Note: Descriptions are shown in the official language in which they were submitted.
wo g6ios867 2 l 9 7 4 6 ~ PC'T/GB951019(i9
COMPOS ITIONS
The present invention relates to 1 ,_~v~ nts in
~ and relating to magnetic resonance imaging (MRI) and in
particular to compositions for use as or in the
preparation of MRI contrast media for imaging of the
stomach, intestine, liver, bile duct and gall bladder.
MRI is now well established as a medical diagnostic
tool. The ability of the technique to generate high
quality images and to differentiate between soft tissues
without requiring the patient to be exposed to ioni~ing
rA~;Rt;o" has contributed to this success.
Although MRI can be performed without using added
contrast media, it has been found that substances which
affect the nuclear spin reequilibration of the nuclei
~hereinafter the ~'imaging nuclei" - generally water
protons in body fluids and tissues) responsible for the
magnetic resonance ~MR) signals from which the images
are generated may be used to enhance image contrast and,
accordingly, in recent years, many such materials have
been suggested as MRI contrast agents.
The P"h~nrP~ contrast obtained with the use of
contrast agents enables particular organs or tissues to
be vi~UAl i7P~ more clearly by increasing or by
decreasing the signal level of the particular organ or
tissue relative to that of its ~ULlU""~; rg~ . Contrast
agents raising the signal level of the target site
relative to that of its ~LLV"~; ngC are termed
~positive~ contrast agents whilst those lowering the
signal level relative to ~uLL~ dings are termed
~negative~ contrast agents.
The majority of materials now being proposed as MRI
contrast media achieve a contrast effect because they
contain paramagnetic, superparamagnetic or ferromagnetic
species.
WOg6~8G7 PCT/GB95/01~ -
21~746~ 2 -
For ferromagnetic and superparamagnetic contra6t
agents, which are negative MRI contrast agents, the
enhanced image contrast derives primarily from the
rG~nctinn in the spin reequilibration parameter known as
T2 or as the spin-spin relaxation time, a reduction
arising from the effect on the imaging nuclei of the
fields generated by the feLL~ ~"etic or
SUpeLl _ ?t~ C particles.
PaI gn~tic contrast agents on the other hand may
be either positive or negative MRI contrast agents. The
effect of paramagnetic substances on magnetic resonance
signal intensities is ~opPn~Pnt on many factors, the
most important of which are the rnrr~n~ration of the
paL. 3n~tl C gubstance at the imaged site, the nature of
the paL -_ ~tic substance itself and the pul~e sequence
and magnetic field strength used in the imaging routine.
Generally, however, paramagnetic contrast agents are
positive MRI contrast agents at low cnnr~ntrations where
their T1 lowering effect dominates and negative MRI
contrast agents at higher cnnrrntrations where their T2
lowering effect is d ~n~nt, In either event, the
relaxation time r~dnrt;~n results from the effect on the
imaging nuclei of the magnetic fields g~n~r~tod by the
paramagnetic centres.
The use o~ tic, f~ gn~tic and
superp~rnm-~n~t;r materials as MRI contrast agents has
been widely advocated and broad ranges of suitable
materials have been suggested in the literature.
An example of a physiologically tolerable
paramagnetic material known for use as an MRI contrast
agent is r~ J~ e ion, which may conveniently be used
in the form of its salts or rh~l~t~s. Indeed, even at
very low i.v. dosages (about S-10 ~mol/kg bodyweight)
manganese has been found to be particularly effective as
a contrast agent for imaging of the liver.
~ owever m~ng~n~se~ when administered intravenously
as a contrast agent, may be teratogenic at clinical
096/05~7 ~1~ 7 4 6 ~ '.il569
dosages. Administered intravenously, ~-ng~nPRe is also
known to interfere with the normal functioning of the
heart by replacement of calcium in the calcium pump of
the heart.
In order to reduce the direct effect on the heart,
~ oral administration has been proposed. This ensures
passage of the contrast agent through the liver before
going to the heart.
