Note: Descriptions are shown in the official language in which they were submitted.
~:~5~i2~9
The invention relates to diagnostic preparations and
the use thereof in diagnosis.
Complex compounds and their sal-ts have been used for a
long time in medicine, for example as auxiliaries for the admin-
istration of sparingly soluble ions (for example iron) and as
antidotes (calcium or zinc complexes being preferred in this
case) for detoxication in the case of lnadvertent incorporation
of heavy metals or their radioactive isotopes.
We have now ~ound that certain physiologically
tolerable complex salts containing one or more central elements
having the atomic numbers of from 21 to 29, 42, 44 and from 57 to
83 can be used for the manufacture of preparations that are sur-
prisingly outstandingly suitable for use in NMR, ultra-sound and
X-ray diagnostics.
The present invention provides a diagnostic preparation
which comprises (i) a physiologically tolerable complex salt
which contains (a) a central element selected from elements
haYing atomic numbers of from 21 to 29 inclusive, 42, 44 and from
57 to 83 inclusive, and (b) a radical of a physiologically
tolerable complex-forming acid and, if desired, (c) a radical
selected from radicals of inorganic and organic bases and amino
acids, and (ii) a physiologically tolerable carrier, especially
an a~ueous carrier.
The physiologically tolerable complex salt (i) may
containe more than one of the radicals (b) and (c).
-
For the intended use of the diagnostic agent according
to the invention, the element or elements having an atomic number
mentioned above, which forms the central element or elements of
the physiologically tolerable complex salt, must not, of course,
be radioactive.
-- 1 --
.
~' ~
, :
~ 2S ~ 2~ ~
In the case where a preparation of the invention is to
be used in NMR diagnostics (see European Patent Application 71
564 published February 9, 19~3, the central element of the
complex salt must be paramagnetic. Such elements are especially
the divalent and trivalent elements having an atomic number of
from 21 to 29, 42, 44 and from 58 to 70. Suitable elements are,
for example, chromium (III), manganese (II), iron (III), iron
(II), cobalt (II), nickel (II), copper (II), praseodymium (III),
neodymium (III), samarium (III) and ytterbium (III). Especially
preferred, owing to their strong magnetic moment, are gadolinium
(III), terbium (III), dysprosium (III), holmium (III) and erbium
(III).
If the prepara-tion of the invention is to be used in X-
ray diagnostics, the central element must be one having a rela-
tively high atomic number in order to o~tain sufficient absorp-
tion of the X-rays. It has
- 2 -
`.~
.
.
- ~ . - . .. .
~ 2S~
--3--
been found that diagnostic preparations that comprise a
physiologically tolerable complex salt containing a
central element or elements having an atomic number of
from 57 to 83 are suitable for this purpose; such
elements are, for example, lanthanum(III), the above-
mentioned elements of the lanthanide series, gold(III),
lead(II) or, especially, bismuthtIII). Especially
- - suitable are physiologically tolerable complex salts in
which the central element (a) has an atomic number of
from 71 to 83.
The preparations of the invention that are to be
used in NMR diagnostics and those that are to be used
in X-ray diagnostics are also suitable for use in
ul~ra-sound diagnostics.
Suitable complex-forming acids are those which
are customarily used for complex formation of the
above-mentioned central elements. Suitable complex-
; forming acids are, for example, those which contain
from 3 to 12, preferably from 3 to 8, methylenephos-
phonic acid groups, methylenecarbohydroxamic acid
groups, carboxyethylidene groups or, especially,
carboxymethylene groups of which at least one, two or
three are bound to a nitrogen atom supporting the
complex formation. If three of the acid groups are
bonded to a nitrogen atom, then the complex-forming
acids in question are those which form the basis of
the complex salts of the general formula
.
-
- ' , . . .
~L256;2
--4--
2 ) 3 t
in which
X represents the radica].s -COOY, -PO3HY or
-CONHOY wherein Y represents a hydrogen atom,
a metal equivalent andl/or a physiologically
tolerable cation of an inorganic or organic base
- or amino acid,
with the proviso that at least two of the substituents
Y are metal equivalents in which the metal has an
atomic number of from 21 to 29, 42, 44 or from 57
to 83.
If in each case only one or two of the acid
groups are bonded to a nitrogen atom, then the nitrogen
atom is bonded to a further nitrogen atom by way of
optionally substituted ethylene or by way of up to four
ethylene units each of which is separated by a
nitrogen, oxygen or sulphur atom supporting the complex
formation. Preferred complex-forming acids of that
type zre those forming the basis of complex salts of
the general formula
X-CH2 CH -X
/
N~A-N \ [I)
V-CHR1 CHR~-V
.. . . . .
.' ': : .
-
~7~S~f~ 9
in which
X represents the radicals -COOY, -PO3HY or
-CONHOY wherein Y represents a hydrogen atom,
a metal equivalent ancl~or a physiologically
tolerable cation of an inorganic or organic base
or amino acid,
and in which
A represents the group -CHR2-CHR3-,
-CH2,~CH2 (ZcH2 C~2)m
N(CH2X)2 CH2-~H2-N(CH2x)2
1 or
-CH2-CH-CH2~ -CH2CH2-~ 1-CH2-CH
in which
X has the meanings given above,
Rl represents in each case a hydrogen atom
or methyl group,
15R2 and R3 together represent a trimethylene group
or a tetramethylene group, or each
-represents a hydrogen atom, lower alkyl
group, phenyl group or benzyl group, or
R2 represents a hydrogen atom
and
R3 represents a group -(CH2)~-C6H4-W-protein
in which
represents 0 or 1,
W represents -NN-, -NHCOCH2- or
-NHCS-
. .
,' ' ' . '
. . , , ~ . .
.
-,
~6~4~
and
~protein represents a protein radical
and
m represents the integer 1, 2 or 3,
Z represents an oxygen atom or a sulphur
atom or the group
CH2X or ~ CH2CH2OR4
~: in which
X has the meanings given above
and
R4 represent a lower alkyl group
and in which
V has the same meaning as X or represents the
group -CH OH -CONH~CH ) X COB
in which
X has the meanings given above,
B represents a protein or lipid radical
and
n represents the integers from 1 to 12
or if Rl, R2 and R3 are hydrogen atoms
both V's together represent the group
CH2X CH2X
-(cH2)wN-cH2-c~2-N (CH2)w
. i . .. . : :-
.
:
~25~2~
in which
X has the meanings given above
and
w represents the integer 1, 2 or 3,
with the proviso that at least two of the
substituents Y are metal e~uivalents in which
the metal has an atomic number of from 21 to
29,42, 44 or from 57 to 83.
The complex-forming acids can, as conjugates, be
bonded to biomolecules that are known to become
especially concentrated in the organ or organ part
under examination. Such biomolecules are, for example,
hormones, such as insulin, prostaglandins, steroid
hormones, amino sugars, peptides, proteins or lipids.
There come into consideration more especially
conjugates with albumens, such as human serum albumen,
antibodies, such as, for example, monoclonal antibodies
specific to tumour-associated antigens, or antimyosin.
The diagnostic preparations formed therefrom are
suitable, for example, for use in tumour and infarct
diagnosis For examinations of the liver there are
suitable, for example, conjugates or inclusion
compounds with lipsomes, which are used, for example,
as unilamellar or multilamellar phosphatidylcholine-
cholestero] vesicles. The conjugates are ~ormed either
- by way of a carboxy group of the complex-~orming acid
., ~
", .
- . :
.. . .
~2S~24~
or, in the case of proteins or peptides, also by way of
a (CH2)~-C6H4-W- group as defined above under R3. In
the conjugate formation of some complex-forming acids
with proteins, peptides or lipids, several acid
radicals may be bonded to the macromolecular
biomolecule. In that case, each complex-forming acid
radical may carry one central elementu If the
complex~forming acids are not bonded to biomolecules,
they ca{ry optionally two central elements, and
10 especially one central element.
Suitable complex salts of the general formula I
above are, for example, those of the general formula Ia
X-CH2 ~H2-X
N-CHR -CHR -~ (Ia~
V-CHRl 1
in which X, V, Rl, R2 and R3 have the meanings given
above.
The following complex-forming acids, inter alia,
are suitable for the manufacture of the complex salts
of the general formula Ia:-
ethylenediaminetetraacetic acid, ethylenediaminetetra-
acetohydroxamic acid, trans-1,2-cyclohexylenediamine-
- tetraacetic acid, DL-2,3-butylenediaminetetraacetic
~ .:
.
.' , ~ '
:
~s~
acid, DL-1,2-butylenediaminetetraacetic acid, DL-1,2-
propylenediaminetetraacetic acid, 1,2-diphenylethylene-
diaminetetraacetic acid, ethylenedinitrolotetrakis-
(methane-phosphonic acid~ and N-(2-hydroxyethyl)-
ethylenediaminetriacetic acid.
Other suitable complex salts of the general
formula I are, for example, those of the general
formula Ib
: X-CH2 CH -X
N-cH2-cH2-(z CH2 CH2)_ \ (Ib)
V-CHR1 1
-'
in which X, V, Z, R1 and m have the meanings given
above. If Z represents an oxygen atom or a sulphur
atom, complex salts in which m represents 1 or 2 are
preferred.
The following complex-forming acids, inter alia,
are suitable for the manufacture of the complex salts
of the general formula Ib:
diethylenetriaminepentaacetic acid, triethylenete-
traminehexaacetic acid, tetraethylenepentaminehepta-
acetic acicl, 13,23-dioxo-15,18,21-tris(carboxymethyl)-
12,15,18,21,24-pentaazapentatriacontanoic diacid, 3,9-
bis-(1-carboxyethyl)-3,6,3-triazaundecanoic diacid,
,
.. - : .
: . .
' ~ ~
~25~
--1 o--
diethylenetriaminepentakis(methylenephosphonic acid),
1,10-diaza-4,7-dioxadecane-1,1,10,10-tetraacetic acid
and 1,10-diaza-4,7-dithiadecane-1,1,10,10-tetraacetic
acid.
