Note: Descriptions are shown in the official language in which they were submitted.
CA 02418790 2003-02-07
WO 02/14309 PCT/EPO1/08499
PerJluoroalkyl-Containing Complexes with Sugar Radicals,
Process for their Production and their Use
Description
The invention relates to the subjects that are characterized in the claims,
namely
perfluoroalkyl-containing metal complexes with sugar radicals of general
formula I, process for
their production and their use in NMR diagnosis and x-ray diagnosis,
radiodiagnosis and
radiotherapy, in MRT-lymphography and as blood-pool agents. The compounds
according to the
invention are quite especially suitable for intravenous lymphography, for
tumor diagnosis and for
infarction and necrosis imaging.
In nuclear magnetic resonance, the element fluorine is second in importance to
the
element hydrogen.
1) Fluorine has a high sensitivity of 83% of that of hydrogen.
2) Fluorine has only one NMR-active isotope.
3) Fluorine has a resonance frequency that is similar to hydrogen -- fluorine
and
hydrogen can be measured with the same system.
4) Fluorine is biologically inert.
5) Fluorine does not occur in biological material (exception: teeth) and can
therefore
be used as a probe or contrast medium against a background that is free of
interfering signals.
The effect of these properties is that fluorine occupies a broad space in
diagnostic patent
literature with magnetic nuclear resonance as a basis: fluorine-19-imaging,
functional diagnosis,
spectroscopy.
U.S. Patent 4,639,364 (Mallinckrodt) thus proposes
trifluoromethanesulfonamides as
contrast media for fluorine-19-imaging:
CF3 S 02NHz
CF3S02NH-CH2-(CHOH)a-CHZOH
German Patent DE 4203254 (Max-Planck-Gesellschaft), in which an aniline
derivative is
proposed:
CF3
CF3 ~ ~ NHCOCHz I
CF3
also relates to fluorine-19-imaging.
Fluorine-19-imaging is the subject of Application WO 93/07907 (Mallinckrodt),
in which
phenyl derivatives are also claimed as contrast media:
HO OH
O
CF3 CF3 HO O CF3 H
HO C ~ ~ CO H
z z N CF3
CF3 CF3 HO CF3 ~ H
O
HO OH
For fluorine-19-imaging, compounds of considerably simpler structure are also
claimed.
Thus, Patent US 4,586,511 (Children's Hospital Medical Center) mentions
perfluorooctylbromide
CF3(CFz)7-Br,
European Patent EP 307863 (Air Products) mentions perfluoro-15-crown-5-ether
CA 02418790 2003-02-07
y~ .<r
F F F F
F O O F
F F
F F
F O O F
F F
O
F ~ \
F F
F F F
and U.S. Patent US 4,588,279 (University of Cincinnati, Children's Hospital
Research
Foundation) mentions perfluorocarbon compounds such as perfluorocyclononane or
-octane,
perfluorinated ethers such as tetrahydrofuran
F F F
F
~F
/ \F
F O F
or diethers such as perfluoropropylene glycol-diether
CF3 CF3
F~O-CFZCF2CF2 O---~-~F
CF3 CF3
The compounds that are mentioned in Application WO 94/22368 (Molecular
Biosystems), e.g.,
O-CO-(CHZ)s-R
CH3(CH2)~-CH=CH,(CHZ)7-CO-O
O-CO-(CHZ}g-R
~CF3
-CF3
v
CF3
R= , die als
CF3
~CF3
CF3 which as
CA 02418790 2003-02-07
fluorine-containing radicals have the perfluorine-1 H group or 1 H-neopentyl
group, are also used
for fluorine-19-imaging.
U.S. Patent US 5,362,478 (VIVORX) indicates another structural type with
expanded
diagnostic use. in which the fluorocarbon/polymer shell combination is claimed
for imaging
purposes. Perfluorononane and human serum albumin are mentioned. This
combination proves
suitable, moreover, for using the fluorine atom as a probe for local
temperature measurement and
for determining the partial oxygen pressure.
Perfluorocarbons are also claimed in U.S. Patent 4,586,511 for oxygen
determination.
In German Patent DE 4008179 (Schering), fluorine-containing
benzenesulfonamides are
claimed as pH probes:
CF3
F ~ ~ NHSOzCHzCHzCOZH
For NMR diagnosis, compounds that contain iodine and fluorine atoms are also
claimed
as contrast-enhancing agents in WO 94/05335 and WO 94/22368 (both molecular
biosystems):
I
CHZ-CH-COzH
Et
I I
NHCOCF3
I / \ (CHZ)6-COZ C(CF3)s
I
/ \ (
I CHz)6-COZ C(CF3)s
HZN I
OCO-(CHZ)e
I
HzN
CA 02418790 2003-02-07
CA 02418790 2003-02-07
The fluorine-paramagnetic metal ion combination is also claimed for fluorine-
19-
imaging, specifically for open-chain completes in WO 94/22368 (Molecular
Biosystems) with,
e. g.:
COZ
N
HZC-HNOC~N~ ~N~CONH-CHz
~CO ~CO
2
\ /
C(CF3)3 C(CF3)3 Gd3+ C(CF3)3 C(CF3)3
and in EP 292 306 (TERUMO Kabushiki Kaisha) with, e.g.:
R
I
N
OZC~N~ ~N~COZ
Gd3+
~O CJ ~CO
z z
F F
-CHZCF3 , CHz ~ \ C(CF3)3 . --CHz ~ ~ F
CFs F F
/ \ CF3
-CH2CH2 N CF3 , -CHz
CF3
\ CF3
CF3
but also for cyclic compounds, as they are mentioned in EP 628 316 (TERUMO
Kabushiki
Kaisha)
-oc~~~
N N COZ OzC N N COZ
Gd3+ Gd3+
~N N\R , ~ U \R
CO ~ CO
2 Z
l
CF3 CF3 CF3
/ \
CF3 CF3
-CHz ~ ~ CF3 . CHZCHZ N CF
3
CF3 / \
CFA
The combination of fluorine atom and rare-earth metal is also claimed for NMR-
spectroscopic temperature measurements in DE 4317588 (Schering):
OzC
N N
Ln3+
C ~ off
~N
CF3
COz
Ln: Seltene Erde: La, Pr, Dy, Eu
[Key: Seltene Erde =_ Rare Earths]
While no interactions occur between the two nuclei in compounds that contain
the
elements fluorine and iodine, intensive interaction does occur in compounds
that contain fluorine
and paramagnetic centers (radicals, metal ions) and that are expressed in a
shortening of the
relaxation time of the fluorine nucleus. The extent of this effect depends on
the number of
CA 02418790 2003-02-07
unpaired electrons of the metal ion (Gd'+ > Mn2+ > Fe3+ > Cu2+) and on the
removal between the
paramagnetic ion and the 19F-atom.
The more unpaired electrons of the metal ion are present and the closer the
latter are
brought to the fluorine, the greater the shortening of the relaxation time of
the fluorine nucleus.
The shortening of the relaxation time as a function of the distance from the
paramagnetic
ion becomes apparent in all nuclei with an uneven spin number, thus also in
the case of protons,
and gadolinium compounds are therefore widely used as contrast media in
nuclear spin
tomography (Magnevist~Ry, Prohance~R~, Omniscan~R~, and Dotarern~R~).
In 1H-MR imaging (1H-MRI), however, relaxation time TI or T2 of the protons,
i.e.,
mainly the protons of water, and not the reaction time of the fluorine nuclei,
is measured and
used for imaging. The quantitative measurement for the shortening of the
relaxation time is
relaxivity [L/mmol~s]. Complexes of paramagnetic ions are successfully used
for shortening
relaxation times. In the following table, the relaxivity of several commercial
preparations is
indicated:
T1-Relaxivity in WaterTl-Relaxivity in Plasma
[L/mmol~s, 39C, 0.47 [L/mmol~s, 39C, 0.47
T] T]
MAGNEVIST~Rt 3.8 4.8
DOTAREMtR~ 3.5 4.3
OMNISCAN~R~ 3.8 4.4
i
PRO HANCE~R~ 3.7 4.9
Only interactions between protons and the gadolinium ion are found in these
compounds.
For these contrast media in water, a relaxivity of about 4 [L/mmol s] is thus
observed.
Both fluorine compounds for fluorine-19-imaging, in which the shortened
relaxation time
of the fluorine nucleus is used, and non-fluorine-containing compounds, in
which the relaxation
time of protons of water is measured, are thus used successfully for MR
imaging.
In the introduction of a perfluorocarbon-containing radical into a
paramagnetic contrast
CA 02418790 2003-02-07
medium, i.e., in the combination of properties that were previously known as
suitable only for
fluorine-imaging compounds, the relaxivity that relates to the protons of
water also quickly
increases, surprisingly enough, with compounds that were used for proton
imaging. It now
reaches values of 10-50 [L/mmol~s) in comparison to values of between 3.5 and
3.8 [L/mmol~s)
as they were already cited for a few commercial products in the table above.
Perfluoroalkyl-containing metal complexes are already known from DE 196 03
033.1.
These compounds, however, cannot be used satisfactorily for all applications.
Thus, there is still
a need for contrast media for the visualization of malignant tumors, lymph
nodes and necrotic
tissue.
Malignant tumors metastasize in clusters in regional lymph nodes, whereby
multiple
lymph node stations may also be involved. Lymph node metastases thus are found
in about 50-
69% of all patients with malignant tumors (Elke, Lymphographie (Lymphography),
in:
Frommhold, Stender, Thurn (eds.), Radiologische Diagnostik in Klinik and
Praxis [Radiological
Diagnosis in Clinical Studies and in Practice], Volume IV, Thieme Verlag
Stuttgart, 7th Ed.,
434-496, 1984). The diagnosis of a metastatic attack of lymph nodes is of
great importance with
respect to the treatment and prognosis of malignant types of diseases. With
modern imaging
methods (CT, US and MRS, lymphogenous evacuations of malignant tumors are
detected only
inadequately, since in most cases only the size of the lymph node can be used
as a diagnostic
criterion. Thus, small metastases in non-enlarged lymph nodes (< 2 cm) cannot
be distinguished
from lymph node hyperplasias without a malignant attack (Steinkamp et al.,
Sonographie and
Kernspintomographie: Differentialdiagnostik von reaktiver Lymphknoten-
vergrol3erung and
Lymphknoten-metastasen am Hals [Sonography and Nuclear Spin Tomography:
Differential
Diagnosis of Reactive Lymph Node Enlargement and Lymph Node Metastasis on the
Neck),
Radiol. Diagn. 33:158, 1992).
It would be desirable if a distinction could be made when using specific
contrast media
lymph nodes with metastatic attack and hyperplastic lymph nodes.
Direct x-ray lymphography (injection of an oily contrast medium suspension
into a
prepared lymph vessel) is known as an invasive method that is used only very
rarely and that can
CA 02418790 2003-02-07
9
visualize only small lymph drainage stations.
Fluorescence-labeled dextrans are also used experimentally in animal
experiments to be
able to observe lymphatic drainage after their interstitial administration.
All commonly used
markers for the visualization of lymph tracts and lymph nodes after
interstitial/intracutaneous
administration have in common the fact that they are substances with
particulate character
("particulates," e.g., emulsions and nanocrystal suspensions) or large
polymers (see above, WO
90/14846). Based on their inadequate local and systemic compatibility as well
as their small
lymphatic passageway, which causes inadequate diagnostic efficiency, the
previously described
preparations still do not prove optimally suitable for indirect lymphography,
however.
Since the visualization of lymph nodes is of central importance for the early
detection of
metastatic attack in cancer patients, there is a great need for lymph-specific
contrast medium
preparations for diagnosis of corresponding changes of the lymphatic system.
The highest possible contrast medium concentration and high stability are just
as
desirable as the diagnostically relevant, most uniform possible lymphatic
concentration over
several lymph stations. The burden on the overall organism should be kept low
by quick and
complete excretion of the contrast medium. A quick start-up, if possible as
early as within a few
hours after the administration of contrast media, is important for the
radiological practice. Good
compatibility is necessary.
Largely for this reason, it is desirable to have available lymph-specific
contrast media that
in a diagnostic session allow both the primary tumor and a possible lymph node
metastasizing to
be visualized.
Another important area in medicine is the detection, localization and
monitoring of
necroses or infarctions. Thus, myocardial infarction is not a stationary
process, but rather a
dynamic process, which extends over a long period (weeks to months). The
disease proceeds in
about three phases, which are not strictly separated from one another, but
rather are overlapping.
The first phase, the development of myocardial infarction, comprises the 24
hours after the
infarction, in which the destruction from the subendocardium to the myocardium
progresses like
a shock wave (wave front phenomenon). The second phase, the already existing
infarction,
CA 02418790 2003-02-07
10
comprises the stabilization of the area in which fiber formation (fibrosis)
takes place as a healing
process. The third phase, the healed infarction, begins after all destroyed
tissue is replaced by
fibrous scar tissue. During this period, an extensive restructuring takes
place.
Up until now, no precise and reliable process is known that enables the
current phase of a
myocardial infarction to be diagnosed in a living patient. To evaluate a
myocardial infarction, it
is of decisive importance to know how large the proportion of the tissue that
is lost in the
infarction is and at what point the loss took place, since the type of therapy
depends on this
knowledge.
Infarctions take place not only in the myocardium, but rather also in other
tissues,
especially in the brain.
While the infarction can be healed to a certain extent, in a necrosis, locally
limited tissue
death, only the harmful sequelae for the residual organism can be prevented or
at least reduced.
Necroses can develop in many ways: by traumas, chemicals, oxygen deficiency or
by radiation.
As in infarction, the knowledge of the extent and type of a necrosis is
important for further
medical treatment.
Tests to improve the localization of infarctions and necroses by using
contrast media in
non-invasive processes, such as scintigraphy or nuclear spin tomography,
therefore already took
place earlier. The literature is full of reports on attempts to use porphyrins
for necrosis imaging.
The results that are achieved, however, paint a contradictory picture.
Winkelman and Hoyes thus
describe in Nature, 2QQ, 903 ( I 967) that manganese-5,10,15,20-tetrakis(4-
sulfonatophenyl)-
porphyrin (TPPS) selectively accumulates in the necrotic portion of a tumor.
Lyon et al. (Magn. Res. Med. 4, 24 ( 1987)) observed, however, that manganese-
TPPS is
dispersed in the body, specifically in the kidney, liver, and tumor and only
in a small portion of
the muscles. In this case, it is advantageous that the concentration in the
tumor reaches its
maximum only on the fourth day and only after the authors had increased the
dose from 0.12
mmol/kg to 0.2 mmol/kg. The authors therefore also speak of a non-specific
take-up of TPPS in
the tumor. Bockhurst et al. in turn report in Acta Neurochir fiQ, 347 (1994,
Suppl.) that MnTPPS
binds selectively to tumor cells.
CA 02418790 2003-02-07
11
Foster et al. (J. Nucl. Med. 2fi, 756 ( 1985)) in turn found that "'In-
5,10,15,20-tetrakis-(4-
N-methyl-pyridinium)-porphyrin (TMPyP) does not accumulate in the necrotic
portion, but rather
in the living edge layers. It follows from the above that a porphyrin-tissue
interaction exists and
is obvious but not necessary.
In Circulation Vol. 90, No. 4, part 2, page 1468, Abstract No. 2512 (1994), Ni
et al.
report that they can visualize infarction areas with a manganese-tetraphenyl-
porphyrin (Mn-TPP)
and a gadolinium-mesoporphyrin (Gd-MP). In International Patent Application WO
95/31219,
both substances were used in infarction and necrosis imaging. Authors Marchal
and Ni write
(see Example 3) that for the compound Gd-MP, the metal content of the
infarction-kidney was
high, similar to that of the non-infarcted organ, but that it was nine times
as large for the
myocardium in the case of infarcted tissue (Example 1). It was surprising that
the ratio of the
signal intensities in MRI for infarcted patients was comparatively high in
comparison to healthy
tissue in both cases with 2.10 or 2.19. Other metalloporphyrins have been
described in
Application DE 19835082 (Schering AG).
Porphyrins tend to be stored in the skin, which results in a
photosensitization. The
sensitization can last for days, and even weeks. This in an undesirable side
effect in using
porphyrins as diagnostic agents. In addition, the therapeutic index for the
porphyrins is only very
small, since, e.g., for Mn-TPPS, an action is used only at a dose of 0.2
mmol/kg, but LDso is
already approximately 0.5 mmol/kg.
Contrast media for necrosis and infarction imaging that are not derived from
the
porphyrin skeleton are described in DE 19744003 (Schering AG), DE 19744004
(Schering AG)
and WO 99/17809 (EPIX). To date, however, there are still no compounds that
can be used
satisfactorily as contrast media in infarction and necrosis imaging.
The object of the invention was therefore to make available contrast media
that can be
used in particular for MRT-lymphography, but also for tumor diagnosis and
necrosis and
infarction imaging.
The object of the invention is achieved by the perfluoroalkyl-containing
complexes with
sugar radicals of general formula I
CA 02418790 2003-02-07
12
(K)1-G - (Z-Rf)m
(Y_R) p
(I)
in which
R represents a monosaccharide or oligosaccharide radical that is bonded via
the 1-
OH position or 1-SH position,
Rt- is a perfluorinated, straight-chain or branched carbon chain with the
formula
-C~Fz"E, in which E represents a terminal fluorine, chlorine, bromine, iodine
or
hydrogen atom, and n stands for numbers 4-30,
K stands for a metal complex of general formula II
COOR'
~'N R~ R3
I
7
R O~
COOK' ( I I )
in which
Rl means a hydrogen atom or a metal ion equivalent of atomic numbers 21-29, 31-
33, 37-39, 42-44, 49 or 57-83,
provided that at Least two R' stand for metal ion equivalents,
Rz and R3, independently of one another, represent hydrogen, Ci-C~ alkyl,
benzyl, phenyl,
-CHzOH or -CHzOCH3, and
U represents -C6Ha-O-CHz-w-, -(CHz)i.s-w, a phenylene group, -CHz-NHCO-CHz-
CH(CH2COOH)-C6H4-co-, -C6Ha-(OCHzCHz)o-i-, N(CHzCOOH)-CHz-w or a Ci-
CIZ-alkylene group or a CrClz-C6Ha-O group that is optionally interrupted by
one
or more oxygen atoms, 1 to 3 -NHCO groups or 1- to 3 -CONH groups and/or is
substituted with 1 to 3 -(CHz)o.sCOOH groups, whereby w stands for the binding
site to -CO-,
CA 02418790 2003-02-07
13
or
of general formula III
COOR'
4
N CtJOR Q
s.--~ ~ f-~... u,.r-~.,~,,
R ooC
CQOR' ( I I I )
in which Rl has the above-mentioned meaning, R4 represents hydrogen or a metal
ion equivalent
that is mentioned under R~, and Ui represents -C6Ha-O-CH2-c~-, whereby w means
the binding
site to -CO-
or of general formula IV
CaoR'
~"I~ R
R'OOC'~
W C
COOK' tIV)
in which Rl and RZ have the above-mentioned meaning
or of general formula V A or V B
-CO~
~~'-CC~OR
R'O QC ~~~
~, --C~ aR'
~N'~-CaaR'
(V A)
CA 02418790 2003-02-07
14
' ~ w-c0 aR'
N~-cOOR'
~ G~ ~N
~ .~COOR'
~N '~COOR'
(V B)
in which R1 has the above-mentioned meaning,
or of general formula VI
R'oac--.~ ~--~, "~-co-
R'OOC--~N N~--COOK' (VI)
in which Rl has the above-mentioned meaning,
or of general formula VII
R'O OC --~ 0
R'o0e--~N u'--~,,-
R'OOC-~,'
R'OOC-~N
(VII)
in which Rl has the above-mentioned meaning, and
U' represents -CsH~-O-CH2-cu-, whereby c~ means the binding site to -CO-,
or of general formula VIII
COOK'
~N
R'OOC~N N N~
~N~ OH H
COOR' (VIII)
in which Rl has the above-mentioned meaning,
and in radical K, optionally present free acid groups optionally can be
present as salts of
CA 02418790 2003-02-07
15
organic and/or inorganic bases or amino acids or amino acid amides,
G for the case that K means metal complexes II to VII represents a radical
that is
functionalized in at least three places and that is selected from radicals a)
to j)
(a)
(b)
(c)
below
H
~,~-H-~CH2)ø e-GO--~ Y
I~ H
Y ~-~0-C-~CH2)~ H ~ ~
I~H
a
~ ~-IV fV-~-~. Y
CA 02418790 2003-02-07
16
(d)
(e)
(f)
~~f
ccv~--- ~
Y
CQ
t
~~H H~
H
NH-Ca- i -(CH~)~ H'~~~~-c~
I~ H
H
H-CO-C-(CH~~~ H~~a
NH
CA 02418790 2003-02-07
17
g)
(h)
(i)
I~ H
I
NH-CO-CH-(CH~)4 I~H-~~
~7-CC~-CH-(CH~)~ (~H-~'cx
NH
Y
NH-CO-CH-(CH2)~-~!H-~~~'a
NH
a~.--H-(CH2)~ C-H-CO-C-(CHZ)4 N---"~I~
GO ~H H
Y
a
H
(~'~"~~-~'--H-(CHz)~_a C-C ~N ~----,~,,"Y
f~H '~~W
CA 02418790 2003-02-07
18
)
oc~'~f'dH-CH -CO -~t
and
G for the case that K means metal complex VIII represents a radical that is
functionalized in at least three places and that is selected from k) or 1),
(k)
~~~-NH-~CH2)4 GH-CO-~'cx
NH
(1)
cc~-CO-CHz CH-CHI Ca-~"Y
I
NH
whereby a means the binding site of G to complex K,13 is the binding site of G
to radical Y
and ~y represents the binding site of G to radical Z,
Y means -CHz-, 8-(CHz)"CO-(3 (whereby n = 1-5), 8-CHz-CHOH-CO-(3 or 8-
CH(CHOH-CH20H)-CHOH-CHOH-CO-(3, whereby $ represents the binding site
to sugar radical R and (3 is the binding site to radical G
Z stands for
Y-N.,~~ N-SOZ E
y-COCHz-N(CzHs)-S02-~,
CA 02418790 2003-02-07
19
'y-COCHz-O-(CHz)z-SOz-~,
O t~
0 ~ ~ ~ ~-SOz E
or
'y-NHCHzCHz-O-CH2CHz-~,
whereby y represents the binding site of Z to radical G, and ~ means the
binding site of Z
to perfluorinated radical Rf,
and
l, m, independently of one another, mean the whole numbers 1 or 2,
and
p means the whole numbers 1 to 4.
If the compound according to the invention is intended for use in NMR
diagnosis, the
metal ion of the signal-transmitting group must be paramagnetic. These are
especially the
divalent and trivalent ions of the elements of atomic numbers 21-29, 42, 44
and 58-70. Suitable
ions are, for example, the chromium(III) ion, iron(II) ion, cobalt(II) ion,
nickel(II) ion, copper(II)
ion, praseodymium(III) ion, neodymium(III) ion, samarium(III) ion and
ytterbium(III) ion.
Because of their strong magnetic moment, gadolinium(III), terbium(III),
dysprosium(III),
holmium(III), erbium(III), iron(III) and manganese(II) ions are especially
preferred.
For the use of the compounds according to the invention in nuclear medicine
(radiodiagnosis and radiotherapy), the metal ion must be radioactive. For
example, radioisotopes
of the elements with atomic numbers 27, 29, 31-33, 37-39, 43, 49, 62, 64, 70,
75 and 77 are
suitable. Technetium, gallium, indium, rhenium, and yttrium are preferred.
If the compound according to the invention is intended for use in x-ray
diagnosis, the
metal ion is preferably derived from an element of a higher atomic number to
achieve a sufficient
absorption of x-rays. It was found that diagnostic agents that contain a
physiologically
CA 02418790 2003-02-07
20
compatible complex salt with metal ions of elements of atomic numbers 25, 26
and 39 as well as
57-83 are suitable for this purpose.
Manganese(II), iron(II), iron(III), praseodyrnium(III), neodymium(III),
samarium(III),
gadolinium(III), ytterbium(III) or bismuth(III) ions, especially
dysprosium(III) ions and
yttrium(III) ions, are preferred.
Acidic hydrogen atoms that are optionally present in R', i.e., those that have
not been
substituted by the central ion, can optionally be replaced completely or
partially by cations of
inorganic and/or organic bases or amino acids or amino acid amides.
Suitable inorganic cations are, for example, the lithium ion, the potassium
ion, the
calcium ion and especially the sodium ion. Suitable canons of organic bases
are, i.a., those of
primary, secondary or tertiary amines, such as, for example, ethanolamine,
diethanolamine,
morpholine, glucamine, N,N-dimethylglucamine and especially N-methylglucamine.
Suitable
cations of amino acids are, for example, those of lysine, arginine, and
ornithine as well as the
amides of otherwise acidic or neutral amino acids.
Especially preferred compounds of general formula I are those with macrocyclic
compound K of general formula II.
Radical U in metal complex K preferably means -CHz- or C6H4-O-CHz-w, whereby w
stands for the binding site to -CO-.
Alkyl groups R2 and R3 in the macrocyclic compound of general formula II can
be
straight-chain or branched. By way of example, methyl, ethyl, propyl,
isopropyl, n-butyl, 1-
methylpropyl, 2-methylpropyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-
methylbutyl, and 1,2-
dimethylpropyl can be mentioned. R2 and R3, independently of one another,
preferably mean
hydrogen or CI-C4 alkyl.
In a quite especially preferred embodiment, R2 Stands for methyl and R3 stands
for
hydrogen.
The benzyl group or the phenyl group Rz or R3 in macrocyclic compound K of
general
formula II can also be substituted in the ring.
Radical R in general formula I means a monosaccharide or oligosaccharide
radical or thin
CA 02418790 2003-02-07
21
sugar radical bonded via the 1-OH position or 1-SH position, whereby in this
connection
according to the invention, this can also be deoxy sugars that contain an H
atom instead of one or
more OH groups. In a preferred embodiment of the invention, R means a
monosaccharide
radical with 5 or 6 C atoms, preferably glucose, mannose, galactose, ribose,
arabinose or xylose
or their deoxy sugars, such as, for example, 6-deoxygalactose (fucose) or 6-
deoxymannose
(rhamnose) or their thio sugars, whereby glucose, mannose and galactose are
especially preferred.
Of the compounds of general formula I according to the invention, in addition
those are
preferred in which Rrmeans -C"F2~+i. n preferably stands for the numbers 4-15.
Quite especially
preferred are radicals -CaF9, -C6F~3, -CsFn, -C1zF25 and -CiaF29 as well as
the radicals of the
compounds that are mentioned in the examples.
Radical G that is functionalized in at least three places in general formula
I, which
represents the "skeleton," means lysine radical (a) or (b) in a preferred
embodiment of the
invention.
Y and Z mean the linkers indicated in general formula I, whereby independently
of one
another, the radical
'Y-NON-SOz E
is preferred for Z and the radical 8-CHzCO-(3 is preferred for Y.
The perfluoroalkyl-containing metal complexes with sugar radicals of general
formula I
(K)~-G - (Z-Rf)m
(z)
(Y-R)p
with K in the meaning of a metal complex of general formulas II to VII and G
in the meaning of
formulas a) to j),
whereby Y, Z, R, Rf, m. p and I have the above-mentioned meaning, are
produced, in a way that
is known in the art, by a carboxylic acid of general formula IIa
CA 02418790 2003-02-07
22
Co 0 RS
/~'N ~ Rz R' O
R~00C~N N N''U~OH
0
Ny
s
Coon
(IIa)
in which RS means a metal ion equivalent of atomic numbers 21-29, 31-33, 37-
39, 42-44, 49 or
57-83 or a carboxyl protective group, and R2, R3 and U have the above-
mentioned meaning,
or a carboxylic acid of general formula IIIa
COOR$
~~ ~
/~''N ~ COOK p
s .r'-' N N ~ U'"'~ O H
R oOC - f
~N.~~
CO o R (IIIa)
in which R4, R5, and U1 have the above-mentioned meaning
or a carboxylic acid of general formula IVa
C0o R$
~I~J ~ R2
OH
R~OOC"~/N N
0
s
COOK
(IV a)
CA 02418790 2003-02-07
23
in which RS and R2 have the above-mentioned meaning
or a carboxylic acid of general formula Va or Vb
.~-C 0-0 H _ s
~~'~CO 0Rs ~~ °~-C00 RS
RSOOC''~ /~J -~'-N
~ -COORS HOOD / s
~N~-C00R5 ~~-C00R
-GOORS
(Va)
(Vb)
in which RS has the above-mentioned meaning
or a carboxylic acid of general formula VIa
R$OOC-~N/~~~..--CO-OH
R OOC--~ ~-COORS (VIa)
in which R' has the above-mentioned meaning
or a carboxylic acid of general formula VIIa
RSOOC-..~ 0
Rs00G--~N U'~OH
Rs0 OC ~,,4. ,
Rs00G-~'~
(VIIa)
in which RS and U' have the above-mentioned meanings,
being reacted in optionally activated form with an amine of general formula IX
H _ G _ ( Z-Rf ) m
(Ix)
(Y-R) P
in which G has the meaning of formulas a) to j),
and R, Re, Y, Z, m and p have the indicated meaning, in a coupling reaction
and optionally
subsequent cleavage of optionally present protective groups to form a metal
complex of general
CA 02418790 2003-02-07
24
formula I,
or
if RS has the meaning of a protective Qroup, being reacted after cleavage of
these
protective groups in a subsequent step in a way that is known in the art with
at least one metal
oxide or metal salt of an element of atomic numbers 21-29, 31-33, 37-39, 42-
44, 49 or 57-83,
and then, if desired, optionally present, acidic hydrogen atoms being
substituted by cations of
inorganic and/or organic bases, amino acid or amino acid amides.
The compounds of general formula I according to the invention with K in the
meaning of
a metal complex of general formula VIII and G in the meaning of formulas k) or
1) are produced
in a way that is known in the art by an amine of general formula VIIIa
BOORS
H
R$GOC~~ ~ H~-
GH
s
~~~R (VIIIa)
in which RS means a metal ion equivalent of atomic numbers 21-29, 31-33, 37-
39, 42-44, 49 or
57-83, or a carboxyl protective group,
being reacted with an optionally activated carboxylic acid of general formula
X
HO - G - ( Z-Rf)
(x>
(Y-R)P
in which G has the meaning of formulas k) or 1) and R, Rf, Y, Z, m and p have
the indicated
meanings, in a coupling reaction and optionally subsequent cleavage of
optionally present
protective groups to form a metal complex of general formula I
CA 02418790 2003-02-07
25
or
if RJ has the meaning of a protective group, being reacted after cleavage of
these
protective groups in a subsequent step in a way that is known in the art with
at least one metal
oxide or metal salt of an element of atomic numbers 21-29, 31-33, 37-39,42-44,
49 or 57-83,
and then, if desired, optionally present acidic hydrogen atoms being
substituted by cations of
inorganic and/or organic bases, amino acids or amino acid amides.
The carboxylic acids of general formulas IIa to VIIa that are used are either
known
compounds or are produced according to the process described in the examples.
Thus, the
production of carboxylic acids of general formula IIa is known from DE 196 52
386. The
production of carboxylic acids of general formula IIIa can be carried out
analogously to Example
3 of this application. The production of the carboxylic acids of general
formula IVa can be
derived from DE 197 28 954.
A precursor for compounds of general formula VA is N3-(2,6-
dioxomorpholinoethyl)-N6-
(ethoxycarbonylmethyl)-3,6-diaza-octanedioic acid, which is described in EP
263 059.
The compounds of general formula VB are derived from the isomeric
diethylenetriamine-
pentaacetic acid, which binds via the acetic acid that is on the center N
atom. This DTPA is
described in Patents DE 195 07 819 and DE 195 08 058.
Compounds of general formula VI are derived from N-(carboxymethyl)-N-[2-(2,6-
dioxo-
4-morpholinyl)-ethyl]-glycine, whose production is described in J. Am. Oil.
Chem. Soc. ( 1982),
SQ (2), 104-107.
Compounds of general formula VII are derived from 1-(4-carboxymethoxybenzyl)-
ethylenediamine-tetraacetic acid, whose production is described in Patent US
4,622,420.
The production of amines of general formula IX and carboxylic acids of general
formula
X is described in detail in the examples of this application and can be carned
out analogously to
the processes described in the examples. The amine of general formula VIIIa is
a known starting
compound.
The perbenzylated sugar acids- used as starting substances can be produced
analogously to
Lockhoff, Angew. Chem. [Applied Chem.] 1998, 110, No. 24, p. 3634 f~ Thus,
e.g., the
CA 02418790 2003-02-07
26
production of the 1-O-acetic acid of perbenzyl-glucose is carried out over 2
stages, via
trichloroacetamidate and reaction with hydroxyacetic acid ethyl ester, BF3-
catalysis in THF and
subsequent saponification with NaOH in MeOH/THF.
In a more advantageous process, the perbenzylated sugar acids that are used as
starting
substances can also be produced by the perbenzylated 1-OH-sugar being
dissolved in a water-
immiscible organic solvent and being reacted with an alkylating reagent of
general formula XI
Nu-L-COO-Sg (XI),
in which Nu means a nucleofuge, L is -(CHz)-", (whereby n = 1-5), -CHz-CHOH-, -
CH(CHOH-
CH20H)-CHOH-CHOH-, and Sg represents a protective group,
in the presence of a base and optionally a phase transfer catalyst. As
nucleofuges, for example,
the radicals -Cl, -Br, -I, OTs, -OMs, -OSOzCF3, -OSOZCaF9 or -OSOzCsF» can be
contained in
the alkylating reagent of general formula XI.
The protective group is a conventional acid protective group. These protective
groups are
well known to one skilled in the art (Protective Groups in Organic Syntheses,
Second Edition, T.
W. Greene and P. G. M. Wuts, John Wiley & Sons Inc., New York 1991).
The reaction according to the invention can be carried out at temperatures of
0-50°C,
preferably 0°C to room temperature. The reaction times are from 10
minutes to 24 hours,
preferably 20 minutes to 12 hours.
The base is added either in solid form, preferably in fine-powder form, or as
10-70%,
preferably 30-50% aqueous solution. NaOH and KOH are used as preferred bases.
As organic, water-immiscible solvents, for example, toluene, benzene, CF3-
benzene,
hexane, cyclohexane, diethyl ether, tetrahydrofuran, dichloromethane, MTB or
their mixtures can
be used in the alkylating process according to the invention.