Oral administration of MnCl2 as a liver imaging MR
contrast agent has been proposed and orally administered
MnCl2 has not been found to be teratogenic. ~owever, the
absorption of MnCl2 through the gut is poor, and as a
result the dosage required for clinical efficacy is of
the order of 100-1000 ~mol/kg bodyweight. In the event
of damage to the gut resulting in increased uptake, such
a high dosage level still has the potential for causing
undesired adverse effects, eg. cardiac effects.
We have now surprisingly found that
gastrointestinal tract m=ng=nPRe contrast agents
suitable for imaging of the liver may be produced by the
incorporation of an uptake promoter capable of PnhAnr;ng
m-ng~nP~e transport across the ' anes of the g.i.
tract.
C ,_' 'R which have been found to be suitable for
use as uptake promoters include reducing compounds
cnntA;n;ng an ~-hydroxy ketone group (-C(OH)-CO-), acids
crntA;n;ng ~- and/or ~-hydroxy or amino groups, as well
as vitamin D.
Thus, viewed from one aspect the present invention
provides a contrast medium composition comprising a
physiologically tolerable ~-ng~n~Re c~ o~d, an uptake
promoter and a physiologically tolerable carrier or
~r; piPnt, having a m-ng=nP.~e concentration of at least
0.3mM or being in a dosage unit form rrntA;n;ng at least
300 ~mol r-ng~nPRe, wherein the uptake promoter
comprises a physiologically tolerable reducing compound
rrnt=;n;ng an ~-hydroxy ketone group, a phys;olsg;rAlly
W096/0s8~7 i~ sS~ -
21 97466 4 _
tolerable acid r~n~;n;ng ~- and/or ~-hydroxy or amino
groups, or a salt thereof, and/or vitamin D.
As used herein, the expression "acid containing ~-
and/or ~-hydroxy or amino groups" is intended to include
aromatic acids c~n~ining ortho-hydroxy or ortho-amino
group~.
The contrast medium composition according to the
invention may comprise a r-ng~n~se compound together
with a mixture of several uptake promoters.
The ~J~l~p~ce c , -~, which prefera~ly is soluble
in gastrointestinal fluid may for examp~e be a chelate
or a salt, or may be a mixture of different salts and/or
chelates. Particularly preferred are metal rhPl~t~ and
salts in which the r~ng~n~e is present as MnlII) rather
than Mn(III) since the former has a higher magnetic
moment and thus is more effective as an MR contrast
agent.
The reducing nature of the uptake promoter is
important since normal uptake of ~ Re by the gut
tends to favour Mn(II) rather than Mn(III).
Pre~erred compositions ~co~;ng to the invention
are those in which the reducing _luulld ~urther
~nt~ i n~ an oxygen atom in a heterocyclic ring
structure.
Particularly pre~erred as an uptake promoter in the
compositions of the invention is ascorbic acid which has
been found to increase the uptake of manganese in the
liver about 5-fold compared with oral administration of
MnCl2 alone. This surprising increase is demonstrated in
Figure 2 of the accompanying drawings. Moreover
ascorbic acid (vitamin C) is particularly preferred as
an uptake promoter since it is cheap, readily available
and particularly well tolerated by the body.
Yet more particularly preferred compositions in
accordance with the invention are those in which the
uptake promoter is kojic acid. The dramatic increase in
the uptake of ~-ng~ e in the liver following
~ W09610s867 2 1 q7466 PCT/GB95/01969
administration of MnCl2 ~ kojic acid can be seen from
Figure 5 of the Acc ying drawings.
R~Ample~ of acids which have been found to be
particularly effective as uptake promoters in the
compositions of the invention include carboxylic acids,
e.g. gluconic and salicyclic acid. The effect of the
addition of salicylic acid to MnClz on MRI ~nhAn~ ~ of
the liver can be seen in Figure 8 of the accompanying
drawings. ~- and ~- amino acids have also been found to
be useful as uptake promoters, in particular ~-amino
acids, e.g. glycine, valine, glutamine, aspartic acid,
glutamic acid, lysine, arginine, cysteine and
methionine, especially arginine, lysine and aspartic
acid. The effect of addition of various ~-amino acids
to MnCl2 on MRI ~nhAn~ of the liver is shown in
accu,..~a.1ying Figure 9.
Other preferred compositions in accordance with the
invention are those which comprise vitamin ~ as an
uptake promoter.
