Monday, July 15, 2013

What De Meirleir has been working on....

Source: European Patent Office
Date: February 27, 2013
URL: http://www.freepatentsonline.com/EP2561879.html
https://data.epo.org/publication-server/rest/v1.0/publication-dates/20130227/patents/EP2561879NWA1/document.pdf


European Patent Application EP2561879
-------------------------------------

Abstract

The present invention relates to Macrophage Activating Factors such as
GcMAF and compositions thereof, for use in the treatment of a patient
suffering from CFS/ME and/or XMRV infection.


Inventors:

Roelant, Christiaan ('s Hertogenlaan 115, B-3000 Leuven, BE)
De Meirleir, Kenny (Stuivenberglaan 89, B-2800 Mechelen, BE)


Application Number:

EP20110178511


Publication Date:

02/27/2013


Filing Date:

08/23/2011

(...)


Assignee:

Protea Biopharma N.V. (De Tyraslaan 111, 1120 Neder-Over-Heembeek, BE)


International Classes:

A61K35/407; A61K31/714; A61K38/19; A61K38/20; A61K38/21; A61P31/12; A61P43/00


View Patent Images:

http://www.freepatentsonline.com/EP2561879.pdf


Attorney, Agent or Firm:

Paemen, Liesbet R. J. (De Clercq & Partners Edgard Gevaertdreef 10 a,
9830 Sint-Martens-Latem, BE)



Claims:

1. A composition comprising a Macrophage Activating Factor for use in
the treatment of a patient suffering from chronic fatigue
syndrome/myalgic encephalomyelitis (CFS/ME) and/or an infection by
xenotropic murine leukemia virus-related virus (XMRV).

2. The composition according to claim 1, wherein said patient is
characterized by reduced NK cell activity and increased inflammatory
mediators.

3. The composition according to claim 1 or 2, for reducing at least
one symptom chosen from the group consisting of substantial impairment
in short-term memory or concentration, sore throat, tender lymph
nodes, muscle pain, multi-joint pain without swelling or redness,
headaches of a new type, pattern, or severity; unrefreshing sleep,
post-exertional malaise lasting more than 24 hours, widespread pain,
fatigue, feeling run down, sluggish muscle cramps and pains,
unexplained or excessive weight gain, inability to lose weight,
gastrointestinal problems, irritable bowel syndrome, poor sleeping,
headaches and migraines, constipation, orthostatism and exhaustion.

4. The composition according to any one of claims 1 to 3, wherein said
patient is diagnosed with aberrant and/or increased metabolite
production.

5. The composition according to claims 1 to 4, wherein said Macrophage
Activating Factor is GcMAF.

6. The composition according to any one of claims 1 to 5, in which
said Macrophage Activating Factor is the only active ingredient.

7. The composition according to any one of claims 1 to 5, further
comprising one or more other active ingredients.

8. The composition according to claim 7, where one of said one or more
other active ingredients is Vitamin B12.

9. The composition according to claim 7 or 8, where at least one of
said one or more other active ingredients is selected from the group
consisting of interferon alpha, interferon beta, interferon gamma and
interleukin-7.

10. The composition according to any one of claims 7, 8 or 9, wherein
at least one of said one or more other active ingredients is selected
from a viral inhibitor and/or an ecto-nox-modifying agent.

11. The composition according to any one of claims 7 to 10, further
comprising a mammalian liver extract.

12. The composition according to any one of claims 1 to 11, wherein
said composition is an enteral composition.

13. The composition according to any one of claims 1 to 12, wherein
said patient is diagnosed with CFS/ME and with XMRV infection.




Description:

FIELD OF THE INVENTION

The present invention relates to the therapeutic and/or prophylactic
use of Macrophage Activating Factors.


BACKGROUND OF THE INVENTION

Macrophage Activating Factors (MAFs) are lymphokines that prime
macrophages to become cytotoxic to tumors. These factors also control
the expression of la antigens on the macrophage cell surface. There
are indications that the use of MAFs, particularly the GcMAF protein,
is effective in the treatment of certain cancers and HIV (EP0837932,
Yamamoto).

Chronic fatigue syndrome (CFS) is a difficult to diagnose, ubiquitous
disorder characterized by extreme fatigue, lymph gland enlargement and
constitutional symptoms such as weight loss, loss of appetite, memory
deterioration and loss of intelligence in some patients. Some CFS
patients manifest neuropsychiatric changes such as depression, loss of
memory and similar derangements. Thus, chronic fatigue syndrome is
sometimes difficult to distinguish from entirely neurological
disorders, particularly situational depression. An accumulating body
of evidence suggests that CFS is associated with disregulation of both
humoral and cellular immunity, including mitogen response,
reactivation of viruses, abnormal cytokine production, diminished
natural killer cell function and changes in intermediary metabolites.
CFS is also referred to by some as CFS/ME (myalgic encephalomyelitis).
The term myalgic encephalomyelitis refers to muscle aches or pains and
inflammation of the brain and spinal cord. While inflammation of the
brain is not observed in all CFS patients, the group of patients
referred to under CFS/ME typically suffer from the same symptoms and
lack a clear etiology

A number of further diseases have been associated with CFS/ME because
of the similarity of the symptoms observed with CFS/ME patients and
patients suffering from such CFS/ME-related diseases. For instance,
fibromyalgia, autoimmune thyroid disease, xenotropic murine leukemia
virus-related virus (XMRV) and several other diseases have been
suggested as being related to CFS/ME and present similar symptoms of
widespread pain, fatigue, feeling run down, sluggish muscle cramps and
pains, unexplained or excessive weight gain, inability to lose weight,
gastrointestinal problems, irritable bowel syndrome, poor sleeping,
headaches and migraines, constipation and exhaustion.

