Note: Descriptions are shown in the official language in which they were submitted.
CA 02478137 2004-08-17
I
Metal Ion Chelate Complexes and U~~e Thereof
Technical Field
The present invention relates to processes for forming metal ion chelate
complexes,
formulations comprising one or more metal ion chelate complexes, methods of
s preparation, and use thereof. More particularly, the preseztt invention
relates to
formulations comprising copper complexes, in parCicular, formulations
compzising
copper-IrDTA.
Background of the tnvention
Minerals such as metal ions play an important physiological role in animals,
io humans and plants. For example, minerals such as zinc, iron, calcium,
copper and the
like, are important for bone development, energy production and fatty acid
oxidation. Tn
addition, metal ions may be incorporated into enzymes and proteins, and may
also
function as co-factors (eg, copper is a co-factor associated with the enzyme
superoxide
dismutase).
~s Administration of metal ions may be therapeutically useful. Metal ions,
such as
c-opper ions, have been adnninistered in the form of mineral salts, for
example, as metal
sulfides, chlorides and nitrates. However, a major limitation of this form of
administration
is toxic side effects or tissue irritation easing from the high concentration
of metal ion
released into the system. '
:.n The present invention provides processes for producing metal ioo, chelate
complexes and methods of using such complexes.
Summary of the Invention
According to a first aspect of the invention there is provided a process for
preparing
one or more metal ion chelate eomplex(es), comprising
zs mixing a chelating agent with an oxidising agent in solution, to obtain a
reaction
mixture;
adding at Ieast one metal capable of foaming a metal ion ehelate complex to
said
reaction mixture so as to form said one or more metal ion chelate complex(es),
or a
mixture thereof.
~o According to a second aspect of the invention there is provided a process
for
preparing at least one metal ion chelate complex(es), comprising:
dissolving a chelating agent in water to obtain a solution;
adding at least ot~e metal capable of forming a metal ion chelate complex with
said
chelating agent to said solution, and
CA 02478137 2004-08-17
2
adding an oxidizing agent to said solution, so as to form at least one metal
ion
chelate complex, or a mixture thereof.
In one embodiment the chelating agent is dissolved in a suitable solvent, such
as
water, prior to addition of the oxidising agent.
s The process according to the invention may further include a separation
step, e.g., a
filtration step, to remove. residual or unreacted metal.
The process according to the invention may further include all additional step
of
isolating said metal ion chelate complex from solution. Suitable methods of
isolation
include solvent evaporation, recrystallisation, solvent extraction, and other
general
ro methods known to those sk-filled in the art.
In one embodiment, the resultant mixture may comprise at Ioast one metal ion
chelate complex, unreacted oxidising agent, metal or chelating agent. In one
embodiment,
the process is continued until at least one of said oxidising agent, said
ehelating agent, and
said metal is consumed. In one embodiment all of the oxidising agent is
consumed.
~s In one embodiment the chelating agent is multidentate. In one embodiment
the
chelating agent comprises N andlor O donor atoms. In one embodiment the
chelating
agent comprises at least one carboxylic acid (or carboxylate) group. In one
embodiment
the chelating agent comprises at Least one amino group, wherein the amino
group may be
a primary, secondary or tertiary amino group, In one embodiment the chelaring
agent may
zo comprise at least one., e.g., 1, 2, 3, 4, 5 or G, imidodiacetic acrd [-
N((:H.~COZH)i] groups,
wherein one or both of the methylene (-CHZ) hydrogen atoms may be replaced
with
another substituent, such as a Ci-Ca alkyl gxoup. In one embodiment the
chelating agent is
selected from EDTA, DTPA, HEDTA, ethylenediaminedisuccinic acid (EDDS),
salicylic
acid, acetyl salicylic. acid, amino acids such as glycine, histidine, Lysine,
arginine,
as cysteine, methionine, and peptides comprising those amines, and salts
thereof. The
peptide comprise two, three, four or five amino acids. The respective amino
acids may be
the same or different. In one embodiment, the peptide is a dipeptide or a
tripeptide. In
one embodiment the. chelating agent is EDTA or a salt thereof. In one
embodiment, the
chelating agent is EDTA disodium salt. In another embodiment the chelating
agent is
~u DTPA or a salt thereof, e.g., a sodium salt thereof.
In alternate embodiments the oxidising agent may be selected from hydrogen
peroxide, ozone, and aqueous solutions, including water and aqueous salt
solutions, acidic
solutions. Other oxidising agents are well known to those skilled in the art
and include,
for example, permmganate salts, and dichromate salts. In one embodiment the
oxidising
agent is an aqueous solution of hydrogen peroxide. For example, the oxidising
agent may
CA 02478137 2004-08-17
3
be a 35wC%, 30wt%, 25wt%, 20wt%, l5wt%, l0wt%, Swt%, 3wt%, lwt% or 0.5 wt%
aqueous solution of hydrogen peroxide.
In one embodiment, the metal is capable of being oxidisc;d to a metal ion
capable of
forming a metal ion chelate complex with a chelating agent. In one embodiment,
the
s metal is a transition metal. In one embodiment the metal is selected from
Mn, Fe, Co; Ni,
Cu, Zn, Cr, Al, Cd, Ag and Se. In one embodiment, the metal is elemental
metal. In one
embodiment, the metal is a solid. For example, the metal may be in the form of
wire,
granules, powder, a sold bar having any shape, or any other suitable solid
form. The
metal may be added to the reaction mixture comprising the c.helating agent
andlor
~o oxidising agent all at once. Alternatively, the metal may be added to the
reaction mixture
comprising the chelating agent and/or the oxidising agent in a portionwise
fashion over a
period of time. For example, the metal may be added portionwise over about 5
seconds to
about 1 hour. Those skilled in the art would be able to ga~xge an appropriate
rate of
addition of the metal taking into consideration factors such as
<;oncentration, temperature,
is reagents, etc.