Suitable complex salts of the general formula I
are also those of the general formula Ic
X-CH2 ` / 2 X
~ 2 CH2 p
CH2 ~CH2
(fH2)W ( 2)w
- ¦ - (Ic)
N-CH -CH -N
X-CH2 2 X
in which X and w have the meaninqs given above.
The following complex-forming acids, inter alia,
l0 are suitable for the manufacture of the complex salts
of the general formula Ic:
1,4,8,11-tetraazacyclotetradecanetetraacetic acid and,
especially, 1,4,7,10-tetraazacyclododecanetetraacetic
acid.
Other complex-forming acids that are suitable for
the manufacture of the complex salts of the general
formula I are, for example:
1,2,3-tris-[bis-(carboxymethyl)-amino]-propane and
: nitrolotris-(ethylenenitrolo)-hexaacetic acid. As an
- ,
' : . . :
.
'
.
- . .
s3
example of a complex-forming acid lFor the manufacture
of complex salts of the general formula II there may be
mentioned nitrolotriacetic acid.
If not all of the acid hydrogen atoms of the
5 complex-forming acid are substituted by the central
element or elements, it is advantageous, for the
purpose of increasing the solubility of the complex
salt, to substitute the remaining hydrogen atoms by
physiologically tolerable cations of inorganic and/or
- lO organic bases or amino acids. Suitable inorganic
cations are for example, lithium, potassium or,
especially, sodium. Suitable cations of organic bases
are, inter alia, those of primary, secondary or
- tertiary amines, such as, for example, ethanolamine,
15 diethanolamine, morpholine, glucamine, N,N-dimethyl-
glucamine or, especially, N-methylglucamine. Suitable
cations of amino acids are, for example, those of
lysine, arginine or ornithine.
The complex-forming acids required for the
20 diagnostic preparations of the invention are known or
can be manufactured in a manner known per seO
For example, 13,23-dioxo-15,18,21-tris~carboxy-
methyl)-12,15,18,21-24-pentaazapentatriacontanoic
diacid is manufactured in the following manner, which
25 is an improvement to the method proposed by
R. A. Bulman et al. in Naturwissenschaften 68, ( 1981 ) 483:
17.85 9 ( = 50 mmol) of 1,5-bis-l2,6-dioxomor-
.,: . - ' . " : ,
. , . - .
- ~, . ' . .
, ~' ' ' ' ' .: .
2~
-12-
pholino)-3-azapentane-3-acetic acid are suspended in
400 ml of dry dimethylformamide and, after the addition
of 20.13 g ( = 100 mmol) of 11-aminoundecanoic acid,
the whole is heated at 70~ for 6 hours. The clear
solution is concentrated in vacuo. The yellow oily
residue is stirred with 500 ml of water at room
temperature. In so doing, an almost white, voluminous
solid precipitates which is suction-filtered and washed
several times with water. For further purification,
the resulting product is introduced into 200 ml of
acetone and the whole is stirred for 30 minutes at room
temperature. After suction-filtering and drying in
vacuo at 50C, 3~.9 9 ( = 97 % of the theoretical
yleld) of a white powder of melting point 134-138C
15 are obtained.
Conjugation of the complex-forming acids with
biomolecules is likewise effected according to methods
known E~ se, for example by reacting the
nucleophilic groups of the biomolecule, such as, for
20 example, amino, hydroxy, thio or imidazole groups, with
an activated derivative of the complex-forming acid.
Activated derivatives of the complex-forming acid
which come into consideration are, for example, acid
chlorides, acid anhydrides, activated esters, nitrenes
25 or isothiocyanates. Conversely, it is also possible to
react an activated biomolecule with the complex-forming
acid.
.. . .
-
.~
' '
:
For conjugation with proteins, substituents of
the structure -C6~4N2 or -C6H4NHCOCH2 halogen may also
be considered.
The manufacture of some of the complex salts is
likewise known or can be carried out in a manner known
per se by dissolving or suspending the metal oxide or
metal salt (for example the nitrate, chloride or
sulphate) of the element havi-ng an atomic number of
from 21 to 29, 42, 44 or from 57 to 83 in water and/or
~ lower alcohol (such as methanol, ethanol or
isopropanol) and adding a solution or suspension of the
equivalent amount of the complex-forming acid in water
; and/or a lower alcohol, and stirring, if necessary
while warming or heating to boiling point, until the
reaction is complete. If the complex salt formed is
insoluble in the solvent used, it is isolated by
filtration. If it is soluble, it can be isolated by
concentrating the solution to dryness by evaporation,
for example by means of spray-drying.
If acid groups are still present in the resulting
complex salt, it is often advantageous to convert the
acid complex salt into a neutral complex salt or salts
by means of inorganic and/or organic bases or amino
acids that form physiologically ~olerable cations and
to isolate the neutral salt. In many cases, this is
indeed unavoidable since the dissociation of the
complex salt is so suppressed by the shift in the pH
: . . . . : . .
'''' ' - .
:iL2562
--1 4--
value to neutral that only in that manner can uniform
products be at all isolated or at least purified.
The manufacture is advantageously carried out
with the aid of organic bases or basic amino acids.
It can, however, also be advantageous if the
neutralisation is carried out by means of inorganic
bases (hydroxides, carbonates or bicarbonates) of
sodium, potassium or lithium.
For the manufacture of the neutral salts there
may, for example, be added to the acid complex salts
- in aqueous solution or suspension as much of the
desired base as is necessary to obtain the neutral
point. The resulting solution can subsequently be
concentrated to dryness in vacuo. It is frequently of
lS advantage to precipitate the resulting neutral salts by
adding water-miscible solvents, such ~s, for example,
lower alcohols (methanol, ethanol, isopropanol, etc.) r
;~ lower ketones (acetone, etc.), and polar ethers (tetra-
hydrouran, dioxane, 1,2-dimethoxyethane, etc.~, and
thus obtain crystallisates that are easily isolated and
readily purified. It has been found especially
advantageous to add the desired base to the reaction
mixture during the complex formation and thereby
dispense with one process step.
~5 If the acid complex salts contain several free
acid groups, it is often advantageous to produce
neutral mixed salts that contain both inorganic and
organic physiologically tolerable cations as ions of
" ' : ' '
- .
- , .
~25~i2~
opposite charge. This can be effected, for example, by
reacting the complex~forming acid in aqueous suspension
or solution with the oxide or salt of the element
supplying the central element and with half the amount
of organic base required for neutralisation, isolating
the complex salt formed, if desired purifying it, and
then adding to it the amount of inorganic base required
for complete neutralisation. The or~er in which the
bases are added can also be reversed.
The manufacture of the diagnostic preparations
according to the invention is likewise effected in a
manner known ~ se by suspending or dissolving the
complex salts in an aqueous medium, optionally with the
addition of the additives customary in galenical
pharmacy, and subsequently sterilising the solution or
suspension. Suitable additives are, for example,
physiologically tolerable buffers (such as, for
example, tromethamine hydrochloride), small additions
of complex formers (such as, for example,
diethylenetriaminepentaacetic acid) or, if necessary,
electrolytes ~such as, for example, sodium chloride).
In principle, it is also possible to manufacture
the diagnostic preparations of the invention even
without isolating the complex sal~s. In each case,
particular care must be taken to effect the chelate
formation in such a manner that the salts and salt
solutions according to the invention are virtually free
.
.' - .' ' .: . :
' ~
. ~ . . . - , . .
' - ' ': , ' ' . '' " : -
~5
-16-
of non-complexed toxically acting metal ions. This can
be ensured, for example, with the aid of colour
indicators, such as xylenol orange, by test titrations
during the manufacturing process. The invention also
therefore provides processes for the manufacture of the
complex salts and of the aforesaid preparations
containing them. As a final safeguard, there is always
purification of the isolated complex salt.
If suspensions of the complex salts in water or
physiological salt solution are desired for oral
administration or other purposes, a sparingly soluble
complex salt is mixed with one or more auxiliaries
customary in galenical pharmacy and/or surfactants
and/or aromatic substances for taste correction.
The diagnostic preparations of the invention
contain preferably from 1 ~mol to 1 mol per litre of
the complex salt and are, as a rule, administered in
doses of from 0.001 to 5 mmol/kg. They are intended
for oral, and especially parenteral, administration.
The diagnostic preparations of the invention meet
the many requirements for suitability as contrast
agents for nuclear spin tomography. For example, after
oral or parenteral administration, they are
outstandingly suitable for improving the information
that can be provided by the image obtained with the aid
of nuclear spin tomography, as a result of increasing
the signal intensity. They also exhibit the high
. ~ ' ~ -
. .
~2~
-17-
activity necessary to keep to a minimum the amount of
foreign substances introduced into the body and the
good tolerability necessary to maintain the non-
invasive character of the examination (the compounds
mentioned in J. Comput. Tomography S,6: 543-46 (1981),
in Radiology 144, 343 (1982) and in Brevet 5pecial de
Medicament No. 484 M (1960) are, for example, too
toxic). The ready water-solubility of the complex
salts used in the preparations of the invention enables
the preparation of highly concentrated solutions,
so that the volume introduced into the circulation can
be kept within reasonable limits and the dilution by
body fluid can be compensated, that is to say the NMR
diagnostic preparations must be 100 to 1000 times more
water-soluble than is necessary for NMR spectroscopy.
Furthermore, the diagnostic preparations of the
invention are not only highly stable in vitro but
also exhibit a surprisingly high stability in vivo,
so that the per se toxic ions that are not covalently
bonded in the complexes are released or exchanged only
extremely slowly over the 24 hours in which, as
pharmacological studies have shown, the novel contrast
agents are completely eliminated. The conjugates with
proteins and antibodies which are used, for example,
for the diagnosis of tumours bring about a surprisingly
high intensification of the signal at such a low dosage
that it is possible to use in this case solutions of
: ' - ' : : ' - ' .- , :
-. . : . : -
., ~ ,
: ' ' ' , ' ~ '
.
~25~
-18-
correspondingly low concentration.
