatidic acid;
b. lyso-phospholipids with headgroups phosphatidyl glycerol, phosphatidyl serine
or phosphatidic acid, and
c. cardiolipin, dilyso-cardiolipin, monolyso-cardiolipin
In one aspect, contemplated herein is using as a replacement for anionic
lipids in the liposomal membrane, cationic lipids selected from the group consisting
a. diacyl-phospholipids with headgroups 3-trimethylammonium-propane ,3-
dimethylammonium-propane, 3-ethylphosphocholine or 3-phosphatidylethanol-
amine; and
b. D-erythro-sphingosine, dimethyldioctadecylammonium bromide, N-[1-(2, 3-
dimyristyloxy)propyl]-N,N-dimethyl-N-(2-hydroxyethyl) ammonium bromide,
N,N,N-trimethylbis[(1-oxooctadecenyl)oxy]-(Z,Z)propanaminium methyl
sulfate or 3β-[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol hydro-
chloride.
In one embodiment, the lipid chains attached to the above headgroups
a. be saturated or unsaturated,
b. vary in length from (CH ) wherein n is between 3 and 24, and
be symmetrically or asymmetrically substituted.
In one embodiment, the antigenic composition as described herein,
comprises a liposome preparation and an adjuvant, particularly lipid A, alum,
calcium phosphate, interleukin 1, and/or microcapsules of polysaccharides and
proteins, but particularly a lipid A, more particularly a detoxified lipid A, such as
monophosphoryl or diphosphoryl lipid A, or alum.
The liposomal composition comprising an Aβ peptide coupled with
lipophilic moieties embedded into liposomes as described herein may be prepared
according to the methodology disclosed herein.
When liposomes are used as a carrier/adjuvant, the antigenic peptide as
described herein may further be modified by coupling to a lipophilic or hydrophobic
moiety that facilitates insertion into the lipid bilayer of the liposome carrier/adjuvant.
Said hydrophobic moiety may be a fatty acid, a triglyceride, a diglyceride, a steroid,
a sphingolipid, a glycolipid or a phospholipid.
In a specific embodiment, the the lipophilic or hydrophobic moiety is an
alkyl group or a fatty acid with a carbon backbone of at least 1 carbon atom,
particularly of at least 2 carbon atoms, particularly of at least 3 carbon atoms,
particularly of at least 4 carbon atoms, particularly of at least 6 carbon atoms,
particularly of at least 8 carbon atoms, particularly of at least 12 carbon atoms,
particularly of at least 16 carbon atoms.
The lipophilic or hydrophobic moieties may be fatty acids, triglycerides
and phospholipids, wherein the fatty acid carbon back bone has at least 4 carbon
atoms particularly lipophilic moieties having fatty acids with a carbon backbone of at
least approximately 14 carbon atoms and up to approximately 24 carbon atoms,
more particularly hydrophobic moieties having a carbon backbone of at least 14
carbon atoms. Examples of hydrophobic moieties include, but are not limited to,
palmitic acid, stearic acid, myristic acid, lauric acid, oleic acid, linoleic acid, linolenic
acid and cholesterol or DSPE. In a specific embodiment of the invention the
hydrophobic moiety is palmitic acid.
In a specific embodiment, the antigenic composition as described herein
comprises a peptide antigen comprising two palmitic acid moieties, particularly four
palmitic acid moieties.
Palmitoylation, while providing an anchor for the peptide in the liposome
bilayer, due to the relative reduced length of the C fatty acid moiety leads to the
16:0
peptide being presented exposed on or in close proximity to the liposome surface.
Therefore, the cells processing the antigen will have to take up the entire liposome
with the peptide.
The antigenic constructs described herein may, in one embodiment,
comprise peptides modified via pegylation (using polyethylene glycol (PEG) or
modified polyethylene glycol), or modified via other methods such by palmitic acid
as described herein before, poly-amino acids (eg poly-glycine, poly-histidine), poly-
saccharides (eg polygalacturonic acid, polylactic acid, polyglycolide, chitin,
chitosan), synthetic polymers (polyamides, polyurethanes, polyesters) or co-
polymers (eg. poly(methacrylic acid) and N-(2-hydroxy) propyl methacrylamide) and
the like.
If PEG is used in the preparation of the antigenic construct, the free PEG
terminus may be covalently attached to a molecule of phosphatidylethanolamine
(where the fatty acid can be: myristic, palmitic, stearic, oleic etc. or combination
thereof). This supramolecular structure may be reconstituted in liposomes consisting
of phospholipids and cholesterol (phosphatidylethanol amine, phosphatidyl glycerol,
cholesterol in varied molar ratios). Other phospholipids can be used. Lipid A may be
used at a concentration of approximately 40 µg/pmole of phospholipids.
In certain embodiments, the antigenic constructs comprise an antigenic peptide
sequence as described herein before, covalently attached to pegylated lysine at
least one at each terminus, but particularly 1 or 2 at each terminus. The length of
the PEG (polyethylenglycol) chain may vary from n = 8 to n = 150.000 or more,
particularly from n = 10 to n = 80.000, more particularly from n = 10 to n = 10.000.
In a specific embodiment of the invention the length of the PEG chain is not more
than n = 45, particularly between n = 5 and n = 40, more particularly between n = 10
and n = 30, and even more particularly n = 10.
In a further embodiment of the invention the Aβ peptide antigen is
modified by two palmitic acids bound to the N- and/or C-terminus of the peptide
molecule.
In certain embodiment, the present disclosure contemplates post-insertion
of different peptide (e.g. antigen) types and/or adjuvant types to the external layer of
preformed liposomes in different concentrations as described in EP application no
18 8832, the disclosure of which is incorporated herein in its entirety. This post-
insertion method comprises pre-forming of liposomes in solution and modification of
antigenic peptides through hydrophobic moieties such that the modified antigenic
peptide is available in a micellar form. The method further comprises releasing of
the antigenic peptides from the micelles by inducing micellar breakdown followed by
integration into the pre-formed liposome. This integration process is driven by
hydrophobic interactions of the modified antigen and/or the adjuvant with the
(phospho)lipid bilayer of the liposomes. In particular, the solubilizing of the modified
antigenic peptide and/or adjuvant into the external layer of liposomes is
accomplished without the aid of any chemical reaction or additional molecule
modification, by diluting the solubilized antigenic peptide or adjuvant (initially
presented in micellar form), below the critical micellar concentration of the
surfactant. The free form of the antigenic peptide or adjuvant is then integrated in
the external layer of the liposomes due to the solubilization of their hydrophobic
domains in the acyl moiety of the phospholipids. Thus, the method provides for a
stock of “empty liposomes” being disposable for loading according to the respective
needs.
In particular, this post-insertion method for preparing a liposome-based
antigenic construct comprising an antigenic peptide as described herein in the
various embodiments modified through hydrophobic moieties reconstituted in a
liposome, comprises the steps of i) preparing liposomes in solution; ii) preparing a
modified antigenic peptide by adding to the N- and/or C-terminus of the peptide
molecule at least one hydrophobic moiety; iii) solubilizing the modified antigenic
peptide in the presence of a surfactant; iv) diluting the solubilized peptide and,
optionally, an adjuvant below the critical micellar concentration (CMC) of the
surfactant; and v) loading the preformed liposomes with the diluted, solubilized
antigenic peptide and, optionally, the adjuvant, by adding said antigenic peptide
and, optionally, said adjuvant to the preformed liposomal preparation and
solubilizing the added peptide and, optionally, the added adjuvant into the external
layer of the liposomes, particularly without the aid of any chemical reaction or
additional molecule modification.
As used herein, the term “critical micellar concentration”, also known as
CMC, is defined as the concentration of surfactants above which micelles are
spontaneously formed. Upon introduction of surfactants (or any surface active
materials) into a system the surfactants will initially partition into the interface, thus
reducing the systems free energy by a) lowering the energy of the interface
(calculated as area x surface tension) and b) by removing the hydrophobic parts of
the surfactant from contacts with water. Subsequently, when the surface coverage
by the surfactants increases and the surface free energy (surface tension)
decreases and the surfactants start aggregating into micelles, thus again decreasing
the system´s free energy by decreasing the contact area of hydrophobic parts of the
surfactant with water. Upon reaching the CMC, any further addition of surfactants
will just increase the number of micelles. (IUPAC. Compendium of Chemical
Terminology, 2nd ed. Blackwell Scientific Publications, Oxford (1997)).
Advantageously, this method leads to high yields of peptide and/or
adjuvant incorporation with a unique molecular display on the liposome facing the
external layer of the liposome bilayer. In particular, at least 75%, 76%, 77%, 78%,
79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 89%, 90%, 91%, 92%, 93%,
94%, 95%, 96%, 97%, 98%, 99% and up to 100% of the reconstituted antigenic
peptide is present on the surface of the liposome, inserted into the lipid bilayer
through its hydrophobic moieties.
The method further results in liposome preparations which show a
homogenous size distribution with a polydispersity index in the range of between 0.4
and 0.6, particularly of 0.45 to 0.55, particularly of 0.5. Further, the method and
constructs allow for a high bioavailability of peptide and/or adjuvant for the immune
system and, as a consequence, an improved immune response. Adjuvant
degradation, e.g. MPLA degradation, is minimized or not present at all and, thus, an
increased batch reproducibility is provided. The constructs prepared by the above
method are stable, capable of sterile filtration and do not induce side immune
responses.
Accordingly, in certain embodiments an antigenic construct is described
comprising an antigenic peptide of the invention for use as described herein in the
various embodiments reconstituted in a liposome, wherein at least 75%, 76%, 77%,
78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 89%, 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, 99% and up to 100% of the reconstituted
antigenic peptide is present on the surface of the liposome, inserted into the lipid
bilayer through its hydrophobic moieties.
In still other embodiments, a liposomal preparation is described
comprising said antigenic construct, which preparation shows a homogenous size
distribution with a polydispersity index in the range of between 0.4 and 0.6,
particularly of 0.45 to 0.55, particularly of 0.5.
In certain embodiments, the antigenic peptide is presented on the surface
of the carrier molecule in a highly repetitive array, particularly a repetitive array
comprising at least 10 repetitive antigenic units/carrier molecule, particularly at least
50 repetitive antigenic units/carrier molecule, particularly at least 100 repetitive
antigenic units/carrier molecule, particularly at least 200 repetitive antigenic
units/carrier molecule, particularly at least 300 repetitive antigenic units/carrier
molecule; particularly at least 400 repetitive antigenic units/carrier molecule,
particularly at least 500 repetitive antigenic units/carrier molecule.
In certain embodiments, an antibody may be used in the methods as
described herein for the treatment, and/or alleviation and/or prevention of memory
and/or cognitive impairments or abnormalities, particularly of AD-like memory and/or
cognitive impairments or abnormalities, particularly memory and/or cognitive
impairments originating in the hippocampus and/or the prefrontal cortex and/or the
entorhinal cortex of the brain, particularly impairments of the recognition memory
and/or impairments of the contextual associative memory and/or impairments of the
associative learning and/or impairments of the declarative memory for facts and
events and/or episodic memory impairments and/or language dysfunction such as
aphasia and/or visuospatial impairments, such as misplacement of items and
difficulty navigating in unfamiliar and familiar terrain and/or decreased executive
functions, such as apathy, disinhibition, social isolation, poor judgment, difficulties
with planning and/or poor abstract reasoning and/or personality changes and/or
emotional changes such as apathy, agitation and psychosis and/or apraxia and/or
impairments of performing learned motor tasks and/or other neurological signs
including pyramidal and extrapyramidal findings as well as myoclonus or seizures, in
children and young to middle-aged subjects with Down’s syndrome, which antibody
has been generated in response to any one of the antigenic constructs disclosed
herein. This antibody may be a polyclonal antibody, a monoclonal antibody, a
humanized antibody, a fully human antibody, a diabody, a camelid antibody or a
functional fragment of any of the foregoing antibodies, which fragment has
substantially the same biological activity in terms of treatment of memory and/or
cognitive impairments or abnormalities as the antibody from which said fragment is
derived.
In certain embodiments, an antibody may be used in the methods as
described herein for the treatment, and/or alleviation and/or prevention of memory
and/or cognitive impairments or abnormalities, particularly of AD-like memory and/or
cognitive impairments or abnormalities, particularly memory and/or cognitive
impairments originating in the hippocampus and/or the prefrontal cortex and/or the
entorhinal cortex of the brain, particularly impairments of the recognition memory
and/or impairments of the contextual associative memory and/or impairments of the
associative learning and/or impairments of the declarative memory for facts and
events and/or episodic memory impairments and/or language dysfunction such as
aphasia and/or visuospatial impairments, such as misplacement of items and
difficulty navigating in unfamiliar and familiar terrain and/or decreased executive
functions, such as apathy, disinhibition, social isolation, poor judgment, difficulties
with planning and/or poor abstract reasoning and/or personality changes and/or
emotional changes such as apathy, agitation and psychosis and/or apraxia and/or
impairments of performing learned motor tasks and/or other neurological signs
including pyramidal and extrapyramidal findings as well as myoclonus or seizures, in
children with Down’s syndrome, which antibody has been generated in response to
any one of the antigenic constructs disclosed herein. This antibody may be a
polyclonal antibody, a monoclonal antibody, a humanized antibody, a fully human
antibody, a diabody, a camelid antibody or a functional fragment of any of the
foregoing antibodies, which fragment has substantially the same biological activity in
terms of treatment of memory and/or cognitive impairments or abnormalities as the
antibody from which said fragment is derived.