In one embodiment the metal is copper which is oxidised to Cuz+ or Cu+ ions,
or a
mixture thereof. In another embodiment the metal is iron which is oxidised to
Fe2+ or
Fe3+ ions, or a mixture thereof. In a further embodiment the metal is zinc
which is
oxidised to Zn~+ ions. In another embodiment the metal is Ni which is oxidised
to Niz+
ao ions, Tn a further embodiment the metal is cobalt which is oxidised to Go2+
or Co3+ ions,
or a mixture thereof. In another embodiment the metal is silver which is
oxidised to Ag
a ons.
Mixtures of different metals may be added to the oxidising solution to produce
mixtures of metal ion complexes or salts thereof.
is The metal ion complex or salt thereof may be water soluble.
The amount of metal added may be sufficient to consume substantially all of
the
oxidising agent. The amount of chelating agent added may be sufficient to
complex
substantially all of the metal ions.
tn various embodiments, the stoichiometric ratio of the metal to the chelating
agent
3o is selected from about 4:1, about 3:1, about 2.8:1, about 2.6:1, about
2.5:1, about 2.4:1,
about 2.2:1, about 2:1, about 1.9:1, about 1.8:1, about 1.7:1, about l.b:l,
about 1.5:1,
about 1.4:1, about 1.3:1, about 1.2:1, about l.I:l, about 1:1, about 1:1.1,
about 1:I.2,
about 1:I.3, about 1:1.4, about I:1.5, about 1:1.6, about 1:1.7,, about 1:1.8,
about 1:1.9,
about 1:2, about I:2,1, about 1:2.2, about 1:2.4, about 1:2.5, about 1;2.6,
about 1.:2.8,
3s about I:3, and about 1:4.
CA 02478137 2004-08-17
4
In one embodiment the metal ion forms a 1:1 metal ion chelate complex with the
chelating agent. In another embodiment the metal ion forms a 2:1 metal ion
chelate
complex with the chelating agent.
In one embodiment the solution is an aqueous solution. 'The pH of the solution
may
s be between about 2 and about 14. In another embodiment the solution is a
basic aqueous
solution wherein the pH of the solution is between about 7.5 .and about I4,
for example,
pH about 8, 9, 10, 11, I2, or 13. In one embodiment, the pH o:f the solution
at completion
of the reaction is about 6 to about 8, e.g, about 7.
Modification of the temperature may be used to control the rate of reaction.
ao Accordingly, the process may optionally include one or more cooling steps.
For example,
the solution may optionally be cooled prior to addition of the metal, after
addition of the
metal, during the oxidation step, ox at completion of the oxidation reaction.
The
temperature may be maintained less than about 100°C, less than about
80°C, 60°C, for
example, between about 0°C and about 50°C, ox between about
10°C and about 40°C. In
a one embodiment, the mixture may be cooled to a temperature selected from
about 40°C,
about 35°C, about 30°C, about 25°C, about 20°C,
about 15°C, .about 10°C, and about S°C.
Alternatively, the process may include one or more heating steps.
According to a third aspect of the invention theze is provided a process for
pzeparing copper-EDTA, comprising
~a mixing EDTA or a salt thereof with an aqueous solution of hydrogen peroxide
to
obtain a mixture; and
adding copper to said mixture so as to form copper-EDTA or a salt thereof.
Tn one embodiment the process further comprises a separation step, e.g., a
filtration
step, for removing residual or unreacte.d copper solid(s).
zs rn one embodiment the process further compzises cooling the mixture of
hydrogen
peroxide and E.DTA or a salt thezeof.
Tn another embodiment, the process may further comprise cooling the mixture
after
copper has been added. Tn one embodiment the temperature is maintained at a
tennperature below about 40°C, eg, between about 5°C and about
40°C.
3a The salt of EDTA may Comprise one or more of sodium ions, potassium ions,
lithium ions, calcium ions, magnesium ions, or mixtures thereof In one
embodiment the
chelating agent is EDTA disodium salt.
In one erzzbodiment the copper is oxidised to Cu2+.
CA 02478137 2004-08-17
In one embodiment the resultant metal chelate complex is a salt, such as an
alkali
earth or alkali metal salt. For example, the metal ion chelat.e complex may be
Na+, K~,
Li+, IvZg2+, or Ca2+ salt. In another embodiment the metal chelate complex is
Cu[EDTA]Na2. In one embodiment the metal chelate complex is Cu2[EDTA]. In a
s further embodiment the product is a mixture of Cu2[EDTA] and Cu[EDTA]Na2.
In a further embodiment the metal chelate complex or salt thereof, may be a
hydrate.
With reference to the first or second aspect of the invent:ian, in one
embodiment the
resultant mixture may be evaporated or lyophilized to obtain a solid
comprising the metal
~o ion cl~elate complex or salt thereof. The metal ion chelate complex may be
purired e.g,
by recrystallization from a suitable solvent.
According to a fourth aspect of the invention there is provided a metal ion
complex
or salt thereof prepared according to the. process of the first, second or
third aspect of the
invention.