The diagnostic preparations of the invention
particularly those in which the physiologically complex
salt contains an element having a relatively high
atomic number that is from 57 to 83, for example 71 to
83, are also outstandingly suitable as X-ray contrast
agents; it should be especially emphasised that, with
these, none of the symptoms of anaphyla~y-type
reactions known in the case of iodine-containing
contrast agents can be detected in biochemical-
pharmacological tests. They are especially valuable by
virtue of their advantageous absorption properties in
regions of relatively high tube voltages for digital
substraction techniques.
Further, the diagnostic preparations of the
invention are also suitable as ultra-sound diagnostics
; owing to their property of favourably influencing the
ultra-sound speed.
In contrast to conventional X-ray diagnostics
with shadow-producing X-ray contrast agents, in NM~
diagnostics with paramagnetic contrast agents there is
no linear relationship between the signal
intensification and the concentration used. As control
studies have shown, increasing the dose administered
does not necessarily result in the signal being
intensified, and, in the case of a high dose of
paramagnetic contrast agent, the signal can even be
, '
,. :
; ' -. - , . . .
4~
--1 9--
extinguished. It was, for that reason, surprising
that some patholoqical processes become visible only
after the administration of doses higher than those
specified in EP 71 564 (which may be from
0.0001 mmol/kg to 5 mmol/kg~ of a preparation of the
invention containing a strongly paramagnetic contrast
agent. Thus, for example, Zl defective blood-brain
barrier in the region of a clanial abscess can be
demonstrated only after giving 0.05 to 2.5 mmol/kg,
preferably 0.1 - 0.5 mmol/kg, of paramagnetic complex
salts such as, for example, gadolinium diethylene-
triaminepentaacetic acid or manganese 1,2-cyclohexy-
lenediaminetetraacetic acid in the form of its readily
water-soluble salts. For a dose of more than
0.1 mmol/kg, solutions of higher concentrations of up
to 1 mol/1, preferably from 0.25 to 0.75 mol/1, are
required since only in this way is the volume reduced
and the ease of handling the injection solution
- ensured.
Especially low doses (under 1 mg/kg) and
therewith solutions of lower concentrations (1~mol/1 to
- 5 mmol/1) than are specified in EP 71 564 are required
for organ-specific NMR diagnostics, for example for
detecting tumours and coronary infarcts.
The invention also provides physiologically
tolerable complex salts containing (a) a central
element selected from elements having atomic numbers of
- -
.. : ' .
:: . . - :
-
. - . '~
:....................... . . '
-20-
from 21 to 29, 42, 44 and from 57 to 83, for example
of from 71 to 83, and (b) a radical of a
physiologically tolerable complex-forming acid, and, if
desired, (c) a radical selected from radicals of
inorganic and organic bases and amino acids, for example a
physiologically tolerable complex salt of the general
formula I given above, in which X, A, V and R1 have
the meanings given above, with the proviso that it
contains from 3 to 12 substituents Y of which at least
two are metal equivalents in which the metal has an
.; atomic number of from 21 to 29, 42, 44 or from 57 to 83
and, in addition, at least one of the substituents Y is
a physiologically tolerable cation of an organic base
or amino acid, any substituents Y which may remain
being hydrogen atoms or cations of an inorganic base.
The present invention further provides a method
of diagnosis using NMR, X-rays or ultra-soundf wherein
a preparat~on of the present invention is administered
to a human or animal body.
`
. .
- . , .
.
. .: ': ' ' - ' :
' ' ~ .' ' .
. . .
~s~
-21-
The following Examples illustrate the invention:-
Exam~le ?Preparation of the gadolinium(lII) complex of
nitrGlo-N,N,N-triacetic acid, C6H6GdNO6
A suspension of 36.2 g ( = 100 m1nol) of
gadolinium oxide (Gd2O3) and 38.2 g ( = 200 mmol) of
nitrolotriacetic acid in 1.2 litres of water is heated,
while stirring, to 90C to 100C and is stirred at this
temperature for 48 hours. The undissolved material is
; lO filtered off over active carbon and the filtrate is
concentrated to dryness by evaporation. The amorphous
residue is pulverised.
Yield: 60 g (87 ~ of the theoretical yield)
m~.p. 300C
15 gadolinium: calculated 45.5 %, found 44.9 ~.
The iron(III) complex of nitrolo-N,N,N-triacetic
acid is obtained in analogous manner with the aid of
iron(III) chloride, FeCl3.
Exam~le ~
20 Preparation of the disodium salt of the gadolinium(IlI)
complex of 13,23-dioxo-15,18,21-tris-(carboxymethyl)-
: ,'
- : - , -: ,:
: . ' .
. ' ~' ' ' ~, - ~ .
' . ~ ' . : . -
.
.:' ': :
~2SG;2~ ~3
~ -22-
12,15,18,21,24-pentaazapentatriacontanoic diacid,
C36H60GdN5O12 . 2 Na.
15.2 g ( = 20 mmol) of 13,23~dioxo-15,18,21-tris-
(carboxymethyl)-12,15,18,21,24-pentaazapentatriacontanoic
diacid are suspended in 400 ml of water and the
suspension is heated to 95C. 7.43 g ( = 20 mmol~ of
gadolinium(III) chloride hexahydrate, dissolved in 60 ml
of water, are slowly added dropwise. The whole is
maintained at this temperature for 2 hours and then, in
order to neutralise the hydrochloric acid formed, 60 ml
of lN sodium hydroxide solution are added.
When the reaction is complete (testing with
xylenol orange) the precipitate obtained is filtered
and washed free of sodium chloride with water. 17.60 g
(96 ~ of the theoretical yield) of a water-insoluble,
white powder of melting point 290-292C are obtained.
Gadolinium(III) complex of 13,23-dioxo-15,18,21-tris-
(carboxymethyl)-12,15,18,21,24-pentaazapentatriacontanoic
diacid.
20 Analysis
(calculated) C 47.30 H 6.84 N 7.66 Gd 17.20
(found) C 47.13 H 6.83 N 7.60 Gd 17.06
.
- .
.
,~ . .
-23~
14.6 9 ( = 16 mmol) of the gadolinium(III)
complex so obtained are suspended in 200 ml of water,
and 31.4 l of lN sodium hydroxide solution are added
dropwise thereto. After 1 hour a clear solution is
obtained which is filtered and then concentrated in
vacuo. After drying in vacu_ at 80C 13.2 9 (87 % of
the theoretical yield) of a readily water-soluble,
- white powder of melting point 279-285C are obtained.
Analysis:
(calculated) C 45.13 H 6.31 N 7.31 Gd 16.41 Na 4.80
(found) C 45.20 H 6.12 N 7.28 Gd 16.26 Na 4.75
In analogous manner there is obtained, using
N-methylglucamine in place of sodium hydroxide
solution,
the di-N-methylglucamine salt of the gadolinium(III)
complex of 13,23-dioxo-15,18,21-tris~carboxymethyl)-12,
15,18,21,24-pentaazapentatriacontanoic diacid,
C50~96GdN722 -
Example 3
Preparation of the disodium salt of the
- gadolinium~III) complex of 3,9-bis(1-carboxyethyl)-6-
-
, :; ~ , .
',, ~
~5;~
-24-
carboxymethyl-3,6,9-triazaundecanoic diacid,
C16H22GdN3O1o 2 Na
. . .
36.2 g ( = 0.1 mol3 of gadolinium(III) oxide and
84.2 g ( = 0.2 mol) of 3,9-bis(1-carboxyethyl)-6-
carboxymethyl-3,6,9-triazaundecanoic diacid are
suspended in 250 ml of water and the whole is refluxed
for 1 hour. The small amount of undissolved material
is filtered off and the solution is concentrated to
dryness ln vacuo. The residue is pulverised and dried
in vacuo at 60C. 112.8 g ( = 98 % of the theoretical
yield) of the complex salt (chelate) is obtained in the
form of a white powder.
Y 16 2~GdN3O1o
(calculated) C 33.39 H 4.20 Gd 27.32 N 7.30
(found) C 47.13 H 6.83 Gd 27.42 N 7.21
57.6 g ( = 0.1 mol) of the complex salt are
introduced into a solution of 0.1 mol of caustic soda
in tO0 ml of water. By adding a further 0.1 ml of
caustic soda powder a pH of 7.5 is established in the
20 solution, the solution is heated to boiling point and
ethanol is added dropwise until the reaction mixture
remains turbid. After stirring for several hours in an
ice bath, the crystallisate is suction-filtered, washed
.
- . .
~' ' ' . ~ ' . : ' ' '
, : ' . ' :
.
.
~2,~
-25-
with ethanol and dried in vacuo. The disodium salt is
obtained in guantitative yield in the form of a white
powder.
Analysis:
5 (calculated) C 31.02 ~l 3.58 Gd 25.38 N 6.78
(found) C 31.10 H 3.71 Gd 25.50 N 6.61
Example 4
Preparation of the dimorpholine salt of the
- gadolinium(III) complex of 3,9-bis~ carboxyethyl)-6-
carboxymethyl-3,6,9-triazaundecanoic diacid,
C24H42GdN5O12
'
17.4 g ( = 0.2 mol) of morpholine are dissolved
in 50 ml of water. 42.1 9 ( = 0.1 mol) of 3,9-bis(1-
carboxyethyl)-6-carboxymethyl-3,6,9-triazaundecanoic
diacid and then 18.2 g ( - 0.05 mol~ of gadolinium(III)
oxide are added and the whole is maintained at reflux
temperature until a clear solution has appeared.
Acetone is then added dropwise until the reaction
mixture remains turbid. After stirring for several
hours in an ice bath, the crystallisate is suction-
filtered, washed with acetone and dried in vacuo.
The dimorpholine salt is obtained in quantitative yield
in the form of a white powder.
'' - '
': ' ' ' .