For many CFS/ME patients, cognitive behavioural therapy and graded
exercise therapy have shown only moderate effectiveness (Whiting et
al. JAMA 286: 1360-1368, 2001). Many patients do not fully recover
from CFS/ME even with treatment (Rimes et al. Occupational Medicine
55: 32-39, 2005). Other treatments of CFS/ME have been proposed but
their effectiveness has not been confirmed (Prins et al. Lancet 367:
346-355, 2006).

XMRV is an infectious human gamma-retrovirus, which has been linked to
both CFS/ME and human prostate cancer (Kearney et al. JID 202:
1463-1466, 2011). Although in vitro studies have indicated that
raltegravir (Trade name Isentress(r)) can act as an inhibitor for XMRV
replication (Singh et al., PLoS ONE 5: e9948, 2010), its effect on the
in vivo XMRV replication is not yet documented.

There remains a need for compounds for use in the treatment and/or
prevention of CFS/ME and CFS/ME-related diseases, such as XMRV
infection. More particularly, there is a need for novel and improved
compounds that are capable of reducing, inhibiting or decreasing the
particular symptoms that are observed in patients suffering from
CFS/ME and/or CFS/ME-related diseases and disorders, such as for
example XMRV infection.


SUMMARY OF THE INVENTION

The present invention relates to a Macrophage Activating Factor (MAF)
for use in the treatment of a patient suffering from chronic fatigue
syndrome/myalgic encephalomyelitis (CFS/ME) and/or one or more
CFS/ME-related diseases and/or disorders, such as infection by
xenotropic murine leukemia virus-related virus (XMRV). In particular
embodiments, the present invention provides a composition comprising a
MAF for use in the treatment of a patient suffering from CFS/ME and/or
an infection by XMRV. In a further aspect the present invention
provides use of a MAF for the manufacture of a medicament for the
treatment of CFS/ME and/or CFS/ME related diseases and disorders, such
as infection by XMRV in a mammal.

Indeed, the inventors found that the administration of a MAF leads to
a significant decrease of the manifestation of the symptoms of CFS/ME
and CFS/ME-related diseases, more particularly of XMRV. For instance,
the inventors have found that administration with a MAF, more
particularly GcMAF, is capable of suppressing or reducing at least one
of the biological parameters associated with CFS/ME and CFS/ME-related
diseases. More particularly, it has been established that indicators
of Natural Killer (NK) cell activity and inflammation, are affected
when patients suffering from such diseases are treated with MAF. Thus,
in particular embodiments, the patient is characterized by reduced NK
cell activity and increased inflammatory mediators. In certain
embodiments, the patient is diagnosed with CFS/ME and with XMRV
infection.

It is further considered that treatment with MAF ensures important
clinical improvement, for example as measured on the Karnofsky scale
or the Bell disability scale (D.S. Bell, The Doctor's Guide to Chronic
Fatigue Syndrome, Reading, MA: Addison-Wesley, 1994). Specifically,
the MAF or compositions according to the present invention are useful
for reducing at least one symptom chosen from the group consisting of
substantial impairment in short-term memory or concentration, sore
throat, tender lymph nodes, muscle pain, multi-joint pain without
swelling or redness, headaches of a new type, pattern, or severity;
unrefreshing sleep, post-exertional malaise lasting more than 24
hours, widespread pain, fatigue, feeling run down, sluggish muscle
cramps and pains, unexplained or excessive weight gain, inability to
lose weight, gastrointestinal problems, irritable bowel syndrome, poor
sleeping, headaches and migraines, constipation, orthostatism and
exhaustion.

In particular embodiments, the present invention provides a MAF as
described above provided in a conjugate or in a pharmaceutical
composition comprising a pharmaceutically acceptable excipient and a
therapeutically effective amount of said MAF.

In certain embodiments, the MAF as described herein is the only active
ingredient of the composition as described herein. In further
embodiments, the present invention provides a MAF as described above
provided in a conjugate or in a pharmaceutical composition consisting
of a pharmaceutically acceptable excipient and a therapeutically
effective amount of said MAF.

In particular embodiments, the present invention relates to methods
for treating a patient having been diagnosed with CFS/ME and/or one or
more CFS/ME-related disease(s) based on aberrant ecto-nox functioning
as described herein. In particular embodiments, the patient described
above is diagnosed with aberrant and/or increased metabolite production.

In particular embodiments, the MAF in the present invention is GcMAF protein.

A further aspect of the present invention relates to the compositions
as described above, wherein said MAF according to the invention is
provided in combination with one or more other pharmacologically
active compounds or ingredients. In particular embodiments, one of the
one or more other active ingredients is Vitamin B12. In certain
embodiments, at least one of the one or more other active ingredients
is selected from the group consisting of interferon alpha, interferon
beta, interferon gamma and interleukin-7. In certain embodiments, at
least one of the one or more other active ingredients is selected from
a viral inhibitor and/or an ecto-nox-modifying agent. In certain
embodiments, the compositions according to the present invention
further comprise a mammalian liver extract.

In particular embodiments, the present invention relates to
compositions as described above, for enteral administration. In
another embodiment, the present invention relates to compositions as
described above, for parenteral administration.


DETAILED DESCRIPTION OF THE INVENTION

In the following passages, different aspects of the invention are
described in more detail. Each aspect so described may be combined
with any other aspect or aspects unless clearly indicated to the
contrary. In particular, any feature indicated as being preferred or
advantageous may be combined with any other feature or features
indicated as being preferred or advantageous.

In the context of the present invention, the terms used are to be
construed in accordance with the following definitions, unless a
context dictates otherwise.

As used herein, the singular forms 'a', 'an', and 'the' include both
singular and plural referents unless the context clearly dictates
otherwise.