~s According to a fifth aspect of the invention there is provided an aqueous
solution
comprising at least one metal ion complex or salt thereof prepared according
to the
process of the first, second or third aspect of the invention. In one
embodiment the
aqueous solution may comprise uncomplexed chelating agent, or residual
oxidizing agent.
According to a sixth aspect of the invention there is provided a
pharmaceutical
Zo formulation comprising a metal ion complex or salt thereof according to the
fourth aspect
of the invention, or a solution according to the fifth aspect of the
invention, together with
a suitable adjuvant, diluent or carrier.
According to a seventh aspect of the invention there is provided a process for
preparing a pharmaceutical formulation e.omprising mixing at least one metal
ion complex
Zs or salt thereof according to the fourth aspect of the invention with a
suitable carrier,
diluent or adjuvant.
According to an eighth aspect of the invention there is provided a process for
preparing a pharmaceutical formulation comprising mixing an ;aqueous solution
of at least
one metal ion comple;~ or salt thereof according to the fifth aspect of the
invention with a
:o suitable acceptable carrier, diluent or adjuvant.
With reference to any one of the sixth, seventh or eighth aspects of the
invention,
the carrier, diluent or adjuvant may be pharmaceutically aca~table andlor
veterinarily
acceptable.
According to a ninth aspect of the invention there is pro ided a method of
treafi~nenc
3s of a disease or condition in a mammal, comprising administezing to said
mammal an
CA 02478137 2004-08-17
6
effective amount of at least one metal ion cheIate complex or a salt thereof,
or a
pharmaceutical composition thexeof.
According to a tenth aspect of the invention there is provided at least one
metal ion
chelate complex or salt thereof, or a pharmaceutical composition thereof, when
used for
s treating a disease or condition in a mammal.
According to an eleventh aspect of. the invention there is provided the use of
a metal
ion chelate complex or salt thereof for the manufacture of a medicament for
treating a
disease or condition.
With reference to any one of the ninth to eleventh aspects of the invention,
in one
to embodiment the metal ion chelate complex may be a metal ion chelate
connplex according
to the fourth aspect of the invenkion. In another embodirnent the metal ion
cixelate
complex may be in the form of an aqueous solution, for example, an aqueous
solution
according to the fifth aspect of the invention. In one embodiment the metal
ion chelate
complex is Cuz[EDTA], Cu[EDTA]Na2, Cu[EDTA-HZ], or a m~i~cture thereof.
a s In one embodiment, the pharmaceutical corraposition is a topical
composition
according to the sixth aspect of the invention. The composition may be in the
form of a
cream, lotion, spray or gel.
The disease or condition to be treated may be selected from reducing
inflammation,
such as inflammation associated with arthritis, including osteoarthritis and
rheumatoid
'o arthritis; promoting growth or regrowth of hair; heating psoriasis;
increasing blood flow;
treating chill blains; treating varicose veins; promoting wound healing;
promoting healing
of scar tissue, including scar tissue resulting from bums; alleviating or
reducing joint or
muscular pain; tzeating sinus inflammation andlor sinus pair; treating
bacterial andJor
fungal infection.
zs Other conditions that may be treated in accordance with the present
invention
include eczema, wrinkles, bruises, joint degeneration, including cartilage
degeneration,
acne, burns, and onychoschizia tsplit nails).
In alternative embodiments of the invention, th.e metal ion complexes) or
salts)
thereof, or compositions comprising said metal ion complexes) or salts)
thereof may be
3o used to as a means of prow ding mineral supplements, e.g, copper, cobalt,
zinc, selenium,
etc, to animals, e.g., humans, sheep, cattle, goats, horses, pigs, etc. In
various
embodiments, compositions may comprise mixtures of different metal ion chelate
complexes, or salts thereof.
In further embodiments, the metal ion chelate complex according to the
invention,
~s or pharmaceutical compositions comprising the complex, may be used as an
alternative to
CA 02478137 2004-08-17
7
conventional sun screen, eg, for humans or animals suffering from conditions
such as
vitiligo (lack of pigment in the skin).
In one embodiment the present invention may enable a non-toxic amount of a
metal
ion to be administered.
s With reference to the eighth or ninth aspects of the invention, the mammal
may be.
human, non-human primate, murine, bovine, ovine, equine, caprine, leporine,
avian,
feline, porcine, or canine. In one embodiment the mammal is human.
The present invention provides a convenient process for prepaxing metal
chelate
complexes and salts thereof. Metal ion chelate complexes in accordance with
the present
io invention, including copper complexes such as copper-BDTA complexes or
salts thereof,
may have a range of beneficial therapeutic properties.
Brief ~escription of the Figures
Figure 1 is a Negative ion MALDI mass spectrum of a. sample solution prepared
according to the process of the invention shoeing major copper containing ions
consistent
is with the species Cu[EDTA~~' or Cu[EDTA]~ (m.w 351 Da).
Figure 2 is an e.lectrospray mass spectrum of a sample solution prepared
according
to the process of the present invention showing a major copper containing ion
consistent
with a sodium ion of Cu[EDTA]Na2.
Figure 3 is a comparison photograph of a male's hands after treatment of one
hand
Zo with a cream according to the invention.
p'iguxe 4 is a comparison photograph of a male's hands after treatment of the
left
hand with a cream according to the. invention.
Figuxe S is a photograph of the hands of an 84 year old woman suffering
arthritis
before treatment.
is Figure 6 is a photograph of the hands of an 84 year old woman suffering
arthritis
after treatment with a cream according to the invention for one week.