~256~4~
-26-
Analysis:
(calculated) C 38.44 H 5.65 Gd 20.97 N 9.34
(found) C 3~.31 H 5.72 Gd 20.76 N 9.32
Example 5
S Preparation of the di-N-methylglucamine salt of the
gadolinium(III) complex of diethylenetriamine-N,N,N',
N ,N -pentaacetic acid, C28H54GdN5O
__ _ _ _
39.3 g ( = 100 mmol) of diethylenetriamine-N,N,N',
N",N"-pentaacetic acid are suspended in 200 ml of
lO water, and 19.5 g ( = 100 mmol) of N-methylglucamine
are added. 18.12 g ( = 50 mmol) of gadolinium(III)
oxide, Gd2O3, are then added in portions and the
resultiny suspension is heated to 95C. After
approximately 1 hour, a further 19.5 g ( = 100 mmol) of
15 N-methylglucamine are added and, after heating for a
- further 2 hours, a clear solution is obtained. When
the reaction is complete (testing with xylenol orange?,
the small amount of undissolved material is filtered
off and the filtr`ate is concentrated to dryness in
20 vacuo. The residue is again dissolved in 100 ml of
water and stirred into 250 ml of ethanol. After
cooling for several hours, the crystallisate is suction-
filtered, washed with cold ethanol and dried at 60C
n vacuo. 92.7 g (99 ~ of the theoretical yield) of a
- . - .
.
:,
' ' -. ' ~ .
l:~S~
-27-
white powder of indeterminate melting point is
obtained.
Analysis:
(calculated) C 35.85 H 5.80 N 7.47 Gd 16.77
(found) C 35.50 H 5.72 N 7.20 Gd 67.54
For purifica~ion of the complex salt, it is
possible to use, in place of ethanol, also acetone,
propanol or isopropanol.
In analogous manner, there are obtainedo
with dysprosiumlIII) oxide, Dy2O3,
the di-N-methylglucamine salt of the dysprosium(III)
complex of diethylenetriamine-N,N,N',N",N"-pentaacetic
acid, C28Hs4DYN5O20;
with lanthanum(III) oxide, La?O3,
the di-N-methylglucamine salt of the lanthanum(III)
complex of diethylenetriamine-N,N,N',N",N"-pentaacetic
acid, C24H54LaN5 20;
with ytterbium (III) oxide, Yb203,
the di-N-methylglucamine salt of the ytterbium(III)
complex of diethylenetriamine-N,N,N',N",N"-pentaacetic
acid, C28Hs4YbN5O20;
with samarium(III) oxide, Sm2O3,
the di-N-methylglucamine salt of the samarium(III)
.
.. . - :, - . . .
:. ,. ., - ' :
,
5~i~4~
-28-
complex of diethylenetriamine-N,N,N',N",Nn-pentaacetic
acid, C28H54SmN520'
with holmium(III) oxide, Ho203,
the di-N-methylglucamine sa].t of the holmium~III)
- 5 complex of diethylenetriamine-N,N,N',N",N"-pentaacetic
acid, C28H54HN5 20
with bismuth(III) oxide, Bi~03,
the di-N-methylglucamine salt of the bismuth(III)
complex of diethylenetriamine-N,N,N',N",Nn-pentaacetic
acid, C28Hs4BiNs20;
with gadolinium(III) oxide, Gd203,
the tri-N-methylglucamine salt of the gadolinium(III)
complex of triethylenetetramine-N,N,N',N",Nn',N"'-
hexaacetic acid, C39H78GdN70~7.
There are also obtained in analogous manner:
with holmium(III) oxide, Ho203, and ethanolamine in
place of N-methylglucamine,
the diethanolamine salt of the holmium(III) complex of
diethylenetriamine-N,N,N',N",Nn-pentaacetic acid,
- 20 C18H34HoN5012
with gadolinium(III) oxide, Gd203, and lysine in place
of N-methylglucamine,
the dilysine salt of the gado~inium(III) complex of
diethylenetriamine-N,N,N',N",N"-pentaacetic acid,
25 C22H42HoN50l4
The salts are obtained in the form of a white
~25~
-29-
powder o~ indeterminate melting point. They are very
readily soluble in water.
Example 6
Manufacture of the disodium salt of the gadolinium(III)
complex of diethylenetriamine-N,N,N',N",N"-pentaacetic
' 14 18GdN310 2Na
.
18.2 g ( = 0.05 mol) of gadolinium~III) oxide and
39.3 g ( = 0.1 mol) of diethylenetriaminepentaacetic
acid are suspended in 110 ml of water and refluxed for
1 hour. The clear solution is cooled and adjusted to
pH 7.5 by the addition of approximately 80 ml of 5
sodium hydroxide solution. The solution is again
heated to boiling point and 250 ml of ethanol are added
dropwise. After stirring for several hours in an ice
15 bath, the crystallisate is suction-filtered, washed
with ice-cold ethanol and dried at 60C in vacuo.
There is obtained in quantitative yield a white powder
which does not melt until 300C.
Analysis:
20 (calculated) C 28.43 ~ 3.07 N 7.10 Gd 26.58
(found) C 28~35 H 2.95 N 7.05Gd 26.37
In analogouc; manner, there are obtained:
- :
.
~ . . . .
.
~25~i2~
--30--
i
with dysprosium(III) oxide, Dy203,
the disodium salt of the dysprosium(III) complex of
diethylenetriamine-N,N,N',N",N"-pentaacetic acid,
C14H18DyN3O10 2 Na;
5 with lanthanum(III) oxide, La2O3,
the disodium salt of the lanthanum(III) complex of
diethylenetriamine-N,N,N',NI',N''-pentaacetic acid,
14 18 3 10 . 2 Na;
with holmium(III) oxide, E~o2O3,
-: lO the disodium salt of the holmium(III~ complex of`
diethylenetriamine-N,N,N',N",N"-pentaacetic acid,
14 18 3 10 2 Na;
with ytterbium(III) oxide, Yb2O3,
the disodium sal~ of the ytterbium(III) complex of
l5 diethylenetriamine-N, N ,N ', N', N" -pentaacetic acid,
14 18 3 10 2 Na;
with samarium(III) oxide, Sm2O3,
the disodium salt of the samarium(III) complex of
diethylenetriamine-N,N,N',N",N"-pentaacetic acid,
20 C14H18SmN3O10 . 2 Na;
with erbium(III) oxide, Eb2O3,
the disodium salt of the erbium(III) complex of
diethylenetriamine-N,N,N',N",N"-pentaacetic acid,
14 18 3 10
25 with ~adolinium(III) oxide, Gd2O3,
the sodium salt of the digadolinium(III) complex of
-~
. .
: ,
~2S6;~
-31-
tetraethylenepentamine-N,N,N',N'',N''',N'V,N'v-
~ C22~30Gd2N514 Na-
These salts are obtaiined as a white powder of
indeterminate melting point: and are very readily
soluble in water~
Example 7
Manufacture of the N-methylglucamine salt of the
iron(III) complex of diethylenetriaminepentaacetic
21 37 415
. .
.
- lO 35.4 g ~ = 90 mmol) of diethylenetriaminepenta-
acetic acid are suspended in 100 ml of water, and
24.3 g ( = 90 mmol) of iron(III) chloride hexahydrate
(FeC13 . 6 H2O), dissolved in 100 ml of water, are
added thereto. The-initially dark brown suspension is
heated to 95C. After approximately 1 hour, the
colour changes to a light yellow. 270 ml of lN sodium
hydroxide solution are added to neutralise the
hydrochloric acid formed and the whole is heated for a
further 3 hours at 95C. The resulting light yellow
precipitate is suction-filtered, washed free of
chloride with water and dried at ~0C ln vacuo.
17.85 g (45 ~ of the theoretical yield) of a light
yellow powder is obtained the melting point of which
is > 300C.
-- ,
.
'' '.
~L256~
-32-
17.85 g ( = 40 mmol) of the iron(III) complex
salt obtained are suspended in 200 ml of water, and
7.8 g ( = 40 mmol) of solid N-methylglucamine are added
in portions. The whole is heated for approximately 3
hours at 50C and an almost clear, red-brown solution
is obt~ined which is filtered and then concentrated to
dryness in vacuo. The residue is dried at 50C in
vacuo. 24.3 g (95 % of the theoretical yield) of a
red-brown powder of melting point 131-1 33C are
lO obtained.
:
Analysis:
(calculated) C 39.82 H 5.89 N 8.85 Fe 8.81
(found) C 39 . 70 H 6 . 00 N 8.65 Fe 9.01
By using sodium hydroxide solution in place of
the ~-methylglucamine there are obtained in analogous
manner~
the sodium salt of the iron(III) complex of ethylene-
diaminetetraacetic acid, C1OH12FeN2O8 . Na;
the sodium salt of the iron(III) complex of trans-1,2-
cyclohexylenediaminetetraacetic acid, C14H18FeN2O8 . Na
the disodium salt of the iron(III) complex of
,'
.
- ' . ' . - . '
. ~ ................................... .
~25~
33-
diethylenetrinitrolopenta(methanephosphonic acid),
CgH23FeN30l5P5 .2 N ;
the sodium salt of the iron(III) complex of 1,10-
diaza-4,7-dioxadecane-1,1,10,10-tetraacetic acid,
14 20FeN210 . Na;
- the sodium-salt of the iron(III) complex of ethylene
diaminetetraacetohydroxamic acid, C1 oH 16~eN608 . Na.
In analogous manner, there are obtained with N-methyl-
glucamine:
the di-N-methylglucamine salt of the iron(III) complex
of diethylenetriamine-N,N,N',N",N"-pentaacetic acid,
20;
the N-methylglucamine salt of the iron(III) complex of
trans-1,2-cyclohexylenediamine-N,N,N',N'-tetraacetic
acid, C21H36FeN3 13'
the N-methylglucamine salt of the iron(III) complex of
ethylenediamine-N,N,N',NI-tetraacetic acid, C17H30Fe3013;
~ .
the tri-N-methylglucamine salt of the iron(III) complex
:
- ':
. . : :
. ' '
~s~
-3~-
of triethylenetetramine-N,N,N',N",N"',N"'-hexaacetic
acid, C39H78FeN7 27
By using diethanolami:ne in place of N-methyl-
glucamine there is obtained in analogous manner:
the di-diethanolamine salt of the iron(III) complex of
diethylenetriamine-N,N,N",N",N"-pentaacetic acid,
22H42Fe 5 14 ~ .