The terms 'comprising', 'comprises' and 'comprised of' as used herein
are synonymous with 'including', 'includes' or 'containing',
'contains', and are inclusive or open-ended and do not exclude
additional, non-recited members, elements or method steps.

The recitation of numerical ranges by endpoints includes all numbers
and fractions subsumed within the respective ranges, as well as the
recited endpoints.

The term 'about' as used herein when referring to a measurable value
such as a parameter, an amount, a temporal duration, and the like, is
meant to encompass variations of +/-10% or less, preferably +/-5% or
less, more preferably +/-1% or less, and still more preferably +/-0.1%
or less of and from the specified value, insofar such variations are
appropriate to perform in the disclosed invention. It is to be
understood that the value to which the modifier 'about' refers is
itself also specifically, and preferably, disclosed.

This invention relates to the use of Macrophage Activating Factors
('MAFs') in the treatment of CFS/ME and/or in the treatment of
infection by xenotropic murine leukemia virus-related virus (XMRV) in
a mammal.

The term 'Macrophage activating factor' or 'MAF' as referred to herein
designates a lymphokine protein that stimulates monocytes and/or
macrophages (designated as macrophages hereafter). This stimulation
results in the macrophages to become cytotoxic to tumors, and/or
primes the phagocytosis of pathogens by macrophages.

In particular, the present invention relates to a GcMAF protein for
use in the treatment of a mammal suffering from CFS/ME and/or one or
more CFS/ME-related diseases, such as for example infection by XMRV.

The term 'GcMAF' as referred to herein designates a Gc (group specific
component) protein derived macrophage activating protein, or a domain
III (natural or cloned) derived macrophage activating protein (e.g.

US patent 6410269, Yamamoto, which is included herein by reference).
The term 'domain III' as referred to herein designates the domain III
region of the Gc protein. The term 'Gc protein' as referred to herein
designates a Vitamin D binding protein (DBP); DBP is also known as
gc-globulin.

A first aspect of the invention relates to a composition comprising a
Macrophage Activating Factor (MAF) for use in the treatment of
patients diagnosed with chronic fatigue syndrome/myalgic
encephalomyelitis (CFS/ME) and/or CFS/ME-related diseases and
disorders. Accordingly, the present invention provides for a treatment
that reduces, inhibits or decreases the symptoms of CFS/ME and
CFS/ME-related diseases, such as XMRV infection.

It is known that CFS/ME and CFS/ME related diseases seriously
interfere with the immune system, causing a dysfunctioning thereof.
The present inventors have found that by using a composition
comprising a MAF, the activity of natural killer cells (or NK cells),
i.e. cytotoxic lymphocytes that play a major role in the regulation of
the innate immune system, is increased. Thus, use of MAF in patients
suffering from CFS/ME or CFS/ME-related diseases causes a positive
stimulation of the immune system. Furthermore, the inventors have
found that by using a MAF in the treatment of patients suffering from
CFS/ME and/or CFS/ME-related diseases, inflammation is reduced.

The term 'chronic fatigue syndrome' or 'CFS' as referred to herein
designates a condition which is diagnosed based on the following
criteria (as developed by the U.S. Centers for Disease Control and
Prevention in 1994):
1. Clinically evaluated, unexplained persistent or relapsing chronic
fatigue that is of new or definite onset (i.e., not lifelong), is not
the result of ongoing exertion, is not substantially alleviated by
rest, and results in substantial reduction in previous levels of
occupational, educational, social, or personal activities.
2. The concurrent occurrence of four or more of the following
symptoms: substantial impairment in short-term memory or
concentration; sore throat; tender lymph nodes; muscle pain;
multi-joint pain without swelling or redness; headaches of a new type,
pattern, or severity; unrefreshing sleep; and post-exertional malaise
lasting more than 24 hours. These symptoms must have persisted or
recurred during 6 or more consecutive months of illness and must not
have predated the fatigue.

The term 'CFS/ME and CFS/ME-related diseases and disorders' refers to
diseases and/or disorders with having an underlying biological
mechanism of action and/or signaling pathways that are the same or
similar to those known to underlie CFS and/or presenting the same or
similar symptoms as observed with patients suffering from CFS.

Several standardized tests exist to follow up patients suffering from
CFS/ME, such as the Activities of Daily Living Questionnaire (ADL)(see
e.g. Collin et al. Int. Disabil. Stud. 10:61-63, 1988), the (Cognitive
Deficit Subset of the) Symptom Checklist Questionnaire (SCL-90-R) (see
e.g. Diaz-Mitoma et al. Journal of Chronic Fatigue Syndrome 11:71-95,
2003) and the Karnofsky Performance Score (KPS) (Karnofsky D and
Burchenal JH. Clinical evaluation of chemotherapeutic agents in
cancer. In Macleod CM. (ed) Evaluation of chemotherapeutic agents.
Columbia University Press, New York 1949; 199-205). An alternative
performance score specifically developed for CFS/ME patients is the
Bell disability scale (D.S. Bell, The Doctor's Guide to Chronic
Fatigue Syndrome, Reading, MA: Addison-Wesley, 1994).

Thus, in particular embodiments, the present invention provides for
compositions comprising a MAF, for use in the treatment of CFS/ME and
CFS/ME related diseases, which are capable of reducing the clinical
symptoms of CFS/ME and/or CFS/ME related diseases as determined by one
or more of the above-described evaluation methods. More particularly
the compositions of the invention ensure a 10% improvement, more
particularly a 20%, 30%, 40%, 50%, 60%, 70% or 80% or more improvement
of clinical symptoms compared to prior to administration of the
composition of the invention.