Definitions
The following ate some definitions that may be. helpful in understanding the
description of the present invention. These are intended as general
definitions and should
:o in no way limit the scope of the present invention to those terms alone,
but are put forth
for a better understanding of the following description.
Unless the context requires otherwise or specifically stated to the contrary,
integers,
steps, or elements of the invention recited herein as singular integers, steps
or elements
CA 02478137 2004-08-17
i
clearly encompass both singular and plural forms of the recited integers,
steps or
elements.
Throughout this specification, unless the context requires otherwise, the word
"comprise", or variations such as "comprises" or "comprising", will be
understood to
s imply the inclusion of a stated step or element or integer or group of steps
or elements ox
integers, but not the exclusion of any other step or element or integer or
group of elements
or integers. Thus, in the context of this specification., the term
"comprising" meats
"including principally, but not necessarily solely".
In the context of the present specification, the. term "metal ion complex"
should be
m understood to include metal ion complexes, as well as salts, protonated
forms, and
hydrates thereof.
In the context of this specification, the term "copper-EDTA" should be
understood
to include within its meaning Cu[EATA]2- andlor salts thereof, e.g,
Cu[EDTA]Na2,
Cu[EDTA-Hi2]and Cuz[EDTA].
~s Tn the contexi of this specification "Ci-Ca allcyl" refers to straight or
branched
saturated alkanes having one to four carbon atoms, for example, methyl, ethyl,
propyl,
isopropyl, butyl, tent-butyl,
Those skilled in the art will appreciate that the invention described herein
is
susceptible to variations and modifications other than those specifically
described. It is to
~o be understood that the. invention includes all such variations and
modifications. The
invention also includes all of the steps, features, compositions and compounds
referred to
or indicated in this specit5cati.on, individually or collectively, and any and
all
combinations or any two or more of said steps or features.
AlI the references cited in this application are specifically incorporated by
reference
z: and are incorporated herein in their entirety.
The. language "therapeutically effective amount" is intended to include within
its
meaning a non-toxic but sufficient amount of a. compound or composition of the
invention
to provide the desired therapeutic effect. The exact therapeutically effective
amount of
the compound or composition will vary according to factors such as the type of
disease or
3o condition to be treated, the age, sex, and weight of the animal, mode of
administration,
and the ability of the compound or composition to permeate cell membranes.
Dosage
regima may be adjusted to provide the optimum therapeutic; response. For
examples
several divided doses may be administered one or several times daily or the
dose can be
proportionally reduced or increased as indicated by the exigencies of the.
therapeutic
~s situation. '
CA 02478137 2004-08-17
9
As used herein the term "treatment", refers to any and all uses which remedy a
disorder or disease state or symptoms, prevent the establishment of a disorder
or disease,
or otherwise prevent, hinder, retard, or reverse the progression of a.
disorder or disease or
other undesirable symptoms in any way whatsoever.
s Abbreviations
EDTA - ethylenediaminetetraacetic acid
DTPA - diethylenetriaminepentaacetic acid
HEDTA- hydroxyethylethylenediaminetria.cetac acid
NSAI:D - non-steroidal anti-inflammatory drug
io GOX-2 - cyclooxygenase-2
Detailed pescription ~f Preferred Embodiment: of the Invention
The present invention relates to processes for fom~ing metal ion chelate
complexes
and salts and protonated forms thereof, compositions comprising one or more
metal ion
chelate complexes, methods of preparation, and use thereof. More particularly,
the
is present invention relates to compositions comprising copper complexes,
including
compositions comprising copper-EDTA, copper-DTPA and eopper-HEDTA, and salts
and protonated forms thereof.
The process for preparing metal ion chelate complexes according to the
invention,
comprises oxidising a metal which is in its elemental state to a
correspoaiding cation in
'0 S~h~ ncino~ an ntiriising agent. In one embodiment, the process of the
invention may be
carried out as a "one pot" process. Alternatively, the reaction can be carried
out i.n a
stepwise fashion, including stepwise addition of reagents.
Whilst not seeking to be limited to any one proposed mechanism of action, the
process may involve an i~a sits redox reaction between the: elemental metal
and the
zs oxidising agent, whereby the. metal is oxidised to one or more
corresponding cations. The
metal cations thus produced may then chelate to the chelating agent to form a
metal ion
chelate complex, or metal complex anion. Suitable combinations of oxidising
agent and
metal can be selected based on the reduction potential of the respective
reagents.
Reduction potentials would be known to those skilled in the art and are
reported, fox
?o example, irt the CRC Hahdboo& of Chemistry and Physics, Weast, R., Ed. SS'h
Edition,
1974-1975, the entire contents of which are incorporated herein by cross-
reference.
By way of illustration, the following reaction ntay be envisaged, where the
oxidising agent is hydrogen peroxide and M is a suitable metal:
CA 02478137 2004-08-17
M~S~ + HZO~ + 2H* M2* + 2H20
When M is copper, the. follor ing redox reaction may be envisaged:
Cu(s) + Hx02 + 2H+ Cu2' * 2H20
s The resultant MZ+ ions, eg, Cu~* ions, may then form a~ metal i.on complex
with a
chelating agent, such as EDTA, DTPA, HEDTA, or salt or protonated form
thereof.
One or more steps of the process of the present invention may be exothermic
arid '
the reaction vessel may optionally be cooled. variation of temperature,
reagent
concentration, rate of addition of reagents, surface area of the metal (e.g,
larger surface
~o areas may lead to an increased reaction rate), and pH may be used as a
means of
controlling the rate of the eomplexation reaction.