Example 8
Manufacture of the N-methylglucamine salt of the
gadolinium(III) complex of trans-1,2-cyclohexylene-
diamine-N,N,N',N'-tetraacetic acid, C21~36GdN3O13
....
20.78 g ( = 60 mmol) of trans-1,2-cyclohexylene-
diamine-N,N,N',N'~tetraacetic acid are suspended in
150 ml of water. After the addition of 11.7 g
( = 60 mmol) of N-methylglucamine, an almost clear
solution is obtained to which 10.88 g ( = 30 mmol) of
gadolinium oxide (Gd2O3) are added. The suspension
again obtained is heated for 6 hours at 95C. The
small amount of undissolved material is filtered off
and the filtrate is concentrated to dryness in vacuo.
.
.
. ' '. ~ ` ' ''
.
.
~ ~dS~:~4~
-35-
The residue is dried in vacuo at 60C and pulverised.
38.6 g (92 % of the theoretical yield) of a white
powder of melting point 258-261~C are obtained.
Analysis:
(calculated~ C 36.25 H 5.22 N 6.04 Gd 22.60
(found) C 36.40 H 5.50 N 5.98 Gd 22.52
In analogous manner, by using sodium hydroxide
solution in place of N-methylglucamine, the sodium salt
of the gadolinium(III) complex of trans-1,2-cyclo-
10 hexylenediamine-N,N,N',N'-tetraacetic acid,
14 18 2 8 a, is obtained.
By using freshly precipitated chromium(III)
hydroxide, Cr(OH)3, the sodium salt of the chromium(III)
complex of ethylenediamine-N,N,N',N'-tetraacetic acid,
15 Cl0Hl2crN2o8 Na, is obtained-
Example 9Preparation of the disodium salt of the manganese(II)
complex of trans-1,2-cyclohexylenediamine-N,N,N',N'-
tetraaCetic acid, Cl4Hl6MnN2o8 . 2 Na
.
Under nitrogen, 34.6 g ( = 100 mmol) of
trans-1,2-cyclohexylenediamine-N,N,N',N'-tetraacetic
.
' ' ' - ' -
- . .
. . ,
. - - ~
. .
~ ,2
-36-
acid are suspended in 100 ml of water, and 11. 5 9
~ = 100 mmol) of manganese(II~ carbonate, MnCO3, are
added. The whole is heated to 95C and 200 ml of lN
sodium hydroxide solution are added dropwise thereto.
The clear solution is concentrated in vacuo and the
residue is dried in vacuo at 60C. 40.8 g t92 % of
the theoretical yield) of a pink-coloured powder are
obtained.
Analysis:
(calculated) C 37.94 H 4.09 N 6.32 Mn 12.40
(found) C 37.78 H 4.12 N 6.20 Mn 12.31
In analogous manner, there are obtained:
from copper(II) carbonate the disodium salt of the
copper(II) complex of trans-1,2~cyclohexylenediamine-
15 tetraacetic acid, Cl4Hl8CuN2O8 . 2 Na;
from cobalt(II) carbonate the disodium salt of thecobalt(II) complex of trans-1,2-cyclohexylenediamine-
tetraacetic acid, C14H18~oN2O8 2 Na;
from nickel(II) carbonate the disodium salt of the
nickel(II) complex of trans-1,2-cyclohexylenediamine-
tetraaCetic acid, Cl4Hl8NiN2o8 . 2 Na.
.
,'' ' ', :
~J~
37-
By using N-methylglucamine in place of sodium
hydroxide solution there are obtained:
the di-N-methylglucamine salt of the manganese(II)
complex of trans-1,2-cyclohexylenediaminetetraacetic
5 acid, C28H54MnN4O18;
the di-N-methylglucamine salt of the manganese(II)
complex of DL-2,3-butylenediaminetetraacetic acid,
C26B52MnN4ol8;
the di-N-methylglucamine salt of the manganese(II)
lO complex of ethylenediamine-N,N,N',N'-tetraacetic acid,
C24H48MnN4ol8;
the di-N-methylglucamine salt of the manganese(II)
complex of DL-1,2-butylenediamine-N,N,N',N'-
tetraacetic acid, C26H52MnM4O18;
15 the di-N-methylglucamine salt of the manganese(II)
complex of DL-1,2-propylenediamine-N,N,N',N'-
~etraacetic acid, C25H50MnN4O 8;
the tri-N-methylglucamine salt of the manganese(II)
complex of diethylenetriaminepentaacetic acid,
20 C35H72MnN62s
.~ '; ' .
- : -
~S~
-38-
with nickel(II) carbonate,. NiC03, there is
obtained:
the di-N-methylglucamine salt of the nickel(II) complex
of ethylenediamine-N,N,N',N'-tetraacetic acid,
C24H48NiN~018;
with cobalt(II) carbonate, CoC03, there is
obtained:
the diethanolamine salt of the cobalt(II) complex of
ethylenediamine-N,N,N',N'-tetraacetic acid,
C14H28CN410;
with copper(II) carbonate, CuC03, and
ethanolamine there is obtained:
'.
the diethanolamine salt of the copper(II) complex of
ethylenediamine-N,N,N',N'-tetraacetic acid,
C14~28CUN410;
with manganese(II) carbonate, MnC03, and
.
diethanolamine there is obtained:
the tri-diethanolamine salt of the manganese~II)
,
.
. . . : . .
.: ,
- - -, : . . . : .
., ~ . :
~i62~
--39--
complex of diethylenetriamine-N,N,N',N",N"-pentaacetic
acid, C26H54MnN616;
with manganese(II) carbonate, MnCO3, and
morpholine there is obtained:
',
5 the dimorpholine salt of the manganese(II) complex of
ethylenediamine-N,N,N",N"-tetraacetic acid,
Cl8H32MnN4o1o.
Example 10
Preparation of the N-methylglucamine salt of the
lO gadolinium(III) complex of ethylenediamine-N,N,N',N'-
tetraacetic acid, C17H30GdN3O13
29.2 g ( = 100 mmol) of ethylenediamine-N,N,N',N'-
tetraacetic acid are suspended in 100 ml of water and
heated to 95C wi~h 18.1 g ( = 50 mmol) of
15 gadolinium(III) oxide, Gd2O3. During the heating-up
process, 19.5 g ( = 100 mmol) of N-methylglucamine are
added in portions. After approximately 3 hours, a
clear solution is obtained which is filtered and
concentrated to dryness ln vacuo. The residue is
20 dried at 60C in vacuo. 61.3 g (95 % of the
theoretica]L yield) of a white powder having an
indeterminate melting point are obtained.
:, : . -
.
- . . - . . : '
- : .
'~', . ' , ~ ' ' . ' ' -
.
,
-40-
Analysis:
(calculated) C 31.B2 H 4.71 N 6.55 Gd 24.51
(found) C 31.65 ~ 4.59 N 6.52 Gd 24.56
In analogous manner, there are obtained:
with dysprosium(III) oxide Dy2O3:
the N-methylglucamine salt of the dysprosium(III)
complex of ethylenediamine-N,N,N',N'-tetraacetic acid,
Cl7H30DyN3ol3.
By using 1,10-diaza-4,7-dioxadecane-1,1,10,10-
tetraacetic acid in place of ethylenediamine-~,N,N',N'-
tetraacetic acid there is obtained:
the N-methylglucamine salt of the gadolinium~III)
complex of 1,10-diaza-4,7-dioxadecane-1,1,10,10-tetra-
acetic acid, C21H38GdN315;
Similarly, by using 1,2-diphenylethylenediamine-
tetraacetic acid there is obtained- -
the N-methylglucamine salt of the gadolinium(III)
complex of 1,2-diphenylethylenediaminetetraacetic acid,
291~38N3O13Gd;
By using lead(II) oxide, PbO, and sodium
chloride, there is obtained:
- ' , . .
'' '' " ~ '
~5~
-41-
the disodium salt of the lead(II) complex of ethylene-
diaminetetraacetic acid, C1 oH 12N2O~Pb . ~ Na;
By using freshly precipitated chromium(III)
hydroxide, Cr(OH)3, there is obtained:
the sodium salt of the chromium(III) complex of
ethylenediaminetetraacetic acid, C10H12Cr~2O8 . Na; and
analogously:
the sodium salt of the gadolinium(III) complex of
ethylenediaminetetraacetohydroxamic acid,
10 16 6 8 a; and
the sodium salt of the gadolinium(III) complex of
ethylenediamine-N,N,N',N'-tetraacetic acid,
c 1 oH 12GdN2O8 . Na.
Example 11
Preparatio-n of the sodium salt of the gadolinium(III)
complex of 1,4,7,10-tetraazacyclododecane-N,N',N",N"~-
tetraacetic acid, C16H24GdN4O8 . Na
. .... .. ..
4.0 g ( = 10 mmol) of 1,4,7,10-tetraazacyclo-
dodecane-N,N',N",N"'-tetraacetic acid are suspended in
20 ml of water, and 10 ml of lN sodium hydroxide
: solution are added. 1.8 9 ( = 5 mmol) of
- ~ . , . ~ , .
.. . . . .
., . :
- . : ~ . .. : . .
~ ~ .. ... .. .
.
:: . . '
?L 25~,L~
--42--
gadolinium(III~ oxide, Gd2O3, are added and the
suspension is heated for 2 hours at 50C. The clear
solution is filtered and concentrated ln vacuo. The
residue is dried and pulverised. 5.5 g (95 % of the
theoretical yield) of a white powder are obtained.
Analysis:
(calculated) C 33.10 H 4.17 N 9.65 Gd 27.08
(found) C 33.01 H 4.20 N 9.57 Gd 27.16
In analogous manner there are obtained:
the N-methylglucamine salt of the gadolinium(III)
coMplex of 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-
tetraacetic acid, C~3H42GdN5O13;
the sodium salt of the gadolinium(III) complex of
1,4,8,11-tetraazacyclotetradecane-N,N',N",N"'-tetra-
' 18H28GdN48 Na.
Example 12
Preparation of the tetra-N-methylglucamine salt of the
gadolinium(III) complex of ethylenedinitrolo-
tetrakis(methanephosphonic acid), C34H85GdN6O32P4
9. 11 g ( = 20~ mmol) of ethylenedinitrolo-
:
,
', ,- ~ .' ' .'