In certain embodiments, an antibody may be used in the methods as
described herein for the treatment, and/or alleviation and/or prevention of memory
and/or cognitive impairments or abnormalities, particularly of AD-like memory and/or
cognitive impairments or abnormalities, particularly memory and/or cognitive
impairments originating in the hippocampus of the brain, particularly impairments of
the recognition memory and/or impairments of the contextual associative memory
and/or impairments of the associative learning and/or impairments of the declarative
memory for facts and events and/or episodic memory impairments and/or language
dysfunction such as aphasia and/or visuospatial impairments, such as
misplacement of items and difficulty navigating in unfamiliar and familiar terrain
and/or decreased executive functions, such as apathy, disinhibition, social isolation,
poor judgment, difficulties with planning and/or poor abstract reasoning and/or
personality changes and/or emotional changes such as apathy, agitation and
psychosis and/or apraxia and/or impairments of performing learned motor tasks
and/or other neurological signs including pyramidal and extrapyramidal findings as
well as myoclonus or seizures, in children subjects with Down’s syndrome, which
antibody has been generated in response to any one of the antigenic constructs
disclosed herein. This antibody may be a polyclonal antibody, a monoclonal
antibody, a humanized antibody, a fully human antibody, a diabody, a camelid
antibody or a functional fragment of any of the foregoing antibodies, which fragment
has substantially the same biological activity in terms of treatment of memory and/or
cognitive impairments or abnormalities as the antibody from which said fragment is
derived.
In another certain embodiment the antigenic peptide fragment derived
from amyloid protein or amyloid-like protein is used in a method as disclosed herein
for the treatment and/or alleviating and/or prevention of memory and/or cognitive
impairments originating in the hippocampus and/or the prefrontal cortex and/or the
entorhinal cortex of the brain.
In one embodiment, described is a method for treating and/or alleviating
and/or preventing memory impairments or abnormalities in a subject with Down’s
syndrome comprising administering to said subject an antigenic peptide or a
composition as described herein in the various embodiments.
In another embodiment, described is a method for treating and/or
alleviating and/ or preventing cognitive impairments or abnormalities in a subject
with Down’s syndrome comprising administering to said subject an antigenic peptide
or a composition as described herein in the various embodiments.
In particular said impairment or abnormality concerns the recognition
memory.
In a specific embodiment, said antigenic peptide for use in a method of
enhancing or restoring recognition memory in subjects with Down’s syndrome is
derived from the N-terminal part of the Aβ peptide, particularly said antigenic peptide
consists of all or part of amino acid residues Aβ1-15, Aβ1-16, Aβ1-17, Aβ1-18,
Aβ1-19, Aβ1-20, Aβ1-22, or Aβ1-23.
In particular said antigenic peptide consists of amino acid residues Aβ1-
In another specific embodiment, said antigenic peptide for use in a
method of enhancing or restoring recognition memory in subjects with Down’s
syndrome is derived from the N-terminal part of the Aβ peptide, particularly said
antigenic peptide consists of all or part of amino acid residues
In particular said impairment or abnormality concerns the contextual
associative memory.
In a specific embodiment, said antigenic peptide for use in a method of
enhancing or restoring contextual associative memory in subjects with Down’s
syndrome is derived from the N-terminal part of the Aβ peptide, particularly said
antigenic peptide consists of all or part of amino acid residues Aβ1-15, Aβ1-16, Aβ1-
17, Aβ1-18, Aβ1-19, Aβ1-20, Aβ1-22, or Aβ1-23.
In particular said antigenic peptide consists of amino acid residues Aβ1-
In particular said impairment or abnormality concerns the associative
learning.
In a specific embodiment, said antigenic peptide for use in a method of
enhancing or restoring associative learning in subjects with Down’s syndrome is
derived from the N-terminal part of the Aβ peptide, particularly said antigenic peptide
consists of all or part of amino acid residues Aβ1-15, Aβ1-16, Aβ1-17, Aβ1-18, Aβ1-
19, Aβ1-20, Aβ1-22, or Aβ1-23.
In another exemplary embodiment, the active agent is an immunotherapy
agent. Non-limiting examples of immunotherapy agents, include inflammatory
agents, biological factors, immune regulatory proteins, human and humanized
antibodies, and immunotherapy drugs, such as AZT and other derivatized or
modified nucleotides. Small molecules can also be employed as agents in the
present invention.
In particular said antigenic peptide consists of amino acid residues Aβ1-15.
In another specific embodiment, said antigenic peptide for use in a method
of enhancing or restoring associative learning in subjects with Down’s syndrome is
derived from the C-terminal part of the Aβ peptide, particularly said antigenic peptide
consists of all or part of amino acid residues Aβ20-36, Aβ20-40, Aβ20-42, Aβ21-36,
Aβ21-40, Aβ21-42, Aβ22-36, Aβ22-40 or Aβ22-42.
In particular said antigenic peptide consists of amino acid residues Aβ22-
Antigenic peptides which are particularly useful for enhancing and/or
restoring associative learning in subjects with Down’s syndrome such as, for
example antigenic peptide consisting of amino acid residues Aβ22-35, may be
combined with one or more biologically active compounds, which have a positive
effect on recognition memory and/or contextual associative memory.
In certain embodiments said subject with Down’s Syndrome, is a young to
middle-aged subject.
In certain embodiments said subject is a young-aged subject.
In still other certain embodiments said subject is an children-aged subject.
In particular, said subjects are below age 60, particularly below age 55,
particularly below age 50, particularly below age 45, particularly below 40,
particularly below 35, particularly below 30, particularly below 25, particularly below
, particularly below 15, particularly below age 10, particularly below age 5,
particularly below age 3.
The terms "antibody” or “antibodies" as used herein is an art recognized
term and is understood to refer to molecules or active fragments of molecules that
bind to known antigens, particularly to immunoglobulin molecules and to
immunologically active portions of immunoglobulin molecules, i.e molecules that
contain a binding site that immunospecifically binds an antigen. The immunoglobulin
according to the invention can be of any type (IgG, IgM, IgD, IgE, IgA and IgY) or
class (IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) or subclasses of immunoglobulin
molecule.
“Antibodies” are intended within the scope of the present invention to
include monoclonal antibodies, polyclonal, chimeric, single chain, bispecific,
simianized, human and humanized antibodies as well as active fragments thereof.
Examples of active fragments of molecules that bind to antigens include Fab and
F(ab') fragments, including the products of an Fab immunoglobulin expression
library and epitope-binding fragments of any of the antibodies and fragments
mentioned above.
These active fragments can be derived from an antibody described herein
by a number of techniques. For example, purified monoclonal antibodies can be
cleaved with an enzyme, such as pepsin, and subjected to HPLC gel filtration. The
appropriate fraction containing Fab fragments can then be collected and
concentrated by membrane filtration and the like. For further description of general
techniques for the isolation of active fragments of antibodies, see for example,
Khaw, B. A. et al. J. Nucl. Med. 23:1011-1019 (1982); Rousseaux et al. Methods
Enzymology, 121:663-69, Academic Press, 1986.
A "humanized antibody" refers to a type of engineered antibody having its
CDRs derived from a non-human donor immunoglobulin, the remaining
immunoglobulin-derived parts of the molecule being derived from one (or more)
human immunoglobulin(s). In addition, framework support residues may be altered
to preserve binding affinity. Methods to obtain “humanized antibodies” are well
known to those skilled in the art. (see, e.g., Queen et al., Proc. Natl Acad Sci USA,
86:10029-10032 (1989), Hodgson et al., Bio/Technoloy, 9:421 (1991)).
A "humanized antibody" may also be obtained by a novel genetic
engineering approach that enables production of affinity-matured humanlike
polyclonal antibodies in large animals such as, for example, rabbits.
The term “monoclonal antibody” is also well recognized in the art and
refers to an antibody that is mass produced in the laboratory from a single clone and
that recognizes only one antigen. Monoclonal antibodies are typically made by
fusing a normally short-lived, antibody-producing B cell to a fast-growing cell, such
as a cancer cell (sometimes referred to as an “immortal” cell). The resulting hybrid
cell, or hybridoma, multiplies rapidly, creating a clone that produces large quantities
of the antibody.
The term “memory and/or cognitive impairments and abnormalities” refers
mainly to clinical symptoms associated with amyloid-related pathology in subjects
with Down’s syndrome (DS), but particularly to impairments and abnormalities
originating in the hippocampus, the prefrontal cortex and/or the entorhinal cortex of
the brain. Examples of such memory and/or cognitive impairments are i.e., but
without being limited thereto, impairments of recognition memory and/or
impairments of the contextual associative memory and/or impairments of the
associative learning and/or impairments of the declarative memory for facts and
events and/or episodic memory impairments and/or language dysfunction such as
aphasia and/or visuospatial impairments, such as misplacement of items and
difficulty navigating in unfamiliar and familiar terrain and/or decreased executive
functions, such as apathy, disinhibition, social isolation, poor judgment, difficulties
with planning and/or poor abstract reasoning and/or personality changes and/or
emotional changes such as apathy, agitation and psychosis and/or apraxia and/or
impairments of performing learned motor tasks and/or other neurological signs
including pyramidal and extrapyramidal findings as well as myoclonus or seizures.
The term “memory impairment” further refers to the cardinal feature of DS
and is often its earliest manifestation. Even when not the primary complaint,
memory deficits can be elicited in most patients with DS at the time of presentation.
The pattern of memory impairment in DS is quite distinctive. Declarative memory for
facts and events, which depends on medial temporal structures such as the
entorhinal cortex and hippocampus, is profoundly affected in DS, while subcortical
systems supporting procedural memory are relatively spared until quite late in the
course of the disease.
Episodic memory is more profoundly impaired in young DS patients,
compared with memory for facts such as vocabulary and concepts (semantic
memory), which often becomes impaired somewhat later. Within episodic memory,
there is a distinction between immediate recall (eg, mental rehearsal of a phone
number), memory for recent events (which comes into play once material that has
departed from consciousness must be recalled), and memory of more remote
events. Memory for recent events, served by the hippocampus, entorhinal cortex in
the medial temporal lobe, is prominently impaired in young DS patients. In contrast,
immediate memory (encoded in the prefrontal cortices) is spared early on, as are
memories that are consolidated for long periods of time (years), which can be
recalled in the absence of hippocampal functioning.
Impairments of procedural memory appear only in middle-aged DS
patients. Dysfunction in language, executive function and other cognitive domains
develop at variable rates of over the course of the disease. The heterogeneity of
the clinical presentation of DS presumably reflects a variable topographical
distribution of the burden of brain pathology. Language dysfunction, that is aphasia,
is a common symptom of DS, and the first manifestations of language dysfunction
usually include word-finding difficulties, circumlocution, and reduced vocabulary in
spontaneous speech and with anomia on confrontational naming tests. This can
progress to include paraphasic errors, impoverished speech content, and impaired
comprehension. However, patients can usually repeat phrases verbatim until the
disease is quite advanced. The language difficulties in DS have often been
described as Wernicke’s and Broca’s type of aphasia. When asked to generate
word lists in one minute's time, patients with DS perform significantly worse on a
category fluency test (eg, lists of animals) than on a letter fluency test (eg, lists of
words beginning with F). This reflects the specific deficit in semantic memory.
Loss of visuospatial skills is another early feature of DS that can
sometimes be quite prominent at presentation. Visuospatial impairments manifest
as misplacement of items and difficulty navigating in first unfamiliar then familiar
terrain. Visual agnosia (inability to recognize objects) and prosopagnosia (inability to
recognize faces) are later features.
Impairment in executive function is also seen in patients with DS. These
symptoms include poor insight, and a reduced ability for abstract reasoning. As the
disease progresses, personality changes (such as apathy, social disengagement,
and disinhibition), poor judgment and planning, and an inability to complete tasks
typically emerges. Superimposed depression, which can be difficult to diagnose in
the setting of dementia can also present in this manner.
Reduced insight into deficits (anosognosia) is another characteristic
feature of DS. It is not uncommon for patients to deny or underestimate their deficits
and offer explanations for them when they are pointed out. Interviewing a collateral
historian, such as a spouse is critical to obtain an accurate history. In fact, it is often
the family member who brings the complaint of cognitive impairment to medical
attention. Loss of insight increases over time along with overall disease severity,
and can be associated with behavioral disturbances; those with relatively preserved
insight are more likely to be depressed, while those with more impaired insight are
likely to be agitated, disinhibited, and exhibit psychotic features. The emergence of
behavioral disturbances, including agitation, aggression, wandering, and psychosis
(hallucinations, delusions, misidentification syndromes) can lead to significant
distress for the patient and family members, and is typically seen in later stages of
Younger patients with DS generally have a normal neurologic examination
except for the cognitive examination. While pyramidal and extrapyramidal motor
signs, myoclonus, and seizures do occur in patients with DS, these are typically
late-stage findings . Similarly, frontal release signs (grasp, snout reflexes,
gegenhalten) and incontinence are late, rather than early, features of DS.
The immunogenic composition described herein, which, in one
embodiment, may be a therapeutic vaccine, comprises the antigenic construct as
described herein before in a therapeutically or prophylactically effective amount and
may be prepared as a liquid solution, or as an injectable suspension, or else in a
solid form suitable for solubilization prior to injection in the context of, for example, a
kit for making use of the composition, as described below.