Suitable cheiating agents include multidenta.te ligands capable of forming
stable '
metal ion complexes. Examples of suitable chelating agents include
ethylenediaminetetra,acetic (EDTA), diethylenetriaminepentaacetic acid (DPTAj
and
is (HEDTA) hydrvxyethylethyienediaminetriacetic acid, the structures of which
are shown
below:
Ho 0 0 off o off
HO~O 0 OH HOCH
~N ~ 2
~N~~N~ \N N
0~ ~ O O ''0 O
~O'H OH H HO 'O~H H ON
EDTA DTPA HEDTA
Multidentate ligands such as EDTA, DTPA, HEDTA may be zwitterionic.
ao EDTA is a hexadentate chelating ligand with 6 possible coordination sites.
The
pKa of the carboxylic acid residues of EDTA are 1.70, 2.60, 6.30 and 10.60,
respectively.
Neutral EDTA can exist as a zwitterion with t'vo of the acid protons being
located on the
nitrogen atoms.
EDTA, DTPA, and HEDTA have a strong affinity for a wide range of metal ions,
zs and, for example, may form complexes with metal ions including Ag+, Caz+,
Coat, Cu2+,
Fe3~, Fe2+, Mg~'~, Ni2*, and Zn~'-. Metal ion complexes prepared according to
the present
invention may be stable at low, neutral aridlor high pH. For example, metal
ion chelate
complexes according to the present invention may be stable at a phl in the
range from
about 4 to about 12. The metal ion complex may comprise the chelate and metal
ion in. a
3e 1:1 stoichiometric ratio or 1:2 stoichiometric ratio. For example, Cu2+ may
chelate to
EDTA in a I:1 ratio to form the anionic species Cu[EDTAJZ', or a salt or
protonated form
CA 02478137 2004-08-17
11
thereof. Suitable counterions may include sodium ions, pots~ssium ions,
calcium ions,
magnesium ions, etc.
In accordance with the process of the present invention, appropriate molar
ratios of
reagents can be determined by those skilled in the art. Thus, it is possible
to manipulate
molar ratios to ensure any one or more reagents are consumed in the course of
the
reaction. For example, 1 mole of copper may be oxidised by one mole of
hydrogen
peroxide to produce one mole of Cu2y ions rfz siW which may then be chelated
by one
mole of EDTA. Accordingly, to ensure all of the hydrogen peroxide is consumed,
an
excess of copper can be used. Alternatively, a molar excess of any oa~e or
more reagents
~o may be used whereby residual excess reagent Will remain at completion of
the reaction
process. In one embodiment, excess chelating agent, e.g., EDT A or a salt
thereof, may be
used. In one embodiment, excess metal, e.g., copper, may be used. Residual
reagents
may be removed upon completion of the reaction.
One advantage of the present invention is that metal ions, such as iron, zinc,
nickel,
2 s cobalt, magnesium, and copper ions (or mixtures thereof), may be
administered to a
subject in the form of a metal ion complex or salt or hydrate thereof, or a
pharmaceutical
composition Thereof.
Techniques known to those skilled in the art, such as ion exchange
chromatogaphy
and reerystallization, may be used to exchange counterions, thus, for examgle,
a salt or
Zo protonated form of a metal ion ch.elate complex (or mixtures thereof) rt2ay
be converted
into another salt of choice.
Aqueous solutions of metal ion chelate complexes produced according to the
process of the present invention may have a relatively stable shelf life, for
example, up to
years, 4 years, 3 years, 2 years, 12 months, 6 months, or 3 months. Buffer
solutions,
zs e.g., ammoniaJammonium chloride, znay be used to stabilise solutions of
metal ion chelate
complexes.
In accordance with the present invention, metal ion complexes or salts oz
hydrates
thereof (e.g., Copper-fiDTA and salts thereof), as well of pharmaceutical
formulations of
metal ion complexes or salts thereof, may be useful in treating a range of
physiological
yo conditions or disorders in humans and animals. Examples of therapeutic uses
include
mineral supplementation, treatment of inflammation, treatment of arthritis,
inducing hair
growth or regrowth, promoting wool growth in sheep azrd goats, treating
psoriasis,
alleviation of pain including sinus pain, joint pain, and muscular pain,
promoting wound
healing including healing of scar tissue, increasing blood flow, treating
chill Mains,
;s treatment of varicose veins, and treatment of microbial infection including
bacterial,
CA 02478137 2004-08-17
12
fungal infection, treating eczema, reducing wrinkles, bruises, preventing or
alleviating
joint degeneration and pain associated therewith, treatizt.g acne, promoting
healing of
burns, and treating onychoschizia (split nails).
For example, for treatment or alleviation of inflammation., a topical
formulation (eg,
a cream, lotion, spray or gel) comprising a metal ion complex according to the
invention
may be rubbed into an inflamed area, e.g, one to three times per day, until
inflammation is
reduced. Treatment may continue for as long as necessary to reduce or
continually
alleviate inflammation.
For promoting hair regrowth, a topical formulation (eg, a cream, lotion, gel,
spray
~o or shampoo) comprising a metal ion eorrxplex according to the invention may
be applied
to the scalp, with rubbing or massaging, e.g., once or twice per day for a
period of time
which is at least sufficient to stimulate or promote hair grov~~th o~r
regrowth.
For use as a sunscreen, a topical formulation (eg, a cream, lotion, gel or
spray)
comprising a metal ion complex according to the invention rtia.y be applied to
an area of
is skin prior to exposure to the sun or ~V. Additional applications may be
made as required.
For relief of sinus pain, a topical farmulation (eg, a cream, lotion, gel or
spray)
comprising a metal ion complex according to the invention ma.y be applied to
an area of
skin around the nose andlor under the eyes or across the forehead when pain
starts,
resulting in pain relief
zo To reduce scarring, a topical formulation (eg, a cream, lotion, gel or
spray)
comprising a metal ion complex according to the. invention may be applied,
generally one
to three times per day, e.g., twice a day, across the scar tissue with aubbing
to work it into
the scar tissue and surrounding skin.
For healing of bruising a topical formulation (eg, a cream, lotion, gel or
spray)
zs comprising a metal ion complex according to the invention may be. applied
to and around
the bruised area once a day until bruising disappears.
For treatment of onychosc.hizia (split nails) a suitable formulation (eg, a
liquid,
cream, tincture, or lotion) comprising a metal ion complex according to the
invention may
be applied once a day until the desired nail quality is attained.
3u For treatment of acne and other skin conditions such as psoriasis, a
suitable
formulation (eg, a liquid, cream, gel, spray, tincture, or lotion) comprising
a metal ion
complex according to the invention may be applied to the appropriate area, one
to three
times per day, e.g, twice per day, in a manner similar to conventional
therapies.
CA 02478137 2004-08-17
13
Pharmaceutical anc)<lor Therapeutic Formulations
In accordance with the present invention, metal ion chelate complexes, such as
copper-EDTA or salts thereof, and pharmaceutical formulations thereof, may be
administered alone or may be used in combination with other known treatments
or
s therapeutic agents, including for example, anti-inflammatory agents (eg,
NSATDs,
cartisones, etc), analgesics and pain killers (eg, NSAIDs, CO~i-2 inhibitors),
antibacterial
and antifungal agents, hair growth stimulants (eg, radical scavengers,
antiandxogens),
sunscreens, and antibacterial and antifungal treatments known ao those skilled
in the art,
including, for example, cephalosporins, penicillins, macrolides, miconazole,
etc.
n Pharmaceutical formulations may optionally include a buffer.
Metal ion chelate complexes or salts or hydrates thereof, or fozmulations
comprising metal ion chelate complexes in accordance with the present
invention may
also he used with conventional wound dressings, e.g., as a means of promoting
wound
healing, zeducing inflammation, promoting healing of scar tissue, preventing
or treating
is microbial infection, etc, Metal ion ehelate complexes according to the
present invention,
or formulations thereof, may be applied to one or snore surfaces of a wound
dressing. For
example, the metal ion chelate complex or salt thereof may be. applied in the
form of a
liquid, cream, spray, or lotion, etc. Alternatively, the wound dressing may be
impregnated with the metal ion complex or formulation thereof according to the
~o invention. In some embodiments, the means of applying the metal ion complex
or salt
thereof to the wound dressing may include using a carrier medium which is
capable of
evaporating after application, e.g, an alcohol. Examples of vvound dressings
include
bandages, swabs, adhesive dressings e.g., BandAid~ adhesive strips, skin
replacement
dressings, burn dressings, blister dressings, and the like.
is In accordance with the present invention, metal ion chelate complexes may
be
incorporated into pharmaceutical formulations in the form of their
pharmaceutically
acceptable salts. By pharmaceutically acceptable salt it is meant those salts
which, within
the scope of sound medical judgement, are suitable for use in contact with the
tissues of
humans and lower animals without undue toxicity, irritation, allergic response
and the
30 like, and are commensurate with a reasonable benefitlrisk ratio.
.Pharmaceutically
acceptable salts are well known in the. azt and include alkali and all:.ali
earth salts.
In the context of the present invention, the language "pharmaceutically
acceptable
carrier" is intended to ine.lude solvents, dispersion media, coatings, anti-
bacterial and anti-
fungal agents, isotonic and absorption delaying agents, and the like. The use
of such
3s media and agents for pharmaceutically active substances is well known in
the art. Except
CA 02478137 2004-08-17
14
insofar as any conventional media or agent is incompatible with the metal ion
chelate
complex, use thereof in the therapeutic compositions and methods of treatment
is
contemplated. Supplementary active compounds may also be incozporated into the
compositions according to the present invention. Dosage unit fozm as used
herein refers
s to physically discrete units suited as unitary dosages for the individual to
be treated; each
unit containing a predetermined quantity of metal ion complex. is calculated
to produce
the desired therapeutic effect in association with the required pharmaceutical
carriez. The
specification for dosage unit foams of the invention rnay be dependent on (a)
the unique
characteristics of the metal ion chelate complex and the particular
therapeutic effect to be
m achieved, and (b) the limitations inherent in the art. The principal metal
ion chelate
complex is compounded for convenient and effective administration in effective
amounts
with a suitable pharmaceutically acceptable carrier in an acceptable dosage
unit. In the
case of compositions containing supplementary active ingredients, the dosages
may be
determined by reference to the usual dose and manner of administration of the
said
~ s ingredi ents.
Convenient modes of administration include topical transdermal application.
For
example, formulations may corztprise a cream, lotion, a liquid, a. spray, a
paste, or a gel.
Dispersions of metal ion chelate complexes may also be prepared in glycerol,
liquid
polyethylene glycols, and mixtures thereof and in oils. Under ordinary
conditions of
;.o storage and use, pharmaceutical preparations may contain a preservative to
prevent the
growth of microorganisms.
Other modes of administration may include injection (subcutaneous,
intravenous,
etc.), oral administration, aid rectal administration. Depending on the route
of
administration, the formulation andlor metal ion chelate complex may be coated
with a
zs material to protect the compound from the action of enzymes, acids and
other natural
conditions which may inactivate the therapeutic activity of the compound. The.
fozznulation andlor metal ion chelate complex may also be administered
parenterally or
intraperiloneally.
Pharmaceutical compositions suitable for injection include sterile aqueous
solutions
so (where water soluble) or dispersions and sterile powders for the
extemporaneous
preparation of sterile injectable solutions or dispersions. Ideally, the
composition is stable
under the conditions of manufacture and storage and ma.y include a
preservative to
stabilise the composition against the contaminating action of microorganisms
such as
bacteria and fungi.
CA 02478137 2004-08-17
In one embodiment of the invention, the formulation and/or metal ion chelate
complex according to the invention may be administered orally, for example,
with an.
inert diluent or an assimilabIe edible carrier, The formulation and/or metal
ion chelate
complex and other ingredients can also be enclosed in a hard or soft shell
gelatin capsule,
s compressed into tablets, or incorporated directly into an in.dividual's
diet. For oral
therapeutic administration, the formulation andlor metal ion chelate complex
can be
incorporated with excipients and used in the form of ingestible tablets,
buccal tablets,
troches, capsules, elixirs, suspensions, syrups, wafers, and the like.
Suitably, such
compositions and preparations may contain at least 1°,% by weight of
active compound.
a The percentage of the active compound in pharmaceutical compositions and
preparations
can, of course, be varied and, for example, may convenienthr range from about
2% to
about 90%, about 5°~° to about 80°f°, about 10% to
about 75%, about 15°l° to about 65%;
about 20~/o to about 60%, about 25% to about 50%, about 30'% to about
4S°~o, or about
35°,% to about 45%, of the weight of the dosage unit. The amount of
active compound in
~s therapeutically useful compositions is such that a suitable dosage will be
obtained.
Single or multiple administrations of a pharmaceutical composition in
accordance
with the present invention may be carried out. One skilled in the art would be
able, by
routine experimentation, to determine effective, non-toxic dosage levels of
the metal ion
complexes andlor compositions thereof according to the invention and an
administration
zo pattern which would be suitable for treating the disorders or diseases to
which the
compounds and compositions are applicable.
Further, it will be apparent to one of ordinary skill in the art that the
optimal course
of treatment, such as the number of applications or administrations per day
for a defined
number of days, can be ascertained using conventional course of treatment
determination
?= tests.
By way of illustration, a topical formulation in the form of a cream, lotion,
spray or
gel for treating or reducing inflammation may be applied locally and rubbed
into the skin
at the site of inflammation. This process could be repeated as necessary over
a suitable
tin3e period, eg, once, twice, three or four times per day for 1 day, 2-3
days, 4-7 days, 7-
30 14 days, up to 3 weeks, up to d. weeks, ete, until the inflammation is
suitably reduced.
Application rnay be ongoing or may be discontinued for z period of time and
then
recommenced.
Generally, an effective dosage per 24 hours may be in the range of about
0.0001 mg
to about 1000 mg per kg body weight; suitably, about 0.001 rrng to about 750
mg per kg
~s body weight; about 0.01 mg to about 500 mg per kg body weight; about 0.1 mg
to about
CA 02478137 2004-08-17
16
500 mg per kg body weight; about 0.1 mg to about 250 mg per kg body weight; or
about
1.0 mg to about 250 mg per kg body weight. Still suitably, an effective dosage
per ~
hours may be in the range of about 1.0 mg to about 200 mg per kg body weight;
about 1.0
mg to about 100 mg per kg body weight; about 1.0 rng to about 50 mg per kg
body
s weight; about 1.0 mg to about 25 mg per kg body weight; about 5.0 mg to
about 50 mg
per kg body weight; about 5.0 mg to about 20 mg per kg body weight; or about
5.0 mg to
about 15 mg per kg body weight.
It will be appreciated by persons skilled in the art that numerous variations
and/or
modifications may be made to the invention as shown in 'the specific
embodiments
m without departing from the spirit or scope of the invention as broadly
described. The
present embodiments are, therefore, to be considered in all respects as
illustrative and not
restrictive.
The. invention will now be described in greatez detail by reference to
specific
examples, which should not be construed as in any way limiting the scope of
the
is mventron.
Example 1
Preparation of Copper-EDTA compalex
Method 1:
EDTA disodium salt (140 g, 0.43 rnol) ~~as added tc> an aqueous solution of
zo hydrogen peroxide (10 wt%, l L), and the mixture stirred until all of the
EDTA disodium
salt had dissolved. The solution was cooled to maintain the temperature below
25 °C.
Copper wire (60g, 0.94 mot) was added and the resultant rnixtuze stirred and
cooled. A
blue solution was produced. The solution was evaporated to obtain a blue
powder
comprising copper-EDTA disodium sail. Metal ion complex n'ay optionally be
purified
zs by recrystallisation. Mass spec (mlz) (Negative Ion MALDI): ?.51 (100%),
Method 2;
EDTA disodium salt was added to water and the resultant mixture. heated with
stirring to promote dissolution. When all of the EDTA salt vvas dissolved, the
solution
was cooled to room temperature. Hydrogen peroxide (10 wt°.~o) was then
added, followed
ac> by coppers) and the mixture stirred, optionally with wazrning but
maintaining the
temperature to less than 40'C to produce a blue solution comprising copper-
EDTA
disodium salt. In a variation of this method the order of addition of the
hydrogen
peroxide and copper may be reversed.