Using the compositions of the invention, the liver
can be effectively MR imaged with a significant
reduction in the dosage of --ng~n~Re otherwise required.
Thus, for example, a 50~ ~nhAn~ of the liver can be
obtained by oral administration of lOO ~mol r ng~n~se/kg
body weight and l mmol ascorbic acid/kg. Such a dosage
results in the same degree of ~nhAns t of the liver
as 5 ~mol Mn(II)~kg body weight (MnCl2, i.v.) or as 500
~mol Mn(II)/kg body weight (MnCl2, p.o.).
Figure l hereto demonstrates the effect of p.o.
administration of MnCl2 and ascorbic acid on MR liver
~nh~n~ nt compared with p.o. administration of MnCl2
alone.
Increase in the ratio of ascorbic acid to MnCl2
results in an increase in the ~nh~n, t effect
obtained. This dose-L~u-1se relationch; r can be seen
from Figure 2 hereto.
The gradual increase in ~nhAn~-~nt of the liver
WO 96/115867 ~ '~ 9 ~ ~ 6 r ~ I/I~D7~ S69
- 6 -
with time following administration oE a composition in
accordance with the invention enables the dynamics of
uptake of the contrast agent by the liver to he
monitored (see for example Figure 2). This is of
particular importance in enabling identification of
areas of healthy tissue and areas of possible tumor
growth.
In the compositions Accor~;ng to the invention, the
preferred molar ratio of manganese to uptake promoter is
from 1:0.2 to 1:50, eg. 1:1 to 1:20, especially 1:3 to
1:6, particular preferably about 1:5.
The uptake promoter may if desired be present in
whole or in part as the counterion to the r~ng~n~e
ions. Thus in one c~~n~; t the composition of the
invention comprises as both ~-J~Pqe - ~ and
uptake promoter a --ng~n~qe salt of a reducing
cnnt~;n;ng an ~-hydroxy ketone group or a ~ngAneqe salt
of an acid ~nnt~;n;ng ~- and/or ~- hydroxy or amino
group~, eg. r ngAnPse ~II) ascorbate or r-ng~n~e
salicylate.
m e compositions according to the invention may be
used to achieve a 60-called ~doublc contrast effectn by
increasing the signal level from the liver whilst at the
same time decreasing that ~rom the surrounding tissues,
in particular ~rom the gut. Such an ef~ect enableq yet
further ~nh~n, ' of the liver.
A double contrast effect and margin definition can
be achieved with the compositions of the invention since
the resulting manganese ion cnnn~ntration within the
g.i. tract will generally be such as to create a signal
~u~p~ssing effect there. In this case, to avoid image
artefacts resulting from pockets of the gut being
contrast agent free, it is desirable to incorporate in
the compcsitions a viscosity ~nh~nn;ng agent and
desirably also an osmoactive agent. Examples of
suitable viscosity Pnh~rr~rs and osmoactive agents are
described in W0 91/01147 and W0 91/01148.
~ W096105867 2 1 ~ 7 4 b b PCTIGBsS/01969
In a particularly preferred embodiment, the
compositions of the invention may be used in ~ ~;n~tion
with a second contrast agent having either a positive or
negative contrast effect. Preferably the compositions
of the invention are used in cn~;n~tion with a second
contrast agent having an opposing contrast effect. This
results in a "double contrast effect" enabling
visllRl;c~t;on and margin definition of the liver to be
particularly PnhRn~P~.
As ~;nnpd above, paramagnetic materials such as
~-ng~nP~e ions may act as either positive or negative
MRI contrast agents ~rPn~;ng upon a number of factors,
including the ronrPntration of the ions at the imaging
site and the magnetic field strength used in the imaging
procedure. At the cnncpntrations of m~ng~n~se
rnnt~ lated for use in the compositions of the
invention, the ~-ng~n~se-cnnt~;n;ng contrast agent will,
in general, function as a positive contrast agent. The
second contrast agent is therefore conveniently a
negative contrast agent and may be any negative MRI
contrast agent suitable for oral administration.
However, as indicated above, any MR contrast agent,
negative or positive, may be used.