A "therapeutically or prophylactically effective amount" refers to the
amount of antibody, peptide, compound or pharmaceutical composition which, when
administered to a human or animal, leads to a therapeutic or prophylactic effect in
said human or animal. The effective amount is readily determined by one of skill in
the art following routine procedures.
The immunogenic composition described herein may be administered to a
human or animal, particularly to a human or animal with Down’s syndrome suffering
from AD-like cognitive impairments or abnormalities, particularly impairments or
abnormalities originating in the hippocampus and/or the prefrontal cortex and/or the
entorhinal cortex of the brain, to induce an immune response in said human or
animal to alleviate said AD-like symptoms associated with the disease or to restore
a condition found in healthy individuals which are unaffected by the disease.
Since virtually all Down’s Syndrome people will eventually suffer from AD-
like cognitive impairments at some point in life, it will also be beneficial to administer
the said vaccine to Down’s Syndrome people before the manifestation of AD-like
impairments. The said vaccine would then act as a preventive treatment.
The immunogenic composition described herein may be administered to a
human or animal by any appropriate standard routes of administration. In general,
the composition may be administered by topical, oral, rectal, nasal or parenteral (for
example, intravenous, subcutaneous, or intramuscular) routes. In addition, the
composition may be incorporated into sustained release matrices such as
biodegradable polymers, the polymers being implanted in the vicinity of where
delivery is desired, for example, at the site of a tumor. The method includes
administration of a single dose, administration of repeated doses at predetermined
time intervals, and sustained administration for a predetermined period of time.
In a specific embodiment the antigenic construct described herein,
particularly an immunogenic composition comprising said antigenic construct in a
therapeutically effective amount, is administered in repeated doses, in particular in 1
to 15 doses, more particularly in 2 to 10 doses, more particularly in 3 to 7 doses and
even more particularly in 4 to 6 doses, in time intervals of between 1 and 10 weeks,
particularly in time intervals of between 1 and 6 weeks, more particularly in time
intervals of between 1 and 4 weeks, and even more particularly in time intervals of
between 2 and 3 weeks. The immune response is monitored by taking sera samples
at a suitable time after boosting, particularly 3 to 10 days after boosting, more
particularly 4 to 8 days after boosting and more particularly 5 to 6 days after
boosting and determining the immunogenicity of the antigenic construct using
known methodology, particularly one of the commonly used immunoassays such as,
for example, an ELISA assay.
In particular, the antigenic peptide composition described herein is
administered by parenteral, particularly by intra-peritoneal, intravenous,
subcutaneous and intra-muscular injection.
The dosage of the composition will depend on the condition being treated,
the particular composition used, and other clinical factors such as weight, size and
condition of the patient, body surface area, the particular compound or composition
to be administered, other drugs being administered concurrently, and the route of
administration.
The immunogenic composition described herein may be administered in
combination with other biologically active substances and procedures for the
treatment of symptoms associated with Down’s syndrome. The other biologically
active substances may be part of the same composition already comprising the
immunogenic composition described herein, in form of a mixture, wherein the
immunogenic composition and the other biologically active substance are intermixed
in or with the same pharmaceutically acceptable solvent and/or carrier or may be
provided separately as part of a separate compositions, which may be offered
separately or together in form of a kit of parts.
The immunogenic composition described herein may be administered
concomitantly with the other biologically active substance or substances,
intermittently or sequentially. For example, the immunogenic composition described
herein may be administered simultaneously with a first additional biologically active
substance or sequentially after or before administration of said composition. If an
application scheme is chosen where more than one additional biologically active
substance are administered together with the at least one immunogenic composition
described herein, the compounds or substances may partially be administered
simultaneously, partially sequentially in various combinations.
Thus also described are mixtures of an immunogenic composition
described herein and, optionally, one or more further biologically active substances
in a therapeutically or prophylactically effective amount, as well as to methods of
using such a composition described herein, or mixtures thereof for the prevention
and/or therapeutic treatment and/or alleviation of the effects of amyloid related
pathology in children, or in young to middle aged subjects with Down’s syndrome,
particularly for amelioration or restoration of memory impairments or abnormalities,
particularly impairments and abnormalities originating in the hippocampus and/or
the prefrontal cortex and/or the entorhinal cortex of the brain, particularly for
amelioration or restoration of the impairments of recognition memory and/or
impairments of the contextual associative memory and/or impairments of the
associative learning and/or impairments of the declarative memory for facts and
events and/or episodic memory impairments and/or language dysfunction such as
aphasia and/or visuospatial impairments, such as misplacement of items and
difficulty navigating in unfamiliar and familiar terrain and/or decreased executive
functions, such as apathy, disinhibition, social isolation, poor judgment, difficulties
with planning and/or poor abstract reasoning and/or personality changes and/or
emotional changes such as apathy, agitation and psychosis and/or apraxia and/or
impairments of performing learned motor tasks and/or other neurological signs
including pyramidal and extrapyramidal findings as well as myoclonus or seizures.
The mixtures described herein may comprise, in addition to an
immunogenic composition described herein, a biologically active substance such as,
for example, known compounds used in the medication of AD-like symptoms in
children, or in young to middle aged subjects with Down’s syndrome.
The other biologically active substance or compound may exert its
biological effect by the same or a similar mechanism as the immunogenic
composition described herein or by an unrelated mechanism of action or by a
multiplicity of related and/or unrelated mechanisms of action.
Generally, the other biologically active compound may include antibodies
raised against and binding to an antigenic peptide as disclosed herein or
compounds used in the medication of neurological disorders such as neutron-
transmission enhancers, psychotherapeutic drugs, acetylcholine esterase inhibitors,
calcium-channel blockers, biogenic amines, benzodiazepine tranquillizers,
acetylcholine synthesis, storage or release enhancers, acetylcholine postsynaptic
receptor agonists, monoamine oxidase-A or –B inhibitors, N-methyl-D-aspartate
glutamate receptor antagonists, non-steroidal anti-inflammatory drugs, antioxidants,
and serotonergic receptor antagonists.
In particular, the mixture described herein may comprise at least one other
biologically active compound selected from the group consisting of compounds
against oxidative stress, anti-apoptotic compounds, metal chelators, inhibitors of
DNA repair such as pirenzepin and metabolites, 3-aminopropanesulfonic acid
(3APS), 1,3-propanedisulfonate (1,3PDS), secretase activators, β- and γ–secretase
inhibitors, β- and γ-secretase modulators, tau proteins, neurotransmitter, β-sheet
breakers, anti-inflammatory molecules, or cholinesterase inhibitors (ChEIs) such as
tacrine, rivastigmine, donepezil, and/or galantamine and other drugs and nutritive
supplements, together with an therapeutic vaccine described herein and, optionally,
a pharmaceutically acceptable carrier and/or a diluent and/or an excipient.
The mixtures described herein may further comprise niacin or memantine
together with an immunogenic composition according to the invention and,
optionally, a pharmaceutically acceptable carrier and/or a diluent and/or an
excipient.
In one embodiment mixtures are described that comprise “atypical
antipsychotics” such as, for example clozapine, ziprasidone, risperidone,
aripiprazole or olanzapine for the treatment of positive and negative psychotic
symptoms including hallucinations, delusions, thought disorders (manifested by
marked incoherence, derailment, tangentiality), and bizarre or disorganized
behavior, as well as anhedonia, flattened affect, apathy, and social withdrawal,
together with an immunogenic composition and/or a therapeutic vaccine described
herein and, optionally, a pharmaceutically acceptable carrier and/or a diluent and/or
an excipient.
Other compounds that can be suitably used in mixtures in combination
with the immunogenic composition are described, for example, in
(see especially pages 16 and 17) including therapeutic drug targets (page 36-39),
alkanesulfonic acids and alkanolsulfuric acid (pages 39-51), cholinesterase
inhibitors (pages 51-56), NMDA receptor antagonists (pages 56-58), estrogens
(pages 58-59), non-steroidal anti-inflammatory drugs (pages 60-61), antioxidants
(pages 61-62), peroxisome proliferators-activated receptors (PPAR) agonists (pages
63-67), cholesterol–lowering agents (pages 68-75); amyloid inhibitors (pages 75-
77), amyloid formation inhibitors (pages 77-78), metal chelators (pages 78-79), anti-
psychotics and anti-depressants (pages 80-82), nutritional supplements (pages 83-
89) and compounds increasing the availability of biologically active substances in
the brain (see pages 89-93) and prodrugs (pages 93 and 94 ), which document is
incorporated herein by reference, but especially the compounds mentioned on the
pages indicated above.
In one embodiment an antigenic construct is described which comprises
an Aβ peptide that does not contain a T-cell epitope and thus is free of potential side
effects such as neurological complications caused by an over-activated complement
system. This can be achieved within the scope of the present disclosure by
administering an Aβ peptide antigen, particularly a palmitoylated Aβ peptide antigen,
more particularly the palmitoylated Aβ peptide antigen, but especially the
1-15
palmitoylated Aβ peptide antigen, Aβ in combination with a complement
1-15 1-15
inhibitor.
The complement inhibitor may be a compound selected from the group
consisting of soluble human complement Receptor 1, anti- human complement
protein C5 such as, for example, a humanized anti C5 monoclonal antibody or a
single-chain fragment of a humanized monoclonal antibody, C1-esterase inhibitor-N
and Natural human C1 Inhibitor.
The modified amyloid peptide antigen such as, for example, the amyloid
beta 1-15 peptide antigen may be synthesized following the method reported in
(Nicolau et al., 2002). The approach reported in Nicolau et al may be modified by
first synthesizing the antigenic peptide which is then further modified by an on-resin
grafting of a lipophilic or hydrophobic moiety, to the terminal amino acid residues of
the pre-formed peptide. In particular, a protected amino acid, particularly a Fmoc-
protected amino acid, is attached to a resin using known coupling chemistry. The
protecting group is removed and a second protected amino acid residue coupled.
Then, standard automated peptide synthesis using known protection chemistry,
particularly Fmoc/tBu chemistry, and standard side-chain protecting groups are then
used to synthesis the Aβ antigenic peptide, particularly the Aβ antigenic peptide
1-15
by coupling on amino acids 1 to 15 of amyloid protein Aβ to produce the peptide
1-42
fragment with a given sequence. In a final step two further protected amino acids
are coupled to the growing peptide fragment. The Mtt groups on the side chains of
first two and last two amino acids can then be selectively cleaved and coupled to
palmitic acid. After washing of the resin, the protecting group is removed and the
resin simultaneously cleaved, followed by side-chain deprotections using standard
methodology. The final product can then be obtained in high purity and its identity
confirmed by methods known in the art such as, for example, electrospray mass
spectrometry.
The modified amyloid Aβ antigenic peptide, particularly the modified Aβ
1-15
antigenic peptide may be reconstituted in a construct consisting of liposomes,
particularly liposomes made of dimyristoyl phosphatidyl choline (DMPC), dimyristoyl
phosphatidyl ethanolamine (DMPEA), dimyristoyl phosphatidyl glycerol (DMPG) and
cholesterol, optionally containing monophosphoryl lipid A.
In one disclosure, contemplated herein is using as a replacement for
cationic lipids in the liposomal membrane, anionic lipids selected from the group
consisting of:
a. diacyl-phospholipids with headgroups phosphatidyl glycerol, phosphatidyl
serine, phosphatidyl inositol, L-α-phosphatidylinositolphosphate or
phosphatidic acid;
b. lyso-phospholipids with headgroups phosphatidyl glycerol, phosphatidyl serine
or phosphatidic acid, and
c. cardiolipin, dilyso-cardiolipin, monolyso-cardiolipin
In one aspect, the disclosure contemplates using as a replacement for
anionic lipids in the liposomal membrane, cationic lipids selected from the group
consisting of:
a. diacyl-phospholipids with headgroups 3-trimethylammonium-propane ,3-
dimethylammonium-propane, 3-ethylphosphocholine or 3-phosphatidylethanol-
amine;
b. D-erythro-sphingosine, dimethyldioctadecylammonium bromide, N-[1-(2, 3-
dimyristyloxy)propyl]-N,N-dimethyl-N-(2-hydroxyethyl) ammonium bromide,
N,N,N-trimethylbis[(1-oxooctadecenyl)oxy]-(Z,Z)propanaminium methyl
sulfate or 3β-[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol hydro-
chloride.
In one embodiment, the lipid chains attached to the above headgroups can
a. be saturated or unsaturated,
b. vary in length from (CH ) wherein n is between 3 and 24, and
be symmetrically or asymmetrically substituted.
In a specific embodiment liposomes with lipid A are used as adjuvant to
prepare the anti-amyloid vaccine. Dimyristoylphosphatidyl-choline, -glycerol and
cholesterol are mixed, particularly in a molar ratio of 0.9:1.0:0.7. A strong
immunmodulator such as, for example, monophosphoryl lipid A is then added at a
suitable concentration, particularly at a concentration of between 30 and 50 mg per
mmol, more particularly at 40 mg per mmol of phospholipids. The modified antigenic
Aβ peptide is then added at a molar ratio peptide to phospholipids of between 1:30
and 1:200, particularly at a molar ratio of between 1:50 and 1:120, more particularly
of 1:100. Solvents are removed, for example through evaporation, and the resulting
film hydrated with sterile buffer solution such as, for example PBS.