CA 02478137 2004-08-17
17
Example 2 -Pharmaceutical Formul:~tions
2(a) - Topical Cream Formulation
(i) 20 mL of the solution prepared in Example. 1 eras mixed into 250 g of
commercially available Vitamin E cream.
s (ii} 10 mL of the solution prepared in Example 1 twos mixed into 200g of
comrzxercially available Sorbolene cream.
2(b) - Topical Cream Formulation
A typical composition formulated as a topical cream is outlined below:
Cu[EDTA]Naz 1.0 g
~o Polawax GP 200 25.0 g
Lanolin Anhydrous 3.0 g
White $eeswax 4.5 g
Methyl hydroxybenzoate 0.1 g
Deionised & sterilised Water to 100.0 g
~s The polawax, beeswax and lanolin are heated together at 60°C, a
solution of methyl
hydroxybenzoate is added and homogenisation achieved using high speed
stirring. The
temperature is then allowed to fall to 50°C. The compound of the
present invention, in
this example being Cu[EDTA]Na2, is then added and dispersed throughout, and
the
composition is allowed to cool with slow speed stirring.
~0 2(c) - 'l: opical Lotion Formulation
A typical composition formulated as a topical lotion is outlined below:
Cu(E.DTA]Nay 1.2 a
Sorbitan Monolaurate 0.8 g
Polysorbate 20 0.7 g
is Cetostearyl Alcohol 1.5 g
Cvlycerin 7.0 g
Methyl ~iydroxybenzoate 0.4 g
Sterilised Water about to 100.00 ml
The methyl hydroxybenaoate a~~d glycerin are dissolved in 70 rnl of the. water
at
3n 75oC. The sorbitan monolaurate, polysorbate. 20 and cetostearyl alcohol are
melted
together at 75°C and added to the aqueous solution. The resulting
emulsion is
homogenised, allowed to cool with continuous stirring and the Cu[EDTA]Nay is
added as
a suspension in the remaining eater. The whole suspension is stirred until
homogenised.
3S
CA 02478137 2004-08-17
18
2(d) - Ointment Formulation
A typical composition for delivery as an ointment includes l.Og of
Cu[EDTA~Na2,
together with white soft paraffin to 100.0 g, dispersed to produce a smooth,
homogeneous
product.
s 2(e) - Liquid Formulation
An example of a liquid formulation can be prepared by diluting ~0 mL of the
solution produced in Example 1 with 200n~L of sterilised water and mixing to
produce a
homogeneous solution.
co 2(f) - Shampoo Formulation
An example of a shampoo formulation can be prepared 'by diluting 40 naL of the
solution produced in Example 1 with 250mL of commercially available shampoo
and
mixing to produce a homogeneous product.
Example 2(g) - Capsule Composition
!s A composition of copper-EDTA disodium in the form of a. capsule may be
prepared
by filling a standard two-piece hard gelatin capsule with 250 mg of
Cu[EDTA]Naa, in
powdered form, 100 mg of lactose, 3S mg of talc and IO mg of magnesium
steatite.
Example 3 - Methods of Treatment
3(a) -'Treatment of inflammation
4o Study (i) A cream containing Goppes-EDTA prepared according to example
(2a)(i) was
applied topically twice per day to an inflamed knee joint of a male aged mid-
40s, then
rubbed in. Treatment was continued for one week to alleviate inflammation: The
duration
of treatment time and number of applications per day may be varied as
necessary.
a~ Study (ii) A cream containing Copper-EDTA prepared according to example
(2a)(i) was
applied topically 3 times per day to the hands of an, 84 yeas old woman having
severe.
arthritis. Initial relief was felt within 24 hours. Substantial relief from
pain and
inflammation was felt within a week.
3n Study (iii) A cream containing Copper-EDTA prepared according to example
(2a)(i) was
applied topically to one hand of a 55 year old male suffering from arthritis.
The cream
was applied twice per day for one tveek. The other hand was not treated. After
one week
CA 02478137 2004-08-17
19
there w as a significant reduction in pain and swelliag in the hand which had
been treated,
in comparison to the hand which had not been treated.
3(b) - Stimulation of Hair Regrowth
A cream containing Copper-EDTA prepared accoxdini; to example. (2a}(i} was
s applied topically once per day to the scalp of a 42 year old male, then
massaged into the
scalp until the cream had been absorbed. Treatment was continued for 1 month
before
fine hairs started to grow. The lairs have continued to grow orrer a. 6 month
period (to
date) without further treatment.
3(c} - Sunscreen
io A female subject (aged early ~lOs) suffering from vitiligo applied a cxeam
prepared
according to Example (2a)(i) to areas of skin lacking pi~nent. The treated
areas regained.
some colour after applying the cream foz about 1 month, After about 1 week of
treatment,
non-pi~nented patches of skin were able to be exposed to sunlight without the
burning
sensation usually experienced by the subject. The. cream is currently applied
once in the
i5 morning to get sun protection for the day.
inc9ustrial Applicability
The present invention relates to a process for preparing metal ion complexes
and
zo plartnaceutical formulations thereof. The invention further relates to
methods of
treatment of inflammatory disorders, pain, promotion of hair growth or
regrowth, and
promotion of wound and scar tissue healing.