On a physiological level, CFS/ME has been associated with a variety of
immune abnormalities. For instance, it has been reported that CFS/ME
patients suffer from a suppression of NK cell activity (Whiteside et
al. Am. J. Med. 105: 27S-34S, 1998). Published results also
demonstrate an association between clinical improvement of patients
suffering from CFS/ME and enhanced NK cell activity (Diaz-Mitoma et
al. Journal of Chronic Fatigue Syndrome 11:71-95, 2003). It is also
reported that CFS/ME is associated with diminished intracellular
perforin, a protein found in the granules of NK cells (Maher et al.,
Clin. Exp. Immunol. 2002, 142, 505). Therefore, NK activity and/or
intracellular perforin levels are suitable indicators for monitoring
the disease state in patients suffering from CFS/ME.

Additionally, CFS/ME is associated with indications of inflammation.
For example, CFS/ME patients often exhibit elevated levels of
pro-inflammatory cytokines such as Interleukin-8 (IL-8). Patients
suffering from CFS/ME also frequently have elevated C4a serum levels
(Sorensen et al. J. Allergy Clin. Immunol. 112: 397-403, 2003). C4a is
an anaphylatoxin generated by cleavage of complement component 4 (C4),
upon activation of the complement system. Anaphylatoxins are able to
trigger degranulation of endothelial cells, mast cells or phagocytes,
which produce a local inflammatory response. Thus, indications of
inflammation such as IL-8 and/or C4a levels are additional parameters
which can be monitored in order to follow up patients suffering from
CFS/ME.

In particular embodiments, the present invention provides for
compositions comprising a MAF, for use in the treatment of CFS/ME and
CFS/ME related diseases, which are capable of reducing the
physiological symptoms of CFS/ME and/or CFS/ME related diseases such
as but not limited to reduced NK cell activity and elevated
inflammatory response. Most particularly, the invention provides
compositions capable of ensuring a 10% improvement, more particularly
a 20%, 30%, 40%, 50%, 60%, 70% or 80% or more improvement of the
physiological symptoms of CFS/ME compared to prior to administration
of the composition of the invention. More particularly, the
compositions of the invention can be used to normalize the NK activity
and/or inflammatory response in a patient suffering from CFS/ME or a
CFS/ME related disorder.

It will be understood that in view of the nature of the disease and
the diagnostic criteria, not all patients suffering from CFS/ME are
characterized by reduced NK cell activity and/or increased
inflammatory responses. Thus, in particular embodiments, the present
invention provides compositions for use in the treatment of patients
suffering from CFS/ME, wherein the patient is diagnosed with increased
C4a serum level, increased IL-8 serum level and/or decreased NK cell
activity. In particular embodiments, the patient is diagnosed with at
least one element selected from the group consisting of increased C4a
serum level, increased IL-8 serum level and decreased NK cell
activity. 'Increased C4a serum level' as used herein refers to a C4a
serum level above 1500 ng/mL, preferably above 2500 ng/mL. 'Increased
IL-8 serum level' as used herein refers to an IL-8 serum level above
15 pg/mL, preferably above 25 pg/mL. 'Decreased NK cell activity' as
used herein refers to a perforin mRNA expression in peripheral blood
mononuclear cells (PBMCs) below.

It has been described that further indicators of CFS/ME include an
aberrant functioning of constitutive ecto-nox proteins.

WO2010109009, which is hereby incorporated by reference, provides
methods for determining the presence of aberrant ecto-nox functioning
in the sample of a patient.

The term 'ecto-nox protein' or 'membrane NADH oxidase' as used herein
refers to a cell-surface protein with both hydroquinone oxidase and
protein-disulfide-thiol interchange activity. Constitutive ecto-nox
proteins or prions are constitutively present in cellular membranes.
They differ in this respect from mutated ecto-nox proteins present
e.g. in cancer cells, such as t-nox.

More particularly the method described in WO2010109009 can be used to
determine whether a patient is suffering from CFS/ME, more
particularly whether the patient is susceptible to treatment with a
compound capable of reducing aberrant ecto-nox functioning.

It is noted in this regard that the term 'normal' when used in the
context of ecto-nox protein or membrane NADH oxidase as used herein
refers to the membrane NADH oxidase identifiable on normal,
non-diseased cells and is characterized by the fact that it is heat
sensitive (loss of activity when subjected to 70 degrees Celsius for
10 min.) and preferentially activated under hypotonic conditions. The
term 'aberrant' when used in the context of ecto-nox protein or
membrane NADH oxidase herein refers to an ecto-nox protein which does
not function properly. In particular embodiments, the ecto-nox protein
is in a permanently activated state and characterized by the fact that
it is heat insensitive (retains activity when subjected to 70 degrees
Celsius for 10 min.) and shows highest activity under isotonic
conditions.

In particular embodiments, the present invention provides for
compositions comprising a MAF, for use in the treatment of CFS/ME and
CFS/ME related diseases, which are capable of reducing aberrant
ecto-nox functioning in patients suffering from CFS/ME and/or CFS/ME
related diseases. Most particularly, the invention provides
compositions capable of ensuring a 10% reduction, more particularly a
20%, 30%, 40%, 50%, 60%, 70% or 80% or more reduction of the aberrant
ecto-nox functioning in patients with CFS/ME compared to prior to
administration of the composition of the invention, more particularly
as determined by the methods disclosed in WO2010109009. More
particularly, the compositions of the invention can be used to
normalize the ecto-nox functioning in a patient suffering from CFS/ME
or a CFS/ME related disorder.

In particular embodiments, the present invention provides compositions
for use in the treatment of patients suffering from CFS/ME, wherein
the patient is diagnosed to have aberrant ecto-nox functioning, more
particularly as determined by the methods disclosed in WO2010109009.

Further indications of CFS/ME also include an increased amount of
metabolites in physiological fluids such as urine. The metabolite
concentration in physiological fluids can be detected and monitored
using reducible dyes, which allows monitoring of the disease
(WO2010142322, which is hereby incorporated by reference).