As phase-transfer catalysts, the quaternary ammonium salts or phosphonium
salts or else
crown ethers, such as, e.g., [15]-crown-5 or [18]-crown-6, that are known for
this purpose are
used in the process according to the invention. Quaternary ammonium salts with
four identical
or different hydrocarbon groups at the cation, selected from methyl, ethyl,
propyl, isopropyl,
butyl or isobutyl, are preferably suitable. The hydrocarbon groups at the
cation must be large
CA 02418790 2003-02-07
27
enough to ensure good solubility of the alkylating reagent in the organic
solvent. According to
the invention, N(butyl)a+-Cl-, N(butyl)al-HSOa , but also N(methyl)a+-Cl- are
especially
preferably used.
It has been shown that the metal complexes according to the invention are
especially
suitable for NMR diagnosis and x-ray diagnosis, but also for radiodiagnosis
and radiotherapy.
The subject of the invention is therefore also the use of the perfluoroalkyl-
containing metal
complexes according to the invention with sugar radicals for the production of
contrast media for
use in NMR diagnosis and x-ray diagnosis, especially for lymphography, for
tumor diagnosis,
and for infarction imaging and necrosis imaging, as well as in radiodiagnosis
and radiotherapy.
The compounds according to the invention are extremely well suited for use in
interstitial
lymphography and especially in intravenous lymphography. In addition, they can
also be used
for visualization of the vascular space (blood-pool agents).
Subjects of the invention are also pharmaceutical agents that contain at least
one
physiologically compatible compound according to the invention, optionally
with the additives
that are commonly used in galenicals.
The compounds of this invention are distinguished by a very good systemic
compatibility
and a high lymph node concentration in three successive lymph node stations
(which is important
especially for i.v. lymphography). They are thus especially well suited for
use in MRT
lymphography.
The compounds according to the invention are also extremely well suited for
detecting
and localizing vascular diseases, since they are dispersed exclusively in the
latter in the
administration in the intravascular space. The compounds according to the
invention make it
possible, with the aid of nuclear spin tomography, to distinguish between
tissue that is well
supplied with blood and tissue that is poorly supplied with blood and thus to
diagnose an
ischemia. Because of its anemia, infarcted tissue can also be distinguished
from surrounding
healthy or ischemic tissue when the contrast media according to the invention
are used. 'This is
of special importance if the point is, e.g., to distinguish a myocardial
infarction from an ischemia.
Compared to the macromolecular compounds previously used as blood-pool agents,
such
CA 02418790 2003-02-07
28
as, for example, Gd-DTPA-polylysine, the compounds according to the invention
also show a
higher T1-relaxivity and thus are distinguished by an increase of signal
intensity in NMR
imaging. Since in addition they have an extended retention in the blood space,
they can also be
administered in relatively small doses (of, e.g., < 50 umol of Gd/l of body
weight). The
compounds according to the invention are primarily quickly and as completely
as possible
eliminated from the body, however.
It was also shown that the compounds according to the invention accumulate in
areas
with elevated vascular permeability, such as, e.g., in tumors; they make it
possible to make
statements on the perfusion of tissues, provide the possibility of determining
the blood volumes
in tissues, to selectively shorten the relaxation times or densities of the
blood and to graphically
visualize the permeability of blood vessels. Such physiological data cannot be
obtained by the
use of extracellular contrast media, such as, e.g., Gd-DTPA (Magnevist~R~).
From these
considerations also arise their uses in the modern imaging processes nuclear
spin tomography
and computer tomography: specific diagnosis of malignant tumors, early therapy
control in
cytostatic, antiphlogistic or vasodilatative therapy, early detection of
underperfused areas (e.g., in
the myocardium), angiography in vascular diseases, and detection and diagnosis
of sterile or
infectious inflammations.
The production of the pharmaceutical agents according to the invention is
carried out in a
way that is known in the art by the complex compounds according to the
invention -- optionally
with the addition of the additives that are commonly used in galenicals --
being suspended or
dissolved in aqueous medium and then the suspension or solution optionally
being sterilized.
Suitable additives are, for example, physiologically harmless buffers (such
as, for example,
tromethamine), additions of complexing agents or weak complexes (such as, for
example,
diethylenetriaminepentaacetic acid or the Ca-complexes that correspond to the
metal complexes
according to the invention) or -- if necessary -- electrolytes such as, for
example, sodium chloride
or -- if necessary -- antioxidants, such as, for example, ascorbic acid.
If suspensions or solutions of the agents according to the invention in water
or
physiological salt solution are desired for enteral or parenteral
administration or other purposes,
CA 02418790 2003-02-07
29
they are mixed with one or more adjuvant(s) that are commonly used in
galenicals [for example,
methyl cellulose, lactose, mannitol] and/or surfactants) [for example,
lecithins, Tween~R~,
Myrj~R~] and/or flavoring substances) for taste correction [for example,
ethereal oils].
Basically, it is also possible to produce the pharmaceutical agents according
to the
invention without isolating the complexes. In any case, special care must be
used to carry out the
chelation so that the complexes according to the invention are practically
free of non-complexed
metal ions that have a toxic effect.
This can be ensured, for example, with the aid of color indicators, such as
xylenol orange,
by control titrations during the production process. The invention therefore
also relates to a
process for the production of the complex compounds and their salts. As a
final precaution, there
remains purification of the isolated complex.
In the in-vivo administration of the agents according to the invention, the
latter can be
administered together with a suitable vehicle, such as, for example, serum or
physiological
common salt solution and together with another protein, such as, for example,
human serum
albumin (HSA).
The agents according to the invention are usually administered parenterally,
preferably
i.v. They can also be administered intravascularly or
interstitially/intracutaneously depending on
whether bodily vessels or tissue are to be studied.
The pharmaceutical agents according to the invention preferably contain 0.1
p.mol - 2
mol/1 of the complex and are generally dosed in amounts of 0.0001 - 5 mmol/kg.
The agents according to the invention meet the many requirements for
suitability as
contrast media for nuclear spin tomography. After oral or parenteral
administration, they are thus
extremely well suited for enhancing the informational value of the image that
is obtained with the
aid of a nuclear spin tomograph by increasing the signal intensity. They also
show the high
effectiveness that is necessary to load the body with the smallest possible
amounts of foreign
substances and the good compatibility that is necessary to maintain the non-
invasive character of
the studies.
The good water solubility and low osmolality of the agents according to the
invention
CA 02418790 2003-02-07
30
make it possible to produce highly concentrated solutions, so as to keep the
volume burden of the
circulatory system within reasonable limits and to offset the dilution by
bodily fluids. In
addition, the agents according to the invention show not only high stability
in vitro, but also
surprisingly high stability in vivo, so that a release or an exchange of the
ions -- which are
inherently toxic -- and which are bonded to the complexes can take place only
extremely slowly
within the time in which the new contrast media are completely excreted again.
In general, the agents according to the invention for use as NMR diagnostic
agents are
dosed in amounts of 0.0001-5 mmol/kg, preferably 0.005-0.5 mmol/kg.
The complex compounds according to the invention also can advantageously be
used as
susceptibility reagents and as shift reagents for in-vivo-NMR spectroscopy.
Based on their advantageous radioactive properties, and the good stability of
the complex
compounds contained therein, the agents according to the invention are also
suitable as
radiodiagnostic agents. Details of such a use and dosage are described in,
e.g., "Radiotracers for
Medical Applications," CRC-Press, Boca Raton, Florida.
The compounds and agents according to the invention can also be used in
positron-
emission tomography, which uses positron-emitting isotopes, such as, e.g.,
43Sc, 44Sc, SZFe, 55Co,
6gGa and 86Y (Heiss, W. D.; Phelps, M. E.; Positron Emission Tomography of
Brain, Springer
Verlag Berlin, Heidelberg, New York 1983).
The compounds according to the invention are also suitable, surprisingly
enough, for
differentiating malignant and benign tumors in areas without blood-brain
barriers.
The contrast media according to the invention can also be used after
intravenous
administration for quantitative determination of changes in capillary
integrity, caused by
hyperoxia (including "acute capillary holes" and restoration of normal
endothelial integrity after
hyperoxic damage).
Histological studies confirm a regional microvascular hyperpermeability.
The contrast media according to the invention can therefore also be used to
visualize
abnormal capillary permeability.
They are also distinguished in that they are completely eliminated from the
body and thus
CA 02418790 2003-02-07
31
are well-tolerated.
Since the substances according to the invention accumulate in malignant tumors
(no
diffusion in healthy tissue, but high permeability of tumor vessels), they can
also support the
radiation therapy of malignant tumors. The latter is distinguished from the
corresponding
diagnosis only by the amount and type of the isotope used. The purpose in this
case is the
destruction of tumor cells with high-energy shortwave radiation with as small
a range of action as
possible. For this purpose, interactions of the metals (such as, e.g., iron or
gadolinium) that are
contained in the complexes are used with ionizing radiation (e.g., x-rays) or
with neutron rays.
By this effect, the local radiation dose at the site where the metal complex
is located (e.g., in
tumors) is significantly increased. To produce the same radiation dose in
malignant tissue, the
radiation exposure for healthy tissue can be considerably reduced when using
such metal
complexes and thus side effects imposing a burden for the patients are
avoided. The metal-
complex conjugates according to the invention are therefore also suitable as
radiosensitizing
substances in radiation therapy of malignant tumors (e.g., use of Mossbauer
effects or in neutron
capture therapy). Suitable (3-emitting ions are, for example, 46Sc, 47Sc,
4gSc, ~2Ga, 73Ga and 9°Y.
Suitably short half lives that have a-emitting ions are, for example, 2l~Bi,
2~2Bi, 2~3B1 and 214Bi,
whereby 212B1 1S preferred. A suitable photon- and electron-emitting ion is
~SgGd, which can be
obtained from ~S~Gd by neutron capture.
If the agent according to the invention is intended for use in the variant of
radiation
therapy proposed by R. L. Mills et al. (Nature Vol. 336, (1988), p. 787), the
central ion must be
derived from a Mossbauer isotope, such as, for example, S~Fe or ~SIEu.
In the in-vivo administration of the agents according to the invention, the
latter can be
administered together with a suitable vehicle, such as, for example, serum, or
physiological
common salt solution and together with another protein, such as, for example,
human serum
albumin. In this case, the dosage depends on the type of cellular disorder,
the metal ion that is
used and the type of imaging method.
The agents according to the invention are usually administered parenterally,
preferably
i.v. They can also -- as already discussed -- be administered intravascularly
or
CA 02418790 2003-02-07
32
interstitially/intracutaneously depending on whether bodily vessels or tissue
are to be studied.
The agents according to the invention are extremely well suited as x-ray
contrast media,
whereby it is especially to be emphasized that no displays of the anaphylaxis-
like reactions
known from the iodine-containing contrast media can be detected with them in
biochemical-
pharmacological studies. Because of the advantageous absorption properties in
the areas of
higher tube voltages, they are especially valuable for digital subtraction
techniques.
In general, the agents according to the invention for use as x-ray contrast
media
analogously to the meglumine-diatrizoate example are dosed in amounts of 0.1-5
mmol/kg,
preferably 0.25-1 mmol/kg.
In particular, higher blood concentrations are achieved with the compounds
according to
the invention than with extracellular contrast media.. They are dispersed
after i.v. administration
only into the intravascular space and thus have a decisive advantage compared
to the
extracellular contrast media.
CA 02418790 2003-02-07
33
F'ramnie l1
a) 2-N-Trifluoroacetyl-6-N-benzyloxycarbonyl-L-lysine
100.0 g (356.7 mmol) of 6-N-benzyloxycarbonyl-L-lysine is dissolved in a
mixture that
consists of 1000 ml of trifluoroacetic acid ethyl ester and 500 ml of ethanol,
and it is stirred for
24 hours at room temperature. It is evaporated to the dry state, and the
residue is crystallized
from diisopropyl ether.
Yield: 128.9 g (96% of theory) of a colorless, crystalline powder.
Melting point: 98.5°C.
Elementary analysis:
Cld: C 51.07 H 5.09 N 7.44 F 15.14
Fnd: C 51.25 H 5.18 N 7.58 F 15.03
b) 2-N-Trifluoroacetyl-6-N-benzyloxycarbonyl-L-lysine [1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide
164.2 g (0.664 mmol) of EEDQ (2-ethoxy-1,2-dihydroquinoline-1-carboxylic acid
ethyl
ester) is added at 0°C to 125.0 g (332.0 mmol) of the title compound of
Example 1 a) and 188.7 g
(332.0 mmol) of 1-perfluorooctylsulfonylpiperazine (produced according to DE
19603033) in
750 ml of tetrahydrofuran, and it is stirred overnight at room temperature. It
is evaporated to the
dry state in a vacuum and chromatographed on silica gel (mobile solvent:
dichloromethane/
methanol=20:1 ).
Yield: 286.0 g (93% of theory) of a colorless solid.
Melting point: 92°C.
Elementary analysis:
Cld: C 36.30 H 2.83 N 6.05 F 41.01 S 3.46
Fnd: C 36.18 H 2.94 N 5.98 F 40.87 S 3.40
c) 6-N-Benzyloxycarbonyl-L-lysine-[1-(4-perfluorooctylsulfonyl)-piperazine]-
amide
Ammonia gas is introduced at 0°C for one hour into a solution that
consists of 280.0 g
CA 02418790 2003-02-07
34
(302.2 mmol) of the title compound of Example 1 b) in 2000 ml of ethanol. It
is then stirred for
four hours at 0°C. It is evaporated to the dry state, and the residue
is absorptively precipitated
from water. The solid is filtered off and dried in a vacuum at 50°C.
Yield: 243.5 g (97% of theory) of an amorphous solid.
Elementary analysis:
Cld: C 37.60 H 3.28 N 6.75 F 38.89 S 3.86
Fnd: C 37.55 H 3.33 N 6.68 F 38.78 S 3.81
d) 6-N-Benzyloxycarbonyl-2-N-[1-O-a-D-carbonylmethyl-(2,3,4,6-tetra-O-benzyl-
mannopyranose]-L-lysine-[ 1-(4-perfluorooctylsulfonyl)-piperazine]-amide
41.27 g (200.0 mmol) of N,N-dicyclohexylcarbodiimide is added at 0°C to
a solution that
consists of 100.0 g ( I 20.4 mol) of the title compound of Example 1 c), 72.1
g ( I 20.4 mol) of 1-O-
a-D-carboxymethyl-2,3,4,6-tetra-O-benzyl-mannopyranose and 13.86 g (120.4 moI)
of N-
hydroxysuccinimide, dissolved in 500 ml of dimethylformamide. It is stirred
for 3 hours at 0°C
and then overnight at room temperature. Precipitated urea is filtered out, the
filtrate is
evaporated to the dry state in a vacuum and chromatographed on silica gel.
(Mobile solvent:
dichloromethane/ ethanol = 20:1 ).
Yield: 136.1 g (87% of theory) of a viscous oil.
Elementary analysis:
Cld: C 57.32 H 4.89 N 4.31 F 24.86 S 2.47
Fnd: C 57.38 H 5.07 N 4.22 F 24.78 S 2.39
e) 2-N-[1-O-a-D-Carbonylmethyl-mannopyranose]-L-lysine-1-[(4-
perfluorooctylsulfonyl)-
piperazine]-amide
130.0 g ( 100.0 mmol) of the title compound of Example 1 d) is dissolved in
2000 ml of
ethanol, and I0.0 g of palladium catalyst (10% Pd/C) is added. It is
hydrogenated for 12 hours at
room temperature. Catalyst is filtered out, and the filtrate is evaporated to
the dry state in a
vacuum.
CA 02418790 2003-02-07
JS
Yield: 9I .7 g (quantitative) of a colorless solid.
Elementary analysis:
Cld: C 34.07 H 3.63 N 6.11 S 3.50 F 35.24
Fnd: C 33.91 H 3.72 N 6.04 S 3.40 F 35.31
f) 6-N-[1,4,7-Tris(carboxylatomethyl)]-1,4,7,10-tetraazacyclododecane-10-N-
(pentanoyl-3-
aza-4-oxo-5-methyl-5-yl)]-2-N-[ 1-O-a-D-carbonylmethyl-mannopyranose]-L-lysine-
[ I -(4-
perfluorooctylsulfonyl)-piperazine]-amide, Gd complex
50.0 g (54.55 mmol) of the title compound of Example 1 e), 6.28 g (54.55 mmol)
of N-
hydroxysuccinimide, 4.62 g (109.0 mol) of lithium chloride and 34.35 g (54.55
mol) of 1,4,7-
tris(carboxylatomethyl)-10-(carboxy-3-aza-4-oxo-5-methyl-pent-5-yI)-1,4,7,10-
tetraazacyclododecane, Gd complex, are dissolved in 400 ml of dimethyl
sulfoxide while being
heated slightly. At 10°C, 16.88 g (8I .8 mol) of N,N-
dicyclohexylcarbodiimide is added, and it is
then stirred overnight at room temperature. The solution is poured into 3000
ml of acetone and
stirred for 10 minutes. The precipitated solid is filtered off and then
purified by chromatography
(RP-18 mobile solvent: gradient that consists of water/ethanol/acetonitrile).
Yield: 75.9 g (91.0% of theory) of a colorless solid.
Water content: 8.6%.
Elementary analysis (relative to anhydrous substance):
Cld: C35.34H4.09N8.24S2.1OF21.12Gd10.28
Fnd: C35.28H4.15N8.19S2.15F21.03Gd10.14
a) 6-N-[1,4,7-Tris(carboxylatomethyl]-1,4,7,10-tetraazacyclododecane-10-N-
(pentanoyl-3-
aza-4-oxo-5-yl)]-2-N-[ 1-O-a-D-carborlylmethyl-mannopyranose]-L-lysine-[ I -(4-
perfluorooctylsulfonyl)-piperazine]-amide, Gd complex
50.0 g (54.55 mmol) of the title compound of Example 1e), 6.28 g (54.55 mmol)
ofN-
hydroxysuccinimide, 4.62 g (109.0 mol) of lithium chloride and 34.35 g (54.55
mol) of 1,4,7-
CA 02418790 2003-02-07
36
tris(carboxylatomethyl)-10-(carboxy-3-aza-4-oxo-5-methyl-pent-5-yl)-1,4,7,10-
tetraazacyclododecane, Gd complex, are dissolved in 400 ml of dimethyl
sulfoxide while being
heated slightly. At 10°C, 16.88 g (81.8 mmol) of N,N-
dicyclohexylcarbodiimide is added and
then stirred overnight at room temperature. The solution is poured into 3000
ml of acetone and
stirred for 10 minutes. The precipitated solid is filtered off and then
purified by chromatography
(RP-18 mobile solvent: gradient that consists of water/ethanol/acetonitrile.
Yield: 76.0 g (92.0% of theory) of a colorless solid.
Water content: 6.88%.
Elementary analysis (relative to anhydrous substance):
Cld: C34.90H3.93N8.32S2.12F21.33Gd10.38
Fnd: C 34.81 H 4.02 N 8.27 S 2.09 F 21.22 Gd 10.19
E~cam,~
a) 2-[4-3-Oxapropionic acid ethyl ester]-phenylacetic acid methyl ester
233.8 g (1400.0 mmol) of 2-bromoacetic acid ethyl ester is added to 200.0 g
(1204.0
mmol) of 4-hydroxyphenylacetic acid methyl ester, 212.0 g (2000.0 mmol) of
sodium carbonate
in 2000 ml of acetone, and it is refluxed for 5 hours. The solid is filtered
off and evaporated to
the dry state in a vacuum. The residue is chromatographed on silica gel
(mobile solvent: n-
hexane/ethyl acetate = 15:1 ).
Yield: 288.5 g (95.0% of theory) of a colorless oil.
Elementary analysis:
Cld: C 61.90 H 6.39
Fnd: C 61.75 H 6.51
b) 2-[4-3-Oxapropionic acid ethyl ester)]-phenyl-2-bromoacetic acid methyl
ester
201.0 g (1130.0 mmol) of N-bromosuccinimide and 100.0 mg of dibenzoyl peroxide
are
added to 285.0 g (1130.0 mmol) of the title compound of Example 3a), dissolved
in 2000 ml of
carbon tetrachloride, and it is refluxed for eight hours. It is cooled in an
ice bath, the precipitated
CA 02418790 2003-02-07
37
succinimide is filtered off, and the filtrate is evaporated to the dry state
in a vacuum. The residue
is purified on silica gel (mobile solvent: n-hexane/acetone = 15:1 ).
Yield: 359.2 g (96.0% of theory) of a colorless, viscous oil.
Elementary analysis:
Cld: C 47.28 H 4.57 Br 24.16
Fnd: C 47.19 H 4.71 Br 24.05
c) 2-[4-(3-Oxapropionic acid ethyl ester)]-phenyl-2-[1-(1,4,7,IO-
tetraazacyclododecan-I-yl]-
acetic acid methyl ester
350.0 g (1057.0 mmol) of the title compound of Example 3b) is added to 603.0 g
(3500.0
mmol) of 1,4,7,10-tetraazacyclododecane, in 6000 ml of chloroform, and it is
stirred overnight at
room temperature. It is extracted 3 times with 3000 ml of water each, the
organic phase is dried
on magnesium sulfate and evaporated to the dry state in a vacuum. The residue
is used without
further purification in the next reaction (Example 3d).
Yield: 448.0 (quantitative) of a viscous oil.
Elementary analysis:
Cld: C 59.70 H 8.1 I N 13.26
Fnd: C 59.58 H 8.20 N 13.18
d) 2-[4-(3-Oxapropionic acid)]-phenyl-2-[1,4,7-tris(carboxymethyl)-1,4,7,10-
tetraazacyclododecan-10-yl]-acetic acid
445.0 g (1053.0 mmol) of the title compound of Example 3c) and 496.0 g (5270.0
mmol)
of chloroacetic acid are dissolved in 4000 ml of water. It is set at a pH of
10 with 30% aqueous
sodium hydroxide solution, and it is stirred for 8 hours at 70°C. Then,
the pH of the reaction
solution is set at 13 by mixing with 30% aqueous sodium hydroxide solution,
and it is refluxed
for 30 minutes. The solution is cooled in an ice bath and set at a pH of 1 by
adding concentrated
hydrochloric acid. It is evaporated to the dry state in a vacuum. The residue
is taken up in 4000
ml of methanol, and it is absorptively precipitated for one hour at room
temperature. Precipitated
CA 02418790 2003-02-07
38
common salt is filtered out, the filtrate is evaporated to the dry state, and
the residue is purified
on RP-18 C (mobile solvent: gradient that consists of
water/ethanol/acetonitrile).
Yield: 403.0 g (69.0% of theory) of a colorless solid.
Water content: 10.2%.
Elementary analysis (relative to anhydrous substance):
Cld: C 51.98 H 6.18 N 10.10
Fnd: C 51.80 H 6.31 N 10.01
e} 2-[4-(3-Oxapropionic acid)]-phenyl-2-[1,4,7-tris(carboxylatomethyl)-
1,4,7,10-
tetraazacyclododecan-10-ylJ-acetic acid, Gd complex
130.73 g (360.65 mmol) of gadolinium oxide is added to 400 g (721.3 mmol) of
the title
compound of Example 3d) in 2000 ml of water, and it is stirred for 5 hours at
80°C. The solution
is filtered, and the filtrate is freeze-dried.
Yield: 511 g (quantitative) of an amorphous solid.
Water content: 11.0%.
Elementary analysis (relative to anhydrous substance):
Cld: C 40.67 H 4.41 N 7.98 Gd 22.19
Fnd: C 40.51 H 4.52 N 8.03 Gd 22.05
f) ' 6-N-[2-[4-(3-Oxapropionyl)-phenylJ-2-[1,4,7-tris{carboxylatomethy1)-
1,4,7,10-
tetraazacyclododecan-10-yIJ-acetic acid)J-2-N-( 1-O-a-D-carbonylmethyl-
mannopyranose)-L-
lysine-[1-(4-perfluorooctylsulfonyl)-piperazineJ-amide, Gd complex, sodium
salt
50.0 g (54.55 mmol) of the title compound of Example 1 e), 6.28 g (54.55 mmol)
of N-
hydroxysuccinimide, 4.62 g (109.0 mmol) of lithium chloride and 38.66 g (54.55
mol) of the title
compound of Example 3e) are dissolved in 400 ml of dimethyl sulfoxide while
being heated
slightly. At 10°C, 16.88 g (81.8 mmol) of N,N-dicyclohexylcarbodiimide
is added and then
stirred overnight at room temperature. The solution is poured into 3000 ml of
acetone and stirred
for 10 minutes. The precipitated solid is filtered off and then purified by
chromatography (RP-
CA 02418790 2003-02-07
39
18; mobile solvent: gradient that consists of water/ethanol/acetonitrile). The
product that is
obtained is dissolved in a little water, and the pH of the solution is set at
7.4 with aqueous
sodium hydroxide solution. Then, the product solution is freeze-dried.
Yield: 79.1 g (89% of theory) of a colorless solid.
Water content: 10.3%.
Elementary analysis (relative to anhydrous substance):
Cld: C 36.86 H 3.77 N 6.88 S 1.97 F 19.82 Gd 9.65
Fnd: C 36.75 H 3.84 N 6.80 S 2.03 F 19.75 Gd 9.57
Fxamnle 44
a) 6-N-[1,4,7-Tris(t butyloxycarbonylmethyl)-1,4,7,10-tetraazacyclododecane-10-
carbonylmethyl]-2-N-( 1-O-a-D-carbonylmethyl-mannopyranose)-L-lysine-[ 1-(4-
perfluorooctylsulfonyl)-piperazineJ-amide
15.0 g (26.19 mmol) of 1,4,7-tris(t-butyloxycarbonylmethyl)-10-carboxymethyl-
1,4,7,10-
tetiaazacyclododecane (produced according to: W091 /05762), 24.0 g (26.19
mmol) of the title
compound of Example 1e) and 3.01 g (26.19 mmol) of N-hydroxysuccinimide are
dissolved in
150 ml of dimethylformamide, and 8.25 g (40.0 mmol) of N,N-
dicyclohexylcarbodiimide is
added at 0°C. It is stirred overnight at room temperature. The
precipitated urea is filtered off,
and the filtrate is evaporated to the dry state in a vacuum. The residue is
chromatographed on
silica gel (mobile solvent: dichloromethane/methanol = 20:1).
Yield: 35.45 g (92.0% of theory) of a colorless solid.
Elementary analysis:
Cld: C 44.08 H 5.69 N 7.62 F 21.95 S 2.18
Fnd: C 44.01 H 5.81 N 7.53 F 21.87 S 2.03
CA 02418790 2003-02-07
40
b) 6-N-[1.4,7-Tris(carboxylatomethyl]-1,4,7,10-tetraazacyclododecane-10-
carbonylmethyl-
]-2-N-[ 1-O-a-D-carbonylmethyl-mannopyranose]-L-lysine-[ 1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide, Gd complex
30.0 g (20.39 mmol) of the title compound of Example 4a) is dissolved in 50 ml
of
chloroform, and 300 ml of trifluoroacetic acid is added. It is stirred for 10
minutes at room
temperature. It is evaporated to the dry state in a vacuum, and the residue is
dissolved in 300 ml
of water. 3.69 g (10.19 mmol) of gadolinium oxide is added, and it is stirred
for 5 hours at 80°C.
The solution is evaporated to the dry state in a vacuum and purified on silica
gel (RP-18; mobile
solvent: gradient that consists of water/ethanol/acetonitrile).
Yield: 11.0 g (37.0% of theory) of a colorless and amorphous solid.
Water content: 11.3%.
Elementary analysis (relative to anhydrous substance):
Cld: C 34.62 H 3.87 N 7.69 F 22.16 S 2.20 Gd 10.97
Fnd: C 34.57 H 3.95 N 7.60 F 22.05 S 2.13 Gd 10.90
Fxamnle 5S
a) 6-N-[3,6,9-Tris(carboxymethyl)-3,6,9-triazaundecanedioic acid-1-carboxy-11-
oyl]-2-N-
[ 1-O-a-D-carbonylmethyl-mannopyranose]-L-lysine-[ 1-(4-
perfluorooctylsuIfonyl)-piperazine]-
amide
12.10 g (30.0 mmol) of 3-N-(2,6-dioxomorpholinoethyl)-6-N-
(ethoxycarbonylmethyl)-
3,6-diazaoctanedioic acid is added to 24.0 g (26.19 mmol) of the title
compound of Example 1 e),
dissolved in 100 ml of dimethylformamide/30 m1 of pyridine, and it is stirred
for 5 hours at 50°C.
It is evaporated to the dry state in a vacuum. The residue is dissolved in 200
ml of water, and
the pH of the resulting solution is set at 13 by adding 20% aqueous sodium
hydroxide solution.
It is stirred for 8 hours at 22°C and a pH of 13. The solution is
brought to a pH of 7.2 by adding
concentrated hydrochloric acid, and then it is evaporated to the dry state in
a vacuum. The
residue is chromatographed on silica gel RP-18 (mobile solvent: gradient that
consists of
water/ethanol/acetonitrile).
CA 02418790 2003-02-07
41
Yield: 17.26 g (51.0% of theory) of a colorless solid.
Water content: 9.3%.
Elementary analysis (relative to anhydrous substance):
Cld: C 37.19 H 4.21 N 7.59 F 25.00 S 2.48
Fnd: C 37.10 H 4.30 N 7.48 F 25.07 S 2.42
b) 6-N-[3,6,9-Tris(carboxylatomethyl)-3,6,9-triazaundecanedioic acid-1-carboxy-
11-oyl]-2-
N-[ 1-O-a-D-carbonylmethyl-mannopyranose]-L-lysine-[ 1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide, Gd complex, sodium salt
1.40 g (3.87 mmol) of gadolinium oxide is added to 10.0 g (7.74 mmol) of the
title
compound of Example 5a) in 100 ml of water, and it is stirred for 2 hours at
70°C. The solution
is filtered. The filtrate is set at a pH of 7.4 with 2N sodium hydroxide
solution, and it is freeze-
dried.
Yield: 1 I .36 g (quantitative) of an amorphous solid.
Water content: 10.5%.
Elementary analysis (relative to anhydrous substance):
Cld: C 32.72 H 3.43 N 6.68 S 2.18 Gd 10.71 Na 1.57 F 22.00
Fnd: C 32.65 H 3.51 N 6.7I S 2.08 Gd 10.61 Na 1.68 F 21.87
F.xan~fle 6
a) 6-N-Benzyloxycarbonyl-2-N-[1,4,7-tris(carboxylatomethyl)-1,4,7,10-
tetraazacyclododecane]-10-(pentanoyl-3aza 4-oxo-5-methyl-5y1)]-L-lysine-[I-(4-
perfluorooctylsulfonyl)-piperazine]-amide, Gd complex
50.0 g (60.20 mmol) of the title compound of Example lc), 6.93 g (60.20 mmol)
of N-
hydroxysuccinimide, 5.09 g (120.0 mmol) of lithium chloride and 37.91 g (60.20
mmol) of 1,4,7-
tris[carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-10-(pentanoyl-3-aza-4-
oxo-5-methyl-5-
yl), Gd complex, are dissolved in 400 ml of dimethyl sulfoxide while being
heated slightly. At
10°C, 20.63 g (100.0 mmol) ofN,N-dicyclohexylcarbodiimide is added, and
it is then stirred
CA 02418790 2003-02-07
42
overnight at room temperature. The solution is poured into 3000 ml of acetone
and stirred for 10
minutes. The precipitated solid is filtered off and then purified by
chromatography (silica gel
RP-18; mobile solvent: gradient that consists of waterlethanol/acetonitrile).
Yield: 75.53 g (87.0% of theory) of a colorless solid.
Water content: 10.1 %.
Elementary analysis (relative to anhydrous substance):
Cld: C 37.48 H 3.84 N 8.74 S 2.22 F 22.39 Gd 10.90
Fnd: C 37.39 H 4.02 N 8.70 S 2.16 F 22.29 Gd 10.75
b) 2-N-[1,4,7-Tris(carboxylatomethyl]-1,4,7,10-tetraazacyclododecane-Gd
complex, 10-
(pentanoyl-3-aza-4-oxo-5-methyl-5-yl)-L-lysine-[ 1-(4-perfluorooctylsulfonyl)-
piperazine]-amide
70.0 g (48.53 mmol) of the title compound of Example 1 d) is dissolved in 500
ml of
water/100 ml of ethanol, mixed with 5.0 g of palladium catalyst (10% Pd/C) and
hydrogenated at
room temperature under a hydrogen atmosphere (1 atm) until no more hydrogen
absorption can
be observed. Then, catalyst is suctioned out, it is thoroughly rewashed with
ethanol (twice with
75 ml each) and evaporated to the dry state in a vacuum. The title compound is
obtained as a
strongly viscous and colorless oil.
Yield: 63.5 g (quantitative).
Water content: 9.8%.
Elementary analysis (relative to anhydrous substance):
Cld: C 37.48 H 3.84 N 8.74 S 2.22 F 22.39 Gd 10.90
Fnd: C 37.39 H 4.03 N 8.65 S 2.20 F 22.31 Gd 10.78
c) 6-N-(1-O-a-D-Carbonylmethyl-2,3,4,6-tetra-O-benzyl-mannopyranose)-2-N-
[1,4,7-
tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane, Gd-complex-10-
(pentanoyl-3-aza-4oxo-
5-methyl-Syl)]-L-lysine-[ 1-(4-perfluorooctylsulfonyl)-piperazine]-amide
50.0 g (38.22 mmol) of the title compound of Example 6b), 4.40 g (38.22 mmol)
of N-
hydroxysuccinimide, 3.39 g (80.0 mmol) of lithium chloride and 22.88 g (38.22
mmol) of 1-O-a-
CA 02418790 2003-02-07
43
D-carboxymethyl-2,3,4,6-tetra-O-benzyl-mannopyranose are dissolved in 400 ml
of dimethyl
sulfoxide while being heated slightly (30 to 40°C). At 10°C,
10.32 g (50.0 mmol) of N,N-
dicyclohexylcarbodiimide is added, and it then is stirred overnight at room
temperature. The
solution is poured into 3000 ml of acetone and stirred for 10 minutes. The
precipitated solid is
filtered off and then purified by chromatography (silica gel RP-18, mobile
solvent: gradient that
consists of water/ethanol/acetonitrile).
Yield: 64.25 g (89.0% of theory) of a colorless solid.
Water content: 10.9%.
Elementary analysis (relative to anhydrous substance):
Cld: C 46.42 H 4.54 N 6.67 S I .70 F 17.10 Gd 8.33
Fnd: C 46.36 H 4.71 N 6.60 S 1.61 F 17.19 Gd 8.21
d) 6-N-(I-O-a-D-Carbonylmethyl-2,3,4,6-tetra-O-benzyl-mannopyranose)-2-N-
[1,4,7-
tris(carboxylatomethyl)-1,4,8,10-tetraazacyclododecane, Gd-complex-10-
(pentanoyl-3-aza-4oxo-
5-methyl-5y1)]-L-lysine-[ 1-(4-perfluorooctylsulfonyl)-piperazine~-amide
60.0 g (31.77 mmol) of the title compound of Example 6c) is dissolved in 500
ml of
ethanol and mixed with 6.0 g of palladium catalyst ( 10% Pd/C). It is
hydrogenated at room
temperature under a hydrogen atmosphere ( 1 atm) until no more hydrogen
absorption can be
observed. Then, catalyst is suctioned out, it is thoroughly rewashed with
ethanol (twice with 150
ml. each) and evaporated to the dry state in a vacuum.