Liposomes may also be prepared by the crossflow injection technique as
described, for example, in (Wagner et al., 2002). During the injection of lipid
solutions into an aqueous buffer system, lipids tend to form “precipitates”, followed
by self arrangement in vesicles. The obtained vesicle size depends on factors such
as lipid concentration, stirring rate, injection rate, and the choice of lipids. The
preparation system may consist of a crossflow injection module, vessels for the
polar phase (e.g. a PBS buffer solution), an ethanol/lipid solution vessel and a
pressure device, but particularly a nitrogen pressure device. While the aqueous or
polar solution is pumped through the crossflow injection module the ethanol/lipid
solution is injected into the polar phase with varying pressures applied. Various
methods of production of the liposome antigenic constructs are described in
WO2007/068411.
In a specific embodiment, the liposomal composition for use in the method
described herein is prepared to comprise a palmitoylated Aβ 1-15, particularly a
tetrapalmitoylated Aβ 1-15 together with monophosphoryl lipid A (MPLA) as an
adjuvant.
Four to ten, particularly five to six doses of the liposomal composition may
be administered subcutaneously and weekly or bi-weekly to a subject to be
vaccinated. Plasma probes may be obtained and analyzed for IgG titers periodically.
The effectiveness of the liposomal composition described herein as an
immunogen in children and young to middle aged subjects with Down’s syndrome
and the therapeutic potential for treating memory deficits in said subjects could be
demonstrated in a relevant animal model for Down’s syndrome. In particular,
Ts65Dn mice were used that are widely accepted as an animal model for DS.
Ts65Dn mice exert a triplicate of the murine chromosome 16, which hosts the
murine APP gene (Davisson et al., 1993;Netzer et al., 2010). These transgenic mice
show 1.5 fold increased murine Aβ (Hunter et al., 2004) and demonstrate behavioral
deficits in several memory tasks (Belichenko et al., 2009).
It could be demonstrated within the present disclosure that the liposomal
composition described herein induced elevated titers of antibodies and that said
antibodies remain elevated even 40 days after the last immunization. Thus, the
liposomal composition as described herein is capable of inducing a robust immune
response and of breaking Aβ self-tolerance in treated subjects with Down’s
syndrome.
The liposomal composition as described herein was further shown to be
capable to induce in young to middle aged mice with Down’s syndrome, IgG titers
as high as in subjects of a control group without Down’s syndrome. The liposomal
composition described herein induces increased titers of antibodies of the IgG2a
isotype as compared to the control group, whereas antibody titers of the IgG1 and
IgG2b isotype are comparable in the treatment and the control group. The titers of
IgM class antibodies are lower in the treatment group as compared to the control
group. This slightly lower IgM level may be caused by the altered immune system of
mice with Down’s syndrome. The liposomal composition is thus capable of
overcoming the impaired adaptive immune response to Aβ described in DS people
(Monsonego et al., 2001).
The liposomal composition as described herein is also safe and does not
induce unwanted side effects. In particular, treatment with the liposomal composition
does not lead to cell activation the brain, such as activation of astrocytes nor
microglia.
The liposomal composition as described herein was shown to result in a
higher discrimination ratio in the animal model indicating that treatment led to a
significant improvement of memory in the treated animals.
The liposomal composition as described herein was further shown in a test
consisting of training, cued and contextual sessions to result in an enhanced
freezing in the animal model reaching levels comparable to that observed in the
control animals. This suggests that the immunization was efficient and enhanced the
memory capacity of the model animals.
The liposomal composition as described herein is thus capable of rescuing
the memory deficits in children and in young to middle aged subjects suffering from
Down’s syndrome, in particular in children and young to middle aged subjects with
Down’s syndrome, who have not yet developed Aβ-associated plaques in the brain.
Patients, who suffer from Down’s syndrome exhibit cognitive abnormalities such as
memory impairment and abnormal activities.
The liposomal composition as described herein is further capable of
rescuing the memory deficits in children and in young to middle aged subjects
suffering from Down’s syndrome, in particular in children and in young to middle
aged subjects with Down’s syndrome, who have already developed Aβ-associated
plaques in the brain.
In one embodiment, treatment with the liposomal composition can
ameliorate or restore the memory deficit in a child or in a young to middle aged
subject with Down’s syndrome, particularly impairments of recognition memory
and/or impairments of the contextual associative memory and/or impairments of the
associative learning and/or impairments of the declarative memory for facts and
events and/or episodic memory impairments and/or language dysfunction such as
aphasia and/or visuospatial impairments, such as misplacement of items and
difficulty navigating in unfamiliar and familiar terrain and/or decreased executive
functions, such as apathy, disinhibition, social isolation, poor judgment, difficulties
with planning and/or poor abstract reasoning and/or personality changes and/or
emotional changes such as apathy, agitation and psychosis and/or apraxia and/or
impairments of performing learned motor tasks and/or other neurological signs
including pyramidal and extrapyramidal findings as well as myoclonus or seizures.
[0160A] The term “comprising” as used in this specification means “consisting
at least in part of”. When interpreting each statement in this specification that
includes the term “comprising”, features other than that or those prefaced by the
term may also be present. Related terms such as “comprise” and “comprises” are
to be interpreted in the same manner.
BRIEF DESCRIPTION OF DRAWINGS
Figure 1 shows a schematic presentation of the ACI-DS-01 vaccine, which
is a liposome-based vaccine with tetra-palmitoylated mouse Aβ1-15 antigen and
MPLA as adjuvant. Three amino acid difference between human and mouse Aβ1-15
are underlined.
Figure 2 shows anti-mouse Aβ antibody levels in Ts65Dn mice immunized
with the vaccine ACI-DS-01. A and B) Anti-mouse-Aβ40 or 42 IgG titers were
detected in the plasma of mice immunized with ACI-DS-01. The induced titers were
observed following the second immunization and remained high even 40 days after
the 6 injections in comparison to mice immunized with empty-liposome. There was
no difference between the measured titers of 2N and Ts65Dn mice. C to F) IgG
isotypes were detected following the 4 immunization. G) IgM titers were barely
lower in the Ts65Dn mice. H) the same level of anti-MPLA IgG titers were detected
in all mice immunized with ACI-DS-01. Graphs represent the mean ± SD. (n=20 2N-
ACI-DS-01, n=15 Ts65Dn-ACI-DS-01, n= 18 2N-Empty and n= 11 Ts65Dn-Empty.
Graphs represent the mean ± SD).
Figure 3 shows efficacy of immunization on memory performance. The
difference in the spontaneous locomotor activity between 2N and Ts65Dn mice
remained similar following the immunization. B) Mice immunized with ACI-DS-01
showed a significant enhanced recognition index (RI) in the novel object recognition
in comparison to the group treated with Empty vaccine. C) In the fear conditioning,
immunized mice showed border line significance for a higher level of freezing during
the contextual session. Graphs represent the mean ± SD. (n= 20 2N-ACI-DS-01, n=
13 Ts65Dn ACI-DS-01, n= 18 2N-Empty and n= 11 Ts65Dn-Empty).
Figure 4 shows level of Aβ40 in mice treated with ACI-DS-01 vaccine. A)
In the cortex, the level of Aβ40 was barely higher in the Ts65Dn in comparison to
the 2N mice. The vaccine ACI-DS-01 showed a trend to decrease Aβ42 and Aβ40
levels in the cortex and the hippocampus. Aβ42 and Aβ40 levels in the cerebellum
showed a statistical significant decrease following the treatment with ACI-DS-01.
Graphs represent the mean ± SD. (n=10 2N-ACI-DS-01, n=10 Ts65Dn ACI-DS-01,
n= 8 2N-Empty and n= 5 Ts65Dn-Empty). B and C) Correlation between the ratio
Aβ-40/42 in the cortex or the cerebellum with the RI. D) The level of anti-Aβ IgG
titers correlate weakly with Aβ40 level in the plasma of Ts65Dn mice treated with the
ACI-DS-01. E) Significant correlation between the level of anti-Aβ IgG and the RI.
(n=19 2N ACI-DS-01, n=13 Ts65Dn ACI-DS-01, n= 18 2N-Empty and n= 11
Ts65Dn-Empty).
Figure 5 shows studies of inflammatory reaction. A) Body and brain
weight. B) Confocal images of GFAP immunoreactivity (left) and CD45 (right) in
treated 2N and Ts65Dn mice. Arrows point individual CD45-positive microglial cells.
The optical density of GFAP immunoreactivity was quantified and revealed no
difference between groups. n=4 from each group.
Figure 6 shows sections of human DS brain samples immuno-stained with
sera of mice immunized with ACI-DS-02. Positive area indicates that the ACI-DS-02
derived antibody binds to amyloid plaques in the brain of DS people. The staining
was similar to the obtained staining with 6E10, a commercially available anti-
amyloid beta antibody, and with ACI-24 derived antibody. ACI-24 vaccine
corresponds to ACI-DS-01 vaccine with the difference that the antigen used in ACI-
24 is a human Aβ 1-15 sequence, while ACI-DS-01 vaccine contains mouse Aβ 1-
sequence as the antigen (for three amino acid difference in those two antigens
see Figure 1). No staining was observed when a section was incubated with sera of
mice immunized with PBS.
Figure 7 shows the learning of TS65Dn mice immunized with ACI-DS-02
vaccine in the fear conditioning test. TS65Dn mice immunized with ACI-DS-02
showed greater freezing during the acquisition session in comparison to TS65Dn
mice immunized with PBS, particularly following the third conditioned-stimulus (CS).
Graphs represent the mean ± SD. (n= 7 Ts65Dn-ACI-DS-02, n= 7 2N-PBS and n= 4
Ts65Dn-PBS. Graphs represent the mean ± SD).
Figure 8 shows sections of human DS brain samples immuno-stained with
sera of mice immunized with ACI-DS-03. Positive area indicates that the ACI-DS-03
derived antibody binds to amyloid plaques in the brain of DS people. The staining
was similar to the obtained staining with 6E10, a commercial available anti-amyloid
beta antibody, and with ACI-24 derived antibody. ACI-24 vaccine corresponds to
ACI-DS-01 vaccine with the difference that the antigen used in ACI-24 is a human
Aβ 1-15 sequence, while ACI-DS-01 vaccine contains mouse Aβ 1-15 sequence as
the antigen (for three amino acid difference in those two antigens see Figure 1). No
staining was observed when section was incubated with sera of mice immunized
with PBS.
Figure 9 shows the morphological analysis of Ts65Dn mice immunized
with ACI-DS-01 vaccine. A) Number of ChAT+ cells in medial septum and B) optical
density and were similar in all groups. C) area of ChAT+ cell body were increased in
Ts65Dn mice immunized with ACI-DS-01 in comparison to Ts65Dn treated with
Empty vaccine. (n= 4 2N-Empty, n= 4 Ts65Dn-Empty, n= 4 2N-ACI-DS-01 and n= 4
Ts65Dn-ACI-DS-01. Graphs represent the mean ± SD).
EXAMPLES
Example 1 General Methodology
1.1 Animals:
Ts65Dn mice known as a DS mouse model (Davisson et al., 1993) were
used (n =30) and the age matched control 2N (n = 40). At starting date, the used
mice were 5 months old. At the end of the study (immunization and behavioral
testing), mice were 9 months old. Therefore, all the samples collected at sacrifice
are from 9 months old mice.
Ts65Dn newborns have elevated lethality (around 6%) mainly due to
congenital heart malformations (Randall 2006, Moore 2006). However, mice can
survive even up to 18 to 24 months. In our study, a death rate of 15% was
observed. The survived mice are ~20% smaller in size compared to normal
littermates. The most severely affected mice that died at birth, do not survive to be
analyzed, resulting in underestimates of the impact of the trisomic genes on some
phenotypes (i.e heart defect). However, for our concerns, the surviving mice
represent an adequate model of DS for analyzing amyloid and AD-like pathologies.
In the DS mouse model used for these studies the mice show increased
levels of amyloid at the age of 4 months (Hunter 2004). At an age of 9 months, the
level of amyloid beta reaches 3 fold the normal level as is predicted in view of the
three copies of the APP gene. Similar to DS people, aging is an important factor for
amyloid deposition in Ts65Dn brains. Despite the age-related accumulation of
amyloid, the TS65Dn mice do not develop plaques. Nevertheless, they do replicate
faithfully the degeneration of neuronal populations that are seen in AD and in people
with Down syndrome (Salehi et al., 2009)
Taken together, the DS-features in the used mouse-model comprise the
presence of pathological proteins and phenotypical aspects; i.e amyloid load and
cognitive impairment. Therefore, Ts65Dn mice can accurately recapitulate the
pathogenic processes of DS. Since these mice start to accumulate amyloid at a
comparable level to humans at 6 months of age, the age of the model mice
compares to human DS patients at an age range from young to middle age.
Mice used for immunization study with ACI-DS-01 vaccine were the
Ts65Dn mouse colony maintained for more than 10 generations by crossing
B6EiC3Sn-Ts(1716)65Dn females (Jackson Laboratory, Bar Harbor, ME) with
B6EiC3Sn F1/J A/a males (Jackson Laboratory). This breeding scheme was used
because trisomic mice breed very poorly or not at all when inbred; the B6C3
background has been the most successful. To distinguish 2N from Ts65Dn mice,
genomic DNA was extracted from tail samples. A quantitative polymerase chain
reaction (PCR) protocol (provided by the Jackson Laboratory) was used to measure
Mx1 gene expression, which is present in three copies in Ts65Dn. Male mice were
used in all studies. They were 4+/-0.3 months at the beginning of the study.
1.2 Vaccine preparation - Preparation of a liposome-based antigenic construct
Vaccine Antigenic peptide sequence Aβ sequence Antigenic peptide
code conformation within
the vaccine (%)
measured by ATR-IR
ACI-DS- H-Lys(Pal)-Lys(Pal)-Asp-Ala- Mouse Aβ1- 73% beta sheet
01 Glu-Phe-Gly-His-Asp-Ser-Gly- 15 19% random coil
Phe-Glu-Val-Arg-His-Gln- (SEQ ID NO: 0% alpha helix or
Lys(Pal)-Lys(Pal)-OH 2) loops
8% beta turns
ACI-DS- H-Lys(Pal)-Glu-Asp-Val-Gly- Human (SEQ 67% beta sheet
02 Ser-Asn-Lys-Gly-Ala-Ile-Ile-Gly- ID NO: 4) 21% random coil
Leu-Met-Lys(Pal)-OH and mouse 0% alpha helix or
(SEQ ID NO: loops
) Aβ22-35 12% beta turns
ACI-DS- H-Lys(Pal)-Lys(Pal)-His-Gln- Human (SEQ 57% beta sheet
03 Lys-Leu-Val-Phe-Phe-Ala-Glu- ID NO: 6) 19% random coil
Asp-Val-Gly-Ser-Asn-Lys-Gly- and mouse 12% alpha helix or
Lys(Pal)-Lys(Pal)-OH (SEQ ID NO: loops
7) Aβ14-29 11% beta turns
Peptide sequences are presented in three-letter code; Pal stands for palmitoylated residue.;
Three amino acid difference between human and mouse Aβ1-15 sequence are underlined
and shown in Figure 1.
The liposome antigenic construct is produced according to the method
described in WO2007/068411. Palmitoylated peptides were prepared by
PolyPeptide Laboratories (Strasbourg, France). In summary, the liposomal vaccines
were prepared by solubilizing dimyristoylphosphatidylcholine (DMPC),
dimyristoylphosphatodyl-glycerol (DMPG), cholesterol and monophosphryl Lipid A
(MPLA), (all Avanti Polar Lipids, AL, USA), at molar ratios 9:1:7:0.05, respectively, in
ethanol at 60°C. The lipid/ethanol solution was diluted in PBS pH 7.4, allowing the
formation of multi-layer vesicles. The resulting preparation was then concentrated by
ultrafiltration (Vivaflow 200 – 100.000 MWCO Polyethersulphone), with a flow rate of
200 ml/min and the reaction volume was reduced with an ultrafiltration step. The
concentrated solution was further submitted to dialyses in a Vivaflow 200 device
where a 10x volume exchange was performed with PBS pH 7.4. The multilamelar
liposomes were then submitted to homogenization (7 cycles at 15.000-20.000 Psi),
followed by sequential 3 extrusion cycles through polycarbonate filters (Whatman)
with a pore size of 0.2 µm and a diameter of 47 mm. Both the homogenization and
extrusion steps were done in an EmulsiFlex-C5 (Avestin, Canada). The resulting
unilamelar liposomes with no antigen were diluted in PBS pH 7.4 and heated to
60°C prior to peptide addition. The palmitoylated peptide was dissolved in PBS pH
11.4 with 5.0% (ß-OG). For the preparation of vaccines ACI-DS-01, ACI-DS-02 and
ACI-DS-03, the corresponding antigenic peptides were used at 1.33, 0.67 and 1.06
mg/mL respectively. The resulting solution comprised a detergent concentration
above its critical micellar concentration (CMC) of 0.73 % (wt/v). This peptide solution
was then injected into the liposome solutions at 60°C and stirred for 30 min,
resulting in a solution having a final ß-OG concentration far below its critical micellar
concentration. The detergent micellar breakdown, induces the incorporation of the
palmitoylated antigen on the surface of the previously formed liposomes. This
liposomal solution with antigen was then concentrated through ultrafiltration and
submitted to 10x volume exchange with PBS pH 7.4 by diafiltration (both in a
100.000 MWCO Polyethersulphone with a flow of 200 ml/min). The resulting
liposomes were then twice sterile filtered by passing through 0.2 µm polycarbonate
syringe filters and stored at 5°C. The peptide to lipid molar ratio in the vaccine was
1:100.
1.3 Immunization with ACI-DS-01 vaccine
All mice received subcutaneous (s.c.) immunizations with 200µl of ACI-DS-
01 (51.2 µg per dose of Pal1-15, tetrapalmitoylated Aβ peptide 1-15) or empty
liposome vaccine. A total of six injections were done at day 1, 14, 28, 42, 56, 70 and
110. On the same days, tail bleedings were performed just before the first vaccine
injection on day -1 (pre-bleeding), day 56 (14 days after the 4th injection) and at
sacrifice, day 110 (40 days after the 6th injection). Mice were sacrificed at the end of
the study at the age of 9 months and brains were collected.
1.4 Quantification of mouse Aβ-specific antibodies after ACI-DS-01 immunization
Aβ1specific IgG responses were determined by ELISA. Briefly, plates
were coated with 10 µg/mL of mouse Aβ1-42 (Bachem H5966, lot: 1013710,
1028321 or 1033165) or mouse Aβ1-40 (Bachem H5638, lot: 1013645) overnight at
4°C. After washing with PBS-0.05% Tween 20 and blocking with 1% BSA, serial
dilutions of plasma were added to the plates and incubated at 37°C for two hours.
The antibody anti-Aβ 4G8 (Covance SIG-39220, lot: 08EC00905, 1 mg/mL diluted
4000) and serum of a C57BL/6JOlaHsd (Harlan) immunized with three injections of
the same ACI-DS-01 batch were used as positive control. After washing, plates
were incubated with alkaline phosphatase (AP) conjugated anti-mouse IgG antibody
(Jackson Immunoresearch West Grove, PA, USA, Cat. N°115164 Lot 87821,
vial diluted at 1/4000) for two hours at 37°C. After final washing, plates were
incubated during 2 hours and a half with AP substrate (pNPP) and read at 405 nm
using an ELISA plate reader. Results are expressed as optical density (O.D).
1.5 Behavioral testing after ACI-DS-01 immunization
All mice were exposed to the same series of behavioral tests starting at
around 8 months old of age. Each mouse was handled for 10 min, twice a day,
during the 7 days that preceded testing and for 3 days in between tests. The tests
were performed at the indicated days (according to the last immunization) and in the
following order: locomotor activity (starting day 11), novel object recognition task
with 24 h delay (starting day 18), T-maze (starting day 25), and contextual fear
conditioning test (starting day 34). All behavioral tests took place during the day-light
cycle between 7:00 A.M. and 7:00 P.M. and were performed at room temperature
(22°C). On the day of testing, mice were kept in their home cages during the day-
light phase in the same experimental room for 2 h for habituation. As an indicator of
anxiety during each test, the number of fecal pellets and urine drops was also
measured. To minimize olfactory cues from the previous trial, each apparatus was
thoroughly cleaned with 10% ethanol after each animal occupation. Handling and all
behavioral tests were performed in a blinded manner, not revealing the genotype
and treatment to the investigator handling the mice.
1.5.1 Spontaneous locomotor activity test after ACI-DS-01 immunization
Spontaneous locomotor activity was monitored using Plexiglas activity
chambers (model MED-OFA-MS; Med Associates) (27.9 x 27.9 x 20 cm) and
activity monitor software (Activity Monitor, version 4.3.6). As described in Belichenko
et al., (Belichenko et al., 2009;Belichenko et al., 2007), mice were placed into the
center of the chamber under bright ambient light conditions and activity was
monitored for 10 min in three separate trials. Averages were determined for total
distance, velocity, total activity time, total activity counts and vertical activity.
1.5.2 Novel object recognition task after ACI-DS-01 immunization
We used the Bevins and Besheer protocol (Bevins and Besheer,
2006;O'Doherty et al., 2005): a one-trial non-matching-to-sample learning task to
study recognition memory for two sample objects with one environment to study
learning and memory with a 24 h delay. Before testing, mice were habituated in a
black Plexiglas chamber (31 x 24 x 20 cm) during 10 min for 2 consecutive days
under dimmed ambient light conditions. Activity of mice at the age of 8 months
during the object recognition task was recorded with a video camera. First, two
identical objects were placed in the chamber, as previously described (Bevins and
Besheer, 2006;O'Doherty et al., 2005). A mouse was placed at the mid-point of the
wall opposite the sample objects. After 10 min exploring the objects, the mouse was
returned to the colony for 24 h. To test for object recognition, one familiar object and
one novel object were placed in the chamber and the mouse was again placed in
the chamber for 3 min to explore the objects. Object recognition was measured in a
single trial. Data were determined using direct observations of video recordings.
Results were: 1) average total exploration time of the sample objects, both novel
and familiar, and 2) discrimination ratio (novel object interaction/total interaction with
both objects).
1.5.3 T-maze testing after ACI-DS-01 immunization
Mice at age 8 months were used. We used a modified Deacon and
Rawlins protocol (Deacon and Rawlins, 2006) and a continuous alternation task in a
T-maze to evaluate hippocampal function. The maze was made of opaque acrylic
glass (Plexiglas) as described (Deacon and Rawlins, 2006) with an additional sliding
door at the beginning of the start arm. During the test, a mouse was placed at the
beginning of the start arm, with its back to the closed sliding door. After all doors
were opened, the mouse ran down the start arm to choose either the right or left
goal arm. After all four legs of the mouse had entered one goal arm, the sliding door
to another goal arm was closed for 5 s, and then all sliding doors were opened
again, allowing the mice to go back to the start arm. T-maze activity was monitored
until the mice finished 10 alternations. This procedure was repeated for 3
consecutive days, for a total of 30 trials. The spontaneous alternation score was
defined as the number of left–right and right–left alternations, expressed as a
percentage of the total number of possible alternations during the session. Results
were averaged for both alternation scores and time spent.
1.5.4 Contextual fear conditioning test after ACI-DS-01 immunization
Contextual and cued fear conditioning was conducted for evaluation of
fear-dependent learning and retrieval. The test was performed using chambers from
Coulbourn Instruments (Whitehall, PA, USA). On the first day animals were placed
in a chamber (Context A) for 3 minutes for baseline recording, followed by five tone-
shock pairings. The shock (0.5 mA, 2 sec) was delivered following the tone (70 dB,
2 kHz, 20 sec) in each conditional /unconditional stimulus pairing. On the second
day, a novel chamber (Context B; new room, new olfactory environment, new
texture of floor, blue plastic inserts for walls, extra source of blue light, and visual
cues) was used for cued testing. Following a 3-minute pre-tone period, three tones
without shocks were presented to animals during a 3-minute testing period. On the
last day of the experiment, the mice were placed in Context A for 5 minutes without
any conditional or unconditional stimulus (Saxe et al., 2006). Freezing was defined
as the complete lack of motion for a minimum of 0.75 seconds as measured by
FreezeFrame software (Actimetrics, Evanston, IL). The percentage of freezing in
each period was reported.
1.6 Measuring body and brain weights after ACI-DS-01 immunization
After all behavioral tests, mice were deeply anesthetized with sodium
pentobarbital (200 mg/kg i.p.) (Abbott Laboratories), weighed, and transcardially
perfused for 1 min with 0.9% sodium chloride (10 ml) and then for 10 min with 4%
paraformaldehyde in 0.1 M PBS, pH 7.4 (100 ml). After perfusion, the brain was
immediately removed. The weight of the brain (including the olfactory bulbs, cortex,
hippocampus, cerebellum, brainstem and the cervical spinal cord through C1–C2)
was recorded. The brain was then placed in fixative until further use.
1.7 Immunofluorescence for inflammatory reaction after ACI-DS-01 immunization
Brain sections were pre-incubated with 5% non-fat milk in 0.1M PBS with
0.3% Triton X-100. Sections were then incubated overnight at 4 °C with rabbit anti-
cow glial fibrillary acidic protein (GFAP) antibody (DAKO, Glostrup, Denmark) at a
dilution of 1:500, or with polyclonal rat anti-CD45 (Pharmingen) at a dilution of
1:5000. The following incubations (1 h each and at room temperature) were the
done; with a biotinylated donkey anti-rabbit secondary antibody (1:200; Jackson
ImmunoResearch Labs, West Grove, PA, USA) and with fluorescein isothiocyanate
(FITC)-conjugated streptavidin (1:500; Jackson ImmunoResearch Labs). A rinse
with PBS (3 times, each 20 min) was done between the incubations described
above. Sections were mounted onto microscope glass slides and coverslipped using
90% glycerol in 0.1M phosphate buffer, pH 7.4. To control for specificity of antibody
staining, selected sections were submitted to the same protocol but without
including the primary antibodies. Immunofluorescence was not observed in control
sections. Slices were examined and scanned in a Radiance 2000 (Bio-Rad,
Hertfordshire, UK) confocal microscope attached to a Nikon Eclipse E800
fluorescence microscope. The laser was an argon/krypton mixed gas laser with
excitation wavelengths for FITC at 488 nm (λ). LaserSharp software (Bio-Rad) was
used to establish optimal conditions for collecting images. The optimal conditions for
confocal imaging of GFAP- immunoreactivity (IR) or CD45-IR were the following: the
lens was a 20x objective (Nikon; Plan Apo 20x/0.75); laser power was 10% or 20%;
gain was 34.7; offset was 0.0; the zoom factor was 3; scan speed was 500 lines/s;
each optical section was scanned three times and Kalman filtering was then
employed to reduce noise; the size of the image was 512x512 pixels and the pixel
size was 0.48x0.48 um.
1.8 Statistical analysis after ACI-DS-01 immunization
Data are shown as mean ± standard deviation (SD) or standard error of
mean (SEM). Statistical analysis was done by unpaired t-test, two tailed. A
probability of p<0.05 was considered significant.
Example 2 Immunization with ACI-DS-01 produced anti-mouse Aβ antibodies
The liposomal vaccine ACI-DS-01 was prepared according to the
SupraAntigen methodology (, WO2007/068411) with
tetrapalmitoylated mouse Aβ 1-15 peptide (Palm1-15), H-Lys(Pal)-Lys(Pal)-Asp-Ala-
Glu-Phe-Gly-His-Asp-Ser-Gly-Phe-Glu-Val-Arg-His-Gln-Lys(Pal)-Lys(Pal)-OH
embedded into liposomes along with monophosphoryl lipid A (MPLA), (Figure 1).
Six doses of ACI-DS-01 were administered subcutaneously and bi-weekly into
Ts65Dn male mice and age-matched control mice (2N). The analysis of plasma
collected after the 4 dose, showed robust IgG titers against mouse Aβ40 or Aβ42 in
the immunized mice of both Ts65Dn and 2N groups (Figure 2A and 2B). The
vaccine-induced antibodies remained elevated even 40 days after the last
immunization. No titers were detected in mice immunized with empty liposome
without the antigen (Figure 2A and 2B). Thus, ACI-DS-01 was able to break Aβ self-
tolerance in the DS mouse model.
Subclasses of IgG were analyzed in plasma of mice immunized with ACI-
DS-01 following the 4 immunization. The ELISA done with Aβ40 showed that the
Ts65Dn group had higher IgG2a titers than the 2N group (Figure 2D) while there
was no difference between the two groups for IgG1 and IgG2b titers (Figure 2C and
2E) (One-way ANOVA, Tukey posthoc; P = 0.05). The IgG3 titers were lower in the
Ts65Dn group (Figure 2F) (One-way ANOVA, Tukey posthoc; P<0.001). IgM titers
were slightly, but significant, lower in the Ts65Dn mice in comparison to 2N mice
(Figure2G). There (One-way ANOVA, Tukey posthoc; P = 0.05). Similar results
were obtained in ELISA performed with Aβ42 (data not shown).
The lower levels of a few of the titers mentioned above in Ts65Dn mice are
unlikely to be due to the immune response capacity of the Ts65Dn since the anti-
MPLA IgG level was comparable in all mice (Figure2H).
Example 3 Immunization with ACI-DS-01 restored the memory deficit of Ts65Dn
mice
Ts65Dn mice exhibit many features of DS cognitive abnormalities such as
memory impairment and abnormal activities. To investigate the efficacy of the ACI-
DS-01 vaccine, a battery of behavioral tests was conducted two weeks after the last
immunization. Mice performed the tests in the following order; open field, object
recognition, T-maze and fear conditioning.
The analysis of the open field test showed that Ts65Dn mice have a
significant higher spontaneous locomotor activity in comparison to 2N mice (data not
shown). Following immunization with ACI-DS-01, Ts65Dn mice continued to be
significantly more active than ACI-DS-01 treated 2N mice (Fig.3A, t test unpaired
one-tailed; p = 0.0004). In the T-maze, 2N and Ts65Dn mice showed no significant
difference in the alteration ratio or in the alteration duration, neither before treatment
nor post treatment (data not shown).
The spatial memory capacity was measured in the object recognition
(ORT). The Ts65Dn mice exert a weak discrimination ratio and thus recognize
poorly the novel object. ACI-DS-01 treated Ts65Dn mice showed a higher
discrimination ratio indicating that treatment led to a significant improvement of
memory (Fig. 3B, t test unpaired one-tailed; p = 0.03. one way ANOVA, genotype, P
= 0.12, vaccine, P =0.002. Interaction P = 0.54). Interestingly, the same results were
observed in the 2N group.
The fear conditioning test consists of training, cued and contextual
sessions. The same level of freezing was observed in all groups during the training
or the cued session (Figure 3C). During the contextual session, Ts65Dn mice
treated with empty vaccine (only the liposome, no antigen) showed a lower percent
of freezing than the control 2N mice. In contrast, after immunization Ts65Dn mice
demonstrated an enhanced freezing and reached a comparable level to the 2N mice
(t-test unpaired one-tailed; P = 0.04. (one way ANOVA, genotype, P = 0.01, vaccine,
P =0.16. Interaction P = 0.33). This suggests that the immunization was efficient and
enhanced the memory capacity of Ts65Dn mice.
In summary, these results indicate that ACI-DS-01 treatment can restore
the memory deficit of Ts65Dn mice.
Example 4 Mechanism of ACI-DS-01 vaccine for improving the cognitive
deficiency
To address the question about the effect of the ACI-DS-01 induced
antibodies on the level of Aβ, ELISA was performed using protein extracts of the
hippocampus, the cortex and the cerebellum. None of these regions showed a
significant difference on Aβ levels following the treatment with ACI-DS-01; neither
for Aβ40 nor for Aβ42 (Figure 4A). It is worthwhile to note that the ratio Aβ-40/42 in
the cortex and the cerebellum correlated significantly with a higher recognition index
(RI) (Figure 4B and 4C. Cortex; Pearson r correlation = -0.3248, P = 0. 0.03.
cerebellum; Pearson r correlation = -0.4127, P = 0.009). These results suggest that
Aβ-40/42 might be responsible of the cognitive deficit. Further analysis showed that
Aβ42 in the plasma was undetectable. In contrast, an increased level of Aβ40 in the
plasma seemed to be associated with the higher values of anti-Aβ IgG measured in
the group of ACI-DS-01 treated Ts65Dn mice (Figure 4D; Pearson r correlation =
0.51, P =0.09). Moreover, the RI correlated with anti-Aβ IgG titers (Figure 4E;
Pearson r correlation = 0.3154, P =0.007).
These results suggest that ACI-DSinduced antibodies may clear
Aβ from the brain to the plasma which then may lead to memory amelioration.
Example 5 Morphology of neurons in Mouse Models of Down Syndrome
The size, the number and optical density of cholinergic neuron in the basal
forebrain (BFCN) were analyzed. Two sections from BFCN level were stained with
ChAT antibody (Millipore, Cat # AB144P, lot # 2010060). Medial septum (MS) area
was scanned on confocal microscope for quantitative evaluation. The following
parameters were calculated 1) density of ChAT+ cells in MS; 2) area of ChAT+
individual neurons, and 3) average of optical density of ChAT in individual neurons.
Cholinergic neurons in medial septum were imaged and ImageJ software was used
to count number of neurons, reveal optical density of staining and outlining of cell
body area.
Results: Number of ChAT+ cells in medial septum and optical density was similar in
2N and Ts65Dn mice treated with empty vaccine (Figure 9A and 9B).
Number of ChAT+ cells in medial septum was similar in ACI-DS-01 treated
2N and Ts65Dn mice (Figure 9A) (2N-Empty = 27.5 ± 7.2 per 100 microm section;
Ts65Dn- Empty = 23.9 ± 7.0; 2N-ACI-DS-01 = 36.6 ± 4.6; Ts65Dn-ACI-DS-01 =
.9 ± 6.6). Optical density of staining in individual ChAT+ cells in medial septum
was also similar in ACI-DS-01 treated 2N and Ts65Dn mice (Figure 9B) (2N- Empty
= 79.9 ± 3.5 arbitrary unit; Ts65Dn- Empty = 74.7 ± 2.6; 2N-ACI-DS-01 = 71.9 ±
2.6; Ts65Dn-ACI-DS-01 = 75.8 ± 4.5). The area of ChAT+ cell body in medial
septum was lower in Ts65Dn than 2N mice (Figure 9C).The area of ChAT+ cell
body in medial septum of Ts65Dn- ACI-DS-01 treated mice was significantly
increased in comparison to empty treated Ts65Dn-veh (Ts65Dn- Empty = 179 ± 6
microm ; Ts65Dn-ACI-DS-01 = 201 ± 8, p = 0.031). 2N and Ts65Dn ACI-DS-01
treated mice had similar area of cell body (2N-ACI-DS-01 = 208 ± 8 microm ;
Ts65Dn-ACI-DS-01 = 201 ± 8, p = 0.55) (Figure 9C). This point out on recovering of
cell body area in ChAT+ cells in medial septum upon ACI-DS-01 treatment.
Equivalently, these results are an indication of the safety of the ACI-DS-01 vaccine
since the number of ChAT+ cells did not change after immunization with the
vaccine. These morphological studies showed less atrophy of neurons after ACI-
DS-01 immunization indicating that this vaccine could be used for prevention of
neurodegeneration.
Example 6 The safety of the liposomal vaccine ACI-DS-01
To determine if immunization with ACI-DS-01 may induce safety concerns,
body weight, general observation, and inflammation markers in mice were recorded.
Before treatment, Ts65Dn mice had a significantly lower body weight than 2N mice
(ANOVA, Tukey posthoc, p = 0.001). Immunization did not alter significantly the
body weight (Figure 5A). Brain weights were similar in all four groups of mice
(Figure 5A). The immunohistochemistry results showed no difference in the number
of glial fibrillary acidic protein (GFAP)-positive astroglial cells or of the CD45-positive
microglial cells. The obtained results were similar in the cortex and hippocampus
(Figure 5). These results indicate that ACI-DS-01 vaccination did not induce an
inflammatory response.
To verify the specificity of ACI-DS-01 induced antibodies, a cross reaction
study was conducted. The brain sections of 2N and Ts65Dn were stained with the
antiserum of ACI-DStreated 2N mice.
In summary, these findings indicate that ACI-DS-01 is a safe vaccine.
Example 7: General Methodology for experiments with ACI-DS-02 and ACI-DS-03
vaccines
7.1 Immunization with ACI-DS-02 and ACI-DS-03 vaccine
Ts65Dn mice, males (Jax laboratory B6EiC3Sn.BLiA-Ts(1716)65Dn/DnJ)
4±0.3 months old at the start of the study, received subcutaneous (s.c.)
immunizations with 200µl of ACI-DS-02 (18.6 µg per dose of dipalmitoylated Aβ
peptide 22-39) (n= 6 Ts65Dn mice) or PBS (n= 4 Ts65Dn mice). A total of four
injections were done at day 1, 14, 28 and 42. Tail bleedings were performed one
week following each immunization; day 7, 21, 35 and 49.
The same procedure was done for the immunization with the vaccine ACI-
DS-03 (38.6 µg per dose of Pal 14-29, tetrapalmitoylated Aβ peptide 14-29) (n= 7
Ts65Dn mice) or PBS (n= 4 Ts65Dn mice).
Control littermate mice (n = 7 2N-mice) received similar injections but with
PBS.
7.2 Quantification of mouse Aβ-specific antibodies
Aβ1specific IgG responses were determined by ELISA. Briefly, plates
were coated with 10 µg/mL of human Aβ1-42 (Bachem H1368, lot: 1000255)
overnight at 4°C. After washing with PBS-0.05% Tween 20 and blocking with 1%
BSA, serial dilutions of plasma were added to the plates and incubated at 37°C for
two hours. The antibody anti-Aβ 6E10 (6E10 from Covance SIG-39320, lot:
09GC01254 vial a, 1 mg/ml diluted 1000x) was used as positive control. After
washing, plates were incubated with alkaline phosphatase (AP) conjugated anti-
mouse IgG antibody (Jackson Immunoresearch West Grove, PA, USA, Cat. N°115-
055-164 Lot 87821, vial diluted at 1/4000) for two hours at 37°C. After final washing,
plates were incubated during 2 hours and a half with AP substrate (pNPP) and read
at 405 nm using an ELISA plate reader. Results are expressed as optical density
(O.D).
7.3 Behavioral testing
All mice were exposed to the same series of behavioral tests starting at
around 6±0.3 months old of age. All behavioral tests took place during the day-light
cycle between 7:00 A.M. and 7:00 P.M. and were performed at room temperature
(22°C).
7.3.1 Spatial object recognition task
Before testing, mice were habituated in a grey polypropylene chamber
(Diameter 40 cm, height 32.4 cm) during 10 min without any object under dimmed
ambient light conditions. In the following day, and during the learning session, two
identical objects (cube of Lego or glass black bottle) were presented (10 min) on
one side of the maze. The retention test was done 3 hours post learning, one of the
identical objects is moved to the opposite side of the maze and is presented during
min. All sessions were recorded with the video tracking software Ethovision XT
8.0. The measured parameters included the average total exploration time of the
sample objects, both novel and familiar, and discrimination ratio (novel object
interaction/total interaction with both objects).
7.3.2 Water Maze memory task
The water maze was conducted to analyze the long term memory. As a
habituation test, mice were subjected in the first day to a water maze (diameter: 165
cm, height: 64 cm) with a visible, cued platform (diameter 11 cm, height: 37 cm).