In particular embodiments, the present invention provides for
compositions comprising a MAF, for use in the treatment of CFS/ME
and/or CFS/ME related diseases, which are capable of reducing aberrant
metabolite production in patients suffering from CFS/ME and/or CFS/ME
related diseases. Most particularly, the invention provides
compositions capable of ensuring a 10% reduction, more particularly a
20%, 30%, 40%, 50%, 60%, 70% or 80% or more reduction of the aberrant
metabolite production in patients with CFS/ME compared to prior to
administration of the composition of the invention, more particularly
as determined by the methods disclosed in

WO2010142322. More particularly, the compositions of the invention can
be used to normalize the ecto-nox functioning in a patient suffering
from CFS/ME or a CFS/ME related disorder.

In particular embodiments, the present invention provides compositions
for use in the treatment of patients suffering from CFS/ME, wherein
the patient is diagnosed with increased metabolite production, more
particularly as determined by the methods disclosed in WO2010142322.

While the link between both conditions remains controversial, there is
increasing evidence that a significant number of patients suffering
from CFS/ME is infected with xenotropic murine leukemia virus-related
virus.

The term 'xenotropic murine leukemia virus-related virus' or 'XMRV' as
referred to herein designates an infectious gamma-retrovirus as found
in prostate tumors, particularly in prostate tumors of patients
homozygous for RNASEL variant, R462Q (e.g., Urisman et al., PLoS
Pathog. 2(3): e25, 2006; Dong et al., Proc. Natl. Acad. ScL USA
104(5): 655, 2007 and

WO 2006/110589; all of which are incorporated herein by reference in
their entirety). The term 'xenotropic murine leukemia virus-related
virus' or 'XMRV' includes any strain of the virus including XMRV VP35
(GenBank Accession No. DQ241301), XMRV VP42 (GenBank Accession No.
DQ241302) and XMRV VP62 (GenBank Accession No. DQ399707).

In addition, a number of diseases have been associated with CFS/ME
because of the similarity of the symptoms observed with CFS/ME
patients and patients suffering from such CFS/ME-related diseases. For
instance, fibromyalgia, autoimmune thyroid disease, xenotropic murine
leukemia virus-related virus (XMRV) and several other diseases have
been suggested as being related to CFS/ME and presenting similar
symptoms of widespread pain, fatigue, feeling run down, sluggish
muscle cramps and pains, unexplained or excessive weight gain,
inability to lose weight, gastrointestinal problems, irritable bowel
syndrome, poor sleeping, headaches and migraines, constipation,
orthostatism and exhaustion.

Although such CFS/ME-related diseases may contribute in the
pathogenesis of CFS/ME, often a causative link between CFS/ME and such
diseases has not been unequivocally established.
Accordingly, it is envisaged herein that while a subgroup of
patients with CFS/ME may additionally have one or more CFS/ME-related
diseases, such as an infection with XMRV not all patients with CFS/ME
are necessarily suffering from an additional CFS/ME related disease,
such as an infection with XMRV. In addition, not all patients with for
example diagnosed to be infected with XMRV, are additionally diagnosed
with CFS/ME, despite that fact that they may show at least some
symptoms of CFS/ME.

The compositions of the present invention are envisaged to be of use
in the treatment of both CFS/ME and CFS/ME related diseases. In
particular embodiments however, the present invention relates to the
treatment of patients with CFS/ME in which an infection with XMRV has
been identified.

The present invention relates to a MAF for use in the treatment of
CFS/ME and/or CFS/ME related diseases or disorders, such as an
infection by XMRV in a mammal. In particular embodiments, the present
invention relates to use of a MAF for the manufacture of a medicament
for the treatment of CFS/ME and/or infection by XMRV in a mammal. In
particular embodiments, the present invention relates to the a MAF for
use in the treatment of CFS/ME and/or infection by XMRV in a mammal.

In particular embodiments, the present invention relates to the use of
a MAF in the manufacture of a medicament for the treatment of, CFS/ME
and/or infection by XMRV in a mammal.

In particular embodiments, the present invention relates to
compositions comprising a MAF for use in the treatment of a patient
suffering from CFS/ME and/or one ore more CFS/ME-related diseases,
such as an infection by XMRV. In particular embodiments, the patient
is also characterized as having aberrant ecto-nox functioning, e.g. by
the method disclosed in WO2010109009. In particular embodiments, the
patient is also diagnosed with aberrant and/or increased metabolite
production, e.g. by the method disclosed in WO2010142322.

In further particular embodiments, the present invention provides in
compositions as described above, comprising a pharmaceutically
acceptable excipient and a therapeutically effective amount of said
MAF. In particular embodiments, the MAF as described herein, is GcMAF.

In particular embodiments, the compositions of the present invention
comprises MAF as the sole active ingredient or consists of a MAF.
However, treatment regimes are also envisaged wherein the MAF is
administered in combination with one or more other therapeutic
compounds. The present invention further envisages treatment regimes
wherein a patient suffering from CFS/ME or a CFS/ME related disorder
is treated with (Gc)MAF in combination with one or more compounds
selected from a viral inhibitor and/or an ecto-nox modifying agent,

More particularly, the present invention relates to the therapeutic
use of a MAF as described above used in combination with at least one
specific inhibitor of a virus wherein said virus is associated with
CFS/ME, more particularly wherein said virus is XMRV. In particular
embodiments, (one of) said viral inhibitor(s) is Raltegravir(r).