Yield: 48.55 g (quantitative) of a colorless solid.
Water content: 3.9%.
Elementary analysis (relative to anhydrous substance}:
Cld: C 35.37 H 4.02 N 8.25 S 2.10 F 21.13 Gd 10.29
Fnd: C 35.28 H 4.13 N 8.17 S 2.03 F 21.05 Gd 10.20
CA 02418790 2003-02-07
44
F:xamnle 77
a) 1,7-Bis-(benzyloxycarbonyl)-4-[2-(N-ethyl-N-perfluorooctylsulfonyl]-amino]-
acetyl]-
1,4,7,10-tetraazacyclododecane)
49.46 g (200.0 mmol) of EEDQ (2-ethoxy-1,2-dihydroquinoline-1-carboxylic acid
ethyl
ester) is added at 0°C to 50.0 g (113.5 mmol) of 1,7-
bis(benzyloxycarbonyl)-1,4,7,10-
tetraazacyclododecane and 66.42 g ( 113.5 mmol) of 2-(N-ethyl-N-
perfluorooctylsulfonyl)-
aminoacetic acid (produced according to DE 196 03 033) in 300 ml of
tetrahydrofuran, and it is
stirred overnight at room temperature. It is evaporated to the dry state in a
vacuum and
chromatographed on silica gel (mobile solvent: dichloromethane/methanol =
20:1).
Yield: 65.2 g (57% of theory) of a colorless solid.
Elementary analysis:
Cld: C 42.91 H 3.80 N 6.95 F 32.05 S 3.18
Fnd: C 42.85 H 3.90 N 6.87 F 31.98 S 3.1 ~
b) 1,7-Bis-(benzyloxy)-4-[2-(N-ethyl-N-perfluorooctylsulfonyl)-amino]-acetyl-
10-[1-O-a-D-
carbonylmethyl-2,3,4,6-tetra-O-benzyl-mannopyranose]-1,4,7,10-
tetraazacyclododecane
24.73 g (100 mmol) of EEDQ (2-ethoxy-1,2-dihydroquinoline-1-carboxylic acid
ethyl
ester) is added at 0°C to 60.0 g (59.53 mmol) of the title compound of
Example 7a) and 35.64 g
(59.53 mmol) of 1-O-a-D-carboxymethyl-2,3,4,6-tetra-O-benzyl-mannopyranose,
produced
according to DE 19728954, in 300 ml of tetrahydrofuran, and it is stirred
overnight at room
temperature. It is evaporated to the dry state in a vacuum and chromatographed
on silica gel
(mobile solvent: dichloromethane/methanol = 20:1).
Yield: 76.6 g (81.0% of theory) of a colorless solid.
Elementary analysis:
Cld: C 54.44 H 4.70 N 4.41 F 20.33 S 2.02
Fnd: C 54.37 H 4.81 N 4.35 F 20.27 S 1.96
CA 02418790 2003-02-07
45
c) 1-[2-(N-Ethyl-N-perfluorooctylsulfonyl)-amino]-acetyl-7-(1-O-a-D-
carbonyimethyl-
mannopyranose)-1,4,7,10-tetraazacyclododecane
70 g (44.07 mmol) of the title compound of Example 7b is dissolved in 800 ml
of
ethanol, and 8 g of palladium catalyst (10% Pd/C) is added. It is hydrogenated
at room
temperature. Catalyst is filtered out, and the filtrate is evaporated to the
dry state in a vacuum.
Yield: 42.3 g (quantitative) of a colorless solid.
Elementary analysis:
Cld: C 35.04 H 3.99 N 7.30 F 33.65 S 3.34
Fnd: C 35.15 H 4.13 N 7.13 F 33.48 S 3.26
d) 1,7-Bis-[1,4,7-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-Gd-
complex-10-
(pentanoyl-3-aza-4-oxo-5-methyl-Syl)-4-[2-(N-ethyl-N-perfluorooctylsulfonyl)-
amino]-acetyl-
10-( 1-O-a-D-carbonylmethyl-mannopyranose)-1,4,7,10-tetraazacyclododecane
20 g (20.84 mmol) of the title compound of Example 7c, 5.09 g (120 mmol) of
lithium
chloride and 37.78 g (60 mmol) of 1,4,7-tris(carboxylatomethyl)-10-pentanoyl-3-
aza-4-oxo-5-
methyl-Syl)-1,4,7,10-tetraazacyclododecane, Gd complex, are dissolved in 400
ml of dimethyl
sulfoxide while being heated slightly. At 10°C, 29.67 g (120 mmol) of
EEDQ is added, and then
it is stirred overnight at room temperature. The solution is poured into 3000
ml of acetone and
stirred for 10 minutes. The precipitated solid is filtered off and then
purified by chromatography
(silica gel RP-18, mobile solvent: gradient that consists of water/ethanol/
acetonitrile).
Yield: 13.2 g (29.0% of theory) of a colorless solid:
Water content: 11.8%.
Elementary analysis (relative to anhydrous substance):
Cld: C 36.31 H 4.34 N 9.62 S 1.47 F 14.79 Gd 14.41
Fnd: C 36.24 H 4.27 N 9.58 S 1.51 F 14.85 Gd 14.25
CA 02418790 2003-02-07
CA 02418790 2003-02-07
46
a) 1,7-Bis(benzyloxycarbonyl)-4-[2-(N-ethyl-N-perfluorooctylsulfonyl)-amino]-
acetyl-10-
[pentanoyl-3-aza-4-oxo-5-methyl-5yl-[1,4,7-tris(carboxylatomethyl)-Gd complex,
1,4,7,10-
tetraazacyclododecan-10-ylJ-1,4,7,10-tetraazacyclododecane
50.0 g (49.61 mmol) of the title compound of Example 7a), 5.71 g (49.61 mmol)
of N-
hydroxysuccinimide, 4.24 g (100 mmol) of lithium chloride and 31.24 g (49.61
mmol) of 1,4,7-
tris(carboxylatomethyl)-10(pentanoyl-3-aza-4-oxo-5-methyl-5-yl)-1,4,7,10-
tetraazacyclododecane, Gd complex, are dissolved in 350 ml of dimethyl
sulfoxide while being
heated slightly. At 10°C, 15.47 g (75 mrnol) of N,N-
dicyclohexylcarbodiimide is added, and it
then is stirred overnight at room temperature. The solution is poured into
2000 ml of acetone and
stirred for 10 minutes. The precipitated solid is filtered off and then
purified by chromatography
(silica gel RP-18, mobile solvent: gradient that consists of
water/ethanol/acetonitrile).
Yield: 65.1 g (81.0% of theory) of a colorless solid.
Water content: 7.9%.
Elementary analysis (relative to anhydrous substance):
Cld: C 40.79 H 4.11 N 8.65 S I .98 F 19.94 Gd 9.72
Fnd: C 40.71 H 4.20 N 8.58 S 2.03 F 19.87 Gd 9.68
b) 1-[2-(N-Ethyl-N-perfluorooctylsulfonyl)-amino]-acetyl-7{-pentanoyl-3-aza-4-
oxo-5-
methyl-5 yl-[tri s(carboxylatomethyl-1,4, 7,10-tetraazacycl ododecan-10-yl]-Gd-
complex } -1,4,7,10
tetiaazacyclododecane
60.0 g (37.05 mmol) of the title compound of Example 8a) is dissolved in 600
ml of
ethanol, and 6.0 g of palladium catalyst (10% PdIC) is added. It is
hydrogenated at room
temperature. Catalyst is filtered out, and the filtrate is evaporated to the
dry state in a vacuum.
Yield: 50.06 g (quantitative) of a colorless solid.
Water content: 3.9%.
Elementary analysis (relative to anhydrous substance):
Cld: C 34.67 H 4.03 N 10.37 S 2.37 F 23.90 Gd 11.64
CA 02418790 2003-02-07
47
Fnd: C 34.58 H 4.15 N 10.2$ S 2.30 F 23.84 Gd 11.57
c) 1-[2-(N-Ethyl-N-perfluorooctylsulfonyl)-amino]-acetyl-4,10-bis[I-O-a-D-
carbonylmethyl-2,3,4,6-tetra-O-benzyl-rriannopyranose]-7-{pentanoyl-3-aza-4-
oxo-5-methyl-S-
yl-[1,4,7-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-10-yl)-Gd
complex}-1,4,7,10-
tetraazacyclododecane
40.0 g (29.60 mmol) of the title compound of Example 8b), 2.54 g (60.0 mmol)
of
lithium chloride and 44.9 g (75.0 mmol) of 1-O-a-D-carboxymethyl-2,3,4,6-tetra-
O-benzyl-
mannopyranose are dissolved in 300 ml of dimethyl sulfoxide while being heated
slightly. At
10°C, 24.73 g ( 100.0 mrnol) of EEDQ is added, and it then is stirred
overnight at room
temperature. The solution is poured into 3000 ml of acetone and stirred for 10
minutes. The
precipitated solid is filtered off and then purified by chromatography (silica
gel RP-18, mobile
solvent: gradient that consists of water/ethanol/acetonitrile).
Yield: 31.98 g (43.0% of theory) of a colorless solid.
Water content: 3.5%.
Elementary analysis (relative to anhydrous substance):
Cld: C 53.06 H 5.05 N 5.57 S 1.28 F 12.85 Gd 6.26
Fnd: C 52.95 H 5.19 N 5.48 S 1.23 F 12.77 Gd 6.14
d) 1-[2-(N-Ethyl-N-perfluorooctylsulfonyl)-amino)-acetyl-4,I0-bis[1-O-a-D-
carbonylmethyl-2,3,4,6-tetra-O-benzyl-mannopyranose]-7-{pentanoyl-3-aza-4-oxo-
5-methyl-5-
yl-[1,4,7-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-10-yl]-Gd
complex}-1,4,7,10-
tetraazacyclododecane
30.0 g (11.94 mmol) of the title compound of Example 8c) is dissolved in 300
ml of
ethano1/30 ml of water, and 4.0 g of palladium catalyst (10% PdIC) is added.
It is hydrogenated
at room temperature, catalyst is filtered out, and the filtrate is evaporated
to the dry state in a
vacuum.
Yield: 21.39 g (quantitative) of a colorless solid.
4s
Water content: 3.4%.
Elementary analysis (relative to anhydrous substance):
Cld: C 36.87 H 4.39 N 7.82 S 1.79 F 18.03 Gd 8.78
Fnd: C 36.80 H 4.50 N 7.85 S 1.68 F 17.91 Gd 8.70
Framnle 99
a) 6-N-[3,6-Bis(carboxymethyl)-octane-1,8-dicarboxylic acid-1-carboxy-8-oyl]-2-
N-(1-O-a-
D-carboxymethyl-mannopyranose)-lysine-[ 1-(4-perfluorooctylsulfonyl)-
piperazine]-amide
25.62 g (100.0 mmol) o~ethylenediamine-N,N,N',N'-tetraacetic acid dianhydride
is added
to 27.5 g (30.0 mmol) of the title compound of Example 1e), dissolved in 300
ml of
dimethylformamide/100 ml of pyridine, and it is stirred for 5 hours at
50°C. It is evaporated to
the dry state in a vacuum. The residue is dissolved in 300 ml of water, set at
a pH of 10 by
adding 20% aqueous sodium hydroxide solution, and then the basic product
solution is brought
to a pH of 3 by adding concentrated hydrochloric acid, and it is evaporated to
the dry state in a
vacuum. The residue is chromatographed on silica gel RP-18 (mobile solvent:
gradient that
consists of water/ethanol/ acetonitrile).
Yield: 18.22 g (51.0% of theory) of a colorless solid:
Water content: 7.9%.
Elementary analysis (relative to anhydrous substance):
Cld: C 36.31 H 3.98 N 7.06 F 27.12 S 2.69
Fnd: C 36.23 H 4.07 N 6.98 F 27.05 S 2.62
b) 6-N-[3,6-Bis(carboxylatomethyl)-octane-1,8-dicarboxylic acid-1-carboxylato-
8-oyl-Mn
complex, sodium salt]-2-N-( 1-O-a-D-carboxymethyl-mannopyranose)-L-lysine-[ 1-
(4-
perfluorooctylsulfonyl)-piperazine]-amide
g (8.397 mmol) of the title compound of Example 9a) is dissolved in 200 ml of
water.
965 mg (8.397 mmol) of manganese(II) carbonate is added, and it is stirred for
3 hours at 60°C.
The solution is set at a pH of 7.4 with 5% aqueous sodium hydroxide solution,
filtered, and then
CA 02418790 2003-02-07
49
freeze-dried.
Yield: 10.52 g (99.0% of theory) of a colorless solid.
Water content: 7.8%.
Elementary analysis (relative to anhydrous substance):
Cld: C 34.16 H 3.50 N 6.64 S 2.53 F 25.52 Mn 4.34 Na 1.82
Fnd: C 34.06 H 3.61 N 6.58 S 2.47 F 25.47 Mn 4.30 Na 1.97
Fxam~le t (1
a) 1,2,3,4,6-Penta-O-acetyl-a,[i-D-mannopyranose
Analogously to what is described in the literature [M. L. Wolfrom and A.
Thompson in
Methods in Carbohydrate Chemistry (R. L. Whistler, M. L. Wolfrom and J. N.
BeMiller, Eds.),
Academic Press, New York, Vol. II, 53, pp. 211-215, (1963)], the reaction of
150 g (832.5
mmol) of a,/3-D-mannopyranose with a mixture that consists of 1500 ml of
absolute pyridine and
1500 ml of acetic acid anhydride yields, after working-up, 315 g (96.7%) of
the above-mentioned
title compound as a crude product in the form of a viscous and colorless oil.
By 1H-NMR
spectroscopic study of the thus obtained title compound, it was possible to
determine the a to [i
ratio of both anomers at 4:1. Separation of a,(3-anomers of the above-
mentioned title compound
can be eliminated to perform the subsequent reaction step.
Elementary analysis:
Cld: C 49.21 H 5.68
Fnd: C 49.12 H 5.78
b) 1-O-a-D-(5-Ethoxycarbonyl)-pentyl-2,3,4,6-tetra-O-acetyl-mannopyranose
Analogously to what is described in the literature for the synthesis of aryl
glycopyranosides [J. Conchie and G. A. Levvy in Methods in Carbohydrate
Chemistry (R. L.
Whistler, M. L. Wolfrom and J. N. BeMiller, Eds.), Academic Press, New York,
Vol. II, 90, pp.
345-347, (1963)], the reaction of 156.2 g (400 mmol) of the title compound of
Example 10a) as
an a,(i-anomer mixture with 67 ml (400 mmol) of 6-hydroxy-hexanoic acid ethyl
ester and 60.8
CA 02418790 2003-02-07
50
ml (~20 mmol) of tin(IV) chloride results in a total of 600 ml of 1,2-
dichloroethane after column-
chromatographic purification (eluant: hexane/ethyl acetate 2:1) to form 100.05
g (51% of theory)
of the above-mentioned title compound as a colorless and viscous oil. By ~H-
NMR-
spectroscopic study of the thus obtained title compound, it was possible to
show that the above-
mentioned title compound is only the pure a-anomer.
Elementary analysis:
Cld: C 52.94 H 6.77
Fnd: C 52.80 H 6.78
c) 1-O-a-D-(5-Carboxy)-pentyl-2,3,4,6-tetra-O-benzyl-mannopyranose
A stirred suspension of 141.0 g (289 mmol) of the title compound of Example l
Ob) in
200 ml of dioxane is mixed at room temperature and with simultaneous vigorous
stirring in
portions with a total of 238.5 g (4.26 mol) of fine-powder potassium hydroxide
powder. To
make it easier to stir, the reaction mixture is mixed with another 200 ml of
dioxane, and the thus
obtained suspension is subsequently heated to boiling heat and mixed drop by
drop at this
temperature with a total of 372 ml (3.128 mol) of benzyl bromide over a period
of two hours.
After a reaction time of 4 hours at 110°C followed by 12 hours at room
temperature, the reaction
mixture is slowly poured into a total of 2.5 liters of ice water for the
purpose of working-up, and
the aqueous phase is subsequently completely extracted with diethyl ether.
After the thus
obtained ether phase is washed and after the subsequent drying of the same on
sodium sulfate,
salt is suctioned out, and the diethyl ether is drawn off in a vacuum. Excess
benzyl bromide is
then distilled off from the reaction mixture in an oil pump vacuum
quantitatively at an oil bath
temperature of 180°C. The thus obtained, resinous-oily residue is
purified on silica gel with use
of ethyl acetate/hexane ( 1:10) as an eluant.
Yield: 172.2 g (91.0% of theory) of the above-mentioned title compound in the
form of a
colorless and extremely viscous oil.
CA 02418790 2003-02-07
CA 02418790 2003-02-07
51
Elementary analysis:
Cld: C 75.68 H 7.I6
Fnd: C 75.79 H 7.04
d) 6-N-Benzyloxycarbonyl-2-N-[1-O-a-D-(5-carbonyl)-pentyl-2,3,4,6-tetra-O-
benzyl-
mannopyranose]-L-lysine-[ 1-(4-perfluorooctylsulfonyl)-piperazine]-amide
100.0 g (134.0 mmol) of the carboxylic acid that is produced under Example l
Oc) and
32.4 g (281.4 mmol) of N-hydroxysuccinimide are dissolved in 500 ml of
dimethylformamide
and mixed in portions at 0°C with a total of 58.0 g (281.4 mmol) of
N,N'-
dicyclohexylcarbodiimide, and it is stirred for 3 more hours at this
temperature. A solution,
cooled to 0°C, of 111.3 g ( 134.0 mmol) of the title compound of
Example 1 c), dissolved in 300
ml of dimethylformamide, is added drop by drop to the thus produced active
ester solution, and it
is stirred for 2 hours at 0°C and for 1_2 hours at room temperature.
For working-up, precipitated
r dicyclohexylurea is filtered out, and the solvent is then evaporated to the
dry state. The thus
obtained residue is then chromatographed on silica gel (mobile solvent:
dichloromethane/ethanol
20:1; chromatography is carned out with use of a solvent gradient with
continuous increase of
the proportion of ethanol).
Yield: 132.5 g (67.4% of theory) of the title compound in the form of a
colorless and
strongly viscous oil.
Elementary analysis:
Cld: C 54.02 H 4.88 N 3.82 F 22.01 S 2.19
Fnd: C 53.87 H 4.85 N 4.02 F 22.55 S 2.06
e) 2-N-[ 1-O-a-D-(5-Carbonyl)pentyl-mannopyranose]-L-lysine-[ 1-(4-
perfluorooctylsulfonyl)-piperazine]-amide
120.0 g (81.77 mmol) of the compound, produced under l Od), is dissolved in
800 ml of
ethanol, mixed with 4.5 g of Pearlman's catalyst (Pd 20%, C) and hydrogenated
at room
temperature under a hydrogen atmosphere ( 1 atm) until no more hydrogen
absorption can be
52
observed (about 8 hours). Catalyst is suctioned out, it is thoroughly rewashed
with ethanol
(about 200 ml) and evaporated to the dry state in a vacuum. The title compound
is obtained as a
strongly viscous and colorless oil.
Yield: 78.5 g (98.7% of theory)
Elementary analysis:
Cld: C 37.04 H 4.25 N 5.76 F 33.20 S 3.30
Fnd: C 36.96 H 4.85 N 5.41 F 34.13 S 3.22
f) 2-N-[1-O-a-D-(5-Carbonyl)pentyl-mannopyranose]-6-N-[1,4,7-tris-
(carboxylatomethyl)-
10-(-3-aza-4-oxo-5-methyl-5-yl-pentanoyl)-1,4,7,10-tetraazacyclododecane]-L-
lysine-[ 1-(4-
perfluorooctylsulfonyl)-piperazine]-amide, Gd complex
99.8 g (158.4 mmol; 2.2 molar equivalents relative to the amine component of
Example
10e) that is used of the Gd complex, described in Patent Application DE 197 28
954 C 1 under
Example 31h), of 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4,7,10-
tetraazacyclododecane-
1,4,7-triacetic acid and 6.7 g of anhydrous lithium chloride (158.4 mmol) are
dissolved at 40°C in
800 mI of absolute dimethyI sulfoxide while being stirred. At this
temperature, it is subsequently
mixed with a total of 18.2 g ( 158.4 mmol) of N-hydroxysuccinimide and 70.0 g
(71.96 mmol) of
the title compound of Example 10e), dissolved in 250 ml of absolute dimethyl
sulfoxide. After
cooling to room temperature, the reaction solution is mixed with 32.7 g (158.4
mmol) of N,N'-
dicyclohexylcarbodiimide and stirred for 12 hours at room temperature. The
suspension that is
obtained is~then mixed with sufficient acetone until precipitation of the
above-mentioned title
compound is completed, the precipitate is suctioned off, dried, taken up in
water, insoluble
dicyclohexylurea is filtered out, and the filtrate is desalinated with an
AMICON~R~ YM-3
ultrafiltration membrane (cut-off: 3,000 Da), and low-molecular components are
removed. The
retentate is then freeze-dried.
Yield: 93.0 g (81.6% of theory) as a colorless lyophilizate.
H20 content (Karl-Fischer): 9.53%.
Elementary analysis (relative to anhydrous substance):
CA 02418790 2003-02-07
53
Cld: C 37.15 H 4.39 N 7.96 F 20.38 S 2.02 Gd 9.92
Fnd: C 36.92 H 4.50 N 7.68 F 19.77 S 1.91 Gd 10.08
Fxamr ~! a ~ 1
a) 2-N-[1-O-a-D-(5-Carbonyl)pentyl-mannopyranose]-6-N-{2-[4-(3-oxapropionyl)-
phenyl]-
2-[1,4,7-tris-(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-10-yl]-acetic
acid}-L-lysine-[1-
(4-perfluorooctylsulfonyl)-piperazine]-amide, Gd complex, sodium salt
A stirred suspension of 5.0 g (9.06 mmol) of the title compound of Example 3e)
in 15 ml
of absolute dimethyl sulfoxide is mixed at 70°C with 0.68 g (15.9 mmol)
of lithium chloride.
After 30 minutes of stirring at 70°C, the now clear reaction solution
is mixed in portions with a
total of 1.83 g (15.9 mmol) of N-hydroxysuccinimide, and the reaction mixture
is kept at this
temperature for 1 more hour. After cooling to 0°C, it is mixed with
4.52 g (23.85 mmol) of
dicyclohexylcarbodiimide, and the reaction solution is stirred for another
hour at 0°C, followed
by 12 hours at 22°C. The thus obtained reaction solution of the N-
hydroxysuccinimide ester of
the title compound of Example 3e) is now mixed drop by drop at 22°C
with a solution of 4.0 g
(4.12 mmol) of the title compound of Example 10e) in 15 ml of absolute
dimethyl sulfoxide, and
it is stirred for another 12 hours at room temperature. For working-up, the
reaction solution is
added in drops at 22°C in 900 ml of acetone, whereby the title compound
precipitates as a
colorless precipitate. The precipitate is suctioned off, dissolved in 200 ml
of distilled water, and
then the pH of this solution is set at exactly 7.2 with 1 molar sodium
hydroxide solution. The
thus obtained aqueous product solution is ultrafiltered three times with a YM-
3 ultrafiltration
membrane (AMICON~R~: cut-off: 3,000 Da) for the purpose of desalination and
the separation of
low-molecular components. The thus obtained retentate is then freeze-dried.
Yield: 6.33 g (92.4% of theory, relative to the amine component that is used)
as a
colorless lyophilizate with a water content of 7.38%.
Elementary analysis (relative to anhydrous substance):
Cld: C 38.48 H 4.13 N 6.65 F 19.16 S 1.90 Gd 9.33 Na 1.36
Fnd: C 39.52 H 4.12 N 6.67 F 19.70 S 1.89 Gd 9.30 Na 1.41
CA 02418790 2003-02-07
54
Fxam In a 12
a) 3,5-Bis-benzyloxycarbonylamino-benzoic acid-N-{3-oxa-
1H,1H,2H,2H,4H,4H,SH,SH-
perfluorotridecyl)-amide
20 g (47.5 mmol) of 3,5-bis-benzyloxycarbonylamino-benzoic acid (synthesis
according
to the subsequent bibliographic reference: Skulnick, Harvey L; Johnson, Paul
D.; Aristoff, Paul
A.; Morris, Jeanette K.; Lovasz, Kristine D.; et al.; J. Med. Chem.; 40; 7;
1997; 1149-1164) and
4.78 g (47.5 mmol) of triethylamine are dissolved in a solvent mixture that
consists of 125 ml of
dry tetrahydrofuran and 125 ml of dry dioxane. After cooling to -15°C,
a solution of 6.56 g (48
mmol) of isobutyl chloroformate in 30 ml of dry tetrahydrofuran is slowly
added in drops while
being stirred, whereby the internal temperature is to be kept below -
10°C. After a reaction time
of 15 minutes at -15°C, a solution of 58.6 g (47.5 mmol) of 1-amino-
1H,1H,2H,2H,4H,4H,SH,SH-3-oxa-perfluorotridecane and 4.78 g (47.5 mmol) of
triethylamine
in 100 ml of dry tetrahydrofuran is added in drops at -20°C. After a
reaction time of one hour at -
15°C and two hours at room temperature, the reaction solution is
evaporated to the dry state in a
vacuum. The remaining residue is taken up in 300 ml of ethyl acetate and
washed twice with 200
ml each of saturated sodium bicarbonate solution and once with 300 ml of
water. After the
organic phase is dried on sodium sulfate, salt is suctioned out, and the ethyl
acetate is drawn off
in a vacuum. The remaining oily residue is purified on silica gel with-use of
dichloromethane/hexane/2-propanol (10:5:1) as an eluant.
Yield: 36.2 g (82.5% of theory) of the title compound as a colorless oil.
Elementary analysis:
Cld: C 46.82 H 3.27 N 4.55 F 34.97
Fnd: C 47.21 H 3.31 N 4.61 F 34.48
b) 3,5-Di-amino-benzoic acid-N-(3-oxa-1H,1H,2H,2H,4H,4H,SH,SH-
perfluorotridecyl)]-
amide
30.0 g (32.4 mmol) of the amide that is produced under 12a) is dissolved in
300 ml of
CA 02418790 2003-02-07
55
ethanol and mixed with 1.2 g of Pearlman's catalyst (Pd 20%, C). It is
hydrogenated at room
temperature under a hydrogen atmosphere ( 1 atm) until no more hydrogen
absorption can be
observed. Catalyst is suctioned out, it is thoroughly rewashed with ethanol
(about 300 ml) and
evaporated to the dry state in a vacuum. The title compound is obtained as a
strongly viscous,
yellowish oil.
Yield: 20.12 g (94.8% of theory).
Elementary analysis:
Cld: C 36.66 H 2.77 N 6.41 F 49.28
Fnd: C 36.07 H 2.87 N 6.23 F 49.43
c) 3-N-[-(1-O-a-D-Carbonylmethyl-2,3,4,6-tetra-O-benzyl-mannopyrariose)-5-
amino-
benzoic acid-N-(3-oxa-1 H,1 H,2H,2H,4H,4H,5H,5H-perfluorotridecyl)-amide
10.95 g (18.30 mmol) of 1-carboxymethyloxy-2,3,4-tetra-O-benzyl-a-D-
mannopyranoside [production as described in Patent DE 197 28 954 C 1 ] is
dissolved in 150 ml
of dimethylformamide and mixed with a total of 2.09 g ( 18.3 mmol) of N-
hydroxysuccinimide.
It is cooled to 0°C, and 3.78 g (18.3 mmol) of dicyclohexylcarbodiimide
is added. It is stirred for
one hour at 0°C and then for 4 hours at room temperature. It is cooled
to 0°C, and a solution that
consists of 24.0 g (36.6 mmol, 2 molar equivalents relative to the carboxylic
acid that is used) of
the diamino compound, described under Example 12b), dissolved in 350 ml of
dimethyl-
formamide, is slowly added in drops within 3 hours. Then, it is stirred for
one more hour at 0°C,
then overnight at room temperature. It is evaporated to the dry state in a
vacuum, and the residue
is taken up in 300 ml of ethyl acetate. Precipitated urea is filtered out, and
the filtrate is washed
twice with 100 ml each of 5% aqueous soda solution. The organic phase is dried
on magnesium
sulfate and evaporated to the dry state in a vacuum. The residue is
chromatographed on silica gel
(mobile solvent: n-hexane/isopropanol 13:1). 16.8 g (74.3% of theory, relative
to the carboxylic
acid that is used) of the title compound is obtained in the form of a
colorless oil. By increasing
the polarity of the eluant composition to n-hexane/isopropanol 5:1, a total of
10.15 g of unreacted
diamino compound 12b) is recovered in the subsequent chromatography fractions,
which can be
CA 02418790 2003-02-07
56
reacted again according to the above-mentioned reaction instructions.
Elementary analysis:
Cld: C 54.42 H 4.40 N 3.40 F 26.13
Fnd: C 54.32 H 4.49 N 3.48 F 25.94
d) 3-N-[-(1-O-a-D-Carbonylmethyl-mannopyranose)J-5-amino-benzoic acid-N-(3-oxa-
1 H,1 H,2H,2H,4H,4H,5H,5H-perfluorotridecyl)-amide
Analogously to what is described for the synthesis of the title compound of
Example
12b), the hydrogenolysis of 12.0 g (9.70 mmol) of the title compound of
Example 12c), with use
of 0.5 g of Pearlman's catalyst (Pd 20%, C) in an ethanol/water (9:1 ) mixture
after working-up
yields 8.08 g (96.7% of theory) of the above-mentioned title compound in the
form of a
yellowish-colored and viscous oil.
Elementary analysis:
Cld: C 37.64 H 3.28 N 4.88 F 37.49
Fnd: C 37.32 H 3.17 N 4.97 F 37.55
e) 3-N-(1-O-a-D-Carbonylmethyl-mannopyranose)-5-N-{2-[4-(3-oxapropionyl)-
phenyl]-2-
[1,4,7-tris-(carboxylatomethyl)-1,4,7;10-tetraazacyclododecan-10-ylJ-acetic
acid}-benzoic acid-
N-(3-oxa-1H,1H,2H,2H,4H,4H,5H,5H-perfluorotridecyl)-amide, Gd complex, sodium
salt
13.6 g (19.2 mmol; 2.2 molar equivalents relative to the amine component that
is used of
Example 12d)) of the Gd complex that is described under Example 3e) and 0.81 g
of anhydrous
lithium chloride (19.2 mmol) are dissolved at 40°C in 100 ml of
absolute dimethyl sulfoxide
while being stirred, and it is mixed at this temperature with a total of 2.2 g
( 19.2 mmol) of N-
hydroxysuccinimide and 7.5 g (8.7 mmol) of the title compound of Example 12d),
dissolved in
50 ml of absolute dimethyl sulfoxide. After cooling to room temperature, the
reaction solution is
mixed with 3.96 g (19.2 mmol) of N,N'-dicyclohexylcarbodiimide and stirred for
12 hours at
room temperature. The suspension that is obtained is then mixed with
sufficient acetone until
precipitation of the above-mentioned title compound is completed, the
precipitate is suctioned
CA 02418790 2003-02-07
57
off, dried, taken up in water, insoluble dicyclohexylurea is filtered out, and
the filtrate is
desalinated with an AMICON~R~ YM-3 ultrafiltration membrane (cut-off 3,000
Da), and low-
molecular components are removed. The retentate is then freeze-dried.
Yield: I 1.51 g (84.5% of theory) as a colorless lyophilizate.
H20 content (Karl-Fischer): 6.77%.
Elementary analysis (relative to anhydrous substance):
Cld: C 40.05 H 3.94 N 6.29 F 20.71 Gd 10.08 Na I.47
Fnd: C 39.98 H 4.00 N 6.31 F 20.73 Gd 10.11 Na 1.42
FxamT
a) 3,5-Bis-(benzyloxycarbonylamino)-1-{N-[1-(4-perfluorooctylsulfonyl)-
piperazine]}-
benzamide
g (23.75 mmol) of 3,5-bis-benzyloxycarbonylamino-benzoic acid (synthesis
according
to the subsequent bibliographic reference: Skulnick, Harvey L; Johnson, Paul
D.; Aristoff, Paul
A.; Moms, Jeanette K.; Lovasz, Kristine D.; et al.; J. Med. Chem.; 40; 7;
1997; 1149-I 164) and
2.39 g (23.75 mmol) of triethylamine are dissolved in a solvent mixture that
consists of 60 ml of
dry tetrahydrofuran and 70 ml of dry dioxane. After cooling to -15°C, a
solution of 3.28 g (24
mmol) of isobutyl chloroformate in 20 ml of dry tetrahydrofuran is slowly
added in drops,
whereby the internal temperature does not exceed -10°C. After a
reaction time of 15 minutes at
-15°C, a solution of 23.0 g (23.75 mmol) of
perfluorooctylsulfonylpiperazine and 2.39 g (23.75
mmol) of triethylamine in 50 ml of dry tetrahydrofuran is added in drops at -
20°C. After a
reaction time of one hour at -15°C and two hours at room temperature,
the reaction solution is
evaporated to the dry state in a vacuum. The remaining residue is taken up in
200 ml of ethyl
acetate and washed twice with 100 ml each of saturated sodium bicarbonate
solution and once
with 300 ml of water. After the organic phase is dried on sodium sulfate, salt
is suctioned out,
and the ethyl acetate is drawn off in a vacuum. The remaining oily residue is
purified on silica
gel with use of dichloromethane/hexane/2-propanol ( 15:5:1 ) as an eluant.
Yield: 18.35 g (79.6% of theory) of the title compound as a colorless oil.
CA 02418790 2003-02-07
58
Elementary analysis:
Cld: C 43.31 H 2.80 N 5.77 F 33.27 S 3.30
Fnd: C 43.21 H 2.75 N 5.61 F 33.38 S 3.22
b) 3,5-Di-amino-1-~N-[1-(4-perfluorooctylsulfonyl)-piperazine]}-benzamide
9.70 g (10.0 mmol) of the amide that is produced under 13a) is dissolved in
100 ml of
ethanol and mixed with 0.4 g of Pearlman's catalyst (Pd 20%, C). It is
hydrogenated at room
temperature under a hydrogen atmosphere ( 1 atm) until no more hydrogen
absorption can be
observed. Catalyst is suctioned out, it is thoroughly rewashed with ethanol
(about 150 ml) and
evaporated to the dry state in a vacuum. The title compound is obtained as a
strongly viscous,
yellowish oil.
Yield: 6.9 g (98.2% of theory).
Elementary analysis:
Cld: C 32.49 H 2.15 N 7.98 F 45.98 S 4.56
Fnd: C 32.56 H 2.17 N 8.09 F 45.63 S 4.61
c) 5-Amino-3-N-(1-O-a-D-carbonylmethyl-2,3,4,6-tetra-O-benzyl-mannopyranose)-
benzoic
acid-N-[ 1-(4-perfluorooctylsulfonyl)-piperazine]-amide
5.48 g (9.15 mmol) of 1-carboxymethyloxy-2,3,4-tetra-O-benzyl-a-D-
mannopyranoside
[production as described in Patent DE I 97 28 954 C 1 ] is dissolved in 100 ml
of
dimethylformamide and mixed with a total of 1.04 g (9.15 mmol) of N-
hydroxysuccinimide. It is
cooled to 0°C, and 1.89 g (91.5 mmol) of dicyclohexylcarbodiimide is
added. It is stirred for one
hour at 0°C and then for 4 hours at room temperature. After being
cooled to 0°C again, a solution
that consists of 12.85 g (18.30 mmol, 2 molar equivalents relative to the
carboxylic acid that is
used) of the diamino compound, described under Example 13b) and dissolved in
250 ml of
dimethylformamide, is slowly added in drops within 3 hours. Then, it is
stirred for one more
hour at 0°C, then overnight at room temperature. It is evaporated to
the dry state in a vacuum,
and the residue is taken up in 100 ml of ethyl acetate. Precipitated urea is
filtered out, and the
CA 02418790 2003-02-07
59
filtrate is washed twice with 100 ml each of 5% aqueous soda solution. The
organic phase is
dried on magnesium sulfate and evaporated to the dry state in a vacuum. The
residue is
chromatographed on silica gel (mobile solvent: n-hexane/isopropanol 13:1).
8.14 g (69.4% of
theory, relative to the carboxylic acid that is used) of the title compound is
obtained in the form
of a colorless oil. By increasing the polarity of the eluant composition
during chromatography to
6:1 (n-hexane/isopropanol), a total of 4.36 g of unreacted diamino compound
13b) is recovered
in the chromatography fractions below, which can be reacted again according to
the above-
mentioned reaction instructions.
Elementary analysis:
Cld: C 51.49 H 4.01 N 4.37 F 25.17 S 2.50
Fnd: C 51.60 H 4.19 N 4.28 F 25.14 S 2.44
d) 5-Amino-3-N-( 1-O-a-D-carbonylmethyl-mannopyranose)-benzoic acid-N-[ 1-(4-
perfluorooctylsulfonyl)-piperazine]-amide
Analogously to what is described for the synthesis of the title compound of
Example
13b), the hydrogenolysis of 6.4 g (5.0 mmol) of the title compound of Example
13c) with use of
0.3 g of Pearlman's catalyst (Pd 20%, C) in an ethanol/water (8:1) mixture
after working-up
yields 4.43 g (96.2% of theory) of the above-mentioned title compound in the
form of a
yellowish-colored and viscous oil.
Elementary analysis:
Cld: C 35.15 H 2.95 N 6.07 F 35.01 S 3.48
Fnd: C 35.32 H 3.02 N 5.89 F 35.05 S 3.58
e) 3-N-(1-O-a-D-Carbonylmethyl-mannopyranose)-5-N-[1,4;7-
tris(carboxylatomethyl)-10-
(3-aza-4-oxo-5-methyl-5-yl-pentanoyl)-1,4,7,10-tetraazacyclododecane]-benzoic
acid-N-[1-(4-
perfluorooctylsulfonyl)-piperazine]-amide, Gd complex
5.54 g (8.8 mmol; 2.2 molar equivalents relative to the amine component of
Example
13d) that is used) of the Gd complex, described in Patent Application DE 197
28 954 C 1 under
CA 02418790 2003-02-07
CA 02418790 2003-02-07
Example 31h), of 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4,7,10-
tetraazacyclododecane-
1,4,7-triacetic acid and 0.37 g of anhydrous lithium chloride (8.8 mmol) are
dissolved at 40°C in
60 ml of absolute dimethyl sulfoxide while being stirred and mixed at this
temperature with a
total of 1.01 g (8.8 mmol) of N-hydroxysuccinimide and 3.7 g (4.0 mmol) of the
title compound
of Example 13d), dissolved in 40 ml of absolute dimethyl sulfoxide. After
cooling to room
temperature, the reaction solution is mixed with 1.82 g (8.8 mmol) of N,N'-
dicyclohexylcarbodiimide, and it is stirred for 12 hours at room temperature.
The suspension
that is obtained is then mixed with sufficient acetone until precipitation of
the above-mentioned
title compound is completed, the precipitate is suctioned off, dried, taken up
in water, insoluble
dicyclohexylurea is filtered out, and the filtrate is desalinated with an
AMICON~R~ YM-3
ultrafiltration membrane (cut-off 3,000 Da), and low-molecular components are
removed. The
retentate is then freeze-dried.
Yield: 5.36 g (87.4% of theory) as a colorless lyophilizate.
Hz0 content (Karl-Fischer): 6.77%.
Elementary analysis (relative to anhydrous substance):
Cld: C 36.01 H 3.61 N 8.22 F 21.05 Gd 10.25 S 2.09
Fnd: C 35.87 H 3.70 N 8.22 F 20.91 Gd 10.18 S 2:16
Exam In a 14
a) 1,4,7-Triazaheptane-1,7-bis-(2-N-trifluoroacetyl-6-N-benzyloxycarbonyl-L-
lysine)-
diamide
100 g (107.9 mmol) of the carboxylic acid that is produced under Example la)
and 26.1 g
(226.59 mmol) of N-hydroxysuccinimide are dissolved in 500 ml of
dimethylformamide and
mixed in portions at 0°C with a total of 46.7 g (226.59 mmol) of N,N'-
dicyclohexylcarbodiimide,
and it is stirred for 3 more hours at this temperature. A solution, cooled to
0°C, of 5.57 g (53.95
mmol) of diethylenetriamine, dissolved in 60 ml of dimethylformamide, is added
drop by drop to
the thus produced active ester solution, and it is stirred for 2 hours at
0°C and for 12 hours at
room temperature. For working-up, precipitated dicyclohexylurea is filtered
out, and the solvent
61
is drawn off until a dry state is reached. The thus obtained residue is then
chromatographed on
silica gel (mobile solvent: dichloromethane/ethanol 15:1; chromatography was
performed with
use of a solvent gradient with continuous increase of the proportion of
ethanol).
Yield: 26.0 g (58.8% of theory, relative to the amine component that is used}
of the title
compound in the form of a colorless and strongly viscous oil.
Elementary analysis:
Cld: C 52.74 H 5.78 N 11.96 F 13.90
Fnd: C 52.66 H 5.89 N 11.88 F 14.02
b) 1,4,7-Triazaheptane-1,7-bis-(2-N-trifluoroacetyl-6-N-benzyloxycarbonyl-L-
lysine)-
diamide-4-[2-(N-ethyl-N-perfluorooctylsulfonyl)-amino]-acetyl
16.18 g (27.0 mmol) of 2-[N-ethyl-N-perfluorooctylsulfonyl)-aminoacetic acid
(production according to: DE 196 03 033), dissolved in 50 ml of
tetrahydrofuran, is added to a
solution that consists of 20 g (24.4 mmol) of the diamide that is produced
under 14a), dissolved
in a mixture that consists of 150 ml of tetrahydrofuran and 15 ml of
chloroform, at 0°C and under
nitrogen atmosphere. Then, a total of 18.0 g (36.6 mmol) of EEDQ [2-ethoxy-1-
ethoxycarbonyl-
1,2-dihydroquinoline] is added in portions at 0°C, and it is allowed to
stir overnight at room
temperature and then concentrated by evaporation in a vacuum. The remaining
oil is
chromatographed on silica gel (mobile solvent: n-hexane/isopropanol 15:1).
24.74 g (72.4% of
theory, relative to the sec-amine that is used} of the title compound is
obtained in the form of a
colorless oil.
Elementary analysis:
Cld: C 42.01 H 3.96 F 31.19 N 8.00 S 2.29
Fnd: C 41.92 H 4.07 F 31.22 N 7.87 S 2.34
c) 1,7-Bis-(6-N-benzyloxycarbonyl-L-lysine)-diamide-4-[2-(N-ethyl-N-
perfluorooctylsulfonyl)-amino]-acetyl-1,4,7-triazaheptane
22.0 g (15.7 mmol) of the title compound that is produced under Example 14b)
is
CA 02418790 2003-02-07
62
dissolved in 100 ml of ethanol, and ammonia gas is introduced into this
solution at 0°C for 40
minutes. Then, it is stirred for another 4 hours at 0°C and for 3 hours
at room temperature and
evaporated to the dry state in a vacuum at a bath temperature of 40°C.
The remaining oily
residue is purified on silica gel with use of dichloromethane/hexane/2-
propanol (20:10:1 ) as an
eluant.
Yield: 12.92 g (98.4% of theory) of the above-mentioned title compound as a
colorless
and strongly viscous oil.
Elementary analysis:
Cld: C 44.22 H 4.64 N 9.38 S 2.68 F 27.03
Fnd: C 44.31 H 4.72 N 9.30 S 2.74 F 26.99
d) 1,7-Bis-[6-N-benzyloxycarbonyl-2-N-(1-O-a-D-carbonylmethyl-2,3,4,6-tetra-O-
benzyl-
mannopyranose)-L-lysine]-diamide-4-[2-(N-ethyl-N-perfluorooctylsulfonyl)-
amino]-acetyl-1,4,7-
triazaheptane
5.47 g (9.15 mmol) of 1-carboxymethyloxy-2,3,4-tetra-O-benzyl-a-D-
mannopyranoside
[production as described in Patent DE 197 28 954 C 1 ] is dissolved in 80 ml
of
dimethylformamide and mixed with a total of 1.05 g (9.15 mmol) of N-
hydroxysuccinimide. It is
cooled to 0°C, and 1.9 g (9.15 mmol) of dicyclohexylcarbodiimide is
added. It is stirred for one
hour at 0°C and then for 4 hours at room temperature. It is cooled to
0°C, and a solution that
consists of 7.65 g (9.15 mmol) of the amino compound, described under Example
14c) and
dissolved in 50 ml of dimethylformariiide, is slowly added in drops within 3
hours. Then; it is
stirred for one more hour at 0°C, then overnight at room temperature.
It is evaporated to the dry
state in a vacuum, and the residue is taken up in 100 ml of ethyl acetate.
Precipitated urea is
filtered out, and the filtrate is washed twice with 50 ml each of 5% aqueous
soda solution. The
organic phase is dried on magnesium sulfate and evaporated to the dry state in
a vacuum. The
residue is chromatographed on silica gel (mobile solvent: n-hexane/isopropanol
20:1 ). 17.01 g
(78.9% of theory, relative to the carboxylic acid that is-used) of the title
compound is obtained in
the form of a colorless oil.
CA 02418790 2003-02-07
63
Elementary analysis:
Cld: C 59.13 H 5.43 N 4.76 F 13.71 S 1.36
Fnd: C 59.22 H 5.39 N 4.85 F 13.70 S 1.40
e) 1,7-Bis-[2-N-(1-O-a-D-carbonylmethyl-mannopyranose)-L-lysine]-diamide-4-[2-
(N-
ethyl-N-perfluorooctylsulfonyl)-amino]-acetyl-1,4,7-triazaheptane
15.0 g (6.36 mmol) of the amide, produced under I4d), is dissolved in 150 ml
of ethanol,
and it is mixed with 0.5 g of Pearlman's catalyst (Pd 20%, C). It is
hydrogenated at room
temperature under a hydrogen atmosphere (I atm) until no more hydrogen
absorption can be
observed. Catalyst is suctioned out, it is thoroughly rewashed with ethanol
(about 100 ml) and
evaporated to the dry state in a vacuum. The title compound is obtained as a
strongly viscous,
yellowish oil.
Yield: 8.54 g (97.2% of theory)
Elementary analysis:
Cld: C 39.13 H 5.04 N 8.11 F 23.38 S 2.32
Fnd: C 39.07 H 4.98 N 8.18 F 23.40 S 2.30
f) 1,7-Bis-[2-N-( I -O-a-D-carbonylmethyl-mannopyranose)-6-N-[ 1,4,7-
tris(carboxylatomethyl)-10-(3-aza-4-oxo-5-methyl-5-yl-pentanoyl)-1,4, 7,10-
tetraazacyclododecane)-L-lysine]-diamide-4-[2-(N-ethyl-N-
perfluorooctylsulfonyl)-amino]-
acetyl-1,4,7-triazaheptane, digadolinium complex
A stirred suspension of 5.7 g (9.06 mmoI) of the Gd complex, described in
Patent
Application DE 197 28 954 Cl under Example 31h), of IO-(4-carboxy-1-methyl-2-
oxo-3-
azabutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid in 75 ml of
absolute dirnethyl
sulfoxide is mixed at 70°C with 0.68 g ( 15.9 mmol) of lithium
chloride. After 30 minutes of
stirring at 70°C, the now clear reaction solution is mixed in portions
with a total of 1.83 g ( 15.9
mmol) of N-hydroxysuccinimide, and the reaction mixture is kept at this
temperature for 1 more
hour. After cooling to 0°C, it is mixed with 4.52 g (23.85 mmol) of
dicyclohexylcarbodiimide,
CA 02418790 2003-02-07
64
and the reaction solution is stirred for another hour at 0°C, followed
by 12 hours at 22°C. The
thus obtained reaction solution of the N-hydroxysuccinimide ester of the Gd
complex of 10-(4-
carboxy-I-methyl-2-oxo-3-azabutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-
triacetic acid is now
mixed at 22°C drop by drop with a solution of 2.84 g (2.06 mmol) of the
title compound of
Example I4e) in 15 ml of absolute dimethyl sulfoxide, and it is stirred for
another 12 hours at
room temperature. For working-up, the reaction solution is added in drops at
22°C in 500 mI of
acetone, whereby the title compound precipitates as colorless precipitate. The
precipitate is
suctioned off, dissolved in 200 ml ofdistilled water and ultrafiltered three
times with a YM3-
ultrafiltration membrane (AMICON~R~: cut-off: 3,000 Da) for the purpose of
desalination and
the separation of low-molecular components. The thus obtained retentate is
then freeze-dried.
Yield: 4.80 g (89.6% of theory, relative to the amine component that is used)
as a
colorless lyophilizate with a water content of 8.98%.
Elementary analysis (relative to anhydrous substance):
Cld: C 38.28 H 4.84 N 9.68 F 12.40 S 1.23 Gd 12.07
Fnd: C 38.20 H 4.9I N 9.77 F 12.45 S I.19 Gd I2.10
a) I,7-Bis(benzyloxycarbonyl)-4-{3-oxa-pentane-I,5-dicarboxylic acid-1-oyl-5-
[1-(4-
perfluorooctylsulfonyl)-piperazine]-amide]-1,4,7,10-tetraazacyclododecane
16.56 g (24.4 mmol) of the title compound of Example 15e), dissolved in 150 ml
of
tetrahydrofuran, is added at 0°C and under nitrogen atmosphere to a
solution of 10.75 g (24.4
mmol) of 1,7-bis-[benzyloxycarbonylJ-1,4,7,10-tetraazacyclododecane, dissolved
in a mixture
that consists of 150 ml of tetrahydrofuran and 15 ml of chloroform. Then, a
total of 18.0 g (36.6
mmol) of EEDQ [2-ethoxy-1-ethoxycarbonyl-I,2-dihydroquinoline] is added in
portions at 0°C
and allowed to stir overnight at room temperature and then concentrated by
evaporation in a
vacuum. The remaining oil is chromatographed on silica gel (mobile solvent: n-
hexane/isopropanol 12:1). 17.22 g (64.3% of theory, relative to the sec-amine
that is used) of the
monoamide and 3.8 g (8.8% of theory) of the diamide are obtained as by-
products. The title
CA 02418790 2003-02-07
65
compound is isolated in the form of a colorless oil.
Elementary analysis:
Cld: C 43.41 H 3.92 F 29.18 N 7.59 S 2.60
Fnd: C 43.52 H 4.07 F 29.24 N 7.67 S 2.55
b) 1,7-Bis(benzyloxycarbonyl)-4-{3-oxa-pentane-1,5-dicarboxylic acid-1-oyl-5-
[1-(4-
perfluorooctylsulfonyl)-piperazine]-amide } -10-[ 1-O-a-D-(5-carbonyl)-pentyl-
2,3,4,6-tetra-O-
benzyl-mannopyranoseJ-1,4,7,10-tetraazacyclododecane
10.0 g (13.4 mmol) of the carboxylic acid that is produced under Example l Oc)
and 3.24
g (28.1 rrimol) of N-hydroxysuccinimide are dissolved in 100 ml of
dimethylformamide and
mixed in portions at 0°C with a total of 5.8 g (28.1 mmol) of N,N'-
dicyclohexylcarbodiimide, and
it is stirred for 3 more hours at this temperature. A solution, cooled to
0°C, of 14.83 g (13.4
mmol) of the title compound of Example 15a), dissolved in 100 ml of
dimethylformamide, is
added drop by drop to the thus produced active ester solution, and it is
stirred for 2 hours at 0°C
and for 12 hours at room temperature. For working-up, precipitated
dicyclohexylurea is filtered
out, and the solvent is drawn off until a dry state is reached. The thus
obtained residue is then
chromatographed on silica gel (mobile solvent: dichloromethane/ethyl acetate
20:1; the
chromatography was carried out with the use of a solvent gradient with
continuous increase of
the proportion of ethyl acetate).
Yield: 18.3 g (78.2% of theory) of the title compound in the form of a
colorless and
strongly viscous oil.
Elementary analysis:
Cld: C 55.11 H 5.03 N 4.82 F 18.52 S 1.84
Fnd: C 54.87 H 4.85 N 4.92 F 18.55 S 1.86
CA 02418790 2003-02-07
66
c) 1-f 3-Oxa-pentane-l,~-dicarboxyIic acid-1-oyl-5-[1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide } -7-[ 1-O-a-D-(5-carbonyl)-pentyl-mannopyranose]-1,4,7,10-
tetraazacyclododecane
17.0 g (9.75 mmol) of the compound that is produced under 14b) is dissolved in
150 ml
of ethanol, mixed with 1.0 g of Pearlman's catalyst (Pd 20%, C) and
hydrogenated at room
temperature under a hydrogen atmosphere (1 atm) until no more hydrogen
absorption can be
observed. Catalyst is suctioned out, it is thoroughly rewashed with ethanol
(twice with 75 ml
each) and evaporated to the dry state in a vacuum. The title compound is
obtained as a strongly
viscous and colorless oil.
Yield: 10.76 g (99.0% of theory).
Elementary analysis:
CId: C 38.78 H 4.61 N 7.54 F 28.97 S 2.88
Fnd: C 38.86 H 4.65 N '7.41 F 29.02 S 2.92
d) 1,7-Bis-[1,4,7-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-Gd-
complex-10-
(pentanoyl-3-aza-4-oxo-5-methyl-Syl)-4-[2-(N-ethyl-N-perfluorooctylsulfonyl]-
amino]-acetyl-2-
oxa-acetyl]-10-[ 1-O-a-D-6-carbonylpentyl-mannopyranose]-1,4,7,10-
tetraazacyclododecane
24.86 g (39.46 mmol; 4.4 molar equivalents relative to the amine component
15c) that is
used of the Gd complex, described in Patent Application DE 197 28 954 C 1
under Example
31h), of 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4,7,10-
tetraazacyclododecane-1,4,7-
triacetic acid and 1.67 g of anhydrous lithium chloride (39.46 mmol) are
dissolved at 40°C in 200
ml of absolute dimethyl sulfoxide while being stirred and mixed at this
temperature with a total
of 4.53 g (39.46 mmol) of N-hydroxysuccinimide and 10.0 g (8.97 mmol) of the
title compound
of Example 14c), dissolved in 100 ml of absolute dimethyl sulfoxide. After
cooling to room
temperature, the reaction solution is mixed with 8.14 g (39.46 mmol) of N,N'-
dicyclohexylcarbodiimide and stirred for t 2 hours at room temperature. The
suspension that is
obtained is then mixed with sufficient acetone until precipitation of the
above-mentioned title
compound is completed, the precipitate is suctioned off, dried, taken up in
water, insoluble
CA 02418790 2003-02-07
67
dicyclohexylurea is filtered out, and the filtrate is desalinated with an
AMICON~R~ YM-3
ultrafiltration membrane (cut-off 3,000 Da), and low-molecular components are
removed. The
retentate is then freeze-dried.
Yield: 16.37 g (79.3% of theory) as a colorless lyophilizate.
Hz0 content (Karl-Fischer): 7.65%.
Elementary analysis (relative to anhydrous substance):
Cld: C 38.0I H 4.61 N 9.58 F 13.81 S 1.37 Gd I3.45
Fnd: C 37.92 H 4.55 N 9.58 F 13.77 S 1.31 Gd 13.48
e) 3-Oxa-pentane-1,5-dicarboxylic acid-mono-[1-(4-perfluorooctylsulfonyl)-
piperazine]-
amide
25 g (44.0 mmol) of I-perfluorooctylsulfonylpiperazine is dissolved in 150 ml
of
tetrahydrofuran and mixed at room temperature with a total of 5.1 g (44.0
mmol) of diglycolic
acid anhydride, and the thus obtained reaction solution is refluxed for 12
hours. After cooling to
room temperature, it is evaporated to the dry state, and the remaining oily
residue is purified on
silica gel with use of dichloromethane/2-propanol ( 16:1 ) as an eluant.
Yield: 27.94 g (92.8% of theory) of the above-mentioned title compound in the
form of a
colorless and viscous oil.
Elementary analysis:
CId: C 58.52 H 4.27 N I.98 S 2.26 F 22.80
Fnd: C 58.42 H 4.41 N 1.80 S 2.28 F 23.02
Fxamnle 1616
b) 1,7-Bis(benzyloxycarbonyl)-4-{3-oxa-pentane-1,5-dicarboxylic acid-1-oyl-5.-
[1-(4-
perfluorooctylsulfonyl)-piperazine]-amide } -[ 10-[ 1-O-(3-D-6-carbonylpentyl-
2,3,4,6-tetra-O-
benzyl-glucopyranose]-1,4,7,10-tetraazacyclododecane)
68.5 g (91.79 mmol) of 1-carboxymethyloxy-2,3,4-tetra-O-benzyl-a-D-
mannopyranoside
[production as described in Patent DE I 97 28 954 C 1 J is dissolved in 750 mI
of dry
CA 02418790 2003-02-07
68
tetrahydrofuran, and then 9.25 g (91.79 mmol) of triethylamine is added. After
the reaction
solution is cooled to -15°C to -20°C, a solution of 12.64 g
(92.5 mmol) of isobutyl chloroformate
in 150 ml of dry tetrahydrofuran is slowly added in drops at this temperature
while being stirred,
whereby the rate of addition by drops can be selected in such a way that an
internal temperature
of -10°C is not exceeded. After a reaction time of 15 minutes at -
15°C, a solution of 101.6 g
(91.79 mmol) of the title compound of Example 1 Sa) and 9.25 g (91.79 mrnol)
of triethylamine
are then slowly added in drops as solution in 500 ml of dry tetrahydrofuran at
-20°C. After a reaction time of one hour at -15°C and two hours
at room temperature, the
reaction solution is evaporated to the dry state in a vacuum. The remaining
residue is taken up in
450 ml of ethyl acetate and washed twice with 300 ml each of saturated sodium
bicarbonate
solution and once with 400 ml of water. After the organic phase is dried on
sodium sulfate, salt
is suctioned out, and the ethyl acetate is drawn off in a vacuum. The
remaining oily residue is
purified on silica gel with use of dichloromethane/hexane/2-propanol (10:20:1)
as an eluant.
Yield: 130.6 g (81.6% of theory) of the above-mentioned title compound as a
colorless
and strongly viscous oil.
Elementary analysis:
Cld: C 55.11 H 5.03 N 4.82 F 18.52 S 1.84
Fnd: C 55.20 H 5.09 N 4.91 F 18.48 S 1.80
b) 1-{3-Oxa-pentane-1,5-dicarboxylic acid-1-oyl-5-[1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide } -7-[ 1-O-a-D-(5-carbonyl)-pentyl-mannopyranose]-1,4,7,10-
tetraazacyclododecane
110.0 g (63.08 mmol) of the compound, produced under 16a), is dissolved in
1000 ml of
ethanol, mixed with 5.0 g of Pearlman's catalyst (Pd 20%, C) and hydrogenated
until quantitative
hydrogen uptake is reached. Catalyst is suctioned out, it is rewashed with
ethanol and evaporated
to the dry state in a vacuum. The title compound is obtained as a viscous and
colorless oil.
Yield: 92.61 g (99.5% of theory)
Elementary analysis:
CA 02418790 2003-02-07
69
Cld: C 52.10 H 5.12 N 5.70 F 21.89 S 2.17
Fnd: C 52.20 H 5.09 N 5.71 F 21.87 S 2.20
b) 1,7-Bis-[1,4,7-tris(carboxylatomethyl)-10-(3-aza-4-oxo-5-methyl-5-yl-
pentanoyl)-4-{3-oxa-
pentane-1,5-dicarboxylic acid-1-oyl-5-[1-(4-perfluorooctylsulfonyl)-
piperazine]-amide}-10-[1-
O-a-D-(S-carbonyl)-pentyl-mannopyranose]-1,4,7,10-tetraazacyclododecane,
digadolinium
complex
SS.4 g [88.0 mmol; 4.4 molar equivalents relative to the diamine component
that is used
of Example l3dO] of the Gd complex, described in Patent Application DE 197 28
954 C 1 under
Example 31h), of 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-
1,4,7,10=tetraazacyclododecane-
1,4,7-triacetic acid and 3.7 g of anhydrous lithium chloride (88.0 mmol) are
dissolved at 40°C in
500 ml of absolute dimethyl sulfoxide while being stirred and mixed at this
temperature with a
total of 10.1 g (88.0 mmol) of N-hydroxysuccinimide and 29.5 g (20.0 mmol) of
the title
compound of Example 16b), dissolved in 200 ml of absolute dimethyl sulfoxide.
After cooling
to room temperature, the reaction solution is mixed with 18.2 g (88.0 mmol) of
N,N'-
dicyclohexylcarbodiimide and stirred for 12 hours at room temperature. The
suspension that is
obtained is then mixed with sufficient acetone until precipitation of the
above-mentioned title
compound is coFnpleted, the precipitate is suctioned off, dried, taken up in
water, insoluble
dicyclohexylurea is filtered out, and the filtrate is desalinated with an
AMICON~R~ YM-3
ultrafiltration membrane (cut-off 3,000 Da), and low-molecular components are
removed. The
retentate is then freeze-dried.
Yield: 35.96 g (76.9% of theory) as a colorless lyophilizate.
H20 content (Karl-Fischer): 5.98%
Elementary analysis (relative to anhydrous substance):
Cld: C 38.01 H 4.61 N 8.22 F 13.81 Gd 13.45 S 1.37
Fnd: C 37.87 H 4.70 N 8.22 F 13.90 Gd 13.48 S 1.36
CA 02418790 2003-02-07
70
b) S-{Ethoxycarbonyl)pentyl-2,3,4,6-tetra-O-acetyl-a-D-mannopyranoside
Analogously to what is described in the literature for the synthesis of aryl
glycopyranosides [J. Conchie and G. A. Levvy in Methods in Carbohydrate
Chemistry (R. L.
Whistler, M. L. Wolfrom and J. N. BeMiller, Eds.), Academic Press, New York,
Vol. II, 90, pp.
34S-347, {1963)], the reaction of 156.2 g (400 mmol) of D-mannosepentaacetate
as a,(3-(a,(3-ratio
= 4:1)-anomer mixture [for synthesis of 1,2,3,4,6-penta-O-acetyl-a,(3-D-
mannopyranose, cf.: M.
L. Wolfrom and A. Thompson in Methods in Carbohydrate Chemistry (R. L.
Whistler, M. L.
Wolfrom and J. N. BeMiller, Eds.), Academic Press, New York, Vol. II, S3, pp.
211-215, (1963)]
with 67 ml (400 mmol) of 6-hydroxy-hexanoic acid ethyl ester and 60.8 ml (S20
mmol) of
tin(IV) chloride in a total of 600 ml of 1,2-dichloroethane after column-
chromatographic
purification (eluant: hexane/ethyl acetate 2:1) results in the formation of
100.05 g (5l% of
theory) of the above-mentioned title compound as a colorless arid viscous oil.
By 1H-NMR-
spectroscopic study of the thus obtained title compound, it was possible to
show that the above-
mentioned title compound is only the pure a-anomer.
Elementary analysis:
Cld: C 52.94 H 6.77
Fnd: C 52.80 H 6.78
b) S-(Carboxy)pentyl-2,3,4,6-tetra-O-benzyl-a-D-mannopyranoside
A stirred suspension of 141.0 g (289 mmol) of the title compound of Example
17a) in 200
ml of dioxane is mixed at room temperature and with simultaneous vigorous
stirring in portions
with a total of 238.5 g (4:26 mol) of fine-powder potassium hydroxide powder.
To make the
stirring easier, the reaction mixture is mixed with another 200 ml of dioxane,
and the thus
obtained suspension is subsequently heated to boiling heat and mixed drop by
drop at this
temperature with a total of 372 ml (3.128 mol) of benzyl bromide over a period
of two hours.
After a reaction time of 4 hours at 110°C, followed by 12 hours at room
temperature, the reaction
mixture is slowly poured into a total of 2.S liters of ice water for the
purpose of working-up, and
CA 02418790 2003-02-07
7i
the aqueous phase is subsequently completely extracted with diethyl ether.
After the thus
obtained ether phase is washed and after the subsequent drying of the ether
phase on sodium
sulfate, salt is suctioned out, and the diethyl ether is drawn off in a
vacuum. Excess benzyl
bromide is then distilled off from the reaction mixture in an oil pump vacuum
quantitatively at an
oil bath temperature of 180°C. The thus obtained, resinous-oily residue
is purified on silica gel
with use of ethyl acetate/hexane (1:10) as an eluant.
Yield: 172.2 g (91.0% of theory) of the above-mentioned title compound in the
form of a
colorless and extremely viscous oil
Elementary analysis:
Cld: C 75.68 H 7.16
Fnd: C 75.79 H 7.04
b) 5-[(Carboxy)-pentyl-2,3,4,6-tetra-O-benzyl-a-D-mannopyranoside-JN-
hydroxysuccinimide ester
60.0 g (91.5 mmol) of the title compound of Example 17b) is dissolved in 750
ml of
dimethylformamide and mixed with a total of 10.4 g (91.5 mmol) of N-
hydroxysuccinimide. It is
cooled to 0°C, and 18.9 g (91.5 mmol) of dicyclohexylcarbodiimide is
added. It is stirred for one
hour at 0°C and then for 4 hours at room temperature. The solvent is
drawn off in a vacuum, and
the remaining residue is mixed with 100 ml of ethyl acetate and cooled to
0°C. Precipitated urea
is filtered out, and the filtrate that is obtained is evaporated to the dry
state in a vacuum. The
thus obtained, resinous-oily residue is purified on silica gel with use of
ethyl acetate/hexane
(1:20) as an eluant.
Yield: 61.23 g (89.0% of theory) of the above-mentioned title compound in the
form of a
colorless and viscous oil.
Elementary analysis:
Cld: C 70.29 H 6.57 N 1.86
Fnd: C 70.39 H 5.64 N 1.91
CA 02418790 2003-02-07
72
b) 2,6-Bis-{6-NS-2-N«-]-[1-O-a-D-6-carbonyl-pentyl-(2,3,4,6-tetra-O-benzyl)-
mannopyranose}-L-lysine]-methyl ester
A solution of 27.51 g (36.6 mmol) of the title compound of Example 17c) in 150
ml of
dimethylformamide is added in drops to a solution, cooled to 0°C, that
consists of 4.26 g (18.30
mmol; 0.5 molar equivalent relative to the carboxylic acid that is used) of L-
lysine methyl ester-
dihydrochloride (commercially available from the Bachem Company) and 4.05 g
(40.26 mmol)
of triethylamine in 100 ml of dimethylformamide. After the addition is
completed, it is stirred
for one more hour at 0°C and then overnight at room temperature. It is
evaporated to the dry
state in a vacuum, and the residue is taken up in 300 ml of ethyl acetate.
Precipitated urea is
filtered out, and the filtrate is washed twice with 100 ml each of 5% aqueous
soda solution. The
.organic phase is dried on magnesium sulfate and evaporated to the dry state
in a vacuum. The
residue is chromatographed on silica gel (mobile solvent: n-hexane/isopropanol
25:1 ). 39.56 g
(75.4% of theory) of the title compound is obtained in the form of a colorless
oil.
Elementary analysis:
Cld: C 72.88 H 7.31 N 1.95
Fnd: C 72.90 H 7.29 N 2.02
b) 2,6-Bis-[6-Ng-2-N«-[1-O-a-D-6-carbonyl-pentyl-(2,3,4,6-tetra-O-benzyl)-
mannopyranose]]-L-lysine
30.0 g (20.92 mmol) of the compound that is produced under Example 17d) is
dissolved
in 150 ml of ethanol. The solution of 4 g (100.0 mmol) of sodium hydroxide in
10 ml of distilled
water is then added to it, and it is stirred for 3 hours at 50°C.
According to the thin-layer
chromatogram, the saponification is quantitative. It is evaporated to the dry
state in a vacuum,
and the remaining residue is taken up in 300 ml of ethyl acetate, and the
organic phase is
extracted twice with 100 ml each of dilute, aqueous citric acid solution.
After drying on sodium
sulfate, it is filtered and evaporated to the dry state in a vacuum. The
residue is chromatographed
on silica gel (mobile solvent: n-hexane/isopropanol 13:1). 25.56 g (88.5% of
theory) of the title
compound is obtained in the form of a colorless oil.
CA 02418790 2003-02-07
CA 02418790 2003-02-07
Elementary analysis:
Cld: C 72.88 H 7.31 N 1.95
Fnd: C 72.78 H 7.33 N 1.96
b) 2,6-Bis-[6-N~-2-N«-[1-O-a-D-6-carbonyl-pentyl-(2,3,4,6-tetra-O-benzyl)-
mannopyranose]-L-lysine]-N-hydroxysuccinimide ester
14.0 g (9.15 mmol) of the title compound of Example 17e) is dissolved in 100
ml of
dimethylformamide and mixed with a total of 1.04 g (9.15 mmol) of N-
hydroxysuccinimide. It is
cooled to 0°C, and 1.89 g (9.15 mmol) of dicyclohexylcarbodiimide is
added. It is stirred for one
hour at 0°C and then for 4 hours at room temperature. The solvent is
then drawn off in a
vacuum, and the remaining residue is mixed with 100 mI of ethyl acetate and
cooled to 0°C.
Precipitated urea is filtered out, and the filtrate that is obtairied is
evaporated to the dry state in a
vacuum. The thus obtained, resinous-oily residue is purified on silica gel
with use of ethyl
acetate/n-hexane (1:20) as an eluant.
Yield: 12.94 g (92.4% of theory) of the above-mentioned title compound in the
form of a
colorless and viscous oil.
Elementary analysis:
Cld: C 71.40 H 7.05 N 2.74
Fnd: C 71.39 H 7.14 N 2.8I
g) 2,6-N,N'-Bis[1-O-a-D-(6-carbonyl)-pentyl-2,3,4,6-tetra-O-benzyl-
mannopyranose]-L-
lysine-1, 7-( 1,4, 7-triazaheptane)-diamide
A.solution that consists of 14.0 g (9.15 mmol; 2 molar equivalents relative to
the amine
that is used) of the title compound of Example 17f) in 100 ml of
dimethylformamide is slowly
added in drops to a solution, cooled to 0°C, of 0.47 g (4.57 mmol) of
diethylenetriamine in 25 ml
of dimethylformamide. After addition is completed, it is stirred for one more
hour at 0°C and
then overnight at room temperature. It is evaporated to the dry state in a
vacuum, and the residue
is taken up in 200 ml of ethyl acetate. Precipitated urea is filtered out, and
the filtrate is washed
CA 02418790 2003-02-07
74
twice with 50 ml each of 5% aqueous soda solution. The organic phase is dried
on magnesium
sulfate and evaporated to the dry state in a vacuum. The residue is
chromatographed on silica gel
(mobile solvent: n-hexane/isopropanol 25:1). 9.53 g (71.4% of theory) of the
title compound is
obtained in the form of a colorless oil.
Elementary analysis:
Cld: C 72.79 H 7.42 N 3.36
Fnd: C 72.90 H 7.39 N 3.32
h) 2-N-[2-(N-Ethyl-N-perfluorooctylsulfonyl)-amino]-acetyl-6-N-
(benzyloxycarbonyl)-L-
lysine-methylester -
20.8 g (35.6 mmol) of the 2-(N-ethyl-N-perfluorooctylsulfonyl)-aminoacetic
acid and
3.60 g (35.6 mmol) of triethylamine are dissolved in 200 ml of
dimethylformamide, and 4.09 g
(35.6 mol) of N-hydroxysuccinimide is added. It is cooled to 0°C, and
7.34 g (35.6 mmol) of
dicyclohexylcarbodiimide is added. It is stirred for one hour at 0°C
and then for 4 hours at room
temperature. It is cooled to 0°C, and a solution that consists of 11.77
g (35.6 mmol) of 6-N-
benzyloxycarbonyl-L-lysine-methyl ester-hydrochloride and 4.0 g (40.0 mmol) of
triethylamine
in 100 ml of dimethylformamide are added in drops within 10 minutes. It is
stirred for one hour
at 0°C, then overnight at room temperature. It is evaporated to the dry
state in a vacuum, and the
residue is taken up in 100 ml of ethyl acetate. Precipitated urea is filtered
out, and the filtrate is
washed twice with 100 ml each of 5% aqueous soda solution. The organic phase
is dried on
magnesium sulfate and evaporated to.the dry state in a vacuum. The residue is
chromatographed
on silica gel (mobile solvent: n-hexane/ethyl acetate 20:1). 27.43 g (88.0% of
theory) of a
colorless oil is obtained.
Elementary analysis:
Cld: C 38.41 H 3.45 N 4.80 F 36.89 S 3.66
Fnd: C 38.4 H 3.38 N 4.88 F 37.02 S 3.71
CA 02418790 2003-02-07
b) 2-Na-{ [2-(N-Ethyl-N-perfluorooctylsulfonyl]-amino-acetyl } -6-N~-
(benzyloxycarbonyl)-
L-lvsine
25.0 g (28.5 mmol) of the compound that is produced under Example 17h) is
dissolved
in 150 ml of ethanol. The solution of 4 g (100.0 mmol) of sodium hydroxide in
10 ml of distilled
water is then added to it and stirred for 3 hours at 50°C. According to
the thin-layenz
chromatogram, the saponification is quantitative. It 'is evaporated to the dry
state in a vacuum,
and the remaining residue is taken up in 300 ml of ethyl acetate, and the
organic phase is
extracted twice with 100 ml each of dilute, aqueous citric acid solution.
After drying on sodium
sulfate, it is filtered, and it is evaporated to the dry state in a vacuum.
The residue is
chromatographed on silica gel (mobile solvent: n-hexane/isopropanol 10:1).
22.73 g (92.4% of
theory) of the title compound is obtained in the form of a colorless oil.
Elementary analysis:
Cld: C 37.64 H 3.28 N 4.88 F 37.49 S 3.72
Fnd: C 37.65 H 3.38 N 4.88 F 37.52 S 3.73
j) 1,4,7-Triazaheptane-4-{2-N-[2-(N-ethyl-N-perfluorooctylsulfonyl)-amino]-
acetyl-6-N-
benzyloxycarbonyl } -L-lysine-amide-1,7-bis { 2,6-N,N'-bis [ 1-O-a-D-( 5-
carbonyl)-pentyl-2,3,4,6-
tetra-O-benzylmannopyranose]-L-lysine-diamide }
15.33 g (17.8 mmol) of the title compound of Example 17i) and 1.80 g (17.8
mmol) of
triethylamine are dissolved in 250 ml of dry tetrahydrofuran. After the
reaction solution is
cooled to -1 S°C to -20°C, a solution of 4.92 g (35.6 mmol) of
isobutyl chloroformate, dissolved
in 50 ml of dry tetrahydrofuran, is slowly added in drops at this temperature
while being stirred,
whereby the rate of addition by drops can be selected in such a way that an
internal temperature
of -10°C is not exceeded. After a reaction time of 15 minutes at -
15°C, a solution of 52.0 g (17.8
mmol) of the title compound of Example 17g) and 1.80 g (17.8 mmol) of
triethylamine in 300 ml
of dry tetrahydrofuran are then slowly added in drops at -20°C. After a
reaction time of one hour
at -15°C and two hours at room temperature, the reaction solution is
evaporated to the dry state in
a vacuum. The remaining residue is taken up in 500 ml of ethyl acetate and
washed twice with
CA 02418790 2003-02-07
76
200 ml each of saturated sodium bicarbonate solution and once with 200 ml of
water. After the
organic phase is dried on sodium sulfate, salt is suctioned out, and the ethyl
acetate is drawn off
in a vacuum. The remaining oily residue is purified on silica gel with use of
ethyl acetate/n-
hexane ( 1:20) as an eluant.
Yield: 54.6 g (81.6% of theory) of the above-mentioned title compound as a
colorless
and strongly viscous oil.
Elementary analysis:
Cld: C 65.09 H 6.45 N 3.72 F 8.58 S 0.85
Fnd: C 65.13 H 4.41 N 3.69 F 8.52 S 0.90
k) 1,4,7-Triazaheptane-4-{2-N-[2-(N-ethyl-N-perfluorooctylsulfonyl)-amino]-
acetyl}-L-
lysine-amide-1,7-bis { 2,6-N,N'-bis[ 1-O-a-D-(5-carbonyl)-pentyl-
mannopyranose]-L-lysine-
diamide }
50.0 g ( 13.28 mmol) of the compound, produced under 17j), is dissolved in 500
ml of
ethanol, mixed with 4.0 g of Pearlman's catalyst (Pd 20%, C) and hydrogenated
at room
temperature under a hydrogen atmosphere (1 atm) until no more hydrogen
absorption can be
observed. Catalyst is suctioned out, it is thoroughly rewashed with ethanol
(about 400 ml) and
evaporated to the dry state in a vacuum. The title compound is obtained as a
strongly viscous
and colorless oil.
Yield: 26.85 g (93.0% of theory)
Elementary analysis:
Cld: C 45.85 H 6.35 N 6.44 F 14.86 S 1.47
Fnd: C 45.76 H 6.35 N 6.41 F 14.92 S 1.39
CA 02418790 2003-02-07
77
1) 1,4,7-Triazaheptane-4-{2-N-[2-(N-ethyl-N-perfluorooctylsulfonyl)-amino]-
acetyl-6-N-
[ 1,4,7-tris(carboxylatomethyl)-10-(3-aza-4-oxo-5-methyl-5-yl-pentanoyl)-
1,4,7,10-
tetraazacyclododecane } -L-lysine-amide-1,7-bis { 2,6-N,N'-bis [ 1-O-a-D-(5-
carbonyl)-pentyl-
mannopyranose]-L-lysine-diamide}, gadolinium complex
5.54 g (8.8 mmol; 2.2 molar equivalents relative to the amine components of
Example
17k) that are used) of the Gd complex, described in Patent Application DE 197
28 954 C 1 under
Example 31h), of 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4,7,10-
tetraazacyclododecane-
1,4,7-triacetic acid and 0.37 g of anhydrous lithium chloride (8.8 mmol) are
dissolved at 40°C in
60 ml of absolute dimethyl sulfoxide while being stirred and mixed at this
temperature with a
total of 1.01 g (8.8 mmol) of N-hydroxysuccinimide and 1.84 g (4.0 mmol) of
the title compound
of Example 17k), dissolved in 40 ml of absolute dimethyl sulfoxide. After
cooling to room
temperature, the reaction solution is mixed with 1.82 g (8.8 mmol) of N,N'-
dicyclohexylcarbodiimide and stirred for 12 hours at room temperature. The
suspension that is
obtained is then mixed with sufficient acetone until precipitation of the
above-mentioned title
compound is completed, the precipitate is suctioned off, dried, taken up in
water, insoluble
dicyclohexylurea is filtered out, and the filtrate is desalinated with an
AMICON~R~ YM-3
ultrafiltration membrane (cut-off 3,000 Da), and low-molecular components are
removed. The
retentate is then freeze-dried.
Yield: 8.77 g (78.7% of theory) as a colorless lyophilizate.
H20 content (Karl-Fischer): 4.43%.
Elementary analysis (relative to anhydrous substance):
Cld: C 43.98 H 5.97 N 7.54 F 11.59 Gd 5.64 S 1.15
Fnd: C 43.97 H 6.02 N 7.62 F 11.61 Gd 10.18 S 1.15
CA 02418790 2003-02-07
78 '
Example 18
a) 2-Na-6-N~-Bis-[1-O-a-D-carbonylmethyl-2,3,4,6-tetra-O-benzyl-mannopyranose]-
L-
lysine]-methyl ester
10.95 g (18.30 mmol) of 1-carboxymethyloxy-2,3,4-tetra-O-benzyl-a-D-
mannopyranoside [production as described in Patent DE 197 28 954 C 1 ] is
dissolved in I 50 ml
of dimethylformamide and mixed with a total of 2.09 g (18.3 mmol) of N-
hydroxysuccinimide.
It is cooled to 0°C, and 3.78 g (18.3 mmol) of dicyclohexylcarbodiimide
is added. It is stirred for
one hour at 0°C and then for 4 hours at room temperature. It is cooled
to 0°C, and within one
hour, a solution that consists of 2.13 g (9.15 mmol: 0.5 molar equivalent
relative to the
carboxylic acid that is used) of L-lysine methyl ester-dihydrochloride
(commercially available
from the Bachem Company) and 2.02 g (20.13 mmol) of triethylamine in 70 ml of
dimethylformamide is added. After the addition is completed, it is stirred for
one more hour at
0°C and then overnight at room temperature. It is evaporated to the dry
state in a vacuum, and
the residue is taken up in 300 ml of ethyl acetate. Precipitated urea is
filtered out, and the filtrate
is washed twice with 100 ml each of 5% aqueous soda solution. The organic
phase is dried on
magnesium sulfate and evaporated to the dry state in a vacuum. The residue is
chromatographed
on silica gel (mobile solvent: n-hexane/isopropanol 25:1). 10.05 g (82.3% of
theory) of the title
compound is obtained in the form of a colorless oil.
Elementary analysis:
Cld: C 71.94 H 6.79 N 2.10
Fnd: C 71.90 H 6.79 N 2.09
b) 2-Na-6-N~-Bis-[1-O-a-D-carbonylmethyl-2,3,4,6-tetra-O-benzyl-mannopyranose]-
L-
lysine
Analogously to what is described in Example 17e} for the synthesis of the
title compound
that is relevant there, the methyl ester saponification of IS g (1 I.23 mmol)
of the title compound
of Example I 8a) results in the formation of 13.89 g (93.6% of theory) of the
above-mentioned
title compound in the form of a colorless and viscous oil.
CA 02418790 2003-02-07
79
Elementary analysis:
Cld: C 71.80 H 6.71 N 2.12
Fnd: C 71.84 H 6.69 N 2.15
c) 2-Na-6-N~-Bis-[1-O-a-D-carbonylmethyl-2,3,4,6-tetra-O-benzyl-mannopyranose]-
L-
lysine-N-hydroxysuccinimide ester
12.09 g (9.15 mmol) of the title compound of Example 18d) is dissolved in 100
ml of
dimethylformamide and mixed with a total of 1.04 g (9.15 mmol) of N-
hydroxysuccinimide. It is
cooled to 0°C, and 1.89 g (9.15 mmol) of dicyclohexylcarbodiimide is
added. It is stirred for one
hour at 0°C and then for 4 hours at room temperature. The solvent is
then drawn off in a
vacuum, and the remaining residue is mixed with 100 ml of ethyl acetate and
cooled to 0°C.
Precipitated urea is filtered out, and the filtrate that is obtained is
evaporated to the dry state in a
vacuum. The thus obtained resinous-oily residue is purified on silica gel with
use of ethyl
acetate/n-hexane (1:20) as an eluant.
Yield: 12.24 g (94.4% of theory) of the above-mentioned title compound in the
form of a
colorless and viscous oil.
Elementary analysis:
Cld: C 70.27 H 6.47 N 2.96
Fnd: C 70.31 H 6.44 N 3.01
d) 6-N-Benzyloxycarbonyl-2-N-{[2,6-N,N'-bis(1-.O-a-D-carbonylmethyl-2,3,4,6-
tetra-O-
benzyl-mannopyrano se)]-L-lysyl- } -L-lysine-[ 1-(4-perfluorooctylsulfonyl)-
piperazine]-amide
19.0 g ( 13.4 mmol) of the carboxylic acid-N-hydroxysuccinimide ester,
produced under
Example 18c), is dissolved in 75 ml of dimethylformamide and mixed drop by
drop at 0°C with a
solution, cooled to 0°C, of 11.13 g ( 13.4 mmol) of the title compound
of Example 1 c), dissolved
in 50.0 ml of dimethylformamide. The resulting reaction solution is stirred
for 2 more hours at
0°C and for 12 hours at room temperature. For working-up, precipitated
dicyclohexylurea is
filtered out, and the solvent is then drawn off in a vacuum until a dry state
is reached. The thus
CA 02418790 2003-02-07
obtained residue is,chromatographed on silica gel [mobile solvent:
dichloromethane/ethanol
28:1; the chromatography is performed here with use of a solvent gradient with
a proportion of
the polar eluant component (here: ethanol) that is used continuously
increasing in the course of
the chromatography].
Yield: 25.28 g (88.4% of theory) of the title compound in the form of a
colorless and
strongly viscous oil.
Elementary analysis:
Cld: C 59.10 H 5.34 N 3.94 F 15.13 S 1.50
Fnd: C 59.18 H 5.35 N 4.02 F 15.15 S 1.56
e) 2-N-{[2,6-N,N'-Bis(1-O-a-D-carbonylmethyl-mannopyranose)]-L-lysyl-L-lysine-
[1-(4-
perfluorooctylsulfonyl)-piperazine]-amide
20.0 g (9.37 mmol) of the compound that is produced under 18d) is dissolved in
200 ml
of ethanol, mixed with 1.5 g of Pearlman's catalyst (Pd 20%, C) and
hydrogenated at room
temperature under a hydrogen atmosphere ( 1 atm) until no more hydrogen
absorption can be
observed. Catalyst is suctioned out, it is thoroughly rewashed with ethanol
(twice with about 100
ml each) and evaporated to the dry state in a vacuum. The title compound is
obtained as a
strongly viscous and colorless oil.
Yield: 11.62 g (97.0% of theory)
Elementary analysis:
Cld: C 38.50 H 4.65 N 6.57 F 25.25 S 2.51
Fnd: C 38.46 H 4.65 N 6.51 F 25.23 S 2.52
f) 6-N-[1,4,7-Tris(carboxylatomethyl)-10-(3-aza-4-oxo-5-methyl-5-yl-pentanoyl)-
1,4,7,10-
tetraazacyclododecane)-2-N- { [2, 6-N,N'-bis( 1-O-a-D-carbonylmethyl-
mannopyranose)]-L-lysyl } -
L-lysine-[1-(4-perfluorooctylsulfonyl)-piperazine]-amide, Gd complex
9.98 g ( 15.84 mmol; 2.2 molar equivalents relative to the amine component of
Example
18e) that is used) of the Gd complex, described in Patent Application DE 197
28 954 C 1 under
CA 02418790 2003-02-07
81
Example 31h), of 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4,7,10-
tetraazacyclododecane-
I,4,7-triacetic acid and 0.67 g of anhydrous lithium chloride (15.84 mmol) are
dissolved at 40°C
in 100 ml of absolute dimethyl sulfoxide while being stirred and mixed at this
temperature with a
total of 1.82 g ( 15.84 mmol) of N-hydroxysuccinimide and 9.19 g (7.19 mmol)
of the title
compound of Example 18e), dissolved in 50 ml of absolute dimethyl sulfoxide.
After cooling to
room temperature, the reaction solution is mixed with 3.27 g (15.84 mmol) of
N,N'-
dicyclohexylcarbodiimide and stirred for 12 hours at room temperature. The
suspension that is
obtained is then mixed with sufficient acetone until the precipitation of the
above-mentioned title
compound is completed, the precipitate is suctioned off, dried, taken up in
water, insoluble
dicyclohexylurea is filtered off, and the filtrate is desalinated with an
AMICON~R~ YM-3
ultrafiltration membrane (cut-off 3,000 Da) and in this case possible, still
present low-molecular
components are removed at the same time. The retentate is then freeze-dried.
Yield: 11.85 g (87.2% of theory) as a colorless lyophilizate.
Hz0 content (Karl-Fischer): 5.54%
Elementary analysis (relative to anhydrous substance):
Cld: C 38.12 H 4.64 N 8.15 F 20.38 S I.70 Gd 8.32
Fnd: C 38.16 H 4.59 N 8.18 F 20.37 S 1.68 Gd 8.28
Fxam In a 19
a) I,7-Bis(benzyloxycarbonyl)-4-(3-oxa-2H,2H,4H,4H,5H,SH-perfluorotridecanoyl)-
1,4,7,10-tetraazacyclododecane
12.74 g (24.4 mmol) of the title compound of Example 19g), dissolved in 150 m1
of
tetrahydrofuran, is added at 0°C and under nitrogen atmosphere to a
solution of 10.75 g (24.4 .
mmol) of 1,7-bis-[benzyloxycarbonyl]-1,4,7,10-tetraazacyclododecane, dissolved
in a mixture
that consists of 150 ml of tetrahydrofuran and 15 ml of chloroform. Then, a
total of I 8.0 g (36.6
mmol) of EEDQ [2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinolineJ is added in
portions at 0°C,
and it is allowed to stir overnight at room temperature and then concentrated
by evaporation in a
vacuum. The remaining oil is chromatographed on silica gel (mobile solvent: n-
CA 02418790 2003-02-07
82
hexane; isopropanol 16:1 ). 15.89 g (69.0% of theory, relative to the sec-
amine that is used) of the
W onoamide and 3.8 g (8.8% of theory) of the diamide are obtained as by-
products. The title
compound is isolated in the form of a colorless oil.
Elementary analysis:
Cld: C 45.77 H 3.95 F 34.19 N 5.93
Fnd: C 45.72 H 4.01 F 34.22 N 5.88
b) 1,7-Bis(benzyloxycarbonyl)-4-(3-oxa-2H,2H,4H,4H,SH,SH-perfluorotridecanoyl)-
10-[1-
S-a-D-(2-carbonyl)-ethyl-2,3,4,6-tetra-O-acetyl-mannopyranoseJ-1, 4,7,10-
tetraazacyclododecane
7.09 g (13.4 mmol) of 3-(2,3,4,6-tetra-O-acetyl-1-thio-a-D-mannopyranosyl)-
propionic
acid-N-hydroxysuccinimide ester (production according to: J. Haensler et al.,
Bioconjugate
Chem. 4, 85, (1993); Chipowsky, S., and Lee, Y. C. (1973), Synthesis of 1-Thio-
aldosides;
Carbohydrate Research 31, 339-346) are dissolved in 100 ml of
dimethylformamide and mixed
drop by drop at 0°C with a solution, cooled to 0°C, of 12.65 g
(13:4 mmol) of the title compound
of Example 19a), dissolved in 100 ml of dimethylformamide. It is stirred for 2
hours at 0°C then
for 12 hours at room temperature. For working-up, the solvent is drawn off in
a vacuum until a
dry state is reached, and the thus obtained residue is then chromatographed on
silica gel (mobile
solvent: dichloromethane/ethyl acetate 20:1; the chromatography was performed
with use of a
solvent gradient with continuous increase of the proportion of ethyl acetate).
Yield: 16.23 g (88.9% of theory) of the title compound in the form of a
colorless and
strongly viscous oil.
Elementary analysis:
Cld: C 46.70 H 4.36 N 4.11 F 23.69 S 2.35
Fnd: C 46.66 H 4.35 N 4.12 F 23.65 S 2.30
c) 1-(3-Oxa-2H,2H,4H,4H,SH,SH-perfluorotridecanoyl)-7-[1-S-a-D-(2-carbonyl)-
ethyl-
2,3,4,6-tetra-O-acetyl-mannopyranoseJ-1,4,7,10-tetraazacyclododecane
15.0 g ( 11.0 mmol) of the compound that is produced under 19b) is dissolved
in 150 ml
CA 02418790 2003-02-07
83
of ethanol, mixed with 1.0 g of Pearlman's catalyst (Pd 20%, C) and
hydrogenated at room
temperature under a hydrogen atmosphere (1 atm) until no more hydrogen
absorption can be
observed. Catalyst is suctioned out, it is thoroughly rewashed with ethanol
(twice with 75 ml
each) and evaporated to the dry state in a vacuum. The title compound is
obtained as a strongly
viscous and colorless oil.
Yield: 11.56 g (96.0% of theory).
Elementary analysis:
Cld: C 40.59 H 4.33 N 5.12 F 29.50 S 2.93
Fnd: C 40.63 H 4.35 N 5.11 F 29.52 S 2.92
d) 1-(3-Oxa-2H,2H,4H,4H,SH,SH-perfluorotridecanoyl)-7-[1-S-a-D-(2-carbonyl)-
ethyl-
mannopyranose]-1,4,7,10-tetraazacyclododecane
10.0 g (9.13 mmol) of the title compound of Example 19c) is suspended in 100
ml of
absolute methanol and mixed at 5°C with a catalytic amount of sodium
methanolate. After a
reaction time of 3 hours at room temperature, even thin-layer chromatographic
checking (eluant:
chloroform/methanol 4:1 ) of the plot of the reaction indicates a quantitative
reaction. For the
purpose of working-up, the now clear reaction solution is neutralized by
mixing with Amberlite
IR 120 (H+ form)-cation-exchange resin, exchanger is suctioned out, rewashed
with methanol,
and the thus obtained methanolic filtrate is drawn off in a vacuum until a dry
state is reached.
The oily residue that is obtained is purified by column chromatography on
silica gel (mobile
solvent: dichloromethane/n-hexane/ethyl acetate 15:20:1; the chromatography
was performed
with use of a solvent gradient with continuous increase of the proportion of
ethyl acetate). After
1H-NMR-spectroscopic study of the title compound, the presence of the a-
configuration at the
anomeric center of the D-mannopyranose was definitively established based on
the size of the
coupling constant of Jm = 0.9 Hz. This a-configuration is the sole existing
configuration at the
anomeric center, i.e., the amount of the [i-configured anomer of the title
compound that can
possibly be formed thus lies below the'H-NMR-spectroscopic detection limit.
The above-
mentioned title compound was accordingly represented only in the form of the
pure a-configured
CA 02418790 2003-02-07
84
anomer.
Yield: 8.28 g {98.0% of theory) of the title compound in the form of a
colorless and
strongly viscous oil.
Elementary analysis:
Cld: C 37.59 H 4.24 N 6.05 F 34.85 S 3.46
Fnd: C 37.57 H 4.28 N 6.02 F 34.85 S 3.44
e) 1-(3-Oxa-2H,2H,4H,4H,SH,SH-perfluorotridecanoyl)-7-[I-S-a-D-(2-carbonyl)-
ethyl-
mannopyranose]-4,10-bis[ 1,4,7-tris(carboxylatomethyl)-10-(3-aza-4-oxo-5-
methyl-5-yl-
pentanoyl)]-1,4,7,10-tetraazacyclododecane, digadolinium complex
2:48 g [(3.94 mmol); 4.4 molar equivalents relative to the diamine component
of 19d)
that iswsed] of the Gd complex, described in Patent Application DE 197 28 954
C 1 under
Example 31h), of 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4,7,10-
tetraazacyclododecane-
1,4,7-triacetic acid and 167 mg of anhydrous lithium chloride (3.94 mmol) are
dissolved at 40°C
in 40 m1 of absolute dimethyl sulfoxide while being stirred and mixed at this
temperature with a
total of 453 mg (3.94 mmol) of N-hydroxysuccinimide and 980 mg (0.895 mmol) of
the title
compound of Example 19d), dissolved in 10 ml of absolute dimethyl sulfoxide.
After cooling to
room temperature, the reaction solution is mixed with 814 mg (3.946 mmol) of
N,N'-
dicyclohexylcarbodiimide and stirred for 12 hours at room temperature. The
suspension that is
obtained is then mixed with sufficient acetone until precipitation of the
above-mentioned title
compound is completed, the precipitate is suctioned off, dried; taken up in
water, insoluble
dicyclohexylurea is filtered out, and the filtrate is desalinated with an
AMICON~Ry YM-3
ultrafiltration membrane (cut-off 3,000 Da), and low-molecular components are
removed. The
retentate is then freeze-dried.
Yield: 1.32 g (69. I % of theory) as a colorless lyophilizate.
H20 content (Karl-Fischer): 7.65%
Elementary analysis (relative to anhydrous substance):
Cld: C 37.43 H 4.45 N 9.12 F 15.02 S 1.49 Gd 14.63
CA 02418790 2003-02-07
Fnd: C 37.42 H 4.50 N 9.18 F 15.07 S 1.51 Gd 14.58
f) 3-Oxa-2H,2H,4H,4H,5H,5H-perfluorotridecanoic acid-t-butyl ester
25.0 g (53.8 mmol) of 1 H,1 H,2H,2H-perfluoro-1-decanol [commercially
available from
the Lancaster Company] is dissolved in 250 ml of absolute toluene and mixed at
room
temperature with a catalytic amount (about 0.75 g) of tetra-N-butyl-ammonium
hydrogen sulfate.
Then, a total of 7.55 g (134.6 mmol; 2.5 equivalents relative to the alcohol
component that is
used) of fine-powder potassium hydroxide powder is added at 0°C,
followed by 15.73 g (80.7
mmol; 1.5 equivalents relative to the alcohol component that is used) of
bromoacetic acid-tert-
butyl ester, and it is allowed to stir for 2 more hours at 0°C. The
thus obtained reaction solution
is stirred for 12 hours at room temperature and for the purpose of working-up,
it is mixed with a
total of 500 ml of ethyl acetate and 250 ml of water. The organic phase is
separated and washed
twice with water. After the organic phase is dried on sodium sulfate, salt is
suctioned out, and
the solvent is drawn off in a vacuum. The remaining oily residue is purified
on silica gel with
use of ethyl acetate/hexane ( 1:10) as an eluant.
Yield: 26.3 g (84.6% of theory) of the above-mentioned title compound as a
colorless
and strongly viscous oil
Elementary analysis:
Cld: C 33.23 H 2.61 F 55.85
Fnd: C 33.29 H 2.61 F 55.90
g) 3-Oxa-2H,2H,4H,4H,5H,5H-perfluorotridecanoic carboxylic acid
20.0 g (34.58 mmol) of the title compound of Example 19f) is suspended in 200
ml of a
mixture that consists of methanol and 0.5 molar sodium hydroxide solution at a
ratio of 2:1 while
being stirred at room temperature, and then it is heated to 60°C. After
a reaction time of 12 hours
at 60°C, the now clear reaction mixture is neutralized for working-up
by mixing with Amberlite
IR 120 (H+ form)-cation-exchange resin, exchanger is suctioned out, and the
thus obtained
methanolic-aqueous filtrate is evaporated to the dry state in a vacuum. The
amorphous-oily
CA 02418790 2003-02-07
86
residue that is obtained is purified on silica gel with use of ethyl acetate/n-
hexane (1:3) as an
eluant.
Yield: 16.0 g (88.6% of theory) of the above-mentioned title compound as a
colorless
and strongly viscous oil
Elementary analysis:
Cld: C 27.60 H 1.35 F 61.85
Fnd: C 27.58 H 1.36 F 61.90
Fxam In a 20
a) 6-Benzyloxycarbonyl-2-[2-(N-ethyl-N-perfluorooctylsulfonyl)-amino]-acetyl-L-
lysine-
methyl ester
16.18 g (27.0 mmol) of 2-[N-ethyl-N-perfluorooctylsulfonyl)-aminoacetic acid
(production according to: DE 196 03 033), dissolved in SO ml of
tetrahydrofuran, is added drop
by drop to 8.0 g (24.4 mmol) of ~-carbonyloxybenzyl-L-lysine methyl ester
hydrochloride
(commercially available from the Bachem Company), dissolved in a mixture that
consists of 150
ml of tetrahydrofuran, 15 ml of chloroform, and 2.62 g (26.0 mmol) of
triethylamine at 0°C and
under nitrogen atmosphere. Then, a total of 18.0 g (36.6 mmol) of EEDQ [2-
ethoxy-1-
ethoxycarbonyl-1,2-dihydroquinoline] is added in portions at 0°C and
allowed to stir overnight at
room temperature. It is then concentrated by evaporation in a vacuum, and the
remaining oil is
chromatographed on silica gel (mobile solvent: n-hexane/isopropanol 15:1).
1?.0 g (79.6% of
theory, relative to the primary amine that is used) of the title compound is
obtained in the form of
a colorless oil.
Elementary analysis:
Cld: C 38.41 H 3.45 F 36.89 N 4.80 S 3.66
Fnd: C 38.42 H 3.47 F 36.92 N 4.87 S 3.64
b) 2-[2-(N-Ethyl-N-perfluorooctylsulfonyl)-amino]-acetyl-L-lysine-methyl ester
15.0 g (20.23 mmol) of the compound that is produced under Example 20a) is
dissolved
CA 02418790 2003-02-07
87
in 200 ml of ethanol, mixed with 800 mg of Pearlman's catalyst (Pd 20% on
activated carbon)
and hydrogenated until the calculated amount of hydrogen is taken up. Catalyst
is suctioned out,
it is thoroughly rewashed with ethanol and evaporated to the dry state in a
vacuum. The title
compound is obtained as a colorless oil.
Yield: 14.68 g (97.9% of theory)
Elementary analysis:
Cld: C 32.40 H 3.26 F 43.56 N 5.67 S 4.32
Fnd: C 32.42 H 3.27 F 43.60 N 5.67 S 4.34
c) 6-( 1-O-a-D-Carbonylmethyl-2,3,4,6-tetra-O-benzyl-mannopyranose) 2-[2-(N-
ethyl-N-
perfluorooctylsulfonyl)-amino]-acetyl-L-lysine-methyl ester
21.31 g (35.6 mmol) of 1-carboxymethyloxy-2,3,4-tetra-O-benzyl-a-D-
mannopyranoside
[production as described in Patent DE 197 28 954 C I ] and 3.60 g (35.6 mmol)
of triethylamine
are dissolved in 500 ml of dry tetrahydrofuran. After the reaction solution is
cooled to -15°C to
-20°C, a solution of 4.92 g (35.6 mmol) of isobutyl chloroformate in 75
ml of dry tetrahydrofuran
is slowly added in drops at this temperature while being stirred, whereby the
rate of addition by
drops can be selected in such a way that an internal temperature of -
10°C is not exceeded. After
a reaction time of 15 minutes at -15°C, a solution of 26.39 g (35.6
mmol) of the title compound
of Example 20b) and 3.60 g (35.6 mmol) of triethylamine, in 100 ml of dry
tetrahydrofuran, is
then slowly added in drops at -20°C. After a reaction time of one hour
at -15°C and.two hours at
room temperature, the.reaction solution is evaporated to the dry state in a
vacuum. The
remaining residue is taken up in 250 ml of ethyl acetate and washed twice with
100 ml each of
saturated sodium bicarbonate solution and once with 200 ml of water. After the
organic phase is
dried on sodium sulfate, salt is suctioned out, and the ethyl acetate is drawn
off in a vacuum. The
remaining oily residue is purified on silica gel with use of ethyl acetate/n-
hexane (1:10) as an
eluant.
Yield: 38.12 g (81.0% of theory) of the above-mentioned title compound as a
colorless
and strongly viscous oil
CA 02418790 2003-02-07
88
Elementary analysis:
Cld: C 49.92 H 3.92 N 2.53 F 29.18 S 2.90
fnd: C 49.99 H 4.11 N 2.69 F 29.22 S 3.01
d) 6-(1-O-a-D-Carbonylmethyl-2,3,4,6-tetra-O-benzyl-mannopyranose) 2-[2-(N-
ethyl-N-
perfluorooctylsulfonyl)-amino]-acetyl-L-lysine
27.65 g (20.92 mmol) of the compound that is produced under Example 20c) is
dissolved
in 250 ml of methanol. The solution of 4.0 g (100.0 mmol) of sodium hydroxide
in 10 ml of
distilled water is then added to it, and it is stirred for 3 hours at
50°C. After the course of the
reaction is checked by means of thin-layer chromatography, methyl ester
saponification has
already taken place quantitatively. It is evaporated to the dry state in a
vacuum, the remaining
residue is taken up in 300 ml of ethyl acetate, and the organic phase is
extracted twice with 100
ml each of dilute, aqueous citric acid solution. After drying on sodium
sulfate, it is filtered and
evaporated to the dry state in a vacuum. The residue that is obtained is
chromatographed on
silica gel (mobile solvent: n-hexane/chloroform/isopropanol 15:10:1). 24.31 g
(88.9% of theory)
of the title compound is obtained in the form of a colorless and viscous oil.
Elementary analysis:
Cld: C 51.46 H 4.70 N 3.21 F 24.71 S 2.45
Fnd: C 51.49 H 4.71 N 3.19 F 24.72 S 2.41
e) 6-(1-O-a-D-Carbonylmethyl-mannopyranose) 2-[2-(N-ethyl-N-
perfluorooctylsulfonyl)-
amino]-acetyl-L-lysine
20.0 g ( 15.30 mmol) of the title compound of Example 20d) is dissolved in a
mixture that
consists of 250 ml of 2-propanol and 25 ml of water, and it is mixed with 1.0
g of palladium
catalyst (10% Pd on activated carbon). It is hydrogenated for 12 hours at room
temperature and a
hydrogen pressure of one atmosphere. Catalyst is filtered out, and the
filtrate is evaporated to the
dry state in a vacuum. The residue is dissolved in 200 ml of methanol, and the
reaction product
is precipitated by mixing with a total of 800 ml of diethyl ether. After the
thus obtained solid is
CA 02418790 2003-02-07
89
suctioned off, the latter is dried in a vacuum at 50°C.
Yield: 14.32 g (99.0% of theory) of an amorphous solid.
Elementary analysis:
Cld: C 35.56 H 3.84 N 4.44 S 3.39 F 34.15
Fnd: C 35.58 H 3.81 N 4.45 S 3.40 F 34.17
f) 6-(1-O-a-D-Carbonylmethyl-mannopyranose) 2-[2-(N-ethyl-N-
perfluorooctylsulfonyl)-
amino]-acetyl-L-lysine-N- { 2 -hydroxy-prop-3 -yl- [ 1,4, 7-
tris(carboxylatomethyl)-1,4, 7,10-
tetraazacyclododecan-10-yl]}-amide, Gd complex
7.48 g (7.91 mmol) of the title compound of Example 20e) is dissolved at
40°C in 50 ml
of dimethyl sulfoxide, and 1.00 g (8.70 mol) of N-hydroxysuccinimide is added.
It is cooled to
20°C, and 1.795 g (8.7 mmol) of dicyclohexylcarbodiimide is added. It
is stirred for one hour at
20°C and then for 4 hours at 40°C. Then, a solution that
consists of 4.53 g (7.91 mmol) of the
gadolinium complex of 10-(2-hydroxy-3-aminopropyl)-4,7,10-tris(carboxymethyl)-
1,4,7,10-
tetraazacyclododecanine [for production, cf.: WO 97/02051] in 20 ml of
dimethyl sulfoxide is
added in drops at this temperature within 10 minutes. It is stirred for one
hour at 40°C, then
overnight at room temperature. The thus obtained suspension is then mixed with
sufficient
acetone until precipitation of the above-mentioned title compound is
completed, the precipitate is
suctioned off, dried, taken up in water, insoluble dicyclohexylurea is
filtered out, and the filtrate
is desalinated with an AMICON~R~ YM-3 ultrafiltration membrane (cut-off 3,000
Da), and low-
molecular components are removed. The retentate is then freeze-dried.
Yield: 9.71 g (81.7% of theory) as a colorless lyophilizate.
H20 content (Karl-Fischer): 3.97%
Elementary analysis (relative to anhydrous substance):
Cld: C 35.16 H 4.16 N 7.45 F 21.48 Gd 10.46 S 2.13
Fnd: C 35.17 H 4.20 N 7.42 F 21.49 Gd 10.48 S 2.09
CA 02418790 2003-02-07
Fxam In a 21
a) 6-N-[1-O-a-D-(5-Carbonyl)-pentyl-2,3,4,6-tetra-O-benzyl-mannopyranose]-2N-
[2-(N-
ethyl-N-perfluorooctylsulfonyl)-amino]-acetyl-L-lysine-methyl ester
5.23 g (8.0 mmol) of the 5-(carboxy)pentyl-2,3,4,6-tetra-O-benzyl-a-D-
mannopyranoside,
described in Example 10c), 1.3 g (8.0 mmol) of 1-hydroxybenzotriazole, and 2.6
g (8.0 mmol) of
2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyIuronium tetrafluoroborate (TBTU;
Peboc Limited,
UK) are dissolved in 75 ml of DMF and stirred for 15 minutes. This solution is
then mixed with
5.16 ml (30 mmol) of N-ethyldiisopropylamine and with 5.93 g (8.0 mmol) of the
amine that is
described under Example 20b), and it is stirred for 1.5 days at room
temperature. For working-
up, the solvent is drawn off in a vacuum until a dry state is reached, and the
thus obtained residue
is then chromatographed on silica gel (mobile solvent: dichloromethane/ethyl
acetate 30:1; the
chromatography was carried out with use of a solvent gradient with continuous
increase of the
proportion of ethyl acetate).
Yield: 9.70 g (88.0% of theory) of the title compound in the form of a
colorless and
strongly viscous oil.
Elementary analysis:
Cld: C 52.29 H 4.97 N 3.05 F 23.43 S 2.33
Fnd: C 52.33 H 4.95 N 3.12 F 23.50 S 2.30
b) 6-N-[1-O-a-D-(5-Carbonyl)-pentyl-2,3,4,6-tetra-O-benzyl-mannopyranose]-2N-
[2-(N-
ethyl-N-perfluorooctylsulfonyl)-amino]-acetyl-L-lysine
9.0 g (12.40 mmol) of the compound that is produced under Example 21 a) is
dissolved ~n
150 ml of methanol. The solution of 2.48 g (62.0 mmol) of sodium hydroxide in
15 ml of
distilled water is then added to it, and it is stirred for 3 hours at
50°C. After the course of the
reaction is checked by means of thin-layer chromatography, the methyl ester
saponification has
already taken place quantitatively after the above-mentioned reaction time. It
is evaporated to the
dry state in a vacuum, and the remaining residue is taken up in 300 ml of
ethyl acetate, and the
organic phase is extracted twice with 100 ml each of dilute, aqueous citric
acid solution. After
CA 02418790 2003-02-07
91
drying on sodium sulfate, it is filtered and evaporated to the dry state in a
vacuum. The residue
that is obtained is chromatographed on silica gel (mobile solvent: n-
hexane/chloroform/isopropanol 25:10:1). 15.88 g (93.9% of theory) of the title
compound is
obtained in the form of a colorless and strongly viscous oil.
Elementary analysis:
Cld: C 51.95 H 4.88 N 3.08 F 23.67 S 2.35
Fnd: C 51.99 H 4.91 N 3.09 F 23.70 S 2.33
c) 6-N-[1-O-a-D-(5-Carbonyl)-pentyl-mannopyranose]-2N-[2-(N-ethyl-N-
perfluorooctyIsulfonyl)-amino]-acetyl-L-lysine
13.0 g (9.52 mmol) of the title compound of Example 21 b) is dissolved in a
mixture that
consists of 150 ml of 2-propanol and 25 ml of water, and 1.0 g of the
palladium catalyst ( 10% Pd
on activated carbon) is added. It is hydrogenated for 12 hours. at 1
atmosphere of hydrogen
pressure and at room temperature. Catalyst is filtered out, and the filtrate
is evaporated to the dry
state in a vacuum. The residue that is obtained is chromatographed on silica
gel (mobile solvent:
n-hexane/chloroform/isopropanol 15:10:1 ). 9.09 g (95.1 % of theory) of the
title compound is
obtained in the form of a colorless and strongly viscous oil.
Elementary analysis:
Cld: C 37.10 H 4.22 N 4.19 F 32.18 S 3.10
Fnd: C 37.09 H 4.21 N 4.19 F 32.20 S 3.13
d) 6-N-[1-O-a-D-(5-Carbonyl)-pentyl-mannopyranose]-2N-[2-(N-ethyl-N-
perfluorooctylsulfonyl)-amino]-acetyl-L-lysine-N-{ 2-hydroxy-prop-3-yI-[ 1,4,7-
tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-10-yl]}-amide, Gd
complex
7.93 g (7.91 mmol) of the title compound of Example 21 c) is dissolved at
40°C in 75 ml
of dimethyl sulfoxide, and it is mixed with 1.00 g (8.70 mol) of N-
hydroxysuccinimide. It is
cooled to room temperature, and a total of 1.795 g (8.7 mmol) of
dicyclohexylcarbodiimide is .
added. It is stirred for one hour at 20°C and then for 4 hours at
40°C. A solution that consists of
CA 02418790 2003-02-07
92
4.53 g (7.91 mmol) of the gadolinium complex of 10-(2-hydroxy-3-aminopropyl)-
4,7,10-
tris(carboxymethyl)-1,4,7,10-tetraazacyclododecanine [for production, c~: WO
97/02051], in 20
ml of dimethyl sulfoxide, is then added in drops at 40°C within 10
minutes to this solution of the
active ester of the title compound of Example 21 c). It is stirred for one
hour at 40°C, then
overnight at room temperature. The thus obtained suspension is then mixed with
the sufficient
amount of a mixture that consists of acetone/2-propanol (2:1) until
precipitation of the above-
mentioned title compound is completed, the precipitate is suctioned off,
rewashed with ethyl
acetate, dried, taken up in water, insoluble dicyclohexylurea is filtered out,
and the filtrate is
desalinated with an AMICON~R~ YM-3 ultrafiltration membrane (cut-off 3,000
Da), and low-
molecular components are removed. The retentate is then freeze-dried.
Yield: 9.71 g (78.8% of theory) as a colorless lyophilizate.
H20 content (Karl-Fischer): 6.65%
Elementary analysis (relative to anhydrous substance):
Cld: C 36.97 H 4.52 N 7.19 F 20.71 Gd 10.08 S 2.06
Fnd: C 37.02 H 4.50 N 7.22 F 20.69 Gd 10.08 S 2.09
~~tam~pl~..22
a) 6-N-{4-[2,3-Bis-(N,N-bis(t-butyloxycarbonylmethyl)-amino)-propyl]-phenyl}-3-
oxa-
propionyl-2-N-( 1-a-D-carbonylmethyl-mannopyranose) L-lysine-[ 1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide
5.25 g (7.72 mmol) of the tetra-t.bu-ester of 1-(4-carboxymethoxybenzyl)-EDTA
(lit.:
Patent, US 4622420) and 781 mg (7.72 mmol) of triethylamine are dissolved in
50 ml of
methylene chloride. At -15°C, a solution that consists of 1.16 g (8.5
mmol) of isobutyl
chloroformate in 10 ml of methylene chloride is added in drops within 5
minutes, and it is stirred
for another 20 minutes at -15°C. Then, the solution is cooled to -
25°C, and a solution that
consists of 7.07 g (7.72 mmol) of the title compound of Example 10e) and 2.12
g (21.0 mmol) of
triethylamine, in 70 ml of tetrahydrofuran, is added in drops within 30
minutes and subsequently
stirred for 30 more minutes at -15°C, and then stirring is continued
overnight at room
CA 02418790 2003-02-07
93
temperature. For workinb up, the solvent is drawn off in a vacuum, and the
remaining oily
residue is taken up in 250 ml of chloroform. The chloroform phase is extracted
twice with 100
ml each of a 10% aqueous ammonium chloride solution, the organic phase is
dried on
magnesium sulfate, and it is evaporated to the dry state in a vacuum. The
residue is
chromatographed on silica gel (mobile solvent: methylene chloride/ethanol =
20:1).
Yield: 9.60 g (79.0% of theory) of a colorless and very viscous oil.
Elementary analysis:
Cld: C 46.39 H 5.55 N 5.32 F 20.45 S 2.03
Fnd: C 46.42 H 5.51 N 5.29 F 20.49 S 2.09
b) 6-N-{4-[2,3-Bis-(N,N-bis(carboxymethyl)-amino)-propyl]-phenyl}-3-oxa-
propionyl-2-N-
(1-a-D-carbonylmethyl-mannopyranose) L-lysine-[1-(4-perfluorooctylsulfonyl)-
piperazine]-
amide
9.0 g (5.70 mmol) of the compound that is produced under Example 22a) is
dissolved in
150 ml of methanol. The solution of 4.0 g (100.0 mmol) of sodium hydroxide in
25 ml of
distilled water is then added to it, and it is stirred for 6 hours at
60°C. After the course of the
reaction is checked by means of thin-layer chromatography; the saponification
of the tetra-t.-
butyl ester has already taken place quantitatively after the above-mentioned
reaction time. It is
evaporated to the dry state in a vacuum, and the remaining residue is taken up
in 50 ml of
dimethyl sulfoxide in the heat, and then it is mixed with a sufficient amount
of a mixture that
consists of acetone/ethyl acetate (l :l) until the precipitation of the above-
mentioned title
compound is completed, the thus obtained precipitate is suctioned off;
rewashed well with ethyl
acetate, dried, taken up in water, the pH of the product solution is set at
3.5 with 1 molar
hydrochloric acid, possibly existing insoluble components are filtered out,
and the filtrate is
desalinated with an AMICON~R~ YM-3 ultrafiltration membrane (cut-off: 3,000
Da), and low-
molecular components are removed. The retentate is then freeze-dried.
Yield: 6.76 g (87.6% of theory) as a colorless lyophilizate.
Hz0 content (Karl-Fischer): 3.30%
CA 02418790 2003-02-07
94
Elementary analysis (relative to anhydrous substance):
Cld: C 39.89 H 4.09 N 6.20 F 23.84 S 2.37
Fnd: C 39.92 H 4.15 N 6.22 F 23.92 S 2.29
c) 6-N-{4-[2,3-Bis-(N,N-bis(carboxylatomethyl)-amino)-propyl]-phenyl}-3-oxa-
propionyl-
2-N-( 1-a-D-carbonylmethyl-mannopyranose) L-lysine-[ 1-(4-
perfluorooctylsulfonyl)-piperazine]-
amide, Mn complex, disodium salt
3.0 g (2.22 mmol) of the title compound of Example 22b) is dissolved in 150 ml
of a
water/ethanol (3:1) mixture at boiling heat and mixed at 80°C in
portions with 0.25 g (2.22
mmol) of manganese(II) carbonate. Then, the thus obtained reaction solution is
refluxed for 5
hours. After cooling to room temperature, the solvent mixture is completely
drawn off in a
vacuum, and the remaining residue is dissolved in a mixture that consists of
200 ml of distilled
water/n-butanol (1:1). While being stirred vigorously, it is set at a pH of
7.2 by mixing with 1N
sodium hydroxide solution. After the n-butanol is completely drawn off in a
vacuum, the
remaining aqueous phase is desalinated with an AMICON~R~ YM-3 ultrafiltration
membrane (cut-
off 3,000 Da), and low-molecular components are removed. The retentate is then
freeze-dried.
Yield: 3.19 g (99.0% of theory) as a colorless lyophilizate.
H20 content (Karl-Fischer): 5.08%.
Elementary analysis (relative to anhydrous substance):
Cld: C37.23H3.54F22.25Mn3.78N5.79Na3.17S2.21
Fnd: C 37.30 H 3.49 F 22.29 Mn 3.81 N 5.76 Na 3.19 S 2.18
gam 1~ a 2~
a) 3-Benzyloxycarbonylamino-glutaric acid-[1-(4-perfluorooctylsulfonyl)-
piperazine]-
monoamide
A stirred solution of 25.0 g (94.96 mmol) of 3-N-(benzyloxycarbonyl)-glutaric
acid-
anhydride [synthesis according to: Hatanaka, Minoru; Yamamoto, Yu-ichi; Nitta,
Hajime;
Ishimaru, Toshiyasu; TELEAY; Tetrahedron Lett.; EN; 22; 39; 1981; 3883-3886;]
in 150 ml of
CA 02418790 2003-02-07
absolute tetrahydrofuran is mixed drop by drop while being stirred with a
solution of 53.97 g
(95.0 mmol) of 1-perfluorooctylsulfonylpiperazine in 150 ml of
tetrahydrofuran, and the thus
obtained reaction solution is refluxed for 12 hours. After cooling to room
temperature, it is
evaporated to the dry state, and the remaining oily residue is purified on
silica gel with use of
dichloromethane/2-propanol (20:1 ) as an eluant.
Yield: 75.80 g (96.0% of theory) of the above-mentioned title compound in the
form of a
colorless and viscous oil.
Elementary analysis:
Cld: C 36.11 H 2.67 N 5.05 S 3.86 F 38.84
Fnd: C 36.12 H 2.61 N 5.08 S 3.88 F 38.82
b) 3-Amino-glutaric acid-[1-(4-perfluorooctylsulfonyl)-piperazine]-monoamide
31.50 g (37.88 mmol) of the compound that is produced under 23b) is dissolved
in 300
ml of ethanol, mixed with 2.5 g of Pearlman's catalyst (Pd 20%, C) and
hydrogenated at 1
atmosphere of hydrogen pressure until quantitative hydrogen uptake is reached.
Catalyst is
suctioned out, it is rewashed with ethanol and evaporated to the dry state in
a vacuum. The title
compound is obtained as a whitish-yellow, viscous oil.
Yield: 25.22 g (95.5% of theory)
Elementary analysis:
Cld: C 29.28 H 2.31 N 6.03 S 4.06 F 46.31
Fnd: C 29.32 H 2.29 N 6.08 S 4.08 F 46.28
c) 3-N-(1-a-D-Carbonylmethyl-2,3,4,6-tetra-O-benzyl-mannopyranose)-glutaric
acid-[1-(4-
perfluorooctylsulfonyl)-piperazine]-monoamide
21.52 g ( 18.96 mmol) of 1-carboxymethyloxy-2,3,4-tetra-O-benzyl-a-D-
mannopyranoside [production as described in Patent DE 197 28 954 Cl] is
dissolved at room
temperature in 100 ml of absolute dimethylformamide and mixed at 0°C
with 2.56 g (22.2 mmol)
of N-hydroxysuccinimide, followed by 4.55 g (22.2 mmol) of
dicyclohexylcarbodiimide. After a
96
reaction time of 60 minutes at 0°C and 3 hours at 22°C,
insoluble dicyclohexylurea is filtered out,
and the thus obtained clear active ester solution of the above-mentioned title
compound is slowly
added in drops at 0°C to a stirred solution of I 3.22 g (18.96 mmol) of
the compound of Example
23b), dissolved in I00 ml of dimethylformamide. After a reaction time of I2
hours at room
temperature, the solvent is drawn off in a vacuum, and the remaining residue
is taken up in 300
ml of ethyl acetate, urea is filtered out, and the organic filtrate is washed
twice with 100 ml each
of saturated sodium bicarbonate solution and once with 100 ml of I O% aqueous
citric acid
solution and once with 200 ml of water. After the organic phase is dried on
sodium sulfate, salt
is suctioned out, and the ethyl acetate is drawn off in a vacuum. The
remaining oily residue is
purified on silica gel with use of ethyl acetate/n-hexane ( 1:15) as an
eluant.
Yield: 21.39 g (88.3% of theory) of the above-mentioned title compound as a
colorless
and strongly viscous oil.
Elementary analysis:
Cld: C 49.81 H 4.10 N 3.29 F 25.27 S 2.51
Fnd: C 49.89 H 4.11 N 3.32 F 25.22 S 2.51
d) 3-N-( 1-a-D-Carbonylmethyl-mannopyranose)-glutaric acid-[ 1-(4-
perfluorooctylsulfonyl)-
piperazine]-monoamide
19.55 g ( 15.30 mmol) of the title compound of Example 23c) is dissolved in a
mixture
that consists of 250 ml of 2-propanol and 25 ml of water, and it is mixed with
1.5 g of palladium
catalyst (10% Pd on activated carbon). It is hydrogenated for 12 hours at room
temperature and a
hydrogen pressure of one atmosphere. Catalyst is filtered out, and the
filtrate is evaporated to the
dry state in a vacuum. The residue is dissolved ~in 200 ml of methanol, and
the reaction product
is precipitated by mixing with a total of 800 ml of diethyl ether. After the
thus obtained solid is
suctioned off, the latter is dried in a vacuum at 40°C.
Yield: 17.49 g (97.5% of theory) of an amorphous solid.
Elementary analysis:
Cld: C 32.73 H 3.08 N 4.58 S 3.49 F 35.20
CA 02418790 2003-02-07
97
Fnd: C 32.68 H 3.15 N 4.55 S 3.50 F 35.17
e) 3-N-(1-a-D-Carbonylmethyl-mannopyranose)-glutaric acid-[1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide-5-N-{ 2-hydroxy-prop-3-yl-[ 1,4,7-tris(carboxylatomethyl)-
1,4,7,10-
tetraazacyclododecan-10-yl]}-amide, Gd complex
14.43 g (1 x.84 mmol) of the title compound of Example 23d) and 0.67 g of
anhydrous
lithium chloride (15.84 mmol) are dissolved at 40°C in 100 ml of
absolute dimethyl sulfoxide
while being stirred and mixed at this temperature with a total of 1.82 g (
15.84 mmol) of N-
hydroxysuccinimide and a solution of 9.08 g (15.84 mmol) of the gadolinium
complex of 10-(2-
hydroxy-3-aminopropyl)-4,7,10-tris(carboxymethyl)-1,4,7,10-
tetraazacyclododecanine [for
production, cf.: WO 97/02051 ], in 50 ml of dimethyl sulfoxide. After cooling
to room
temperature, the reaction solution is mixed with 3.27 g ( 15.84 mmol) of N,N'-
dicyclohexylcarbodiimide, and it is stirred for 12 hours at room temperature.
The suspension
that is obtained is then mixed with su~cient acetone until precipitation of
the above-mentioned
title compound is completed, the precipitate is suctioned off, dried, taken up
in water, insoluble
dicyclohexylurea is filtered out, and the filtrate is desalinated with an
AMICON~R~ YM-3
ultrafiltration membrane (cut-off 3,000 Da), and in this case possible still
present low-molecular
components are removed at the same time. The retentate is then freeze-dried.
Yield: 18.71 g (80.2% of theory) as a colorless lyophilizate.
H20 content (Karl-Fischer): 4.87%.
Elementary analysis (relative to anhydrous substance):
Cld: C 34.24 H 3.83 N 7.61 F 21.92 S 2.18 Gd 10.67
Fnd: C 34.26 H 3.79 N 7.58 F 21.87 S 2.18 Gd 10.68
CA 02418790 2003-02-07
98
Fx, ample 24
1,7-Bis(benzyloxycarbonyl)-4-{3-oxa-pentane-1,5-dicarboxylic acid-1-oyl-5-[1-
(4-
perfluorooctylsulfonyl)-piperazine]-amide } -10-[2,6-N,N'-bis( 1-O-a-D-
carbonylmethyl-2,3,4,6-
tetra-O-benzyl-mannopyranose)]-L-lysyl-1,4, 7,10-tetraazacyclododecane
33.04 g (25.0 mmol) of the title compound of Example 18c), dissolved in 250 ml
of
tetrahydrofuran, is added to a solution that consists of 27.0 g (24.4 mrnol)
of the sec-amine,
produced under Example 1 Sa), in a mixture that consists of 150 ml of
tetrahydrofuran and 15 ml
of chloroform at 0°C and under nitrogen atmosphere. Then, a total of
18.0 g (36.6 mmol) of
EEDQ [2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline] is added in portions at
0°C and
allowed to stir overnight at room temperature. It is then evaporated to the
dry state in a vacuum,
and the remaining oil is chromatographed on silica gel (mobile solvent: n-
hexane/isopropanol
25:1 ). 45.87 g (78.0% of theory, relative to the sec-amine that is used) of
the title compound is
obtained in the form of a colorless oil.
Elementary analysis:
Cld: C 59.30 H 5.39 F 13.40 N 4.65 S 1.33
Fnd: C 59.32 H 5.37 F 13.37 N 4.70 S 1.34
b) 1-{3-Oxa-pentane-1,5-dicarboxylic acid-1-oyl-5-[1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide } -7-[2,6-N,N'-bis( 1-O-a-D-carbonylmethyl-mannopyranose)]-L-
lysyl-1,4, 7,10-
tetraazacyclododecane
24.I g (10.0 mmol) of the title compound that is produced under Example 24a)
is
dissolved in 250 ml of ethanol and mixed with 1.4 g of Pearlman's catalyst (Pd
20%, C). It is
hydrogenated until quantitative hydrogen uptake is reached, then catalyst is
suctioned out, it is
thoroughly rewashed with ethanol and evaporated to the dry state in a vacuum.
The product is
obtained as a yellowish-colored and extremely viscous oil.
Yield: 12:80 g (90.1 % of theory).
Elementary analysis:
Cld: C 39.72 H 4.89 F 22.73 N 7.88 S 2.26
CA 02418790 2003-02-07
CA 02418790 2003-02-07
99
Fnd: C 39.72 H 4.87 F 22.77 N 7.90 S 2.24
c) 1-{3-Oxa-pentane-1,5-dicarboxylic acid-1-oyl-5-[1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide } -7-[2,6-N,N'-bis( 1-O-a-D-carbonylmethyl-mannopyranose)]-L-
lysyl-4,10-
bis[1,4,7-tris(carboxylatomethyl)-10-(3-aza-4-oxo-5-methyl-5-yl-pentanoyl)J-
1,4,7,10-
tetraazacyclododecane, digadolinium complex
5.54 g [8.8 mmol; 2.2 molar equivalents relative to the amine component of
Example
24b) that is used] of the Gd complex, described in Patent Application DE 197
28 954 C 1 under
Example 3Ih), of 10-(4-carboxy-1-methyl-2-oxo-3=azabutyl)-1,4,7,10-
tetraazacyclododecane-
1,4,7-triacetic acid and anhydrous lithium chloride (0.37 g, 8.8 mmol) are
dissolved at 40°C in 60
ml of absolute dimethyl sulfoxide while being stirred and mixed at this
temperature with a total
of 1.01 g (8.8 mmol) of N-hydroxysuccinimide and 5.68 g (4.0 mmol) of the
title compound of
Example 24b), dissolved in 40 ml of absolute dimethyl sulfoxide. After cooling
to room
temperature, the reaction solution is mixed with 1.82 g (8.8 mmol) of N,N'-
dicyclohexylcarbodiimide and stirred for 12 hours at room temperature. The
suspension that is
obtained is then mixed with sufficient acetone until precipitation of the
above-mentioned title
compound is completed, the precipitate is suctioned off, dried, taken up in
water, insoluble
dicyclohexylurea is filtered out, and the filtrate is desalinated with an
AMICON~R~ YM-3
ultrafiltration membrane (cut-off 3,000 Da), and low-molecular components are
removed. The
retentate is then freeze-dried.
Yield: 8.52 g (80.6% of theory; relative to the diamine component that is
used) as a
colorless lyophilizate.
H20 content (Karl-Fischer): 6.09%.
Elementary analysis (relative to anhydrous substance):
Cld: C 38.61 H 4.76 N 9.53 F 12.21 Gd 11.89 S 1.I2
Fnd: C 38.57 H 4.82 N 9.52 F 12.21 Gd 11.93 S 1.15
i o0
Fxam In a 25
a) 1,7-Bis(benzyloxycarbonyl)-4-{3-oxa-pentane-1,5-dicarboxylic acid-1-oyl-5-
[1-(4-
perfluorooctylsulfonyl)-piperazine]-amide}-10-{ 2,6-N,N'-bis( 1-O-a-D-(5-
carbonyl)-pentyl-
2, 3,4,6-tetra-O-benzyl-mannopyranose) } -L-lysyl-1,4,7,10-
tetraazacyclododecane
35.80 g (25.0 mmol) of the title compound of Example 17e), dissolved in 250 ml
of
tetrahydrofuran, is added at 0°C and under nitrogen atmosphere to a
solution that consists of 27.0
g (24.4 mmol) of the sec-amine, produced under Example 15a), in a mixture that
consists of 150
ml of tetrahydrofuran and 15 ml of chloroform. Then, a total of 18.0 g (36.6
mmol) of EEDQ [2-
ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline] is added in portions at
0°C, and it is allowed to
stir overnight at room temperature. It is then evaporated to the dry state in
a vacuum, and the
remaining oil is chromatographed on silica gel (mobile solvent: n-
hexane/isopropanol 20:1).
49.48 g (80.4% of theory, relative to the sec-amine that is used) of the title
compound is obtained
in the form of a colorless oil.
Elementary analysis:
Cld: C 60.47 H 5.79 F 12.80 N 4.44 S 1.27
Fnd: C 60.52 H 5.77 F 12.77 N 4.50 S 1.30
b) 1-{3-Oxa-pentane-1,5-dicarboxylic acid-1-oyl-5-[1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide}-7-[2,6-N,N'-bis( 1-O-a-D-(5-carbonyl)-pentyl-
mannopyranose)]-L-lysyl-
1,4,7,10-tetraazacyclododecane
25.2 g (10.0 mmol) of the title compound that is produced under Example 25a)
is
dissolved in 250 ml of ethanol and mixed with 1.8 g of Pearlman's catalyst (Pd
20%, C). It is
hydrogenated until quantitative hydrogen uptake is reached, then catalyst is
suctioned out, it is
thoroughly rewashed with ethanol and evaporated to the dry state in a vacuum.
The product is
obtained as a yellow-colored and extremely viscous oil.
Yield: 14.11 g (92.5% of theory)
Elementary analysis:
Cld: C 49.60 H 7.20 F 21.17 N 7.34 S 2.10
CA 02418790 2003-02-07
101
Fnd: C 49.62 H 7.17 F 21.20 N 7.30 S 2.14
c) 1-{3-Oxa-pentane-1,5-dicarboxylic acid-1-oyl-5-[1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide } -7-[2,6-N,N'-bi s( 1-O-a-D-(5-carbonyl)-pentyl-
mannopyranose)]-L-lysyl-4,10-
bis[ 1,4,7-tris(carboxylatomethyl)-10-(3-aza-4-oxo-5-methyl-5-yl-pentanoyl)]-
1,4,7,10-
tetraazacyclododecane, digadolinium complex
5.54 g [8.8 mmol; 2.2 molar equivalents relative to the amine component of
Example
25b) that is used] of the Gd complex, described in Patent Application DE 197
28 954 C 1 under
Example 31h), of 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4,7,10-
tetraazacyclododecane-
1,4,7-triacetic acid and anhydrous lithium chloride (0.37 g, 8.8 mmol) are
dissolved at 40°C in 60
ml of absolute dimethyl sulfoxide while being stirred and mixed at this
temperature with a total
of 1.01 g (8.8 mmol) of N-hydroxysuccinimide and 6.10 g (4.0 mmol) of the
title compound of
Example 25b), dissolved in 40 ml of absolute dimethyl sulfoxide. After cooling
to room
temperature, the reaction solution is mixed with 1.82 g (8.8 mmol) of N,N'-
dicyclohexylcarbodiimide and stirred for 12 hours at room temperature. The
suspension that is
obtained is then mixed with sufficient acetone until precipitation of the
above-mentioned title
compound is completed, the precipitate is suctioned off, dried, taken up in
water, insoluble
dicyclohexylurea is filtered out, and the filtrate is desalinated with an
AMICON~R~ YM-3
ultrafiltration membrane (cut-off 3,000 Da), and low-molecular components are
removed. The
retentate is then freeze-dried.
Yield: 9.26 g (84.0% of theory; relative to the diamine component that is
used) as a
colorless lyophilizate.
Hz0 content (Karl-Fischer): 5.89%.
Elementary analysis (relative to anhydrous substance):
Cld: C 40.52 H 5.16 N 9.15 F 11.72 Gd 11.41 S 1.16
Fnd: C 40.57 H 5.20 N 9.12 F 11.69 Gd 11.43 S 1.18
CA 02418790 2003-02-07
102
Fxam~~)e 26
a) 6-N-t-Butyloxycarbonyl-2-N-benzyloxycarbonyl-L-lysine-[ 1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide
19.02 g (50.0 mmol) of a-N-(benzyloxycarbonyl)-~-N'-(tert-butyloxycarbonyl)-L-
lysine
(commercially available from the Bachem Company) is dissolved in 150 ml of
absolute
tetrahydrofuran. 8.31 g (50.0 mmol) of carbonyldiimidazole and 5.03 g (50.0
mmol) of
triethylamine, dissolved in 75 ml of dry tetrahydrofuran, are added drop by
drop at 0°C, and
stirnng is allowed to continue for 10 minutes at this temperature. Then, a
solution of 48.42 g
(50.0 mmol) of perfluorooctylsulfonyl-piperazine and 5.03 g (50.0 mmol) of
triethylamine in 250
ml of dry tetrahydrofuran is added in drops at 0°C. After stirring
overnight, the tetrahydrofuran is
drawn off in a vacuum, and the remaining oil is chromatographed on silica gel
(mobile solvent:
n-hexane/isopropanol 15:1). 49.48 g (80.4% of theory, relative to the sec-
amine that is used) of
the title compound is obtained in the form of a colorless oil.
Elementary analysis (relative to anhydrous substance):
Cld: C 40.01 H 3.79 N 6.02 F 34.70 S 3.45
Fnd: C 40.07 H 3.82 N 6.02 F 34.67 S 3.48
b} 6-N-t-Butyloxycarbonyl-L-lysine-[1-(4-perfluorooctylsulfonyl)-piperazine]-
amide
30.0 g (32.2 mmol) of the title compound of Example 26a) is dissolved in 300
ml of
isopropanol and mixed with 1.5 g of Pearlman's catalyst (20% palladium
hydroxide on carbon}.
It is hydrogenated for i 0 hours at room temperature, whereby after the course
of the reaction is
checked by thin-layer chromatography, the hydrogenolytic cleavage of the
benzyloxycarbonyl
protective group is carried out quantitatively even after the above-mentioned
reaction time.
Catalyst is filtered out, and the filtrate is evaporated to the dry state in a
vacuum. The remaining
residue is chromatographed on silica gel (mobile solvent: n-hexane/isopropanol
25:1). 25.13 g
(98.0% of theory) of the title compound is obtained in the form of a colorless
oil.
Elementary analysis:
Cld: C 34.68 H 3.67 F 40.55 N 7.03 S 4.03
CA 02418790 2003-02-07
CA 02418790 2003-02-07
103
Fnd: C 34.72 H 3.70 F 40.60 N 7.0I S 3.98
c) 6-N-t-Butyloxycarbonyl-2-N-[1-S-a-D-(2-carbonyl)-ethyl-2,3,4,6-tetra-O-
acetyl-
mannopyranose]-L-lysine-[ 1-(4-perfluorooctylsulfonyl)-piperazine]-amide
15.53 g (35.60 mmol) of the 3-(2,3,4,6-tetra-O-acetyl-1-thin-a-D-
mannopyranosyl)-
propionic acid (production according to: J. Haensler et al., Bioconjugate
Chem. 4, 85, (1993);
Chipowsky, S. and Lee, Y. C. (1973), Synthesis of I-Thio-aldosides;
Carbohydrate Research 31,
339-346 , and 3.60 g (35.60 mmol) of triethylamine are dissolved in 300 ml of
dry
tetrahydrofuran. After the reaction solution is cooled to -15°C to -
20°C, a solution of 4.92 g
(35.60 mmol) of isobutyl chloroformate in 75 ml of dry tetrahydrofuran is
slowly added in drops
at this temperature while being stirred, whereby the rate of addition by drops
can be selected in
such a way that an internal temperature of -10°C is not exceeded. After
a reaction time of 15
minutes at -15°C, a solution of 28.35 g (35.60 mmol) of the title
compound of Example 22b) and
3.60 g (35.60 mmol) of triethylamine, in 200 ml of dry tetrahydrofuran, is
then slowly added in
drops at 20°C. After a reaction time of one hour at -15°C and
two hours at room temperature, the
reaction solution is evaporated to the dry state in a vacuum. The remaining
residue is taken up in
250 ml of ethyl acetate and washed twice with 100 ml each of saturated sodium
bicarbonate
solution and once with 200 ml of water. After the organic phase is dried on
sodium sulfate, salt
is suctioned out, and the ethyl acetate is drawn off in a vacuum. The
remaining oily residue is
purified on silica gel with use of ethyl acetate/n-hexane (1:25) as an eluant.
Yield: 34.21 g (79.1 % of theory) of the above-mentioned title compound as a
colorless
and strongly viscous oil.
Elementary analysis:
Cld: C 39.54 H 4.23 N 4.61 F 26.58 S 5.28
Fnd: C 39.49 H 4.21 N 4.59 F 26.52 S 5.31
104
d) 6-N-t-Butyloxycarbonyl-2-N-[1-S-a-D-(2-carbonyl)-ethyl-mannopyranose]-L-
lysine-[1-
(4-perfluorooctylsulfonyl)-piperazine]-amide
29.93 g (24.64 mmol) of the title compound of Example 26c) is suspended in 400
ml of
absolute methanol and mixed at 5°C with a catalytic amount of sodium
methanolate. After a
reaction time of 3 hours at room temperature, even thin-layer chromatographic
checking (eluant:
chloroform/methanol = 9:1) of the course of the reaction indicates a
quantitative reaction. For
the purpose of working-up, the now clear reaction solution is neutralized by
mixing with
Amberlite~R~ IR 120 (H+ form)-cation-exchange resin, exchanger is suctioned
out, and the thus
obtained methanolic filtrate is drawn off in a vacuum until a dry state is
reached. The amorphous
residue that is obtained is purified by chromatography on silica gel with use
of 2-propanollethyl
acetate/n-hexane (1:1:15) as an eluant.
Yield: 23.42 g (90.8% of theory) of a colorless and viscous oil.
Elementary analysis:
Cld: C 36.72 H 4.14 N 5.35 F 30.85 S 6.13
Fnd: C 36.69 H 4.11 N 5.35 F 30.82 S 6.11
e) 2-N-[1-S-a-D-(2-Carbonyl)-ethyl-mannopyranose]-L-lysine-[1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide
20.93 g (20:0 mmol) of the title compound of Example 26d) is dissolved in a
mixture that
consists of 50 ml of trifluoroacetic acid and 100 ml of dichloromethane at
0°C while being stirred
vigorously, and it is stirred for 10 minutes at this temperature. Then, it is
evaporated to the dry
state in a vacuum; and the residue is taken up in 150 ml of water. The pH of
this aqueous
product solution is set at 9.5 by adding 2 molar aqueous sodium hydroxide
solution drop by drop.
The aqueous product solution is desalinated with an AMICON~R~ YM-3
ultrafiltration membrane
(cut-off: 3,000 Da), and in this case possible, still present, low-molecular
components are
removed at the same time. The retentate is then freeze-dried.
Yield: 17.79 g (94.2% of theory) of the free amine as a colorless
lyophilizate.
H20 content (Karl-Fischer): 3.09%.
CA 02418790 2003-02-07
Los
Elementary analysis (relative to anhydrous substance):
Cld: C 34.26 H 3.73 N 5.92 F 34.12 S 6.77
Fnd: C 34.26 H 3.79 N 5.88 F 34.07 S 6.80
f) 2-N-[1-S-a-D-(2-Carbonyl)-ethyl-mannopyranose]-6-N-[1,4,7-
tris(carboxylatomethyl)-
10-(3-aza-4-oxo-5-methyl-5-yl-pentanoyl)-1,4,7,10-tetraazacyclododecane]-L-
lysine-[ 1-(4-
perfluorooctylsulfonyl)-piperazine]-amide, gadolinium complex
5.54 g [(8.8 mmol, 2.2 molar equivalents relative to the amine component of
Example
26e) that is used] of the Gd complex, described in Patent Application DE 197
28 954 C 1 under
Example 31h), of 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4;7,10-
tetraazacyclododecane-
1,4,7-triacetic acid and 0.37 g of anhydrous lithium chloride (8.8 mmol) are
dissolved at 40°C in
60 ml of absolute dimethyl sulfoxide while being stirred and mixed at this
temperature with a
total of 1.01 g (8.8 mmol) of N-hydroxysuccinimide and 3.78 g (4.0 mmol) of
the title compound
of Example 26e), dissolved in 40 ml of absolute dimethyl sulfoxide. After
cooling to room
temperature, the reaction solution is mixed with 1.82 g (8.8 mmol) of N,N'-
dicyclohexylcarbodiimide and stirred for 12 hours at room temperature. The
suspension that is
obtained is then mixed with sufficient acetone until precipitation of the
above-mentioned title
compound is completed, the precipitate is suctioned off, taken up in water,
insoluble
dicyclohexylurea is filtered out, and the filtrate is desalinated with an
AMICON~R~ YM-3
ultrafiltration membrane (cut-off 3,000 Da), and low-molecular components are
removed. The
retentate is then freeze-dried.
Yield: 5.17 g (83.0% of theory) as a colorless lyophilizate.
Hz0 content (Karl-Fischer): 4.43%.
Elementary analysis (relative to anhydrous substance):
Cld: C 35.45 H 4.07 N 8.09 F 20.72 Gd 10.09 S 4.11
Fnd: C 35.50 H 4.01 N 8.12 F 20.68 Gd 10.13 S 4.14
CA 02418790 2003-02-07
106
a) 6-N-Benzyloxycarbonyl-2-N-(1-O-(3-D-carbonylmethyl-2,3,4,6-tetra-O-
benzylglucopyranose)-L-lysine-[ 1-(4-perfluorooctylsulfonyl)-piperazine]-amide
8.02 g (13.4 mmol) of the title compound [1-carboxymethyloxy-2,3,4,6-tetra-O-
benzyl-~3-
D-glucopyranoside] that is described in Patent Application DE 197 28 954 C 1
under Example
46a) and 3.24 g (28.14 mmol) of N-hydroxysuccinimide are dissolved in 100 ml
of
dimethylformamide and mixed in portions at 0°C with a total of 5.80 g
(28.14 mmol) of N,N'-
dicyclohexylcarbodiimide. It is stirred for 3 more hours at this temperature.
A solution, cooled
to 0°C, of 11.13 g (13.4 mmol) of the title compound of Example lc),
dissolved in 50 ml of
dimethylformamide, is added in drops to the thus produced active ester
solution, and it is stirred
for 2 hours at 0°C and for 12 hours at room temperature. For working-
up, precipitated
dicyclohexylurea is filtered out, and the solvent is then drawn off until a
dry state is reached. The
thus obtained residue is then chromatographed on silica gel (mobile solvent:
dichloromethane/ethanol, 20:1; the chromatography was carried out with use of
a solvent
gradient with continuous increase of the ethanol content).
Yield: 12.67 g (67.0% of theory) of the title compound in the form of a
colorless and
strongly viscous oil.
Elementary analysis:
Cld: C 52.77 H 4.50 N 3.97 F 22.89 S 2.27
Fnd: C 52.75 H 4.61 N 3.98 F 22.94 S 2.26
b) 2-N-(1-O-/3-D-Carbonylmethyl-glucopyranose)-L-lysine-[1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide
11.52 g (8.17 mmol) of the compound that is produced under 27a) is dissolved
in 100 ml
of ethanol, mixed with 0.5 g of Pearlman's catalyst (Pd 20%, C) and
hydrogenated at room
temperature under a hydrogen atmosphere (1 atm) until no more hydrogen
absorption can be
observed. Catalyst is suctioned out, it is thoroughly rewashed with ethanol
(three times with
about 40 ml in each case) and evaporated to the dry state in a vacuum. The
title compound is
CA 02418790 2003-02-07
io7
obtained as a strongly viscous and colorless oil.
Yield: 7.36 g (98.4% of theory).
Elementary analysis:
Cld: C 34.07 H 3.63 N 6.11 F 35.24 S 3.50
Fnd: C 34.11 H 3.59 N 6.08 F 35.23 S 3.52
c) 2-N-(1-O-(3-D-Carbonylmethyl-glucopyranose)-6-N-[1,4,7-
tris(carboxylatomethyl)-10-
(aza-4-oxo-5-methyl-5-yl-pentanoyl)-1,4,7,10-tetraazacyclododecane]-L-lysine-[
1-(4-
perfluorooctylsulfonyl)-piperazine]-amide, Gd complex
9.98 g [( 15.84 mmol; 2.2 molar equivalents relative to the amine component of
Example
27b) that is used] of the Gd complex, described in Patent Application DE 197
28 954 C1 under
Example 31h), of 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4,7,10-
tetraazacyclododecane-
1,4,7-triacetic acid and 0.67 g (15.84 mmol) of anhydrous lithium chloride are
dissolved at 40°C
in 80 ml of absolute dimethyl sulfoxide while being stirred and mixed at this
temperature with a
total of 1.82 g (15.84 mmol) of N-hydroxysuccinimide and 7.25 g (7.19 mmol) of
the title
compound of Example 27b), dissolved in 30 ml of absolute dimethyl sulfoxide.
After cooling to
room temperature, the reaction solution is mixed with 3.27 g (15.84 mmol) of
N,N'-
dicyclohexylcarbodiimide and stirred for 12 hours at room temperature. The
suspension that is
obtained is then mixed with sufficient acetone until precipitation of the
above-mentioned title
compound is completed, the precipitate is suctioned off, taken up in water,
insoluble
dicyclohexylurea is filtered out, the filtrate is desalinated with an
AMICON~R~ YM-3
ultrafiltration membrane (cut-off 3,000 Da), and low-molecular components are
removed. The
retentate is then freeze-dried.
Yield: 9.11 g (83.0% of theory) as a colorless lyophilizate.
H20 content (according to Karl-Fischer): 4.02%.
Elementary analysis (relative to anhydrous substance):
Cld: C 35.37 H 4.02 N 8.25 F 21.13 S 2.10 Gd 10.29
Fnd: C 35.42 H 4.07 N 8.18 F 21.09 S 2.06 Gd 10.34
CA 02418790 2003-02-07
108
~~~$
a) 2-N-Trifluoroacetyl-L-lysine-[1-(4-perfluorooctylsulfonyl)-piperazine]-
amide
10.0 g ( 11.46 mmol) of the compound; produced under 1 b), is dissolved in 100
ml of
ethanol, mixed with 1.0 g of Pearlman's catalyst (Pd 20%/C) and hydrogenated
until quantitative
hydrogen uptake is reached. Catalyst is suctioned out, it is rewashed with
ethanol and evaporated
to the dry state in a vacuum. The title compound is obtained as a viscous and
colorless oil.
Yield: 8.85 g (97.5% of theory).
Elementary analysis:
Cld: C 30.31 H 2.54 N 7.07 F 47.95 S 4.05
Fnd: C 30.36 H 2.50 N 7.11 F 47.99 S 4.00
b) 2-N-Trifluoroacetyl-6-N-[1-O-a-D-(5-carbonyl)-pentyl-2,3,4,6-tetra-O-benzyl-
mannopyranose]-L-lysine-[ 1-(4-perfluorooctylsulfonyl)-piperazine]-amide
A solution of 27.51 g (36.6 mmol) of the title compound of Example 17c} in 150
ml of
dimethylformamide is added in drops to a solution, cooled to 0°C, that
consists of 29.0 g (36.6
mmol) of the title compound of Example 28a) and 4.05 g (40.26 mmol) of
triethylamine in 100
ml of dimethylformamide. After the addition is completed, it is stirred for
one more hour at 0°C
and then overnight at room temperature. It is evaporated to the dry state in a
vacuum, and the
residue is taken up in 300 ml of ethyl- acetate. Insoluble components are
filtered out, and the
filtrate is washed twice with 100 ml each of 5% aqueous soda solution. The
organic phase is
dried on magnesium sulfate and evaporated to the dry state in a vacuum. The
residue is
chromatographed on silica gel (mobile solvent: n-hexane/isopropanol 25:1).
42.05 g (80.4% of
theory} of the title compound is obtained in the form of a colorless oil.
Elementary analysis:
Cld: C 50.42 H 4.51 N 7.96 F 26.59 S 2.24
Fnd: C 50.38 H 4.50 N 7.91 F 26.62 S 2.20
CA 02418790 2003-02-07
109
c) 6-N-[1-O-a-D-(5-Carbonyl)-pentyl-2,3,4,6-tetra-O-benzyl-mannopyranose]-L-
lysine-[1-
(4-perfluorooctylsulfonyl)-piperazine]-amide
20.0 g (14.0 mmol) of the compound, produced under Example 28b), is dissolved
in 150
ml of ethanol. The solution of 2.8 g (70.0 mmol) of sodium hydroxide in 2~ ml
of distilled water
is then added to it, and it is stirred for 0.5 hour at 50°C. According
to the thin-layer
chromatogram, the protective group cleavage at this point has already taken
place quantitatively:
It is evaporated to the dry state in a vacuum, and traces of water are removed
by repeated co-
distillation with ethanol. The residue is chromatographed on silica gel
(mobile solvent: n-
hexane/isopropanol 20:1 ). 16.66 g (89.3% of theory) of the title compound is
obtained in the
form of a colorless oil.
Elementary analysis:
Cld: C 52.25 H 4.91 N 4.20 F 24.22 S 2.41
Fnd: C 52.30 H 4.90 N 4.18 F 24.22 S 2.38
d) 6-N-[1-O-a-D-(5-Carbonyl)-pentyl-mannopyranose]-L-lysine-[1-(4-
perfluorooctylsulfonyl)-piperazine]-amide
15.0 g (11.25 mmol) of the compound, produced under 28c), is dissolved in 150
ml of a
10:1 mixture that consists of ethanol and water, and it is mixed with 1.0 g of
Pearlman's catalyst
(Pd 20%/C). Then, it is hydrogenated until quantitative hydrogen uptake is
reached at room
temperature and under one atmosphere of hydrogen pressure. Catalyst is
suctioned out, it is
rewashed with ethanol/water (10:1) and evaporated to the dry state in a
vacuum. The title
compound is obtained as a viscous and colorless oil.
Yield: 10.77 g (98.4% of theory).
Elementary analysis:
Cld: C 37.04 H 4.25 N 5.76 F 33.20 S 3.30
Fnd: C 37.06 H 4.20 N 5.18 F 33.19 S 3.30
CA 02418790 2003-02-07
CA 02418790 2003-02-07
110
e) 6-N-[1-O-a-D-(5-Carbonyl)-pentyl-mannopyranose]-2-N-[1,4,7-
tris(carboxylatomethyl)-
10-(3-aza-4-oxo-5-methyl-5-yl-pentanoyl)-1,4,7,10-tetraazacyclododecane]-L-
lysine-[ 1-(4-
perfluorooctylsulfonyl)-piperazine]-amide, Gd complex
5.54 g [(8.8 mmol; 2.2 molar equivalents relative to the amine component of
Example
28d) that is used] of the Gd complex, described in Patent Application DE 197
28 954 C1 under
Example 31 h), of 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4,7,10-
tetraazacyclododecane-
1,4,7-triacetic acid and 0.37 g (8.8 mmol) of anhydrous lithium chloride are
dissolved at 40°C in
60 ml of absolute dimethyl sulfoxide while being stirred and mixed at this
temperature with a
total of 1.01 g (8.8 mmol) of N-hydroxysuccinimide and 3.89 g (4.0 mmol) of
the title compound
of Example 28d), dissolved in 60 ml of absolute dimethyl sulfoxide. After
cooling to room
temperature, the reaction solution is mixed with 1.82 g (8.8 mmol) of N,N'-
dicyclohexylcarbodiimide and stirred for 12 hours at room temperature. The
suspension that is
obtained is then mixed with sufficient acetone until precipitation of the
above-mentioned title
compound is completed, the precipitate is suctioned off, taken up in water,
insoluble
dicyclohexylurea is filtered out, and the filtrate is desalinated with an
AMICON~R~ YM-3
ultrafiltration membrane (cut-off: 3,000 Da), and low-molecular components are
removed. The
retentate is then freeze-dried.
Yield: 4.81 g (75.9% o~ theory) as a colorless lyophilizate.
H20 content (Karl-Fischer): 8.98%.
Elementary analysis (relative to anhydrous substance):
Cld: C 37.15 H 4.39 N 7.96 F 20.38 Gd 9.92 S 2.02
Fnd: C 37.27 H 4.40 N 8.02 F 20.31 Gd 10.00 S 1.98
Fxam=le 29
a) 1,7-Bis(benzyloxycarbonyl)-4-( 1-O-[i-D-carbonylmethyl-2,3,4,6-tetra-O-
benzyl-
galactopyranose)-10-{3-oxa-pentane-1,5-dicarboxylic acid-1-oyl-5-[1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide } -1,4,7,10-tetraazacyclododecane
35.80 g (25.0 mmol) of the title compound of Example 17e), dissolved in 250 ml
of
CA 02418790 2003-02-07
111
tetrahydrofuran, is added to a solution that consists of 27:0 g (24.4 mmol) of
the sec-amine,
produced under Example 15a), in a mixture that consists of I50 ml of
tetrahydrofuran and 15 ml
of chloroform at 0°C and under nitrogen atmosphere. Then, a total of
18.0 g (36.6 rnmol) of
EEDQ [2-ethoxy-I-ethoxycarbonyl-1,2-dihydroquinoline] is added in portions at
0°C and
allowed to stir overnight at room temperature. It is then evaporated to the
dry state in a vacuum,
and the remaining oil is chromatographed on silica gel (mobile solvent: n-
hexane/isopropanol
20:1). 32.11 g (78.0% of theory, relative to the sec-amine that is used) of
the title compound is
obtained in the form of a colorless oil.
Elementary analysis:
Cld: C 54.09 H 4.72 F 19.14 N 4.98 S 1.90
Fnd: C 54.12 H 4.77 F 19.17 N 5.03 S 1.90
b) 1-(I-O-/3-D-Carbonylmethyl-galactopyranose)-7-{3-oxa-pentane-1,5-
dicarboxylic acid-1-
oyI-5-[ I -(4-perfluorooctylsulfonyl)-piperazine]-amide } - I ,4, 7,10-
tetraazacyclododecane
30.0 g (17.77 mmol) of the title compound, produced under Example 29a), is
dissolved in
250 ml of ethanol and mixed with 3.0 g of Pearlman's catalyst (Pd 20%/C). It
is hydrogenated
until quantitative hydrogen uptake is reached, catalyst is then suctioned out,
it is thoroughly
rewashed with ethanol and evaporated to the dry state in a vacuum. The product
is obtained as a
yellowish-colored and extremely viscous oil.
Yield: 17.89 g (95.1 % of theory)
Elementary analysis:
Cld: C 36.30 H 4.09 F 30.50 N 7.94 S 3.03
Fnd: C 36.26 H 4.12 F 30.46 N 7.90 S 3.04
112
c) 1-(1-O-(3-D-Carbonylmethyl-galactopyranose)-7-{3-oxapentane-1,5-
dicarboxylic acid-1-
oyl-5-[ I -(4-perfluorooctylsulfonyl)-piperazine]-amide } -4,10-bis[ 1,4,7-
tris(carboxylatomethyl)-
10-(3-aza-4-oxo-5-methyl-5-yl-pentanoyl)-1,4,7,10-tetraazacyclododecane]- I
,4,7,10-
tetraazacyclododecane, di-gadolinium complex
5.54 g [8.8 mmol; 4.4 molar equivalents relative to the amine component of
Example
29b) that is used] of the Gd complex, described in Patent Application DE 197
28 954 C 1 under
Example 31h), of IO-{4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4,7,10-
tetraazacyclododecane-
1,4,7-triacetic acid and 0.37 g (8.8 mmol) of anhydrous lithium chloride are
dissolved at 40°C in
60 m1 of absolute dimethyl sulfoxide while being stirred and mixed at this
temperature with a
total of 1.01 g (8.8 mmol) of N-hydroxysuccinimide and 2.1 I g (2.0 mmol) of
the title compound
of Example 29b), dissolved in 25 ml of absolute dimethyl sulfoxide. After
cooling to room
temperature, the reaction solution is mixed with 1.82 g (8.8 mmol) of N,N'-
dicyclohexyIcarbodiimide and stirred for 12 hours at room temperature. The
suspension that is
obtained is then mixed with sufficient acetone until precipitation of the
above-mentioned title
compound is completed, the precipitate is suctioned off, dried, taken up in
water, insoluble
dicyclohexylurea is filtered out, and the filtrate is desalinated with an
AMICON~R~ YM-3
ultrafiltration membrane (cut-off: 3,000 Da), and low-molecular components are
removed. The
retentate is then freeze-dried.
Yield: 3.29 g (72.2% of theory; relative to the amine component that is used)
as a
colorless lyophilizate.
H20 content (Karl-Fischer): 5.99%.
Elementary analysis (relative to anhydrous substance):
Cld: C 36.84 H 4.37 N 9.82 F 14.15 Gd 19.63 S 1.40
Fnd: C 36.87 H 4.40 N 9.82 F 14.09 Gd 19.59 S 1.38
CA 02418790 2003-02-07
113
T,xam In a 30
a) 3-(1-O-a-D-2,3,4,6-Tetra-O-benzyl-mannopyranose)-2-N-benzyloxycarbonyl-L-
serine-
methyl ester
21.42 g (39.61 mmol) of 2,3,4,6-tetra-O-benzyl-a-D-mannopyranose (production
according to: F. Kong et al., J. Carbohydr. Chem.; 16; 6; 1997; 877-890) is
dissolved in 500 ml
of dry acetonitrile. After the reaction solution is cooled to 5°C, a
solution of 13.23 g (59.52
mmol) of trifluoromethanesulfonic acid-trimethyl silyl ester in 30 ml of
acetonitrile, followed by
a solution that consists of 20.06 g (79.21 mmol) of N-benzyloxycarbonyl-L-
serine methyl ester
(commercially available from the Bachem Company) in 50 ml of acetonitrile, are
slowly added in
drops at this temperature while being stirred, whereby the rate of addition by
drops can be
selected in such a way that an internal temperature of 10°C is not
exceeded. After a reaction time
of 15 hours at room temperature; the reaction solution is evaporated to the
dry state in a vacuum.
The remaining residue is taken up in 250 ml of ethyl acetate and washed twice
with 104 ml each
of saturated sodium bicarbonate solution and once with 200 ml of water. After
the organic phase
is dried on sodium sulfate, salt is suctioned out, and the ethyl acetate is
removed in a vacuum.
The remaining oily residue is purified on silica gel with use of ethyl
acetate/n-hexane (1:5) as an
eluant.
Yield: 23:60 g (76.8% of theory) of the above-mentioned title compound as a
colorless
oil.
Elementary analysis:
Cld: C 71.21 H 6.37 N 1.81
Fnd: C 71.19 H 6.41 N 1.79
b) 3-(1-O-a-D-2,3;4,6-Tetra-O-benzyl-mannopyranose)-2-N-benzyloxycarbonyl-L-
serine
10.0 g (12.90 mmol) of the compound that is produced under Example 30a) is
dissolved
in a mixture that consists of 20 ml of methanol, 20 ml of water and 50 ml of
tetrahydrofuran.
0.47 g (19.35 mmol) of lithium hydroxide, dissolved in 25 ml of distilled
water, is then added at
room temperature, and it is then stirred for 6 hours at 60°C. After the
course of the reaction is
CA 02418790 2003-02-07
CA 02418790 2003-02-07
114
checked by means of thin-layer chromatography (eluant: methylene
chloride/methanol 10:1), the
saponification of the methyl ester of Example 30a) has already taken place
quantitatively after the
above-mentioned reaction time. For the purpose of working-up, the product
solution is
evaporated to the dry state in a vacuum, and the remaining residue is taken up
in 250 ml of ethyl
acetate in the heat (about 60°C). Then, the thus obtained ethyl acetate
phase is washed twice
with 50 ml each of a 15% aqueous hydrochloric acid, and once with 100 ml of
distilled water.
The organic phase is dried on magnesium sulfate, filtered and evaporated to
the dry state in a
vacuum. The residue is chromatographed on silica gel (mobile solvent: n-
hexane/ethyl acetate
5:1). 8.40 g (85.7% of theory) of the title compound is obtained in the form
of a colorless oil.
Elementary analysis:
Cld: C 70.94 H 6.22 N 1.84
Fnd: C 70.97 H 6.30 N 1.78
c) 3-(1-O-a-D-2,3,4,6-Tetra-O-benzyl-mannopyranose-2-N-benzyloxycarbonyl-L-
serine-[1-
(4-perfluorooctylsulfonyl)piperazine]-amide
20.57 g (27.0 mmol) of the carboxylic acid, produced according to Example
30b),
dissolved in 50 ml of tetrahydrofuran, is added drop by drop to 13.86 g (24.40
mmol) of 1-
perfluorooctylsulfonylpiperazine (produced according to DE 19603033),
dissolved in a mixture
that consists of 1 SO ml of tetrahydrofuran and 15 ml of chloroform at
0°C and under nitrogen
atmosphere. Then, a total of 18.0 g (36.60 mmol) of EEDQ [2-ethoxy-1-
ethoxycarbonyl-1,2-
dihydroquinoline] is added in portions at 0°C and allowed to stir
overnight at room temperature.
For the purpose of working-up, the reaction solution is concentrated by
evaporation in a vacuum,
and the remaining, extremely viscous oil is chromatographed on silica gel with
use of an n-
hexane/isopropanol (15:I) mixture as an eluant system. 17.0 g (79.6% of
theory; relative to the
primaryamine that is used) of the title compound is obtained in the form of a
colorless and
viscous oil.
Elementary analysis:
Cld: C S I .53 H 4.23 N 3.15 F 25.65 S 2.41
CA 02418790 2003-02-07
IIS
Fnd: C 51.48 H 4.27 N 3.10 F 25.71 S 2.35
d) 3-(1-O-a-D-Mannopyranose)-L-serine-[1-(4-perfluorooctylsulfonyl)piperazineJ-
amide
15.0 g (11.41 mmol) of the compound, produced according to Example 30c}, is
dissolved
in 200 ml of ethanol and mixed with I .5 g of Pearlman's catalyst (Pd 20%, C).
Then, the reaction
solution is hydrogenated at room temperature under a hydrogen atmosphere ( 1
atm) until no more
hydrogen absorption can be observed (about 8 hours). For the purpose of
working-up, catalyst is
suctioned out, it is thoroughly rewashed with ethanol (twice with about 100 ml
each), and the
product-containing ethanolic filtrate is evaporated to the dry state in a
vacuum. The title
compound is obtained as a strongly viscous and colorless oil.
Yield: 8.79 g (94.0% of theory).
Elementary analysis:
Cld: C 30.78 H 3.20 N 5.13 F 39.41 S 3.91
Fnd: C 30.87 H 3.14 N 5.19 F 39.50 S 3.88
e) 3-(1-O-a-D-Mannopyranose)-2-N-[1,4,7-tris(carboxylatomethyl)-10-(3-aza-4-
oxo-5-
methyl-5-yl-pentanoyl)-1,4,7,10-tetraazacyclododecane]-L-serine-[ 1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide, Gd complex
A stirred suspension of 5.7 g [9.06 mmol; corresponding to 1.5 molar
equivalents relative
to the title compound (primary amine) of Example 30d) that is used] of the Gd
complex,
described in Patent Application DE 197 28 954 C1 under Example 31h), of 10-(4-
carboxy-1-
methyl-2-oxo-3-azabutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid
in 75 ml of
absolute dimethyl sulfoxide is mixed at 70°C with 0.68 g (15.9 mmol) of
lithium chloride. After
30 minutes of stirring at 70°C, the now clear reaction solution is
mixed in portions with a total of
1.83 g (15.9 mmol) of N-hydroyxsuccinimide, and the reaction mixture is held
for 1 more hour at
70°C. After the reaction solution is cooled to 10°C, it is mixed
with 4.52 g (23.85 mmol) of
dicyclohexylcarbodiimide; and the reaction solution is stirred for another
hour at 0°C, followed
by 12 hours at 22°C. The thus obtained solution of N-hydroxysuccinimide
ester of the Gd
116
complex of 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4,7,10-
tetraazacyclododecane-1,4,7-
triacetic acid is now mixed drop by drop at 22°C with a solution of
4.94 g (6.03 mmol) of the title
compound of Example 30d), in 15 ml of absolute dimethyl sulfoxide, and it is
stirred for another
12 hours at room temperature. For working-up, the reaction solution is slowly
added in drops at
22°C into a solvent mixture that consists of 250 ml of acetone and 250
ml of 2-propanol,
whereby after 12 hours at 10°C, the title compound is deposited
completely as a light yellowish-
colored oil. The supernatant eluant mixture is carefully decanted out, and the
oily product is
taken up in 200 ml of distilled water; whereby the latter completely dissolves
so that a light
yellowish-colored aqueous solution of the above-mentioned title compound is
obtained. The
aqueous product solution is subsequently filtered first with a membrane filter
and then, for the
purpose of desalination and separation of low-molecular components, it is
ultrafiltered three
times with a YM3-ultrafiltration membrane (AMICON~R~: cut-off: 3,000 Da). The
thus
obtained retentate is then freeze-dried.
Yield: 8.63 g (80.2% of theory, relative to the title compound of Example 30d)
that is
used) as a colorless lyophilizate with a water content of 7.65%.
Elementary analysis (relative to anhydrous substance):
Cld: C 33.57 H 3.80 N 7.83 F 22.57 Gd 10.99 S 2.24
Fnd: C 33.57 H 3.76 N 7.82 F 22.63 Gd I1.06 S 2.T8
a) 6-N-Benzyloxycarbonyl-2-N-[O-(3-D-galactopyranosyl (1-~4)-gluconosyl]-L-
lysine-[1-
(4-perfluorooctylsulfonyl)-piperazine]-amide
A solution of 13.3 g (37.2 mmol) of O-[i-D-galactopyranosyl-(1-~4)-D-glucono-
1,5-
lactone [lactobionolactone; production according to: (a) Williams, T. 3.;
Plessas, N. R.,
Goldstein, I. J. Carbohydr. Res. 1978, 67, Cl. (b) Kobayashi, K.; Sumitomo,
H.; Ina, Y. Polym. J.
1985, 17; 567, (c) Hiromi Kitano, Katsuko Sohda, and Ayako Kosaka,
Bioconjugate Chem.
1995, 6 131-134) in 40 ml of absolute dimethyl sulfoxide is added drop by drop
to a stirred
solution of 4.98 g (6.0 mmol) of the title compound of Example 1c) in 40 ml of
absolute
CA 02418790 2003-02-07
117
dimethyl sulfoxide at room temperature. The thus obtained reaction solution is
then stirred for
14 hours at 40°C. For working-up, it is mixed at room temperature with
500 ml of absolute 2-
propanol, and the colorless precipitate that is produced is suctioned off
using a G4 frit and
thoroughly rewashed with a total of 250 ml of absolute 2-propanol. The thus
obtained solid is
now dissolved in 300 ml of distilled water and ultrafiltered a total of three
times with a YM3-
ultrafiltration membrane (AMICON~R~: cut-off: 3,000 Da). Both the excess
lactobionolactone
and possibly still present, low-molecular components of the desired product
are separated by the
ultrafiltration process that is repeated three times. The residue that remains
in the ultrafiltration
membrane is subsequently completely dissolved in 300 ml of distilled water and
freeze-dried.
Yield: 6.51 g (92.7% of theory) as a colorless lyophilizate.
Water content: 10.03%.
Elementary analysis (relative to anhydrous substance):
Cld: C 38.98 H 4.05 N 4.79 F 27.58 S 2.74
Fnd: C 39.04 H 4.09 N 4.82 F 27.61 S 2.71
b) 2-N-[O-[i-D-Galactopyranosyl (l.-~4)-gluconosyl]-L-lysine-[1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide
5.0 g (4.27 mmol) of the compound that is produced under 31 a) is dissolved in
100 ml of
ethanol, mixed with 0.5 g of Pearlman's catalyst (Pd 20%, C) and hydrogenated
at 1 atmosphere
of hydrogen pressure until quantitative hydrogen uptake is reached. Catalyst
is suctioned out, it
is rewashed with ethanol and evaporated to the dry state in a vacuum. The
title compound is
obtained as a colorless and viscous oil.
Yield: 4.36 g (98.5% of theory).
Elementary analysis:
Cld: C 34.76 H 3.99 N 5.40 F 31.51 S 3.09
Fnd: C 34.78 H 4.04 N 5.34 F 31.51 S 3.15
CA 02418790 2003-02-07
118
c) 2-N-[O-[3-D-Galactopyranosyl (1-~4)-gluconosyl]-6-N-[1,4,7-
tris(carboxylatomethyl)-10-
(3-aza-4-oxo-5-methyl-5-yl-pentanoyl)-1,4,7,10-tetraazacyclododecane]-L-lysine-
[ 1-(4-
perfluorooctylsulfonyl)-piperazine]-amide, Gd complex
5.54 g [(8.8 mmol; 2.2 molar equivalents relative to the amine component of
Example
31 b) that is used] of the Gd complex, described in Patent Application DE 197
28 954 C 1 under
Example 31 h), of 10-(4-carboxy-1-methyl-2-oxo-3-azabutyl)-1,4,7,10-
tetraazacyclododecane-
1,4,7-triacetic acid and 0.37 g (8.8 mmol) of anhydrous lithium chloride are
dissolved at 40°C in
60 ml of absolute dimethyl sulfoxide while being stirred and mixed at this
temperature with a
total of 1.01 g (8.8 mmol) of N-hydroxysuccinimide and 3.85 g (4.0 mmol) of
the title compound
of Example 31 b), dissolved in 60 ml of absolute dimethyl sulfoxide. After
cooling to room
temperature, the reaction solution is mixed with 1.82 g (8.8 mmol) of N,N'-
dicyclohexylcarbodiimide and stirred for 12 hours at room temperature. The
suspension that is
obtained is then mixed with sufficient acetone/2-propanol (1:1) until
precipitation of the above-
mentioned title compound is completed, and the precipitate is suctioned off.