This test consisted in 5 trials of each of 120 seconds with 30 minutes inter-trial
interval. The final 6th trial was conducted without the visible, cued platform. The
following day, mice were subjected to 4 days training with a hidden platform. On
each day, mice performed 6 trials each of 120 seconds and with inter-trial interval
minutes. At the beginning of a trial, a mouse is placed in the water facing the
edge of the tank, at one of the 4 starting points (North-East, South-East, South-
West, and North-West). The starting points are changed from trial to trial, while the
escape platform is fixed (West quadrant) throughout the experiment. If the mouse
finds the platform within 120 sec, it is allowed to remain on the platform for 15 sec. If
it fails to find the platform within 120 sec, the trial is stopped and the mouse is gently
guided to the platform and left there for 15 sec. The measured parameters included
the escape latency time (s) spent in the different parts of the maze, number of
entries into each quadrant of the maze, distance (cm) travelled into each quadrant
of the maze, distance (cm) to target area, % time in each quadrant, % time in the
extended target platform annulus during probe trial, number of crossing of the
extended target platform annulus during probe trial and velocity (cm/s).
7.3.3 Contextual fear conditioning test
Contextual and cued fear conditioning was conducted for evaluation of
fear-dependent learning and retrieval. The test was performed using chambers from
Med Associates Instruments. On the first day mice were placed in a chamber
(Context A) for 2 minutes for baseline recording, followed by four tone-shock
pairings (conditioned-stimuli, CS). The shock 30 seconds tone stimulus (5000 Hz,
80 dB) co-terminated with 2 seconds foot-shock (0.7 mA) in each conditional
/unconditional stimulus pairing. On the second day, the mice were placed in context
A for 5 minutes without any conditional or unconditional stimulus. On the last day of
the experiment, a novel chamber (Context B; new room, new olfactory environment,
new texture of floor, blue plastic inserts for walls, extra source of blue light, and
visual cues) was used for cued testing. Following a 2-minute pre-tone period, three
tones without shocks were presented to animals during 8-minute testing period.
Freezing was defined as the complete lack of motion for a minimum of 0.75
seconds. The percentage of freezing in each period was reported.
7.4 Immuno-staining of amyloid plaques in human DS brain samples with ACI-
DS-02 and ACI-DS-03 vaccines induced anti-Aβ antibodies
Brain sections from DS people were stained with vaccine derived anti-Aβ
antibodies in sera obtained from immunized mice with the ACI-DS-02 vaccine. A
pool of sera from five c57BL/6 mice immunized with ACI-DS-02 vaccine was used at
the dilution of 1/100 for the procedure of immuno-staining. Briefly, cryosections of
10µm of a DS individual of 59 years old was incubated in 4% paraformaldehyde
(Sigma-Aldrich, 252549) for 20 minutes. Following the incubation in 70% formic acid
(Merck, 1.00264.1000) and 10% Triton X-100 (Sigma, 234729) each for 15 minutes,
sections were blocked for 2 hours in 10% Normal goat Serum (NGS, Invitrogen,
6210072) in PBST (PBS plus 0.5% Triton X-100). The incubation with the primary
antibody (sera of immunized mice) in PBST with 10% NGS was done overnight. The
next day, and after rinsing in PBSA, sections were incubated with Alexa Fluor 488
conjugated AffiniPure Goat Anti-Mouse IgG (Jackson ImmunoResearch, 115
146). Sectiones were mounted in ProLong Gold (Invitrogen, P36931). As positive
control, the primary antibody anti-Aβ 6E10 was used (Covance, SIG-39320, anti- Aβ
against N-terminal end of Aβ using a dilution of 1:10000). Sera of c57BL/6 mice
immunized with PBS were used as negative control. Sera of c57BL/6 mice
immunized with ACI-24 were used as a control. ACI-24 vaccine corresponds to ACI-
DS-01 vaccine with the difference that the antigen used in ACI-24 is a human Aβ 1-
sequence, while ACI-DS-01 vaccine contains mouse Aβ 1-15 sequence as the
antigen (for three amino acid difference in those two antigens see Figure 1).
7.5 Statistical analysis
Data are shown as mean ± standard deviation (SD) or standard error of
mean (SEM). Statistical analysis was done by unpaired t-test, two tailed or ANOVA
using prism pad graph version 5. A probability of p<0.05 was considered significant.
Example 8 Immunization with ACI-DS-02 produced anti-human Aβ antibodies
Four doses of ACI-DS-02 were administered subcutaneously and bi-
weekly into Ts65Dn male mice and age-matched control mice 2N. The analysis of
plasma collected 7 days after each dose, showed IgG titers against human Aβ42 in
the immunized mice of both Ts65Dn and 2N groups (data not shown). The IgG titers
of Ts65Dn mice immunized with ACI-DS-02 were significantly different from those
immunized with PBS at day 21 and 35 (one way ANOVA, Tukey posthoc, P = 0.01
and 0.05 respectively). Thus, ACI-DS-02 was able to break Aβ self-tolerance in the
DS mouse model.
The IgM titers were analyzed in plasma of mice immunized with the ACI-
DS-02 vaccine. The ELISA done with Aβ42 showed that the immunized Ts65Dn
group had higher IgM titers than the group immunized with PBS at day 7, day 35
and day 49 (one way ANOVA, Tukey posthoc, P<0.01, P < 0.05 and P<0.01
respectively, data not shown.)
Example 8 ACI-DS-02 derived anti-human Aβ antibodies recognize DS amyloid
plaques
The immuno-staining experiment showed the ACI-DS-02 derived
antibodies bind to amyloid plaques in the brain samples of DS people (Figure 6).
The staining was similar to the obtained staining with 6E10, a commercial available
anti-amyloid beta antibody, and with ACI-24 derived antibody. ACI-24 vaccine
corresponds to ACI-DS-01 vaccine with the difference that the antigen used in ACI-
24 is a human Aβ 1-15 sequence, while ACI-DS-01 vaccine contains mouse Aβ 1-
sequence as the antigen (for three amino acid difference in those two antigens
see Figure 1). No staining was observed when a section was incubated with sera of
mice immunized with PBS. This result indicates that immunization with the ACI-DS-
02 vaccine induces antibodies that recognize amyloid plaques in the brain of DS
people.
Example 9 ACI-DS-02 enhanced associative learning in the fear conditioning test
During the conditioning test, Ts65Dn mice immunized with ACI-DS-02
vaccine showed a trend to higher levels of freezing after receiving each conditioned
stimulus (Figure 7A). The enhanced freezing was significantly different from the
PBS immunized Ts65Dn group only at the third conditioned stimulus (CS3) (two way
ANOVA, time, P<0.0001, a trend for vaccine (P = 0.09), Bonferroni posttests
showed P = 0.05 at the CS3).
The learning session is known to be mediated by synaptic plasticity-
dependent mechanism (Johansen 2011). However, previously acquired fear
memory (context, cued and extinction sessions) is known to be mediated by a
different neuronal mechanism. This result suggests that the ACI-DS-02
immunization enhanced acquisition/training phase of fear conditioning during which
learning occurs.
Example 10 Immunization with ACI-DS-03 produced anti-human Aβ antibodies
Four doses of ACI-DS-03 were administered subcutaneously and bi-
weekly into Ts65Dn male mice and age-matched control mice 2N. The analysis of
plasma collected 7 days after each dose, showed IgG titers against human Aβ42 in
the immunized mice of both Ts65Dn and 2N groups (data not shown). The IgG titers
of Ts65Dn mice immunized with ACI-DS-03 were significantly different from those
immunized with PBS at day 21 (one way ANOVA Tukey posthoc, P = 0.05). Thus,
ACI-DS-03 was able to break Aβ self-tolerance in the DS mouse model.
The IgM titers were analyzed in plasma of mice immunized with ACI-DS-03
vaccine. The ELISA done with Aβ42 showed that the immunized Ts65Dn group had
higher IgM titers than the group immunized with PBS at day 7 and 49 (one way
ANOVA, Tukey posthoc, P<0.001 and P<0.01 respectively (data not shown).
Example 11 ACI-DS-03 derived anti-human Aβ antibodies recognize DS amyloid
plaques
The immuno-staining experiment showed the ACI-DS-03 derived
antibodies bind to amyloid plaques in the brain of DS people (Figure 8). The staining
was similar to the obtained staining with 6E10, a commercial available anti-amyloid
beta antibody, and with ACI-24 derived antibody. ACI-24 vaccine corresponds to
ACI-DS-01 vaccine with the difference that the antigen used in ACI-24 is a human
Aβ 1-15 sequence, while ACI-DS-01 vaccine contains mouse Aβ 1-15 sequence as
the antigen (for three amino acid difference in those two antigens see Figure 1). No
staining was observed when section was incubated with sera of mice immunized
with PBS. This result indicates that immunization by ACI-DS-03 vaccine induces
antibodies that recognize amyloid plaques in the brain of DS people.
Example 12 Attenuated Total Reflectance InfraRed spectroscopy
Liposomal samples were analyzed as a D O hydrated thin-film from 15 µl
dried on the ZnSe crystal. Spectra were obtained with a BRUKER TENSOR 27
FTIR spectrometer equipped with a liquid nitrogen cooled mercury-cadmium-
telluride detector and coupled to a BioATR-II device. For each spectrum 2000 scans
were collected using the clean ATR crystal as background with a resolution of 2 cm-
1, averaged and FT processed. The upper and the lower frequency folding limits
were 4000 cm-1 and 900 cm-1. The sample chamber was continuously purged with
dried air and all measurements were performed at 25 ºC. Then acquired spectra
were cut in the amide I region, baseline corrected and labeled. Narrowing
techniques [second derivative and fourier self deconvolution (FSD)] were applied in
order to reveal the overlapping components of the broad amide I band. A qualitative
assignment of the major secondary conformation was done from the main detected
bands using the following frequency ranges in D O: b-sheet 1613-1637 cm ,
-1 -1 -1
random coil 1637-1646 cm , a-helix/loops 1646-1662 cm , b-turns 1662-1682 cm ,
anti-parallel b-sheet 1682-1698 cm . A quantitative analysis was done by curve-
fitting with the corresponding application in OPUS software, using the bands
identified in the 2 derivative and FSD spectra as initial values for the individual
components. Component peaks were assumed to be of mixed Gaussian-Loretzian
shape. The iteration process was carried out by fixing the frequencies and allowing
the peak widths, intensities and shapes to vary and then for the second iteration
process, the frequencies were also allowed to vary. Secondary structure estimates
were made by calculating the % area of the total amide I region for each secondary
structure band, assuming equal molar absorptivities of different components.
Reference List
1. Antonarakis,S.E., Lyle,R., Dermitzakis,E.T., Reymond,A., Deutsch,S., 2004.
Chromosome 21 and down syndrome: from genomics to pathophysiology.
Nat. Rev. Genet. 5, 725-738.
2. Ballard,C., Gauthier,S., Corbett,A., Brayne,C., Aarsland,D., Jones,E., 2011.
Alzheimer's disease. Lancet. 377, 1019-1031.
3. Belichenko,N.P., Belichenko,P.V., Kleschevnikov,A.M., Salehi,A.,
Reeves,R.H., Mobley,W.C., 2009. The "Down syndrome critical region" is
sufficient in the mouse model to confer behavioral, neurophysiological, and
synaptic phenotypes characteristic of Down syndrome. J Neurosci. 29, 5938-
5948.
4. Belichenko,P.V., Kleschevnikov,A.M., Salehi,A., Epstein,C.J., Mobley,W.C.,
2007. Synaptic and cognitive abnormalities in mouse models of Down
syndrome: exploring genotype-phenotype relationships. J Comp. Neurol. 504,
329-345.
. Bevins,R.A., Besheer,J., 2006. Object recognition in rats and mice: a one-trial
non-matching-to-sample learning task to study 'recognition memory'. Nat.
Protoc. 1, 1306-1311.
6. Davisson,M.T., Schmidt,C., Reeves,R.H., Irving,N.G., Akeson,E.C.,
Harris,B.S., Bronson,R.T., 1993. Segmental trisomy as a mouse model for
Down syndrome. Prog. Clin. Biol. Res. 384, 117-133.
7. Deacon,R.M., Rawlins,J.N., 2006. T-maze alternation in the rodent. Nat.
Protoc. 1, 7-12.
8. Gilman,S., Koller,M., Black,R.S., Jenkins,L., Griffith,S.G., Fox,N.C., Eisner,L.,
Kirby,L., Boada,R.M., Forette,F., Orgogozo,J.M., 2005. Clinical effects of
A{beta} immunization (AN1792) in patients with AD in an interrupted trial.
Neurology.
9. Hunter,C.L., Bimonte-Nelson,H.A., Nelson,M., Eckman,C.B., Granholm,A.C.,
2004. Behavioral and neurobiological markers of Alzheimer's disease in
Ts65Dn mice: effects of estrogen. Neurobiol. Aging. 25, 873-884.
. Johansen I.P., Cain C.K.,Ostroff L.E and LeDoux J.E., 2011. Molecular
Mechanisms of Fear Learning and Memory. Cell 147
11. Lee,M., Bard,F., Johnson-Wood,K., Lee,C., Hu,K., Griffith,S.G., Black,R.S.,
Schenk,D., Seubert,P., 2005. Abeta42 immunization in Alzheimer's disease
generates Abeta N-terminal antibodies. Ann. Neurol. 58, 430-435.
12. Lemere,C.A., Blusztajn,J.K., Yamaguchi,H., Wisniewski,T., Saido,T.C.,
Selkoe,D.J., 1996. Sequence of deposition of heterogeneous amyloid beta-
peptides and APO E in Down syndrome: implications for initial events in
amyloid plaque formation. Neurobiol Dis. 3, 16-32.
13. Liu,F., Grundke-Iqbal,I., Iqbal,K., Oda,Y., Tomizawa,K., Gong,C.X., 2005.
Truncation and activation of calcineurin a by calpain I in Alzheimer disease
brain. J. Biol. Chem. 280, 37755-37762.
14. Liu,F., Liang,Z., Wegiel,J., Hwang,Y.W., Iqbal,K., Grundke-Iqbal,I.,
Ramakrishna,N., Gong,C.X., 2008. Overexpression of Dyrk1A contributes to
neurofibrillary degeneration in Down syndrome. FASEB J. 22, 3224-3233.
. Moncaster,J.A., Pineda,R., Moir,R.D., Lu,S., Burton,M.A., Ghosh,J.G.,
Ericsson,M., Soscia,S.J., Mocofanescu,A., Folkerth,R.D., Robb,R.M.,
Kuszak,J.R., Clark,J.I., Tanzi,R.E., Hunter,D.G., Goldstein,L.E., 2010.
Alzheimer's disease amyloid-beta links lens and brain pathology in Down
syndrome. PLoS One 5, e10659.
16. Monsonego,A., Maron,R., Zota,V., Selkoe,D.J., Weiner,H.L., 2001. Immune
hyporesponsiveness to amyloid beta-peptide in amyloid precursor protein
transgenic mice: implications for the pathogenesis and treatment of
Alzheimer's disease. Proc. Natl. Acad. Sci. U. S. A 98, 10273-10278.
17. Netzer,W.J., Powell,C., Nong,Y., Blundell,J., Wong,L., Duff,K., Flajolet,M.,
Greengard,P., 2010. Lowering beta-amyloid levels rescues learning and
memory in a Down syndrome mouse model. PLoS One 5, e10943.
18. Nicolau,C., Greferath,R., Balaban,T.S., Lazarte,J.E., Hopkins,R.J., 2002. A
liposome-based therapeutic vaccine against beta -amyloid plaques on the
pancreas of transgenic NORBA mice. Proc. Natl. Acad. Sci. U. S. A 99, 2332-
2337.
19. O'Doherty,A., Ruf,S., Mulligan,C., Hildreth,V., Errington,M.L., Cooke,S.,
Sesay,A., Modino,S., Vanes,L., Hernandez,D., Linehan,J.M., Sharpe,P.T.,
Brandner,S., Bliss,T.V., Henderson,D.J., Nizetic,D., Tybulewicz,V.L.,
Fisher,E.M., 2005. An aneuploid mouse strain carrying human chromosome
21 with Down syndrome phenotypes. Science. 309, 2033-2037.
. Prasher,V.P., 1993. Down's syndrome, longevity, and Alzheimer's disease.
Br. J Psychiatry. 162, 711.
21. Prasher,V.P., 2004. Review of donepezil, rivastigmine, galantamine and
memantine for the treatment of dementia in Alzheimer's disease in adults with
Down syndrome: implications for the intellectual disability population. Int. J
Geriatr. Psychiatry. 19, 509-515.
22. Rafii,M.S., 2010. The pulse of drug development for Alzheimer's disease.
Rev. Recent. Clin. Trials. 5, 57-62.
23. Saxe,M.D., Battaglia,F., Wang,J.W., Malleret,G., David,D.J., Monckton,J.E.,
Garcia,A.D., Sofroniew,M.V., Kandel,E.R., Santarelli,L., Hen,R., Drew,M.R.,
2006. Ablation of hippocampal neurogenesis impairs contextual fear
conditioning and synaptic plasticity in the dentate gyrus. Proc. Natl. Acad.
Sci. U. S. A. 103, 17501-17506.
24. Stanton L.R, Coetzee R.H . Down's syndrome and dementia. 10, 50-58.
2004. Advances in Psychiatric Treatment .
Ref Type: Generic
. Stoltzner,S.E., Grenfell,T.J., Mori,C., Wisniewski,K.E., Wisniewski,T.M.,
Selkoe,D.J., Lemere,C.A., 2000. Temporal accrual of complement proteins in
amyloid plaques in Down's syndrome with Alzheimer's disease. Am. J.
Pathol. 156, 489-499.
26. Tapiola,T., Soininen,H., Pirttila,T., 2001. CSF tau and Abeta42 levels in
patients with Down's syndrome. Neurology. 56, 979-980.
27. Weiner,H.L., Frenkel,D., 2006. Immunology and immunotherapy of
Alzheimer's disease. Nat. Rev. Immunol. 6, 404-416.
28. Fernandez F, Morishita W, Zuniga E, Nguyen J, Blank M, Malenka RC and
Garner CC (2007)
29. Hanney M, Prasher V, Williams N, Jones EL, Aarsland D, Corbett A,
Lawrence D, Yu LM, Tyrer S, Francis PT, Johnson T, Bullock R and Ballard C
(2012) Memantine for Dementia in Adults Older Than 40 Years With Down's
Syndrome (MEADOWS): a Randomised, Double-Blind, Placebo-Controlled
Trial. Lancet 379:528-536.
. Salehi A, Delcroix J-D, Belichenko P.V., Zhan K ; Wu C., Valletta J C,
Takimoto-Kimura R, Kleschevnikov A M, Sambamurti K, Chung PP, Xia
W,Villar A, Campbell AW, Kulnane LS, Nixon RA, Lamb BT, Epstein CJ,
Stokin GB, Goldstein L.S.B, Mobley W.C. (2006) Increased App Expression
in a Mouse Model of Down’s Syndrome Disrupts NGF Transport and Causes
Cholinergic Neuron Degeneration, Neuron, 51, 29-42.
31. Salehi A, Faizi M, Colas D, Valletta J, Laguna J, Takimoto-Kimura R.,
Kleschevnikov A., Wagner S. L., Aisen P., Shamloo M., MobleyW. C (2009)
Restoration of Norepinephrine-Modulated Contextual Memory in a Mouse
Model of Down Syndrome, Sci Transl Med 1, 7ra17.
In this specification where reference has been made to patent specifications, other
external documents, or other sources of information, this is generally for the purpose of
providing a context for discussing the features of the invention. Unless specifically
stated otherwise, reference to such external documents is not to be construed as an
admission that such documents, or such sources of information, in any jurisdiction, are
prior art, or form part of the common general knowledge in the art.
In the description in this specification reference may be made to subject matter that is
not within the scope of the claims of the current application. That subject matter should
be readily identifiable by a person skilled in the art and may assist in putting into
practice the invention as defined in the claims of this application.
Claims (39)
1. Use of an antigenic peptide fragment derived from amyloid protein or amyloid-like protein selected from the group consisting of prion protein, tau protein, alpha-synuclein, huntingtin and amyloid beta for the preparation of a medicament for use in the treatment, alleviation or prevention of memory and/or cognitive impairments or abnormalities in subjects with Down’s syndrome; wherein said subjects with Down’s syndrome have not yet developed amyloid protein-associated or amyloid- like protein-associated, plaques in the brain.
2. The use of claim 1, wherein said treatment leads to a. an amelioration or restoration of memory; and/or b. an increase in the retention or a complete restoration of cognitive capacity in the treated subject.
3. The use of claim 1 for the preparation of a medicament for use in the prevention of memory and/or cognitive impairments or abnormalities in subjects with Down’s syndrome.
4. The use of any one of claims 1-3, wherein said memory and/or cognitive impairments originate in the hippocampus, the prefrontal cortex or the entorhinal cortex of the brain.
5. The use of any one of claims 1 to 4 wherein said memory and/or cognitive impairments are impairments of recognition memory, impairments of the contextual associative memory, impairments of the associative learning, impairments of the declarative memory for facts and events, episodic memory impairments, language dysfunction, visuospatial impairments, decreased executive functions, poor abstract reasoning, personality changes, emotional changes, impairments of performing learned motor tasks, other neurological signs including pyramidal and extrapyramidal findings, myoclonus or seizures or various combinations thereof.
6. The use of claim 5, wherein language dysfunction results in aphasia, visuospatial impairments result in misplacement of items and difficulty navigating in unfamiliar and familiar terrain, decreased executive functions result in apathy, disinhibition, social isolation, poor judgment, and difficulties with planning, emotional changes result in apathy, agitation and psychosis.
7. The use of claim 5, wherein contextual associative memory in subjects with Down’s syndrome is enhanced or restored.
8. The use of claim 5, wherein associative learning in subjects with Down’s syndrome is enhanced or restored.
9. The use of any one of claims 1 to 8, wherein said amyloid protein or amyloid-like protein is an antigenic peptide fragment derived from amyloid-beta protein.
10. The use of any one of the preceding claims, wherein said memory and/or cognitive impairments or abnormalities in subjects with Down’s syndrome are AD-like memory or cognitive impairments or abnormalities.
11. The use of any one of the preceding claims, wherein said subject with Down’s syndrome is a young to middle-aged subject.
12. The use of any one of the preceding claims, wherein said subject with Down’s syndrome is a middle-aged subject.
13. The use of any one of claims 1 to 11, wherein said subject with Down’s syndrome is a young-aged subject.
14. The use of any one of claims 1 to 11, wherein said subject with Down’s syndrome is a children-aged subject.
15. The use of any one of claims 1 to 11, wherein said subject is below age
16. The use of any one of claims 1 to 11, wherein said subject is below age below age 50.
17. The use of any one of claims 1 to 11, wherein said subject is below age
18. The use of any one of claims 1 to 11, wherein said subject is below age
19. The use of any one of claims 1 to 11, wherein said subject is below age
20. The use of any one of claims 1 to 11, wherein said subject is below age
21. The use of any one of the preceding claims, wherein said treatment does not induce unwanted side effects.
22. The use of any one of the preceding claims, wherein said treatment prevents the development of Aβ-associated plaques in the brain.
23. The use of any one of the preceding claims, wherein said Aβ antigenic peptide fragment corresponds to the N-terminal part of the Aβ peptide.
24. The use of claim 23, wherein said antigenic peptide is derived from the N- terminal part of the Aβ peptide consisting of all or part of amino acid residues Aβ1-15, Aβ1-16, Aβ1-17, Aβ1-18, Aβ1-19, Aβ1-20, Aβ1-22, or Aβ1-23.
25. The use of claim 24, wherein said antigenic peptide consists of residues Aβ1-15.
26. The use of any one of the preceding claims, wherein said Aβ antigenic peptide fragment corresponds to the central part of the Aβ peptide.
27. The use of any one of the preceding claims, wherein said Aβ antigenic peptide fragment corresponds to the C-terminal part of the Aβ peptide.
28. The use of claim 26 or 27, wherein said antigenic peptide is derived from the C-terminal part of the Aβ peptide consisting of all or part of amino acid residues Aβ20-36, Aβ20-40, Aβ20-42, Aβ21-36, Aβ21-40, Aβ21-42, Aβ22- 36, Aβ22-40 or Aβ22-42.
29. The use of claim 28, wherein said antigenic peptide consists of residues Aβ22-35.
30. The use of any one of the preceding claims, wherein the Aβ antigenic peptide fragment is presented reconstituted in a liposome.
31. The use of claim 30, wherein at least 75% of the reconstituted antigenic peptide is present on the surface of the liposome, inserted into the lipid bilayer through its hydrophobic moieties.
32. The use of any one of the preceding claims, wherein the Aβ antigenic peptide fragment is modified by a lipophilic or hydrophobic moiety that facilitates insertion into the lipid bilayer of the liposome carrier/adjuvant.
33. The use of claim 32, wherein the Aβ antigenic peptide fragment is modified by a palmitic acid bound to the N- and/or C-terminus of the peptide.
34. The use of claim 33, wherein the Aβ antigenic peptide fragment is modified by four palmitic acids bound to the N- and/or C-terminus of the peptide.
35. The use of claim 33, wherein the Aβ antigenic peptide fragment is modified by two palmitic acids bound to the N- and/or C-terminus of the peptide.
36. The use of any one of the preceding claims, wherein the Aβ antigenic peptide fragment is presented in a repetitive array on the surface of the liposome.
37. The use of claim 36, wherein said highly repetitive array on the surface of the carrier or liposome comprises at least 10 repetitive antigenic units/carrier molecule.
38. The use of any one of the preceding claims, wherein said treatment leads to the induction of an immune response against amyloid protein or amyloid-like protein in a subject with Down’s syndrome.
39. Use of a composition comprising (i) an antigenic peptide fragment derived from amyloid protein or amyloid-like protein selected from the group consisting of prion protein, tau protein, alpha-synuclein, huntingtin and amyloid beta as defined in any one of claims 1, 9 and 23-37, and (ii) a
Applications Claiming Priority (3)
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USPCT/US2011/052992 | 2011-09-23 | ||
USPCT/US2011/052992 | 2011-09-23 | ||
PCT/US2012/056728 WO2013044147A1 (en) | 2011-09-23 | 2012-09-21 | Vaccine therapy |
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NZ622653A NZ622653A (en) | 2015-12-24 |
NZ622653B2 true NZ622653B2 (en) | 2016-03-30 |
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