In particular embodiments, the present invention relates to the
therapeutic use of a MAF as described above used in combination with
an ecto-nox modifying agent. The term 'ecto-nox protein modifying
agent' as used herein refers to any compound capable of converting a
normal ecto-nox protein (or membrane NADH oxidase) into an aberrant
ecto-nox protein (or membrane NADH oxidase) or capable of inducing
aberrant NADH oxidase activity in a sample. In particular embodiments,
the ecto-nox protein modifying agent is a molecule having NADH oxidase
activity capable of competing with the ecto-nox protein, or a molecule
capable of inhibiting the association of an ecto-nox protein with the
plasma membrane, or an NADH oxidase inhibitor. In particular
embodiments, the ecto-nox modifying agent is selected from the group
consisting of a prion molecule (such as NADH oxidase) and a solvent
(such as dimethyl sulfoxide, i.e. DMSO).

In further particular embodiments, the present invention relates to
the therapeutic use of a MAF as described above used in combination
with at least one mammalian liver extract. Mammalian liver extract has
shown NADH oxidase activity and an antiviral activity against certain
viruses. Mammalian liver extract has been commercialized under the
names of Kutapressin(r) and Nexavir(r).

US5,055,296 describes the use of a mammalian liver extract for the
treatment of viral infections and chronic fatigue syndrome. The
extract is described to be thermostable, acetone-insoluble and soluble
in water. Chemical analysis of the liver extract revealed at least
five polypeptides of which one was found to have
bradykinin-potentiating activity. In US5,334,395, which relates to the
use of a mammalian liver extract in the treatment of Epstein Barr
Virus (EBV) infection, 9 peptides are identified in the Kutapressin
extract as having angiotensin converting enzyme inhibitory activity.
One of these peptides is demonstrated to be capable of inhibiting EBV
infection in vitro.

The inventors further discovered positive synergistic effects when the
MAF of the present invention was used in combination with Vitamin B12.
In particular embodiments, the present invention thus relates to the
therapeutic use of a MAF as described above used in combination with
Vitamin B12. In particular embodiments, the MAF of the present
invention and Vitamin B12 are administered to the patient separately.

The inventors also discovered positive synergistic effects when the
MAF of the present invention was used in combination with one or more
compounds selected from interferon alpha, interferon beta, interferon
gamma and/or interleukin-7. In particular embodiments, the present
invention relates to the therapeutic use of a MAF according to the
present invention used in combination with one or more compounds
selected from interferon alpha, interferon beta, interferon gamma
and/or interleukin-7. In particular embodiments, the MAF of the
present invention and interferon alpha, interferon beta, interferon
gamma and/or interleukin-7 are administered to the patient separately.

As described hereabove, in the treatment regimens envisaged in the
context of the present invention the MAF can be administered
separately or simultaneously with another compound of interest. When
administered simultaneously, the compounds may be comprised in the
same or separate formulations. However, in particular embodiments, the
invention provides compositions for combined treatment regimens. Thus,
in particular embodiments, the present invention relates to a MAF or
composition as described above, and/or to the use thereof,wherein said
MAF is provided in combination with at least one other
pharmacologically active compound, for example, a viral inhibitor, an
ecto-nox protein modifying agent, a mammalian liver extract, vitamin
B12, interferon alpha, interferon beta, interferon gamma and/or
interleukin-7.

In particular embodiments, at least one other pharmacologically active
compound is selected from a viral inhibitor, and an ecto-nox protein
modifying agent. In particular embodiments the composition comprises,
or further comprises a mammalian liver extract, preferably Nexavir(r).
In particular embodiments the composition comprises, or further
comprises Vitamin B12. In particular embodiments, the composition
comprises, or further comprises one or more compounds selected from
interferon alpha, interferon beta, interferon gamma and/or
interleukin-7.

The present invention provides a use of a MAF or composition as
described above in the manufacture of a medicament for the treatment
of CFS/ME and/or for inhibiting infection by XMRV in a mammal, wherein
said MAF is provided in a conjugate or in a pharmaceutical composition
further comprising a pharmaceutically acceptable excipient and a
therapeutically effective amount of said MAF.

The term 'therapeutically effective amount' as used herein means that
amount of MAF, compound, conjugate or pharmaceutical agent that
elicits the biological or medicinal response in a tissue, system,
animal or human that is being sought by a researcher, veterinarian,
medical doctor or other clinician, which includes alleviation of the
symptoms of the disease being treated.

The pharmaceutical composition can be prepared in a manner known per
se to one of skill in the art. For this purpose, at least one MAF, one
or more solid or liquid pharmaceutical excipients and, if desired, in
combination with at least one other pharmaceutical active compound,
are brought into a suitable administration form or dosage form which
can then be used as a pharmaceutical in human medicine or veterinary
medicine.

Particular forms of the pharmaceutical composition may be, for
example, solutions, suspensions, emulsions, creams, tablets, pills,
capsules, nasal sprays, liposomes or micro-reservoirs, especially
compositions in orally ingestible or sterile injectable form, for
example, as sterile injectable aqueous or oleaginous suspensions or
suppositories and sterile packaged powders (which are usually
reconstituted prior to use) for administration as a bolus and/or for
continuous administration. The solid carrier may comprise one or more
excipients, e.g. lactose, dextrose, sucrose, sorbitol, mannitol,
starches, gum acacia, calcium phosphate, alginates, tragacanth,
gelatin, calcium silicate, microcrystalline cellulose,
polyvinylpyrrolidone, polyethylene glycol, fillers, disintegrating
agents, binders, e.g. cellulose, carboxymethylcellulose or starch or
anti-stick agents, e.g. magnesium stearate, to prevent tablets from
adhering to tabletting equipment, (sterile) water, methylcellulose,
methyl- and propylhydroxybenzoates, talc, magnesium stearate, edible
oils, vegetable oils and mineral oils or suitable mixtures thereof.
Tablets, pills and boluses may be formed so as to disintegrate rapidly
or to provide slow release of the active ingredient. The formulations
can optionally contain other pharmaceutically active substances (which
may or may not lead to a synergistic effect with the MAF of the
invention) and other substances that are commonly used in
pharmaceutical formulations, such as lubricating agents, wetting
agents, emulsifying, and suspending agents, dispersing agents,
desintegrants, bulking agents, fillers, preserving agents, sweetening
agents, flavoring agents, flow regulators, release agents, etc. The
compositions may also be formulated so as to provide rapid, sustained,
or delayed release of the active compound(s) contained therein, for
example using liposomes or hydrophilic polymeric matrices based on
natural gels or synthetic polymers.