The thus obtained
precipitate is subsequently taken up in 300 m1 of water, and insoluble
dicyclohexylurea is filtered
out. The filtrate is ultrafiltered three times with an AMICON~R~ YM-3
ultrafiltration membrane
(cut-off: 3,000 Da). Both the excess Gd complex and possibly still present,
low-molecular
components are separated from the desired product by the ultrafiltration
process that is performed
three times. The residue that remains in the ultrafiltration membrane is
subsequently completely
dissolved in 500 ml of distilled water and freeze-dried.
Yield: 4.64 g (70.4% of theory) as a colorless lyophilizate.
H20 content (Karl-Fischer): 10.08%.
Elementary analysis (relative to anhydrous substance):
Cld: C 35.70 H 4.22 N 7.65 F 19.59 Gd 9.54 S 1.95
Fnd: C 3 5.77 H 4.17 N 7.71 F 19.61 Gd 9.60 S 1.99
CA 02418790 2003-02-07
CA 02418790 2003-02-07
y , 119
Example 32
a) 6-N-Benzyloxycarbonyl-2-N-(2,3,4,5-pentahydroxy-hexanoyl)L-lysine-[1-(4-
perfluorooctylsulfonyl)-piperazine)-amide
A solution that consists of 21.45 g (120.4 mol) of 5-gluconolactone in 50 ml
of
tetrahydrofuran is added in drops at SO°C to a solution that consists
of 100.0 g (120.4 mol) of the
title compound of Example 1 c), in 500 ml of dry tetrahydrofuran. It is
stirred for 3 hours at 60°C
and then overnight at room temperature. It is evaporated to the dry state in a
vacuum, and the
residue is chromatographed on silica gel (mobile solvent:
dichloromethane/ethanol = 20:1).
Yield: 98.37 g (82% of theory) of a viscous oil.
Elementary analysis:
Cld: C 38.10 H 3.70 F 32.02 N 5.55 S 3.18
Fnd: C 38.22 H 3.79 F 32.02 N 5.42 S 3.29
b) 2-N-(2,3,4,5-Pentahydroxy-hexanoyl)-L-lysine-[1-[(4-perfluorooctylsulfonyl)-
piperazineJ-
amide
100.9 g (100.0 mmol) of the title compound of Example 32a) is dissolved in
2000 ml of
ethanol, and 10.0 g of palladium catalyst (10% Pd/C) is added to it. It is
hydrogenated for I2
hours at room temperature. Catalyst is filtered out, and the filtrate is
evaporated to the dry state
in a vacuum.
Yield: 87.46 g (quantitative) of a colorless solid.
Elementary analysis:
Cld: C 32.96 H 3.57 N 6.41 S 3.67 F 36.93
Fnd: C 32.91 H 3.72 N 6.34 S 3.50 F 36.78
c) 6-N-[1,4,7-Tris(carboxylatomethyl)]-1,4,7,10-tetraazacyclododecane-10-N-
(pentanoyl-3-aza-
4-oxo-5-methyl-5-yl)]-2-N-[ 1-O-a-D-carbonylmethyl-mannopyranose)-L-lysine-[ 1-
(4-
perfluorooctylsulfonyl)-piperazine)-amide, Gd complex
50.0 g (54.55 mmol) of the title compound of Example 1 e), 6.28 g (54.55 mmol)
of N-
120
hydroxysuccinimide, 4.62 g (109.0 mol) of lithium chloride and 34.35 g (54.55
mol) of 1,4,7-
tris(carboxylatomethyl)-10-(carboxy-3-aza-4-oxo-5-methyl-pent-5-yl)-1,4,7,10-
tetraazacyclododecane, Gd complex, are dissolved in 400 ml of dimethyl
sulfoxide while being
heated slightly. At 10°C, 16.88 g (81.8 mol) of N,N-
dicyclohexylcarbodiimide is added, and it
then is stirred overnight at room temperature. The solution is poured into
3000 ml of acetone and
stirred for 10 minutes. The precipitated solid is filtered off and then
purified by chromatography
(RP-18 mobile solvent: gradient that consists of water/ethanol/acetonitrile).
Yield: 75.9 g (91.0% of theory) of a colorless solid.
Water content: 8.6%.
Elementary analysis (relative to anhydrous substance):
Cld: C 35.34 H 4.09 N 8.24 S 2.10 F 21.12 Gd 10.28
Fnd: C 35.28 H 4.15 N 8.19 S 2.15 F 21.03 Gd 10.14
Example 33
a) 6-N-Benzyloxycarbonyl-2-N-(2,3,4,5-pentahydroxy-hexanoyl)L-lysine-[1-(4-
perfluorooctylsulfonyl)-piperazineJ-amide
A solution that consists of 21.45 g (120.4 mol) of 5-gluconolactone in 50 ml
of
tetrahydrofuran is added in drops at 50°C to a solution that consists
of 100.0 g ( 120.4 mmol) of
the title compound of Example 1 c), and 12.18 g ( 120.4 mmol) of triethylamine
in 500 ml of dry
tetrahydrofuran: It is stirred for 3 hours at 60°C and then overnight
at room temperature. 400 ml
of 5% aqueous hydrochloric acid is then added to it, it is stirred for 5
minutes at room
temperature, mixed with sodium chloride, the organic phase is separated, it is
dried on
magnesium sulfate, evaporated to the dry state in a vacuum, and the residue is
chromatographed
on silica gel (mobile solvent: dichloromethane/ethanol = 20:1 ).
Yield: 100.97 g (82% of theory) of a viscous oil.
Elementary analysis:
Cld: C 37.58 H 3.45 F 31.58 N 5.48 S 3.14
Fnd: C 37.72 H 3.59 F 31.72 N 5.42 S 3.29
CA 02418790 2003-02-07
121
b) 2-N-(2,3,4,5-Pentahydroxy-hexanoyl)-L-lysine-1-[(4-perfluorooctylsulfonyl)-
piperazine]-
amide
100.9 g (100.0 mmol) of the title compound of Example 32a) is dissolved in
2000 ml of
ethanol, and 10.0 g of palladium catalyst (10% Pd/C) is added to it. It is
hydrogenated for 12
hours at room temperature. Catalyst is filtered out, and the filtrate is
evaporated to the dry state
in a vacuum.
Yield: 87.46 g (quantitative) of a colorless solid.
Elementary analysis:
Cld: C 32.96 H 3.57 N 6.41 S 3.67 F 36.93
Fnd: C 32.91 H 3.72 N 6.34 S 3.50 F 36.78
c) 6-N-[1,4,7-Tris(carboxylatomethyl)]-1,4,7,10-tetraazacyclododecane-10-N-
(pentanoyl-3-aza-
4-oxo-5-methyl-5-yl)]-2-N-[ 1-O-a-D-carbonylmethyl-mannopyranose]-L-lysine-[ 1-
(4-
perfluorooctylsulfonyl)-piperazine]-amide, Gd complex
50.0 g (54.55 mmol) of the title compound of Example 1 e), 6.28 g (54.55 mmol)
of N-
hydroxysuccinimide, 4.62 g (109.0 mol) of lithium chloride and 34.35 g (54.55
mol) of 1,4,7-
tris(carboxylatomethyl)-10-(carboxy-3-aza-4-oxo-5-methyl-pent-5-yl)-1,4,7,10-
tetraazacyclododecane, Gd complex, are dissolved in 400 mI of dimethyl
sulfoxide while being
heated slightly. At 10°C, 16.88 g (81.8 mol) of N,N-
dicyclohexylcarbodiimide is added, and it
then is stirred overnight at room temperature. The solution is poured into
3000 ml of acetone and
stirred for 10 minutes. The precipitated solid is filtered off and then
purified by chromatography
(RP-18 mobile solvent: gradient that consists of water/ethanol/acetonitrile).
Yield: 75.9 g (91.0% of theory) of a colorless solid.
Water content: 8.6%
Elementary analysis (relative to anhydrous substance):
Cld: C 35.34 H 4.09 N 8.24 S 2.10 F 21.12 Gd 10.28
Fnd: C 35.28 H 4.15 N 8.19 S 2.15 F 21.03 Gd 10.14
CA 02418790 2003-02-07
122
Under the conditions of Example 1 f, mannose was replaced with glucose or
galactose.
Example 34
a) 6-N-Benzyloxycarbonyl-2-N-[1-O-a-D-carbonylmethyl-(2,3,4,6-tetra-O-benzyl
glucopyranose]-L-lysine-[ 1-(4-perfluorooctylsulfonyl)-piperazine]-amide
41.27 g (200.0 mmol) of N,N-dicyclohexylcarbodiimide is added at 0°C to
a solution that
consists of 100.0 g (120.4 mol) of the title compound of Example 1 c), 72.1 g
(120.4 mol) of 1-O-
a-D-carboxymethyl-2,3,4,6-tetra-O-benzyl-glucopyranose and 13.86 g (120.4 mol)
ofN-
hydroxysuccinimide, dissolved in 500 ml of dimethylformamide. It is stirred
for 3 hours at 0°C
and then overnight at room temperature. Precipitated urea is filtered out, the
filtrate is
evaporated to the dry state in a vacuum and chromatographed on silica gel
(mobile solvent: '
dichloromethane/ethanol = 20:1 ).
Yield: 136.1 g (87% of theory) of a viscous oil.
Elementary analysis:
Cld: C 57.32 H 4.89 N 4.31 F 24.86 S 2.47
Fnd: C 57.48 H 5.04 N 4.20 F 24.69 S 2.38
b) 2-N-[1-O-a-D-Carbonylmethylglucopyranose]-L-lysine-1-[(4-
perfluorooctylsulfonyl)-
piperazine]-amide
130.0 g ( 100.0 mmol) of the title compound of Example 34a) is dissolved in
2000 ml of
ethanol, and 10.0 g of palladium catalyst (10% Pd/C) is added to it. It is
hydrogenated for 12
hours at room temperature. Catalyst is filtered out, and the filtrate is
evaporated to the dry state
in a vacuum:
Yield: 91.7 g (quantitative) of a colorless solid.
Elementary analysis:
Cld: C 34.07 H 3.63 N 6.11 S 3.50 F 35.24
Fnd: C 33.92 H 3.71 N 6.02 S 3.42 F 35.33
CA 02418790 2003-02-07
123
c) 6-N-(1,4,7-Tris(carboxylatomethyl)]-1,4,7,10-tetraazacyclododecane-10-N-
(pentanoyl-3-aza-
4-oxo-5-methyl-5-y1)]-2-N-[ 1-O-a-D-carbonyImethyl-glucopyranose]-L-lysine-[ 1-
(4-
perfluorooctylsulfonyl)-piperazine]-amide, Gd complex
50.0 g (54.55 mmol) of the title compound of Example 34b), 6.28 g (54.55 mmol)
of N-
hydroxysuccinimide, 4.62 g (109.0 mol) of lithium chloride and 34.35 g (54.55
mol) of 1,4,7-
tris(carboxylatomethyl)-10-(carboxy-3-aza-4-oxo-5-methyl-pent-5-yl)-1,4,7, I 0-
tetraazacyclododecane, Gd complex, are dissolved in 400 ml of dimethyl
sulfoxide while being
heated slightly. At 10°C, 16.88 g (81.8 mol) of N,N-
dicyclohexylcarbodiimide is added, and it
then is stirred overnight at room temperature. The solution is poured into
3000 ml of acetone,
and it is stirred for 10 minutes. The precipitated solid is filtered off and
then purified by
chromatography (RP-18 mobile solvent: gradient that consists of
water/ethanol/acetonitrile).
Yield: 75.9 g (91.0% of theory) of a colorless solid.
Water content: 8.6%.
Elementary analysis (relative to anhydrous substance):
Cld: C 35.34 H 4.09 N 8.24 S 2.10 F 21.12 Gd 10.28
Fnd: C 35.26 H 4.18 N 8.14 S 2.158 F 21.01 Gd 10.13
Example 35
a) 6-N-Benzyloxycarbonyl-2-N-(1-O-a-D-carbonylmethyl-(2,3,4,6-tetra-O-benzyl-
galactopyranose]-L-lysine-{ 1-(4-perfluorooctylsulfonyl)-piperazine]-amide
20.64 g (100.0 mmol) ofN,N-dicyclohexylcarbodiimide is added at 0°C to
a solution that
consists of 50.0 g (60.2 mol) ofthe title compound of Example lc), 36.05 g
(60.2 mmol) of 1-O-
a-D-carboxymethyl-2,3,4,6-tetra-O-benzyl-galactopyranose and 6.93 g (60.2
mmol) of N-
hydroxysuccinimide, dissolved in 500 ml of dimethylformamide. It is stirred
for 3 hours at 0°C
and then overnight at room temperature. Precipitated urea is filtered out,-the
filtrate is
evaporated to the dry state in a vacuum and chromatographed on silica gel
(mobile solvent:
dichloromethane/ethanol = 20:1 ).
CA 02418790 2003-02-07
124
Yield: 68.1 g (87% of theory) of a viscous oil.
Elementary analysis:
Cld: C 57.32 H 4.89 N 4.31 F 24.86 S 2.47
Fnd: C 57.47 H 5.05 N 4.19 F 24.72 S 2.29
b) 2-N-[1-O-a-D-Carbonylmethyl-galactopyranoseJ-L-lysine-1-[(4-
perfluorooctylsulfonyl)-
piperazine]-amide
65.0 g (50.0 mmol) of the title compound of Example 35a) is dissolved in 1000
ml of
ethanol, and 5.0 g of palladium catalyst (10% Pd/C) is added to it. It is
hydrogenated for 12
hours at room temperature. Catalyst is filtered out, and the filtrate is
evaporated to the dry state
in a vacuum.
Yield: 45.85 g (quantitative) of a colorless solid.
Elementary analysis:
Cld: C 34.07 H 3.63 N 6.11 S 3.50 F 35.24
Fnd: C 33.93 H 3.74 N 6.01 S 3.39 F 35.05
c) 6-N-[ 1,4,7-Tris(carboxylatomethyl)]-1,4,7,10-tetraazacyclododecane-10-N-
(pentanoyl-3-aza-
4-oxo-5-methyl-5-yl)J-2-N-[ 1-O-a-D-carbonylmethyl-galactopyranose]-L-lysine-[
1-(4-
perfluorooctylsulfonyl)-piperazineJ-amide, Gd complex
50.0 g (54.55 mmol) of the title compound of Example 35b), 6.28 g (54.55 mmol)
of N-
hydroxysuccinirriide, 4.62 g (109.0 mol) of lithium chloride and 34:35 g
(54.55 mol) of 1,4,7-
tris(carboxylatomethyl)-10-(carboxy-3-aza-4-oxo-5-methyl-pent-5-yl)-1,4,7,10-
tetraazacyclododecane, Gd complex, are dissolved in 400 ml of dimethyl
sulfoxide while being
heated slightly. At 10°C, 16.88 g (81.8 mol) of N,N-
dicyclohexylcarbodiimide is added, and it
then is stirred overnight at room temperature. The solution is poured into
3000 ml of acetone and
stirred for 10 minutes. The precipitated solid is filtered off and then
purified by chromatography
(RP-18 mobile solvent: gradient that consists of water/ethanol/acetonitrile).
Yield: 37.95 g (91.0% of theory) of a colorless solid.
CA 02418790 2003-02-07
125
Water content: 8.6%.
Elementary analysis (relative to anhydrous substance):
Cld: C 35.34 H 4.09 N 8.24 S 2.10 F 21.12 Gd 10.28
Fnd: C 3 5.22 H 4.17 N 8.18 S 2.19 F 20.91 Gd 10.12
Example 36
a) N-Trifluoroacetyl-L-glutamic acid-mono-benzyl ester
100 g (421.5 mmol) of L-glutamic acid-mono-benzyl ester is dissolved in a
mixture that
consists of 1000 ml of trifluoroacetic acid-ethyl ester/500 ml of ethanol, and
it is stirred for 24
hours at room temperature. It is evaporated to the dry state, and the residue
is crystallized from
diisopropyl ether.
Yield: 140.47 g (96% of theory) of a colorless, crystalline powder.
Elementary analysis:
Cld: C 50.46 H 4.23 F 17.10 N 4.20
Fnd: C 51.35 H 4.18 F 17.03 N 4.28
b) 2-N-Trifluoacetyl-L-glutamic acid-mono-benzyl ester-5-N-(methyl)-N-
(2,3,4,5,6-
pentahydroxyhexyl)-amide
8.25 g (40 mmol) of N,N-dicyclohexylcarbodiimide is added at 0°C to a
solution that
consists of 24.9 g (24.08 mmol) of the title compound of Example 36a, 2x g
(24.08 mmol) of N-
methylglucamine and 2.77 g (24.08 mmol) of N-hydroxysuccinimide, dissolved in
150 ml of
dimethylformamide. It is stirred for 3 hours at 0°C, then overnight at
room temperature.
Precipitated urea is filtered out, the filtrate is evaporated to the dry state
in a vacuum and
chromatographed on silica gel (mobile solvent = dichloromethane/ethanol = 20:1
).
Yield: 109.40 g (89% of theory) of a viscous oil.
Elementary analysis:
Cld: C 51.43 H 5.51 F 13.56 N 6.66
CA 02418790 2003-02-07
126
Fnd: C 51.22 H 5.41 F 13.40 N 6.75
c) N-Trifluoroacetyl-L-glutamic acid-N-(methyl)-N-(2,3,4,5,6-
pentahydroxyhexyl)-amide
77.33 g (15.15 mmol) of the title compound of Example 36b is dissolved in 500
ml of
ethanol, and 3 g of palladium catalyst ( 10% Pd/C) is added. It is
hydrogenated at room
temperature. Catalyst is filtered out, and the filtrate is evaporated to the
dry state in a vacuum.
Yield: 43.0 g (quantitative) of a colorless solid.
Elementary analysis:
Cld: C 40.01 H 5.19 F 17.26 N 8.48
Fnd: C 39.84 H 5.13 F 17.09 N 8.68
d) Trifluoroacetyl-L-glutamic acid-S-N-(methyl)-N-(2,3,4,5,6-
pentahydroxyhexyl)-amide-[1-(4-
perfluorooctylsulfonyl)-piperazine]-amidopiperazine]-amide
16.42 g (66.4 mmol) of EEDQ (2-ethoxy-1,2,-dihydroquinoline-I-carboxylic acid
ethyl
ester) is added at 0°C to 10.96 g (33.2 mmol) of the title compound of
Example 36c and 18.87 g
(33.2 mmol) of 1-perfluorooctylsulfonyl-piperazine (produced according to DE
19603033) in 80
ml of tetrahydrofuran, and it is stirred overnight at room temperature. It is
evaporated to the dry
state in a vacuum and chromatographed on silica gel (mobile solvent:
dichloromethane/methanol
= 20:1).
Yield: 28.67 g (92% of theory) of a colorless solid.
Elementary analysis:
Cld: C 39.61 H 2.89 F 35.66 N 6.19 S 3.54
Fnd: C 39.68 H 2.74 F 35.81 N 6.13 S 3.40
e) L-Glutamic acid-5-N-(methyl)-N-(2,3,4,5,6-pentahydroxyhexyl)-amide-[1-(4-
perfluorooctylsulfonyl)-piperazine]-amide
Ammonia gas is introduced at 0°C for one hour into a solution that
consists of 28.36 g
(30.22 mmol) of the title compound of Example 36d in 200 ml of ethanol. It
then is stirred for 4
CA 02418790 2003-02-07
127
hours at 0°C. It is evaporated to the dry state, and the residue is
absorptively precipitated from
water. The solid is filtered off and dried in a vacuum (50°C).
Yield: 24.19 g (95% of theory) of an amorphous solid.
Elementary analysis:
Cld: C 41.12 H 2.89 F 35.66 N 6.19 S 3.54
Fnd: C 41.15 H 2.83 F 35.78 N 6.28 S 3.71
f) N-[1,4,7-Tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-10-
(pentanoyl-3-aza-4-oxo-
5-methyl-5-yl)]-L-glutamic acid-5-N-(methyl)-N-(2,3,4,5,6-pentahydroxyhexyl)-
amide-5-[1-(4-
perfluorooctylsulfonyl)-piperazine]-amide, Gd complex
20.43 g (24.25 mmol) of the title compound of Example 36e, 2.79 g (24.25 mmol)
of N-
hydroxysuccinimide, 2.12 g (50 mmol) of lithium chloride and 15.27 g (24..25
mmol) of 1,4,7-
tris(carboxylatomethyl)-10-[(3-aza-4-oxo-5-methyl-5-yl}]-pentanoic acid]-
1,4,7,10-
tetraazacyclododecane, Gd complex, are dissolved in 200 ml of dimethyl
sulfoxide while being
heated slightly. At 10°C, 8.25 g (40 mmol) of N,N-
dicyclohexylcarbodiimide is added, and it
then is stirred overnight at room temperature. The solution is poured into
3000 ml of acetone and
stirred for 10 minutes. The precipitated solid is filtered off and then
purified by chromatography
(silica gel RP-18, mobile solvent: gradient that consists of
water/ethanol/acetonitrile).
Yield: 28.45 g (79% of theory) of a colorless solid.
Water content: I 1.0%
Elementary analysis (relative to anhydrous substance):
Cld: C 34.41 H 3.83 F 23.13 N 9.03 S 2.30 Gd 11.26
Fnd: C 34.34 H 3.98 F 23.29 N 9.19 S 2.15 Gd 11.07
Example 37
a) 6-N-Benzyloxycarbonyl-2-N-[1-O-a-D-carbonytmethyl-(2,3,4-tri O benzyl-
glucuronic acid
benzyl ester]-L-lysine-( 1-(4-perfluorooctylsulfonyl)-piperazine]-amide
41.27 g (200.0 mmol) of N,N-dicyclohexylcarbodiimide is added at 0°C to
a solution that
CA 02418790 2003-02-07
128
consists of 100.0 g (120.4 mol) of the title compound of Example lc), 73.77 g
(120.4 mol) of 1-
O-a-D-carboxymethyl-2,3,4-tri-O-benzyl-glucuronic acid benzyl ester and 13.86
g (120.4 mol) of
N-hydroxysuccinimide, dissolved in 500 ml of dimethylformamide. It is stirred
for 3 hours at
0°C and then overnight at room temperature. Precipitated urea is
filtered out, the filtrate is
evaporated to the dry state in a vacuum and chromatographed on silica gel
(mobile solvent:
dichloromethane/ethanol = 20:1).
Yield: 147.58 g (86% of theory) of a viscous oil.
Elementary analysis:
Cld: C 52.25 H 4.31 N 3.93 F 22.66 S 2.45
Fnd: C 52.38 H 4.17 N 4.12 F 22.78 S 2.39
b) 2-N-[1-O-a-D-Carbonylmethyl-glucuronic acid]-L-lysine-1-[(4-
perfluorooctylsulfonyl)-
piperazine]-amide
142.52 g (100.0 mmol) of the title compound of Example 37a) is dissolved in
2000 ml of
ethanol, and 10.0 g of palladium catalyst (10% Pd/C) is added to it. It is
hydrogenated for 12
hours at room temperature. Catalyst is filtered out, and the filtrate is
evaporated to the dry state
in a vacuum.
Yield: 93.06 g (quantitative) of a colorless solid.
Elementary analysis:
Cld: C 33.56 H 3.36 N 6.02 S 3.45 F 34.71
Fnd: C 33.31 H 3.42 N 6.04 S 3.40 F 35.51
c) 6-N-[1,4,7-Tris(carboxylatomethyl)]-1,4,7,10-tetraazacyclododecane-10-N-
(pentanoyl-3-aza-
4-oxo-5-methyl-5-yl)]-2-N-[1-O-a-D-carbonylmethyl-glucuronic acid]-L-lysine-[1-
(4-
perfluorooctylsulfonyl)-piperazineJ-amide, Gd complex, sodium salt
50.76 g (54.55 mmol) of the title compound of Example 37b), 6.28 g (54.55
mmol) of N-
hydroxysuccinimide, 4.62 g (109.0 mol) of lithium chloride and 34.35 g (54.55
mol) of 1,4,7-tris
(carboxylatomethyl)-10-(carboxy-3-aza-4-oxo-5-methyl-pent-5-yl)-1,4,7,10-
CA 02418790 2003-02-07
CA 02418790 2003-02-07
129
tetraazacyclododecane, Gd complex, are dissolved in 400 ml of dimethyl
sulfoxide while being
heated slightly. At 10°C, 16.88 g (81.8 mol) of N,N-
dicyclohexylcarbodiimide is added, and it
then is stirred overnight at room temperature. The solution is poured into
3000 ml of acetone and
stirred for 10 minutes. The precipitated solid is filtered off and then
purified by chromatography
(RP-18 mobile solvent: gradient that consists of water/ethanol/acetonitrile).
Yield: 75.149 g (88.0% of theory) of a colorless solid.
Water content: 8.6%.
Elementary analysis (relative to anhydrous substance):
Cld: C 34.53 H 3.80 N 8.05 Na 1.47 S 2.05 F 20.63 Gd 10.05
Fnd: C 34.38 H 3.95 N 8.19 Na 1.63 S 2.15 F 20.83 Gd 10.14
Example 38
a) 6-N-Benzyloxycarbonyl)-2-[2-(N-ethyl-N-perfluorooctylsulfonyl]-amino]-
acetyl-L;-lysine
49.46 g (200.0 mmol) of EEDQ (2-ethoxy-1,2-dihydroquinoline-1-carboxylic acid
ethyl
ester) is added at 0°C to 31.820 g (113.5 mmol) of 6-N-
benzyloxycarbonyl)-L-lysine and 66.42 g
( 113.5 mmol) of 2-(N-ethyl-N-perfluorooctylsulfonyl)-aminoacetic acid
(produced according to
DE 196 03 033) in 300 ml of tetrahydrofuran), and it is stirred overnight at
room temperature. It
is evaporated to the dry state in a vacuum and chromatographed on silica gel
(mobile solvent:
dichloromethane/methanol = 20:1 ).
Yield: 55.79 g (58% of theory) of a colorless solid.
Elementary analysis:
Cld: C 36.85 H 3.09 N 4.96 F 38.11 S 3.78
Fnd: C 36.85 H 3.19 N 4.87 F 38.28 S 3.95
b) 6-N-Benzyloxycarbonyl-2-N-[2-(N-ethyl-N-perfluorooctylsulfonyl)-amino]-
acetyl-L-lysine-
N-methyl-N-(2,3,4,5,6-pentahydroxy-hexyl)-amide
20.64 g (100.0 mmol) of N,N-dicyclohexylcarbodiimide is added at 0°C to
a solution that
consists of 51.02 g (60.2 mol) of the title compound of Example 38a), 11.75 g
(60.2 mol) of N-
CA 02418790 2003-02-07
130
methyl-glucamine and 6.93 g (60.2 mol) of N-hydroxysuccinimide, dissolved in
250 ml of
dimethylformamide. It is stirred for 3 hours at 0°C and then overnight
at room temperature.
Precipitated urea is filtered out, the filtrate is evaporated to the dry state
in a vacuum, and it is
chromatographed on silica gel (mobile solvent: dichloromethane/ethanol =
20:I).
Yield: 53.05 g (86% of theory) of a viscous oil.
Elementary analysis:
Cld: C 38.68 H 4.03 N 5.47 F 31.52 S 3.13
Fnd: C 38.49 H 4.17 N 5.32 F 31.70 S 3.29
c) 2-N-[2-(N-Ethyl-N-perfluorooctylsulfonyl)-amino]-acetyl-L-lysine-N-methyl-N-
(2,3,4,5,6-
pentahydroxy-hexyl)-amide
102.48 g (100.0 mmol) of the title compound of Example 38b) is dissolved in
2000 ml of
ethanol, and 10.0 g of palladium catalyst (10% Pd/C) is added to it. It is
hydrogenated for 12
hours at room temperature. Catalyst is filtered out, and the filtrate is
evaporated to the dry state
in a vacuum.
Yield: 89.06 g (quantitative) of a colorless solid:
Elementary analysis:
Cld: C 33.72 H 3.96 N 6.29 S 3.60 F 36.26
Fnd: C 33.91 H 3.82 N 6.14 S 3.47 F 36.31
d) 6-N-[1,4,7-Tris(carboxylatomethyl)]-1,4,7,10-tetraazacyclododecane-10-N-
(pentanoyl-3-aza-
4-oxo-5-methyl-5-yl)]-2-N-j2-(N-ethyl-N-perfluorooctylsulfonyl)-amino]-acetyl-
L-lysine-N-
methyl-N-(2,3,4,5,6-pentahydroxy-hexyl)-amide, Gd complex
48.58 g (54.55 mmol) of the title compound of Example 38c), 6.28 g (54.55
mmol) of N-
hydroxysuccinimide, 4.62 g (109.0 mol) of lithium chloride and 34.35 g (54.55
moI) of 1,4,7-
tris(carboxylatomethyl)-10-(carboxy-3-aza-4-oxo-5-methyl-pent-5-yl)-1,4,7,10-
tetraazacyclododecane, Gd complex, are dissolved in 400 ml of dimethyl
sulfoxide while being
heated slightly. At 10°C, 16.88 g (81.8 mol) of N,N-
dicycIohexylcarbodiimide is added, and it
131
CA 02418790 2003-02-07
then is stirred overnight at room temperature. The solution is poured into
3000 ml of acetone and
stirred for 10 minutes. The precipitated solid is filtered off and then
purified by chromatography
(RP-18 mobile solvent: gradient that consists of water/ethanol/acetonitrile).
Yield: 73.27 g (89.4% of theory) of a colorless solid.
Water content: 8.6%.
Elementary analysis (relative to anhydrous substance):
Cld: C 35.18 H 4.23 N 4.23 S 2.13 F 21.50 Gd 10.47
Fnd: C35.28 H4.15 N4.19 S2.18 F21.33 Gd10.61
Fxam~'e 39~
Organ distribution (including tumor and lymph node concentration) after
intravenous
administration of the contrast medium of Example 1 according to the invention
in prostate-
cancer-carrying rats
After intravenous administration of 225 ~.mol of total gadolinium/kg of body
weight of
the title compound of Example 1 in rats (Cop-inbreeding Dunning 83327 MAT-Lu
prostate
cancer i.m.-implanted 12 days earlier), the metal content in various organs,
in tumors and in
lymph nodes (pooled as mesenteral and peripheral lymph nodes) was determined
10 minutes, 1
hour and 24 hours after administration (MW t SD, n = 3).
132
Title ple
Com 1
ound
of
Exam
Gd %
Concentration Dose
per
Total
Tissue
( mol/1]
10 in 1 24 10 1 24
m p.i. min
p.i.
hour hours hour hours
.i. .i. .i. .i.
Liver 387 ~ 26 364 t 8 746 ~34 5.46t 0.165.81t 0.1611.65~ 0.97
Spleen 548 ~ 22 487 t 25 645 t27 0.39~ 0.030.39~ 0.020.57~ 0.03
Pancreas 229 t 27 199 t 30 130 ~13 0.26~ 0.050.21~ 0.050.17~ 0.02
Kidney 2081 ~ 537883 t 94 1178 t1395.02~ 1.292.15t 0.232.97t 0.21
Lung 837 t 32 658 t 29 370 ~34 1.69t 0.061.38~ 0.080.73~ 0.04
Heart 438 t 29 289 ~ 24 131 t9 0.46t 0.010.31f 0.030.14t 0.02
Brain 47 t 13 26 t 5 14 ~2 O.ISt Ø030.08~ 0.020.04t 0.00
Muscle** 99 t 5 78 t 1 36 ~1 0.11~ 0.030.09t 0.030.04f 0.00
Tumor 185 ~ 36 184 f 13 199 f19 0.28t 0.100.21f 0.020.31t 0.01
Femur 184 t 4 I27 t 9 87 ~. 0.65t 0.010.46~ 0.030.31~ 0.03
6
mes. LK 359 t 72 697 f 42 854 t1350.11t 0.040.24t 0.020.32t 0.04
periph. 229 t 15 436 t 44 373 ~24 0.10t 0.010.20t 0.030.18t 0.01
LK
Stomach 23I t 10 219 ~ 46 I38 t9 0.57~ 0.040.54t O.I20.37t 0.06
(emptied)
Intestine342 t 16 409 ~ 67 243 ~22 2.91t 0.183.41~ 1.022.14~ 0.14
(emptied)
Blood 1665 t 1I0825 t 67 214 ~9 42.9~ 2.5921.4t 1.781.78t 0.03
*
5 7
Remainder-- - -- -- - -- 225 t31 -- - -- -- - -- 30.83~ 4.05
of the
Bod ****
Urine -- - -- -- - -- 94 ~20 -- - -- -- - -- 20.20t 4.41
0-24
hours
Feces -- - -- -- - -- 3128 t204-- - -- -- - -- 21.85t 1.46
0-24
hours
~58 ml of blood/kg of body weight
i;Only tissue sample from right lower leg muscle
i»~The sum total of all of the organs at 10 and 60 min. p.i., less the
remainder of the body
..~.
The remainder of the body also contains the remainder of the blood
CA 02418790 2003-02-07
133
Lymph node visualization (MRT) after intravenous administration of the
contrast medium of
Example 1 according to the invention in VX2-tumor-carrying rabbits
The figures in Fig. 1 show MR images of iliac lymph nodes pre-contrast and up
to 24
hours after intravenous administration of 200 ~mol of Gd/kg of body weight of
the title
compound of Example 1 in rabbits with VX2-tumors implanted i.m. The Ti-
weighted gradient-
echo images (1.5 T; sequence: MPRange; TR 11.1 ms, TE 4.3 ms, a 15~ illustrate
the strong
signal rise in healthy lymph node tissue. A zone without a signal rise within
the lymph node was
diagnosed as metastasis and confirmed histologically (H/E-staining of the
lymph node section).
Later (24 hours) after contrast medium administration, however, a signal
reversal was observed,
surprisingly enough. The signal rise in the healthy lymph node tissue was
reduced, while the
metastasis now exhibited a considerable signal rise.
As early as immediately after administration, surprisingly enough, a.
considerable
enhancement of the primary tumor (especially the periphery) was observed.
Later (24 hours p.i.),
this enhancement also propagates out from the center of the tumor.
Infarction visualization (MRT) after intravenous administration of the
contrast medium of
Example 1 according to the invention in rats
The figures in Fig. 2 show MR images of the heart (in vivo and post mortem) 24
hours
after intravenous administration of 100 pmol of Gd/kg of body weight of the
title compound of
Example 1 in rats with acute induced myocardial infarction. The T~-weighted
spin-echo images
(1.5 T; TR: 400 ms, TE: 6 ms; NA: 4; Matrix: 128128; layer thickness: 2.5 mm)
illustrate the
strong signal rise in the infarction area. The successful indication of an
acute myocardial
infarction was confirmed using NBT staining.
CA 02418790 2003-02-07