-
~s~
-43-
tetrakis(methanephosphonic acid) are suspended in
150 ml of water and the suspension is adjusted to a p~
of 5 with the corresponding amount of N-methyl-
glucamine. 3.6 g ( = 10 mmol) of gadolinium(III)
oxide, Gd2O3, are added thereto and the whole i~ heated
to 70~C. After approximately 1 hour, a clear solution
is obtained to which there is added the remaining
- portion of the N-methylgluca~ine. A total of 15.6 g
( = 80 mmol) of N-methylglucamine is used. The
solution is concentrated to dryness in vacuo and the
gel-like residue remaining is introduced into 200 ml of
acetonitrile. The mixture is stirred for approximately
20 hours at 30 and the resultin~ fine precipitate is
suction-filtered. After drying in vacuo at 40C,
23.4 9 (85 % of the theoretical yield) of a white
powder of melting point 115-118C are obtained.
' '
.
Analysis:
- (calculated) C 29.78 H 6.25 N 6.13 P 9.04 Gd 11.47
(found) C 29.85 H 6.57 N 5~98 P 8.78 Gd 11.26
; 20 In analogous manner there are obtained:
the hepta-N-methylglucamine salt of the gadolinium(III)
complex of diethylenetriamine-N,N,N',N",N"-penta(methane-
phosphonic acid)~ C58H144GdN1050 5'
' ~ . ' ' . -:
~"' ' ~ '
1;~5~
-44-
and, by using sodium hydroxicle solution in place
of N-methylglucamine.
the disodium salt of the gadolinium(III) complex of
diethylene-trinitrolo~penta(methanephosphonic acid),
9 23 3 15P5 2 Na.
Example 13
Preparation of the disodium salt of the manganese~
complex of ethylenedinitrolo-tetra(acetohydroxamic
' 10 16 6 8 . 2 Na
2.30 g of manganese(II) carbonate and 7.05 g of
ethylenedinitrolo-tetra(acetohydroxamic acid) are
refluxed in 18 ml of water for 3 hours. The pH is then
ad]usted to 7 by the addition of dilute sodium
hydroxide solution and 40 ml o acetone are added
dropwise. After stirring for several hours in an ice
bath, the crystallisate which has separated is suction-
filtered, washed with acetone and dried at 50C in
vacuo. A dihydrate is obtained in quantitative yield
in the form of a white powder having a melting point
above 300C.
Mn: (calculated) 11.30
(found) 11.12
.
- : : . ~ . . .
-
~' ' : '- ' ': ' , ,
-45-
Example 1g
Preparation of a mixed salt solution comprising the
sodium and the N-methylglucamine salt of the
gadolinium(III) complex of diethylenetriaminepenta-
acetic acid
a) preparation of the mono-N-methylglucamine salt of
the complex, C21H37GdN4O15
. .
195.2 g (1 mol) of N-methylglucamine are
dissolved in 7 litres of water. 393.3 g (1 mol) of
diethylenetriaminepentaacetic acid and 181.3 9 (0.5 mol)
of gadolinium(IlI) oxide, Gd2O3, are added and whole is
refluxed for 2 hours. The filtered clear solution is
spray-dried. A white crystalline powder having a water
content of 2.6 %, which sinters at 133C and melts,
15 with foaming, at 190C is obtained.
Gd: ~calculated) 21.17
(found) 21.34
b) preparation of the neutral mixed salt solution
730.8 g ( = 1 mol) of the salt ohtained in a) are
20 suspended in 630 ml of water p.i. (~ injectione), and
- ~ . ~ , ' , -
.' '' '' ~ ~ ' : '
,
'., ' ' : -
.
~ 25 ~ J
-46-
40 g ( = 1 mol) of caustic soda powdee are added in
portions. The neutral solution is made up to 1000 ml
with water ~ , introduced into bottles over a
pyrogen filter and heat-sterilised. This 1 molar
sol~tion contains 753.8 g of the mixed salt per litre.
Example 15
Preparation of a solution of the di N-methylglucamine
salt of the gadoliniumtIII) complex of die~hylene-
triaminepentaacetic acid
-
535.0 g ( = 730 mmol) of the salt described in
Example 5 are made into a paste in 500 ml of water
p.i. and dissolved by adding 142.4 g ( = 730 mmol) of
N-methylglucamine at pH 7.2. The solution is then made
up to 1000 ml with water p.iol is introduced into
ampoules and heat-sterilised.
Example 16
.
Preparation of a solution of the disodium salt of the
gadolinium(III) complex of diethylenetriaminepenta-
acetic acid
.. _ . _ .. . _ . . . . _ _ _
~o 485.1 9 ( = 820 mmol) of the disodium salt
obtained in Example 6 are made into a paste in 500 ml
- ~ ~ - - ~ ., - '
,
.
.
-:. ~ ~ ' '
'
~z~
-47
of water p.i.. The volume is then made up to 1000 ml
with water p.i. and the solution is introduced into
ampoules and heat-sterilised.
Example 17
Preparation of a solution oE the disodium salt of the
gadolinium(III) complex of 13,23-dioxo-15,18,21-tris-
(carboxymethyl)-12,15,18,21,24-pentaazapentatria-
contanoic diacid
392.0 g ( = 400 mmol) of the sal~ described in
lO Example 2 are made into a paste in 500 ml of water
p.i. and dissolved by making up the volume to 1000 ml
with water p.i. while heating slightly. The solution
is placed in bottles and heat-sterilised.
Example 18
15 Preparation of a solution of the N-methylglucamine salt
of the gadolinium(III) complex of 1/4,7,10-tetraaza-
cyclododecanetetraacetic acid
.. ..
370.9 g ( = 500 mmol) of the complex salt
described in Example 11 are made into a paste in 500 ml
20 of water p.i. and dissolved by making up the volume
to 1000 ml with water p.i.. The solution is
- ~ ~ '~ . ' : ,
. . .
-
-
- ' . .
.~ ~" ~ .
3~ ;2J~
-~8-
introduced into ampoules and heat-sterilised.
Example 19
Preparation of a solution of the di-N-methylglucamine
salt of the manganese(II) complex of trans-1,2-
S cyclohexylenediaminetetraacetic acid
:
395.9 g ( = 500 mmol) of the comple~ salt
described in Example 9 are suspended in 500 ml of water
p i.. 1.3 9 of ascorbic acid are added and the suspension
is dissolved by making up the volume to 1000 ml with
water p The solution is sterile filtered and
placed in ampoules.
.
Example 20
Preparation of a solution of the tri-N-methylglucamine
salt of the manganese(II) complex of diethylenetriamine-
pentaacetic acid
-
514.4 9 ( = 500 mmol) of the complex salt
described in Example 9 are suspended in 600 ml of water
~ . 1.3 g of ascorbic acid are added and the solid
matter is dissolved by making up the volume to 1000 ml
with water p.i.. After being sterile-filtered, the
. . :
.
~ ~ ~ . ' . ' ' ' .'
- ~9 -
solution is placed in ampoules.
Example 21
Preparation of a solution o the di-N-methylglucamine
salt of the iron(III) complex of diethylenetriamine-
5 pentaacetic acid
44.6 g ( = 0.1 mol) of the iron~III) complex of
diethylenetriaminepentaacetic acid obtained in Example
7 are suspended in 40 ml of water ~. After the
addition of 0~18 g of tromethamine hydrochloride and
lO 39.1 g ( = 0.2 mol) of N-methylglucamine, the solid
matter is dissolved at the neutral point, the solution
is made up to 100 ml with water ~ , introduced into
ampoules and heat-sterilised.
Exam~le 22
.
15 Preparation of a solution of the gadolinium(III)
complex of nitrolotriacetic acid
- --
1.9 g ( = 10 mmol) of nitrolotriacetic acid and
1.8 g ( = 5 mmol) of gadolinium(III) oxide are
dissolved in 100 ml of water ~ while heating. The
20 solution is introduced into ampoules and heat-
sterilised.
,
'.''; '' ~ . ~ , ' .: ,' ' '
- .
. .
- - . , . ~ : .
: ~ , , ~ , .. . :
. ~ - -. . .
:. .:' ~ ' ' ~ ,
.~ . . . ..
-50-
Example 23
Preparation of a solution of the N-methylglucamine salt
of the gadolinium(III~ complex of ethylenediaminetetra-
acetic acid
.:
38.52 g ( = 60 mmol) of the substance described
in Example 10 are dissolved in 70 ml of water p.i..
After the addition of 0.12 g of tromethamine, the
solution is made up to lQ0 ml with water p.i. J placed
in ampoules and heat-sterilised.
-
Example 24Preparation of a solution of the di-N-methylglucamine
salt of the dysprosium(III) complex of diethylene-
triaminepentaacetic acid
. .
35.7 g ( = 60 mmol) of the dysprosium(III)
complex of diethylenetriaminepentaacetic acid (water
content 8.0 %) are suspended in 70 ml of water p.i.
and dissolved at pH 7.5 by adding 21.2 g ( = 120 mmol)
of N-methylglucamine. The solution is then made up to
100 ml with water p.i., placed in ampoules and heat-
20 sterilisecl.
' -': - , ' : '
., ~ . .
'~' ':
~ 2
-51-
Exam~le 25
Preparation of a solution of the N-methylglucamine salt
of the gadolinium(III) complex of trans-1,2-cyclo-
hexylenediaminetetraacetic acid
- ---- . ..
555.8 g ( - 0.8 mol) of the salt~described in
Example 8 are dissolved in water ~ to a volume of
1000 ml. After filtration over a pyrogen filter, the
solution is placed in ampoules and heat-sterilised~
Example_26
Preparation of a solution of the N-methylglucamine salt
of the ruthenium(lII) complex of 1,10-diaza-4,7-
dithiadecane-l,l,10,10-tetraacetic acid
-
:
15.6 g ( 2 0.03 mmol) of the ruthenium(III)
complex of 1,10-diaza-4,7-dithiadecane-1,1,10,10-tetra-
acetic acid are suspended in 50 ml of water p.i. ~nddissolved at pH 7.5 by adding 5.9 g ( = 0.03 mol) of
N-methylglucamine. The solution is made up.to 1000 ml
.`................. with water p.i., placed in ampoules and heat-
sterilised.