3xamples of negative MRI contrast agents for use in
cn~inAt;nn with the compositions of the invention
include known ferrn~-gnptic and superparamagnetic
species, such as for example magnetic iron oxide
particles either free or enclosed within or bound to a
non-magnetic matrix material such as a polysaccharide
eg. LUMIREM and 5nlrhnn~tPd polystyrene eg. ABDOSCAN~.
Further ~l~c of contrast agents for use in
combination with the compositions of the invention
include Gd and Dy ions bound to a polymeric matrix, for
example LUMIREM or GADOLITE (~ nl; n; alumina silicate
oral suspension).
When using the compositions of the invention to
achieve a double contrast effect, it is particularly
W096/0586~ 9 7 ~b 6 - 8 - PCT~GR9~0196s
preferable to incorporate a viscosity ~rh~n~; ng agent
which attains its full visco~ity ~nhAnr;ng effect only
after administration of the contrast medium. The
contrast medium is thus able to be inyested in a
relatively tolerable form while yet developing the
desired viscosity at or during passage towards the site
which is to be imaged.
The compositions of the invention are particularly
suited to use, if required after dispersion in aqueous
media, for imaging of the liver. For such a purpose the
compositions may be administered into the
gastrointestinal tract orally, rectally or via a stomach
t.ube.
Thus, viewed from a further aspect the present
invention provides a method of generating a magnetic
resonance image of a human or non-humanl preferably
r~ n, ani~al body which methoa comprises
administering into the gastrn;nt~Rt;n~l tract of a said
body a contrast medium comprising a physiologically
tolerable r-n~n~e ~- _ ' and a physiologically
tolerable reducing ol,nfl ~nnt~;n;ng an a-hydroxy
ketone group or a physiologically tolerable aoid
nt~;n;ng a- and/or ~- hydroxy or amino groups, or a
salt thereof, and~or vitamin D, and generati~g a
magnetic r~nn~nce image of the liver and the gastro-
intestinal tract. of said body.
Viewed from a yet further aspect the invention also
provides a method of generating a magnetic resonance
image of a human or non-human animal body, which method
comprises administering into the gastrointestinal tràct
of a said body an effective amount of a composition
comprising: (a~ a first contrast agent comprising a
physiologically tolerable r-rg~n~Re compound, a
physiologically tolerable reducing ~ fl ~nnt~i n; ng
an a-hydroxy ketone group or a physiologically tol~r~hle
acid cnnt~;n;ng a- and~or ~- hydroxy or amino groups, or
a salt thereof, and/or vitamin D, preferably having a
~ W09610s867 2 1 ~7 4~6 r~ . r~ol~g
g
~ngAn~Se cnnrPntration of at least 0.3mM or being in a
dosage unit form cnnt~;n;ng at least 300 ~mol ~-ngAnPse,
together with (b~ a second contrast agent and generating
a magnetic resonance image of the liver and abdomen of
said body.
It is possible to fu~ te the contrast medium
immediately or shortly prior to administration by mixing
the uptake promoter with the ~-ng~npse species. Thus,
in a further aspect the invention also provides an MRI
contrast agent kit comprising in a first rnntA;n~r a
physiologically tolerable ~-ngAnpse ~o~ u~.d, and in a
second r~ntA;nPr a physiologically tolerable reducing
compound containing an ~-hydroxy ketone group or a
physiologically tolerable acid cnntA;n;ng ~- and/or ~-
hydroxy or amino groups, or a salt thereof, and/or
vitamin D.
Viewed from a further aspect the invention also
provides an MRI contrast agent kit comprising in a first
rnnt~;nPr a first contrast agent comprising a
physiologically tolerable ~-ngAnPre c~ronn~, a
physiologically tolerable reducing c _u-~d Cnnt~;n;ng
an ~-hydroxy ketone group or a physiologically tolerable
acid nnntAin;ng ~- and/or ~- hydroxy or amino groups, or
a salt thereof, and/or vitamin D, preferably having a
~-n~AnP~e ~nn~Pntr~tinn of at least 0.3mM or being in a
dosage unit form cnntAin;"s at least 300 ~mol ~-ng~n~se,
and in a second rnntAinPr a second contrast agent
comprising a particulate ferromagnetic or
superparamagnetic material or Gd or Dy ions bound to a
polymeric matrix.
The contrast agent compositions of the invention
may of course include components other than the uptake
promoter, the ~-ngAnP~e compound, the viscosity
~nh~nc;ng and osmoactive agents, for example
convPn~;nn~l ph~rr~relltical fo~ tion aids such as
wetting agents, buffers, disintegrants, binders,
fillers, flavouring agents and liquid carrier media such
W096/058G7 ~ 1 97466 PCT/GB95~1969 ~
- 10 -
as sterile water, water/ethanol etc.
For oral administration, the pH of the composition
is preferably in the acid range, eg. 2 to 7 and while
the uptake promoter may itself serve to yield a
composition with this pH, buffers or pH adjusting agents
may be used.
The contrast media may be fnr~ t~a, in
conventional pharmaceutical administration forms, such
as tablets, capsules, powders, solutions, dispersions,
syrups, suppositories etc.
The preferred dosage of the composition according
to the present invention will vary according to a number
of factors, such as the administration route, the age,
weight and species of the subject and the particular
uptake promoter used. Conveniently, the dosage of
~~ng~nrRP will be in the range of fr ~ 5 to 500 ~mol/kg
bodyweight, preferably from 5 to 150 ~mol/kg bodyweight,
more preferably from 10 to 100 ~mol/kg bodyweight, while
the dosage of the uptake promoter will be in the range
of from 5 ~mol to 1 mmol/kg bodyweight, preferably from
25 ~mol to 0.5 ~mol/kg bodyweight.
Preferred : ~ a; tR 0~ the invention will now be
described by reference to the following non-1;m;t;ng
~xamples and the ~r~ ~nying drawings, in which:
Figure 1 is a graph illustrating the effect of p.o.
administration of different Mn2~ salts on liver
hAnCr~-~t;
Figure 2 is a graph illustrating the effect of p.o.
administration of MnC12 + ascorbic acid on liver
,- at varying conr~ntrations of ascorbic acid;
and
Figure 3 is a graph illustrating the effect of p.o.
administration of different doses of MnCl2 rontAin;ng 0.1
mmol/kg ascorbic acid on liver ~nh~nr~~~nt
Figure 4 is a graph illustrating the effect of the
~ a.; t; nn of ascorbic acid or ascorbic acid-palmitate to
MnCl2 on ~nh~rc t of the liver.
~ W0961058C7 ~1 97~66 PCTlGBg5101969
Figure 5 is a graph illustrating the effect of the
addition of ascorbic acid or kojic acid to MnCl2 on
~nh~nl L of the liver.
Figure 6 is a graph illustrating the results of a
phar~ro~;nPtic study to determine the variation in
~ c~ncPn~ation of Mn(II) in the blood following
administration of various Mn(II)-c~nt~;n;ng
compositions. ~ ~
Figure 7 is a graph comparing the effect on liver
~nh~nr - of i.v. administration of Mn DPDP (S-095)
with that of p.o. administration of MnCl2 + ascorbic
acid.
Figure 8 is a graph illustrating the effect of the
addition of ascorbic and salicylic acids to MnCl2 on
liver Pnh~rr
Figure 9 is a graph illustrating the effect of the
addition of different amino acids to MnCl2 on liver
Pnh~nr ,
Figure 10 illustrates transversal Tl-weighted (SE
57/13; 2.4 T) liver images from a control rat and from
three rats 2 hours after oral administration of 200
~mol/kg MnCl2 + 1000 ~mol/kg ascorbate. The signal
intensity of the liver is substantially increased after
gavage administration of Mn2t and ascorbate.
Figure 11 illustrates coronal Tl-weighted (SE
90/17; 2.4 T) liver images from two rats 2 hours after
oral administration of 200 ~mol/kg MnCl2 + 1000 ~mol/kg
ascorbate. The signal intensity in the gastrointestinal
lumen is reduced a_ter administration of Mn2+.
Figures 12 and 13 are graphs illustrating the
effect of the addition of ABDOSCAN~ to Mn-ascorbate on
the Pnh~nc ~ of the liver.
Figure 14 illustrates transversal Tl-weighted (SE
57/13; 2.4 T) liver images from a control rat and from
three rats 2 hours after oral administration of 200
~mol/kg MnCl2 + 1000 ~mol/kg ascorbate + ~3DOSCAN~ (21
W096/0s867 21 ~74G6 - 12 - PcT/~B9~/01~69 ~
~mol/kg Pe). The addition of A~3DOSCAN did not influence
the signal intensity of the liver.
Figure 15 illustrates coronal Tl-weig~hted (SE
9O/17; 2.4 T~ liver images from a control rat and from a
rat 2 hours after oral administration of 200 ~mol/kg
MnCl2 + 1000 ~mol/kg ascorbate + ~BDOSCAN~ (21 ~mol/kg
Fe). The signal intensity in the gastrointestinal lumen
is markedly reduced after co-administration of Mn2+ and
AB~OSCAN .
For the measurement of the curves of Figures l to 9
the following materials were used:
Fi~-re 1
Mn - A ccorbate
MnCl2 x ZH,O 6.48 g
Ascor~ic acid 35.2 g
Water ~ 1000 ml
Mn-gluconAte
Mn-gluconate 19.2 g
Water ~ lOOO ml
Mn-citrate
Mncl2 x 2H2O 6.48 g
Na3-citrate x 2H2O 23.5 g
Water ~1 1000 ml
Ficnlre 2
~ 12
MnCl2 x 2~2O 6.48 g
Water ad 1000 ml
Mn~l2 + D.l mmnl /kg ~ccnrhic Arid
MnCl2 x 2X2O 6.48 g
~rorhjc acid 3.52 g
. .
~ W096/05867 ~l 9746b PCT/GB95lolg69
Water ~d 1000 ml
Mn~12 + 0.4 rm~l/kg ascorbic acid
MnCl2 x 2H20 6.48 g
Ascorbic acid 14.1 g
Water ~ 1000 ml
Mn~l ~ + 1.0 1 /kg ~corbic acid
MnCl2 x 2H20 ~ 6.48 g
Ascorbic acid ~ 35.2 g
Water ~d 1000 ml
Figure 3
MnCl2 (0.2 l/kg) + ~qcorbic acid
MnC12 x 2H20 6.48 g
Ascorbic acid 3.52 g
Water ~d 1000 ml
Mn~l 2 (~ . 5 l/kg) + aqc~rhic acid
M~Cl2 x 2H20 16.2 g
Ascorbic acid 3.52 g
Water ~d 1000 ml
Mn~ 2 (2.0 l/kg) + ~r~rh; c acid
MnCl2 x 2H20 64.8 g
Ascorbic acid 3.52 g
nater ad 1000 ml
Fiq~re 4
~2
MnCl2 x 2H20 13.0 g
Water ad 1000 ml
Mn~12 + ~r~rhic acid ~ m;tate (0.4 ~l/kg)
L-ascorbic acid 6-palmitate 66.4 g
W096/~867 ~l 9 74 6 ~ PCT~GB9~01969
- 14 -
Polyethylene glycol 300 ~ 1000 ml
Fi~re 5
Mn~l2 + k~Jlc arid tO.4 1/kq)
MnCl2 X 2~0 6.48 g
Koj ic acid 11. 4 g
Water ~ 1000 ml
Fi~lre 8
Mn~l2 (0. 2 l/kg)
MnCl2 X 2~20 6.4 8 g
Water ~ 1000 ml
~l2 ~0.2 l/kg) + ~rnrhic acid (Q.4 l/k,g~
MnCl2 X 2H~0 6.48 g
Ascorbic acid 14.1 g
Water ~ 1000 ml
Mnrl2 (0.2 l/kg) + 5al;cyllc aci~ (0.4 r~ k,q)
MnCl2 x 2~206.48 g
Salicyclic acid sodium sal~~2. 8 g
Water d 1000
Fi~lre 9
Mn~l 2 (0 .2 ~mn3 /kg)
MnCl2 X 2H2~ 6.48 g
Water ad 1000 ml
Mn~l 2 (0 .2 l/k~) + ~ccnrh;c acid ~o .4 ~mnl ~kg~
MnCl2 X 2H2~ 6.48 g
Ascorbic acid 14.1 g
Water ~ lO00 ml
~ W096/05867 ~ 19 7466 PCTIGB95/01969
- 15 -
Mnrl2 (0.2 ~l/kg) + glycin~ (0.4 l/kg)
MnCl2 x 2H20 6.48 g
Glycine 7.76 g
~ Water ~1000 ml
~ Mnrl2 (0.2 ~l/kg) + v~l;nP (0.4 ~l/kg)
MnCl2 x 2H20 6.48 g
Valine 9.36 g
Water ~1000 ml
Mn~la (0.2 l/kg) + glut~m;n~ (0 4 l/kg)
MnCl2 x 2H20 6.48 g
Glutamine 11.7 g
Water ~1000 ml
MnGl2 (0.2 -l/kg) + ~ rtic acid ~0.4 l/kg)
MnCl2 x 2H20 6.48 g
Aspartic acid 13 .8 g
Water ~ 1000 ml
Mn~l2 (0.2 l~kg) + glutAmic ~id (0.4 l/kg)
MnCl2 x 2H206. 48 g
Glutamic acid monosodium salt
monohydrate15.0 g
Water ~1 1000 ml
Mn~l2 (0.2 l/kg) + lys;n~ (0 4 l/kg)
MnCl2 x 2H20 6.48 g
Lysine monohydrochloride 14.6 g
Water ~ 1000 ml
Mn~l2 (0.2 l/kg) + ~rginin~ (0 4 l/kg)
MnCl2 x 2H20 6.48 g
Arginine monohydrochloride 16.9 g
Water ~ 1000 ml
W096~05X67 2 ~ 9 74 6 6 PCI~/GB95/019b9 ~
- 16 -
MnC-2 ~0.2 l/kg) + c~ste;n~ lo 4 l/kgl
MnCl2 x 2~2O 6.48 g
Cysteine monohydrochloride
monohydrate 14.0 g
Water ad 1000 ml
MnCl2 10.2 l/k~) + ~th;on;n~ (0.4 ~l/kgi
MnC12 x 2~2O 6.48 g
MPth; ~n; ne 11.9 g
Water ad 1000 ml
Por the mea~uL. of the curves of Figures lZ and 13
the following materials were used:
MnCl2 x 2~2O 0.567 g
Ascorbic acid 3.08 g
A}3DOSCAN0 23.4 mg Fe
(one dose-package~
Water ~d 200 ml
r le 1
Or~l C ~~itinn
MnCl2 x 2~20 6.48 g
A~corbic acid 35.2 g
Water ~ 1000 ml
The ~-ng~n~e chloride and ascorbic acid are dissolved
in sterile deionised water. The dose for a 70 kg adult
human would be 350 ml, taken orally.
E le 2
Or~l f O~itinn
MlICl2 X 2~20 6 . 48 g
Kojic acid 11.4 g
W096/05867 2l 9 7 4 6 b PCT/GB9S/01969
- 17 -
Water ~ 1000 ml
The m~ng~nGse chloride and kojic acid are dissolved in
~ sterile deionised water. The dose for a 70 kg adult
human would be 350 ml, taken orally.
E le 3
OrAl Co~osition
MnCl2 x 2~20 13.0 g
Water ~ 1000 ml
B.
L-ascorbic acid 6-palmitate66.4 g
~ Polyethylene glycol 300 ~1000 ml
The dose for a 70 kg adult human would be 175 ml of A
and 175 ml of B, taken orally.
~m~le 4
Orsl Cnm~osit;nn
MnCl2 x 2H20 0.567 g
Ascorbic acid 3.08 g
ABDOSCAN0 23.4 mg Fe
Water ad 200 ml
The doRe for a 70 kg adult human would be 4 x 200 ml,
taken orally.
746~
W096/05~6~ pcrlGB9s~l96
- 18 -
~le 5
Oral t~m,rosition - Mnt~l2 (0.2 r-~-,1/kq) + vi~;~m;n
~ 0 . 4 r~ kq~
A.
MnCl2 x 2E~20 13 . O g
Water ~ lOOO ml
Vitamin D 30 . O g
Polyethylene glycol 300 ~51 1000 ml