For the purposes of the present invention, the MAF or compounds or the
pharmaceutical composition of the present invention may be
administered by any of several routes including but not limited to
oral administration, buccal (e.g. sub-lingual) administration, topical
administration and parenterally injection, i.e. including intravenous,
intraperitoneal, intramuscular, intrasternal or subcutaneous
injection. The MAF or compound of the present invention will generally
be administered in an 'effective amount', by which is meant any amount
of a MAF that, upon suitable administration, is sufficient to achieve
the desired therapeutic or prophylactic effect in the individual to
which it is administered.

Usually, depending on the condition to be prevented or treated and the
route of administration, such an effective amount will usually be
between 0.01 to 100 ng, preferably between 0.1 and 50 ng and even more
preferably between 0.1 and 10 ng of the MAF per kilogram body weight,
which may be administered as a single daily dose, divided over one or
more daily doses, or essentially continuously, e.g. using a drip
infusion.

In particular embodiments, the effective amount is between 25 and 200
ng MAF/dose. In further particular embodiments, the effective amount
is between 25-500ng, more particularly about 10 ng MAF per week.

In particular embodiments, the MAF or pharmaceutical composition of
the present invention are administered once per week, during 2 to 50
weeks, preferably during 5 to 40 weeks, such as during 15 weeks.

The amount(s) to be administered, the route of administration and the
further treatment regimen may be determined by the treating clinician,
depending on factors such as the age, gender and general condition of
the patient and the nature and severity of the disease/symptoms to be
treated.

In accordance with the method of the present invention, said
pharmaceutical composition can be administered separately at different
times during the course of therapy or concurrently in divided or
single combination forms. The present invention is therefore to be
understood as embracing all such regimes of simultaneous or
alternating treatment and the term 'administering' is to be
interpreted accordingly.

For an oral administration form, the compositions of the present
invention can be mixed with suitable additives, such as excipients,
stabilizers or inert diluents, and brought by means of the customary
methods into the suitable administration forms, such as tablets,
coated tablets, hard capsules, aqueous, alcoholic, or oily solutions.
Examples of suitable inert carriers are gum arabic, magnesia,
magnesium carbonate, potassium phosphate, lactose, glucose or starch,
in particular, corn starch. In this case, the preparation can be
carried out both as dry and as moist granules. Suitable oily
excipients or solvents are vegetable or animal oils, such as sunflower
oil or cod liver oil. Suitable solvents for aqueous or alcolholic
solutions are water, glycerol and sugar solutions. Polyethylene
glycols and polypropylene glycols are also useful as further
auxiliaries for other administration forms. As immediate release
tablets, these compositions may contain microcrystalline cellulose,
dicalcium phosphate, starch, magnesium stearate and lactose and/or
other excipients, binders, extenders, disintegrants, diluents and
lubricants known in the art.

The oral administration of a pharmaceutical composition comprising the
MAF according to the invention, is suitably accomplished by uniformly
and intimately blending together a suitable amount of said MAF in the
form of a powder, optionally also including a finely divided solid
carrier, and encapsulating the blend in, for example, a hard gelatin
capsule. The solid carrier can include one or more substances, which
act as binders, lubricants, disintegrating agents, coloring agents,
and the like. Suitable solid carriers include, for example, calcium
phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch,
gelatin, cellulose, polyvinylpyrrolidine, low melting waxes and ion
exchange resins.

Oral administration of a pharmaceutical composition comprising the MAF
according to the present invention can also be accomplished by
preparing capsules or tablets containing the desired amount of said
MAF, optionally blended with a solid carrier as described above.
Compressed tablets containing the pharmaceutical composition of the
invention can be prepared by uniformly and intimately mixing the
active ingredient with a solid carrier such as described above to
provide a mixture having the necessary compression properties, and
then compressing the mixture in a suitable machine to the shape and
size desired. Molded tablets maybe made by molding in a suitable
machine, a mixture of powdered compound or MAF according to the
invention moistened with an inert liquid diluent.

For parenterally administration such as subcutaneous or intravenous
administration, the MAF or compounds of the present invention, if
desired with the substances customary therefore such as solubilizers,
emulsifiers or further auxiliaries, are brought into solution,
suspension, or emulsion. The MAF or compounds of the invention can
also be lyophilized and the lyophilizates obtained used, for example,
for the production of injection or infusion preparations. Suitable
solvents are, for example, water, physiological saline solution or
alcohols, e.g. glycerol, in addition also sugar solutions such as
glucose or mannitol solutions, or alternatively mixtures of the
various solvents mentioned. The injectable solutions or suspensions
may be formulated according to known art, using suitable nontoxic,
parenterally-acceptable diluents or solvents, such as mannitol,
1,3-butanediol, water, Ringer's solution or isotonic sodium chloride
solution, or suitable dispersing or wetting and suspending agents,
such as sterile, bland, fixed oils, including synthetic mono- or
diglycerides, and fatty acids, including oleic acid.