~ ' '
- ' . `.~ ' :
: . . . - '. :
: ' :
:
~ ~l5
-52-
Exam~le 27
Preparation of a solution of the dilysine salt of the
gadolinium(III~ complex of diethylenetriaminepentaacetic
acid
. . _ . _ . . .
273.8 g' ( = 0.5 mol) of the gadolinium(III)
- complex of diethylenetriaminepentaacetic acid are
suspended in 500 ml of water p.i.. 292.4 g ( = 1 mol)
of lysine are added, the whole is stirred for several
hours while heating gently and the volume is then made
up to 1000 ml with water p .. The solution is placed
in bottles and heat-sterilised.
Example 28
Preparation of a solution of the tri-N-methylglucamine
salt of the molybdenum(VI) complex of diethylene-
triaminepentaacetic acid
. _ _ _ . . ...
18.8 g ( = 0.28 mol) of the complex H3[Mo2O2(OH)4 .- C14H23N3Olo] are suspended in 50 ml of water p.i. and
~ dissolved at the neutral point by adding 16.4 9
( = 0.84 mol) of N-methylglucamine. 0.15 9 of
tromethamine is added, the solution is made up to 1 no ml
with water p.i._, s~bjected to sterile filtration and
placed in ampoules.
: . .
: . ~ ~ ' . . .
'
5 ~ ~,4 ~ ~
!
--53--
Example 29
Preparation of a solution of the disodium salt of the
manganese(II) complex of ethylenediaminetetraacetic
acid
. .
343,2 g ( = 1 mol) of the manganese(II) complex
of ethylenediaminetetraacetic acid are suspended in
500 ml of water p.i. and dissolv~ed at the neutral
point by adding, in portions, 80 g ( = 2 mol) of
caustic soda. After the addition of 1.5 g of
tromethamine, the solution is made up to 1000 ml with
water p.i., placed in bottles and heat-sterilised.
Example 30
Preparation of a solution of the sodium salt of the
iron(III) complex of ethylenediaminetetraacetic acid
-
345.7 g ( = 1 mol) of the iron(III) complex of
ethylenediaminetetraacetic acid are suspended in 500 ml
of water p.i. and dissolved at the neutral point by
adding, in portions, 40 g ( = 1 mol) of caustic soda.
After the addition of 1.5 g of tromethamine, the
solution is made up to 1000 ml with water p.i.,
placed in bottles and heat-sterilised.
.
, ` , ~ ' . -
:~2S~
Example 31
Preparation of a solution of the disodium salt of the
iron(III) complex of diethylenetriaminepentaacetic acid
. _ _ . _ ....
334.6 g ( = 0.75 mol) of the iron(III) complex of
diethylenetriaminepentaacetic acid are suspended in
500 ml of water p.i. and dissolved at the neutral
point ~y adding, in portions, 60 g ~ = 1.5 mol) of
caustic soda. The solution is made up to 1000 ml with
water p , placed in bottles and heat-sterilised.
Example 32
Preparation of a solution of the sodium salt of the
gadolinium(III) complex of trans-1,2-cyclohexylene-
diaminetetraacetic acid
_
558.6 g ( = 1 mol) of the sodium salt of the
complex salt listed in Example 8 are dissolved in water
and made up to 1000 ml. The solution is placed
in bottles and heat-sterilised.
Example 33
Preparation of a solution of the N~methylglucamine salt
'
- " , .
~5~
-55-
of the gadolinium(III) complex of 1,2-diphenylethylene-
diaminetetraacetic acid
.
396.9 g ( = 500 mmol) of the N-methylglucamine
salt of the complex salt containing gadolinium and the
1,2-diphenylethylenediaminetetraacetic acid radical
listed in Example 10 are macle into a paste in 600 ml of
water p.i. and dissolved by making up the volume to
1000 ml. The solution is placed in ampoules and heat-
sterilised.
; 10 Example 34
Preparation of a solution of the sodium salt of the
iron(III) complex of ethylenediaminetetraacetic acid
. . _ _ . . _ . .
183.5 g ( = 500 mmol) o the sodium salt of the
complex salt of iron and ethylenediaminetetraacetic
acid listed in Example 7 are made into a paste in
500 ml of water ~ ~. 1.O g of tromethamine are added,
the volume is made up to 1000 ml with water p.i., and
the solution is placed in ampoules and heat-sterilised.
Example 35
Preparation of a solution of the di-N-methylglucamine
.
..
.
': ,, ' ' - ' ~ ': ' ' ~- . . ,
. . . .
~95~2f~3
-56-
salt of the lanthanum(lII) complex of diethylene-
triaminepentaacetic acid
459,8 g ( = 500 mmol) of the di-N-methylglucamine
salt of the complex salt containing lanthanum and the
diethylenetriaminepentaacetic acid radical listed in
Example 5 are made,into a paste in 650 ml of water
E~ and dissolved by making up the volume to 1000 ml
with water p i.. The solution is placed in ampoules
and heat-sterilised.
Example 36
Preparation of a solution of the di-N-methylglucamine
salt of the bismuth(III~ complex of diethylenetriamine-
pentaacetic acid
. . . -- .
692.8 9 ( = 700 ml) of the di-N-methylglucamine
salt of the complex salt containing bismuth and the
diethylenetriaminepentaacetic acid radical listed in
Example 5 are made into a paste in 600 ml of water
and, after ~he addition of 1.8 9 of
tromethamine, dissolved by making up the volume to
1000 ml with water p.i. while heating slightly. The
solution is placed in ampoules and heat-sterilised.
- :
,
' ' ~ ' , ~ '
~2.~
-57-
Example 37
Preparation of a solution of the di-N-methylglucamine
salt of the holmium(III) complex of diethylenetriamine-
pentaacetic acid
662.0 g ~ = 700 mmol) of the di-N-methylglucamine
salt of the complex salt containing holmium and the
diethylenetriaminepentaacetic acid radical listed in
Example 5 are made into a paste with 600 ml of water
p.i. and, after the addition of 1.8 g of
tromethamine, dissolved by making up the volume to
1000 ml with water p.i. while heating slightly. The
solution is placed in ampoules and heat-sterilised.
Example 38
Preparation of a solution of the di-N-methylglucamine
salt of the ytterbiumtIII) complex of diethylene-
triaminepentaacetic acid
.
476.9 g ( = 500 ml) of the di-N-methylglucamine
salt of the complex salt containing ytterbium and the
diethylenetriaminepentaacetic acid radical listed in
20 Example 5 are made into a paste with 650 ml of water
p.i. and, after the addition of 1~5 g of
tromethamine, dissolved by making up the volume to
,
,
s~
-58-
1000 ml with water ~ . The solutic)n is placed in
ampoules and heat-sterilised.
Example_39
Preparation of a solution of the disodium salt of the
lanthanum(III) complex of diethylenetriaminepentaacetic
acid
. . _ _ .
573.2 g ( = 1000 mmol) of the disodium salt of
the complex salt containing lanthanum and diethylene-
triaminepentaacetic acid radical listed in Example 6
10 are made into a paste in 650 ml of water p.i. and
dissolved by making up the volume to 1000 ml with water
p i.. The solution is placed in ampoules and heat-
sterilised.
Example 40
15 Preparation of a solution of the disodium salt of the
dysprosium(III) complex of diethylenetriaminepenta-
acetic acid
. .
477.4 9 ( = 800 mmol) of the disodium salt of the
complex sal~ containing dysprosium and the
20 diethylenetriaminepentaacetic acid radical listed in
Example 6 are made into a paste in 600 ml of water p.i.
. , - . .
, ~ `' ,' '- ' , ~ '
'' ~
~L25~
-59-
and dissolved by making up the volume to 1000 ml with
water p i.. The solution is placed in ampoules and
heat-sterilised.
Exam~le 41
Preparation of a solution of the disodium salt of the
holmium(III) complex of diethylenetriaminepentaacetic
acid
299.6 g ( = 500 mmol) of the disodium salt of the
complex salt containing holmium and the diethylene-
triaminepentaacetic acid radical listed in Example 6are made into a paste in 500 ml of water p and
dissolved by making up the volume to 1000 ml with water
~_~ . The solution is placed in ampoules and heat-
sterilised.
Example 42
Preparation of a solution of the disodium salt of the
ytterbium~III) complex of diethylenetriaminepentaacetic
acid
. . .
.
303.5 9 ( = 500 mmol) of the complex salt
containing ytterbium listed in Example 6 are made into
a paste in 500 ml of water p.i. and dissolved by
.: ' .
. . ~ . - : .
,
~25~
-60-
making up the volume to 1000 ml with water p.i.. The
solution is placed in ampoules and heat-sterilised.
Example 43
Preparation of a solution of the tetra-N-methylglucamine
salt of the gadolinium(III) complex of
ethylenedinitrolo-tetrakis(methanephosphonic acid)
. _ _ _ .., . . _ .
137.1 g ( = 100 mmol) of the complex salt
described in Example 12 are made into a paste in 500 ml
of water p.i. and, after the addition of 0.8 9 of
tromethamine, dissolved by making up the volume to
1000 ml with water E~. The solution is placed in
ampoules and heat-sterilised.
Example 44
Preparation of a solution of the gadolinium(III)
15 complex of N'-(2-hydroxyethyl)-ethylenediamine-N,N,N'-
triacetic acid
_
1.9 g ( = 6.7 mmol) of N'-(2-hydroxyethyl)-ethyl-
enediamine-N,N,N'-triacetic acid and 1.2 g ( = 3.35 mol)
of gadolinium(III) oxide are dissolved in 6 ml of water
20 ~ while heating. The solution is placed in
. , ; .
''- ' ' . ~, . ' ' . ' - ~:
.,
- . ,
'
. , ~ , .
~2S~
-6 1 -
ampoules and heat-sterilised.
Example 45
Preparation of a solution of the disodium salt of the
manganese(II) complex of trans-1,~-cyclohexylene-
diaminetetraacetic acid
--
Under nitrogen, 44.3 g ( = 100 mmol) of the
' complex salt described in Example 9 are made into a
paste in 60 ml of water p.i. and dissolved by making
up the volume to 100 ml with water ~. The solution
is placed in ampoules and heat-sterilised.
Example 46
Preparation of a solution of the sodium salt of the
gadolinium(III) complex of 1,4,8,11-tetraazacyclotetra-
decane-N,N',N",N"'-tetraacetic acid
. _
55?.6 9 ( = 1 mol) of the complex salt containing
gadolinium and the 1,4,8,11-tetraazacyclotetradecane-
tetraacetic acid radical listed in Example 11 are
dissolved in water ~ and made up to 1000 ml. The
solution is placed in bottles and heat-sterilised.
- , , ~ : .,
: : ~ - : .. . .
. ~ ., . , .: . - .
- . . , : ,
., ~ : -- . . - ' :
~. , . - : . . . - :
~25~
-62-
Example 47
Preparation of a solution o the disodium salt of the
bismuthtIII~ complex of diethylenetriaminepentaacetic
acid
_ _
~3.4 g ( = 50 mmol) of bismuth(III) oxide are
suspended in 50 ml of water p After the addition
of 39.3 g ( = 100 mmol) of diethylenetriaminepentaacetic
acid and 4.0 g ( = 50 mmol) of caustic soda, the whole
is refluxed until a clear solution is obtained. The
solution, cooled to room temperature, is neutralised by
adding 4.0 g of caustic soda and made up to 100 ml with
water p.i.. The solution is introduced into ampoules
and heat-sterilised.
Example 48
lS Preparation of a solution of the disodium salt of the
samarium(III) complex of diethylenetriaminepentaacetic
acid
58.5 g ( = 100 mmol) of the complex salt
containing samarium listed in Example 6 are dissolved
in 65 ml of water ~ ~ while heating. Water ~ is
added to make a total volume of 100 ml, and the
. .
'
- ' ~ ' ~ . ' - ~ ,
'' . . ~ , ' . .
.
-63-
solution is introduced into ampoules and heat-
sterilised.
Example 49
Preparation of a solution oE the di-N-methylglucamine
salt of the gadolinium(III) complex of 13,23-dioxo-15,
18,21-tris(carboxymethyl)-12,15,18,21,24-pentaazapenta-
triacontanoic diacid
.
- 130.4 g ( = 100 mmol) of the di-N-methylglucamine
complex salt listed in Example 2 are made into a paste
- 10 in 250 ml of water ~ and dissolved while heating.
The solution is made up to 500 ml with water p.i.,
introduced into ampoules and heat-sterilised.
,: .
Example 50
Preparation of a solution of the di-N-methylglucamine
salt of the manganese(II) complex of ethylenediamine-
tetraacetic acid
. _ .
3.68 g ( = 5 mmol) of the complex salt containing
manganese and the ethylenediaminetetraacetic acid
radical listed in Example 9 are dissolved in 70 ml of
20 water p.i., and 0.4 g of sodium chloride is added to
the solution. The solution is then made up to 100 ml
. ' ' . ~
'
"
-64-
with water p.i. and introduced into ampoules through
a sterile filter. The solution is at 280 m~sm isotonic
with blood.
Example 51
Preparation o~ a solution of the disodium salt of the
gadolinium(IlI) complex of diethylenetrinitrolo-penta-
(methanephosphonic acid)
.~
38.57 g ( = 50 mmol) of the disodium salt of the
complex containing gadolinium and the diethylene-
trinitrolo-penta(methanephosphonic acid) listed in
Example 12 are made into a paste with 50 ml of water
p The pH is adjusted to 7.2 by adding caustic
soda powder and the volume is made up to 100 ml with
water p.i.. The solution is introduced into ampoules
and heat-sterilised.
Examjele 52
Preparation of a solution of the trisodium salt of the
manganese(II) complex of diethylenetriaminepentaacetic
acid
. . _ _ . _
20~nder nitrogen, 39.3 9 ( = 100 mmol) of
- diethylenetriaminepentaacetic acid are suspended in
:
.
.
~s~
-65-
100 ml of water ~, and 11.5 g of manganese(II)
carbonate are added. The whole is heated to 95C and
300 ml of lN sodium hydroxide solution are added
dropwise. The neutral solution is sterile-filtered and
introduced into ampoules.
Example 53
Composition of a powder for the preparation of a
suspension
.
4.000 g gadolinium(III) complex of diethylenetri-
aminepentaacetic acid (water content 8.0 %)
3.835 g saccharose
0.100 g polyoxyethylenepolyoxypropylene polymer
0.005 g aromatic substance
8.000 g
Example 54
Preparation of a solution of the gadolinium(III)
complex of the conjugate of diethylenetriaminepenta-
acetic acid with human serum albumen
~ . ~
,
10 mg of 1,5-bis-(2,6-dioxomorpholino)-3-azapen-
tane-3-acetic acid are added to 20 ml of a solution of
3 mg of the protein in a 0.05 molar sodium bicarbonate
:: . .
-66-
buffer (pH 7-8). The whole is stirred for 30 minutes
at room temperature and is then dialysed against a 0.3
molar sodium phosphate buffer. 50 mg of
gadolinium(III) acetate are then added and purification
is effected by gel chromatrography over a Sephadex G25
column. The fraction obtained is sterile-filtered and
placed in multi~dose phials. Freeze-drying produces a
dry preparation that can be stored.
In an analogous manner, there is obtained with
immunoglobulin a solution of the corresponding complex
conjugate.
Example 55
Preparation of a solution of the gadolinium(III)
complex of the conjugate of diethylenetriaminepenta-
acetic acid (DTPA~ with a monoclonal antibody
_ _ . .......... . . . _ _ . ..
1 mg of a mixed DTPA anhydride (obtained, forexample-, from DTPA and isobutyl chloroformate) is added
to 20 Ul of a solution of 0.3 mg of a monoclonal anti-
body in a 0.05 molar sodium bicarbonate buffer (pH 7-8)
and the whole is stirred for 30 minutes at room
temperature. Dialysis is carried out against a 0.3
molar sodium phosphate buffer, and 2 mg of the
gadolinium(III) complex of ethylenediaminetetraacetic
acid (EDTA) are added to the antibody fraction
. : ,
"~ ~ ' ' .
.~ .
~ ~ 5 ~ ~,,., LÇ4 ~J
-67-
btained. After purification by gel chromatography
ra d ~
over Sephadex G25, the sterile-filtered solution is
placed in multi-dose phials and freeze-dried.
Using the mixed anhydride of trans-1,2-diamino-
cyclohexanetetraacetic acid (CDTA) there is obtained,in analogous manner, a solution of the corresponding
gadolinium(III) complex of the CDTA antibody.
Using the manganese(II) complex of ethylene-
diaminetetraacetic acid there is obtained in an
analogous manner the manganese(II) complexes of the
antibodies coupled with DTPA or CDTA.
Example 56
Preparation of a solution of the gadolinium(III)
complex of the conjugate of 1-phenyl-ethylenediamine-
tetraacetic acid with immunoglobulin
According to the procedure described in J. Med.Chem. 1974, vol. 17~ p. 1307, a 2 ~ solution of the
protein in a 0.12 molar sodium bicarbonate solution
containing 0.01 mol of ethylenediaminetetraacetic acid
20 is cooled to +4C and there is added dropwise the
proportion, equivalent to the protein, of a freshly
prepared ice-cold diazonium salt solution of 1~(p-
aminophenyl)-ethylenediaminetetraacetic acid. The
whole is stirred overnight (pH 8.1) at +4C and is
: : . . - ,
125~i~4~
-68-
then dialysed against a 0.1 molar sodium citrate
solution. When dialysis is complete, an excess of
gadolinium(III) chloride is added to the sol~1tion of
the conjugate and ultra-filtration is carried out to
remove ions. Finally, the sterile-filtered solution is
placed in multi-dose phials and freeze-dried.
Example 57
Preparation of a colloidal dispersion of a Mn(II) CDTA-
lipid conjugate
~ . . _ . _ . .
0.1 mmol of distearoylphosphatidylethanolamine
and 0.1 mmol of the bisanhydride of trans-1,2-diamino-
cyclohexanetetraacetic acid in 50 ml of water are
stirred at room temperature for 24 hours. 0.1 mmol of
manganese(II) carbonate is added and stirring is again
15 carried out at room temperature for 6 hours. After
purification over a Sephadex G50 column, the sterile-
filtered solution is placed in multi-dose phials and
freeze-dried.
A cclloidal dispersion of the gadolinium-DTPA-
20 lipid conjugate can be obtained analogously with
gadolinium(III) oxide.
.. . , : ~
-. ' - ' ~ ' "' .:
~' ' ' ' ' '
~z~
-69-
Example 5B
Preparation of lipsomes charged with gadolinium(III)-
DTPA
~..
According to the procedure described in Proc.
Natl. Acad. Sci. U.S.A. 75, 4194, a lipid mixture
comprising 75 mol % of egg phosphatidylchloline and
25 mol ~ of cholesterol is prepared as a dry substance.
500 mg thereof are dissolved in 30 ml of diethyl ether
and, in an ultrasonic bath, 3 ml of a 0.1 molar
solution of the di-~-methylglucamine salt of the
gadolinium(III) complex of diethylenetriaminepenta-
acetic acid in water p.i. are added dropwise
thereto. When the addition of the solution is
complete, the exposure to ultrasonic waves is continued
for 10 minutes and then concentration is carried out in
a rotary evaporator~ The gel-like residue is suspended
in a 0.125 molar sodium chloride solution and, at
0C, repeatedly freed of non-encapsulated contrast
agent portions by centrifugation (20000 g/29 minutes).
Finally, the lipsomes so obtained are freeze-dried in
multi-dose phials. The preparation is administered as
a colloidal dispersion in a 0.9 ~ by weight sodium
chloride so:Lution.
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