For topical administration, which includes the application of
medicinal substances to the skin or various body orifices, the MAF or
compounds of the present invention may be applied in a variety of
forms such as liquid, semisolid or solid. Some of the possible
components of topical, transdermal and transmucosal formulations and
delivery devices include solubilizing agents, suspending agents,
dispersing agents, preservatives, animal and vegetable fats, oils, or
waxes, emollients, stabilizing agents, thickening or gelling agents,
buffering agents, adhesive agents, adjuvants and additives,
emulsifiers and penetration enhancing agents as known to the person
skilled in the art.

For sub-lingual administration, the MAF or compounds of the present
invention may be formulated in admixture with with at least a proper
excipient typical of sublingual formulations. As non limiting example
for the artisan skilled in the art, said excipients are selected from
the group including: water-soluble inert excipients (such as, for
example, mannitol, sorbitol, lactose); water-insoluble excipients
which help the delivery of the active substance at the sublingual
mucosa level (such as, for example, microcrystalline cellulose);
sweeteners (such as, for example, aspartame, sodium saccharate);
flavourings (such as, for example, peach, apricot, banana, strawberry,
orange, mandarin flavours); lubricants (such as, for example,
magnesium stearate, PEG 6000); taste correctors (such as, for example,
sodium citrate); other excipients, additives, carriers commonly used
in the formulation pharmaceutical art.

In addition to administration with conventional carriers, the MAF or
compounds of the present invention may be administered by a variety of
specialized oligonucleotide or nucleic acid delivery techniques such
as by encapsulation in various encapsulating materials, such as in
unilamellar liposomes (Bayard et al., Eur. J. Biochem. 151: 319, 1985)
or by conjugation to carrier molecules such as poly(L-lysine).
Reconstituted Sendai virus envelopes have been successfully used to
deliver RNA and DNA to cells (Arad et al., Biochem. Biophys. Acta. 85:
88, 1986). Moreover, the virus envelope is not limited to Sendai
virus, but could include encapsulation in any retroviral amphotrophic
particle. These techniques may be utilized for introduction of the
present Macrophage Activation Factors into cells.

The pharmaceutical compositions of this invention can be administered
to humans in dosage ranges specific for each MAF or compound comprised
in said compositions. The MAF or compounds comprised in said
composition can be administered together or separately.

It will be understood, however, that specific dose level and frequency
of dosage for any particular patient may be varied and will depend
upon a variety of factors including the activity of the specific MAF
or compound of the invention employed, the metabolic stability and
length of action of that MAF or compound, the age, body weight,
general health, sex, diet, mode and time of administration, rate of
excretion, drug combination, the severity of the particular condition,
and the host undergoing therapy.


EXAMPLES

1. Effect of GcMAF on a patient suffering from CFS/ME.

The patient, a 29 year old female, has suffered from CFS/ME ever since
an acute Epstein-Barr virus infection at the age of 17. The patient
also tested positive for XMRV in three laboratories with different
diagnostic techniques. The patient was treated with intravenous
injections of GcMAF, once a week during 10 weeks. Upon each injection,
200 ng of GcMAF was administered to the patient.

The patient showed significant clinical improvement after the
treatment with GcMAF. In addition, indicators of NK cell activity and
inflammation were positively affected. The results are summarized in
Table 1.

The GcMAF treatment resulted in a decrease of Interleukin-8 (IL8)
concentration in the serum. IL8 is a protein produced by macrophages
and other cell types such as epithelial cells, and is associated with
inflammation. Thus, the decrease of IL8 concentration indicates
reduced inflammation.

Moreover, also a significant decrease of the patient's C4a serum level
was measured after treatment with GcMAF. The C4a measurement was
performed using a Becton Dickinson OptEIA human C4a elisa kit.

Additionally, an increase of intracellular perforin in PBMCs was
observed. The Perforin mRNA level was measured by real-time PCR, using
gene expression assay reagents from Applied Biosystems.

As a conclusion, the results in Table 1 show that the treatment with
GcMAF on a patient suffering from CFS/ME resulted in increased NK cell
activity and reduced inflammation. As reduced NK cell activity and
inflammation are associated with CFS/ME, this indicates that GcMAF
treatment is effective in the treatment of patients suffering from
CFS/ME.


Table 1: Laboratory results before and after GcMAF treatment.
-------------------------------------------------------------
Before After
-------------------------------------------------------------
IL8 serum (pg/mL) 29 12.6
Perforin PBMC 552 1184
C4a serum level (ng/mL) 4519 2292
-------------------------------------------------------------

2. Clinical study on 108 patients suffering from CFS/ME.

A clinical test fo the effects of GcMAF was run on 108 CFS/ME
patients, aged between 18 and 65. The patients tested positive on
Murine Leukemia Virus infection (34 patients), XMRVc (XMRV detection
in co-culture - 54 patients) or serology (20 patients). The patients
were treated with intravenous injections of GcMAF, once a week. Upon
each injection, 25-100 ng GcMAF in 1 mL physiological serum was
administered. 75% of the patients were treated with 100 ng GcMAF per
week.

Already after fifteen weeks, 68 of the 108 patients showed clear
improvements, as listed in Table 2. These results clearly show that
the administration of GcMAF results in clinical improvements of
patients suffering from CFS/ME.


Table 2: Clinical improvements in 68 after 15 weeks of GcMAF treatment.
-----------------------------------------------------------------------
Clinical improvement Fraction of patients
-----------------------------------------------------------------------
Fatigue reduced or disappeared 44/68
Improvement of sleep quality 39/68
Pain reduced or disappeared 35/68
Memory/concentration improved 27/68
Faster or normalized recovery 42/68
Orthostatism reduced or disappeared 22/68*
Digestive problems reduced or disappeared 22/68
-----------------------------------------------------------------------
* Not all patients suffered from orthostatism prior to GcMAF treatment.

--------
(c) 2013 EPO

No comments: