U.S. patent application number 14/710886 was filed with the patent office on 2015-10-15 for macrocyclic compositions and metal complexes for bioimaging and biomedical applications.
The applicant listed for this patent is National Yang Ming University. Invention is credited to Bhalchandra Vishnu Bhagwat, Cheng Allen Chang, Ren-Shyan Liu.
Application Number | 20150291538 14/710886 |
Document ID | / |
Family ID | 51017861 |
Filed Date | 2015-10-15 |
United States Patent
Application |
20150291538 |
Kind Code |
A1 |
Chang; Cheng Allen ; et
al. |
October 15, 2015 |
Macrocyclic Compositions And Metal Complexes For Bioimaging And
Biomedical Applications
Abstract
The present disclosure provides a novel class of macrocyclic
compounds and their metal complexes formed with transition metal
ion, lanthanide metal ions and other metal ions (e.g., Al, Ga, Y,
In, Sn, Tl, Pb and Bi) and their applications in the fields of
contrast agents, artificial nucleases, fluorescence probes, nuclear
medicines and other biomedical applications in the therapeutics or
diagnostics.
Inventors: |
Chang; Cheng Allen; (Taipei,
TW) ; Liu; Ren-Shyan; (Taipei, TW) ; Bhagwat;
Bhalchandra Vishnu; (Taipei, TW) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
National Yang Ming University |
Taipei |
|
TW |
|
|
Family ID: |
51017861 |
Appl. No.: |
14/710886 |
Filed: |
May 13, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13731545 |
Dec 31, 2012 |
9062009 |
|
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14710886 |
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Current U.S.
Class: |
514/183 ;
540/474 |
Current CPC
Class: |
C07F 5/003 20130101;
A61K 51/0482 20130101; C07D 273/00 20130101; C07D 413/10 20130101;
C07D 257/02 20130101; C07D 401/14 20130101 |
International
Class: |
C07D 257/02 20060101
C07D257/02 |
Claims
1.-20. (canceled)
21. A compound of formula (I): ##STR00020## or an enantiomer, a
tautomer, a pharmaceutically acceptable salt, or a prodrug thereof;
wherein: Ar is an optionally substituted phenyl ring or a pyridine
ring, an aryl or a heterocyclic ring optionally substituted with
R.sub.1; G is N; J is --CH.sub.2--, or --CH.sub.2CH.sub.2--;
L.sub.1 is --CH.sub.2--, or --CH.sub.2CH.sub.2--; K is
--CH.sub.2--, or --CH.sub.2CH.sub.2--; L is --C--, --S(O)--, or
--P(OH)--; each R.sub.1 is independently --OH,
--(CH.sub.2).sub.nOR.sub.4, --(CH.sub.2).sub.n-alkyl,
--(CH.sub.2).sub.n-aryl, --(CH.sub.2).sub.n--N.sub.3; each R.sub.2
is independently H, optionally substituted alkyl,
--(CH.sub.2).sub.n-aryl, --(CH.sub.2).sub.n--N.sub.3; each R.sub.3
is independently --OH, or --NHR.sub.6; each R.sub.6 is
independently H, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted aryl, or optionally substituted heterocyclic; m is
2.
22. The compound of claim 21, wherein the compound of formula (I)
is a compound of formula (III): ##STR00021## wherein R.sub.2 is
independently H, methyl, p-methoxybenzyl, or
6-methyl-pyridien-2-carboxylic acid.
23. The compound of claim 21, further comprising at least one group
of formula (IV): --W--(X.sub.s--Y.sub.t).sub.p-- (IV) that binds to
Ar of the compound of formula (I) and to a biomolecule, wherein: W
is --CH.sub.2--, --SO--, --SO.sub.2--, --CO--, --NHNH--, --CONH--,
--NHCO--, --NHCONH--, --CONHCO--, --NHCONHO--, --COO--,
--COCH.sub.2--, --CH.sub.2CO--, --NHO--, --ONH--, --CH.dbd.CH--,
--NH--, --NR.sub.8, --NH(C.dbd.NH)NH--, --CH.dbd.N--, --N.dbd.CH--,
--SO.sub.2NH--, --NHSO.sub.2--, --NH--CS--, --NHCSNH--,
--NH(C.dbd.NH)NH--O--, --OCO--, --S--S--, or --N.dbd.N--, R.sub.8
is alkyl, cycloalkyl, or aryl; X and Y are same or different and
independently optionally substituted alkyl, cycloalkyl, aryl, or
heterocylic; and s, t, and p are an integer from 0 to 100.
24. The compound of claim 23, wherein X and Y further comprises
negatively charged carboxylate or positively charged ammonium
moieties.
25. The compound of claim 23, wherein the biomolecule is a protein,
an oligopeptide, an oligonucleotide, a modified oligopeptide or
oligonucleotide, or a polymer.
26. The compound of claim 25, wherein the modified oligopeptide or
oligonucleotide is phosphoroamidate, phosphoromonothioate, or
phosphorodithioate, and wherein the polymer is polyethylene glycol
or polylysine, or a nanoparticle.
27. A pharmaceutical composition, comprising a pharmaceutically
acceptable carrier and a compound of formula (I) or a metal complex
of a compound of formula (I): ##STR00022## or an enantiomer, a
tautomer, a pharmaceutically acceptable salt, or a prodrug thereof;
wherein: Ar is an optionally substituted phenyl ring or a pyridine
ring, an aryl or a heterocyclic ring optionally substituted with
R.sub.1; G is N; J is --CH.sub.2--, or --CH.sub.2CH.sub.2--;
L.sub.1 is --CH.sub.2--, or --CH.sub.2CH.sub.2--; K is
--CH.sub.2--, or --CH.sub.2CH.sub.2--; L is --C--, --S(O)--, or
--P(OH)--; each R.sub.1 is independently --OH,
--(CH.sub.2).sub.nOR.sub.4, --(CH.sub.2).sub.n-alkyl,
--(CH.sub.2).sub.n-aryl, --(CH.sub.2).sub.n--N.sub.3; each R.sub.2
is independently H, optionally substituted alkyl,
--(CH.sub.2).sub.n-aryl, --(CH.sub.2).sub.n--N.sub.3; each R.sub.3
is independently --OH, or --NHR.sub.6; each R.sub.6 is
independently H, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted aryl, or optionally substituted heterocyclic; m is
2.
28. The pharmaceutical composition of claim 27, wherein the
compound of formula (I) further comprises at least one group of
formula (IV): --W--(X.sub.s--Y.sub.t)-- (IV) that binds to Ar of
the compound of formula (I) and to a biomolecule, wherein: W is
--CH.sub.2--, --SO--, --SO.sub.2--, --CO--, --NHNH--, --CONH--,
--NHCO--, --NHCONH--, --CONHCO--, --NHCONHO--, --COO--,
--COCH.sub.2--, --CH.sub.2CO--, --NHO--, --ONH--, --CH.dbd.CH--,
--NH--, --NR.sub.8, --NH(C.dbd.NH)NH--, --CH.dbd.N--, --N.dbd.CH--,
--SO.sub.2NH--, --NHSO.sub.2--, --NH--CS--, --NHCSNH--,
--NH(C.dbd.NH)NH--O--, --OCO--, --S--S--, or --N.dbd.N-- R.sub.8 is
alkyl, cycloalkyl, or aryl; X and Y are same or different and
independently optionally substituted alkyl, cycloalkyl, aryl, or
heterocylic; and s, t, and p are an integer from 0 to 100.
29. The pharmaceutical composition of claim 28, wherein X and Y
further comprises negatively charged carboxylate or positively
charged ammonium moieties.
30. The pharmaceutical composition of claim 28, wherein the
biomolecule is a protein, an oligopeptide, an oligonucleotide, a
modified oligopeptide or oligonucleotide, or a polymer.
31. The pharmaceutical composition of claim 28, wherein the
modified oligopeptide or oligonucleotide is phosphoroamidate,
phosphoromonothioate, or phosphorodithioate, and wherein the
polymer is polyethylene glycol or polylysine, or a nanoparticle.
Description
TECHNICAL FIELD
[0001] The present disclosure relates to macrocyclic compounds and
particularly relates to mono- or bi-functional macrocyclic
compounds capable of forming metal complex with metal (e.g.,
transition or lanthanide) ions, processes for their preparation and
their use in the fields of contrast agents, artificial nucleases,
fluorescence probes, nuclear medicines, and other biomedical
applications in the therapeutics or diagnostics.
BACKGROUND
[0002] Applications of macrocyclic metal complexes are numerous. In
the field of artificial metallonucleases, it's a challenge to
cleave DNA or RNA at specific site for further development in
genetic engineering. To achieve this goal, it is very important to
understand interactions of metal complexes with large structured
RNA or DNA molecule, which are dependent on the selective binding
of metal complexes to specific RNA or DNA sites and subsequent
hydrolysis reactions. In this regards, the macrocyclic ligands and
their substituents play a key role in providing specific binding
with nucleic acids (DNA or RNA molecules) and hydrolysis reactions
(Bridger, G. J. et. al, J. Med. Chem. 1999, 42, 3971 -81; New, K.
et al., J. Am. Chem. Soc. 2008, 130, 14861-71; Rossiter, C. S. et
al., J. Inorg Biochem 2009, 103, 64-71; Chang C. A. et al., Inorg.
Chem. 2005, 44, 6646-54). Macrocyclic ligands also play an
important role in the field of nuclear medicine by forming stable
metal complexes with various radionuclei (WO2011031073). Tetraaza
complexes formed between linear or macrocyclic amines and
radioactive rhodium-105 have been used in diagnostics and
therapeutics (Bounsall, E. J. et al., Canadian J. of chemistry
1970, 48, 1481; U.S. Pat. No. 5,489,425). A Cobalt complex,
[Co(III)(cyclen)Cl.sub.2]Cl, has been found to be selectively
cytotoxic to human leukaemia cells (He, Z. et al., J. Pharmacology
2010, 637, 11-15). In medical diagnostics, macrocyclic ligand-metal
complexes have been widely used as contrast agents (WO2011031073;
U.S. Pat. No. 5,482,699; U.S. Patent Application Publication
2011/0177002). For example, a .sup.64Cu-tetraaza complex has been
used for imaging tumor U87MG, and tetraaza-.sup.99mTc complexes
have been used in diagnostics (Cai, W. et al., Eur. J. Nucl. Med.
Mol. Imaging 2007, 34, 850-858; Troutner, D. E. et al., J Nucl Med
1980, 21, 443-448). Furthermore, many gadolinium complexes have
been commercialized with trade names, Magnevist, MultiHance,
Omniscan, OptiMARK, ProHance, and Dotarem, as magnetic resonance
imaging (MRI) contrast agents for diagnostics in daily practice
(Caravan, P. et al., Chem. Rev. 1999, 99, 2293-2352; Kumar. K. et
al., Inorg. Chem. 1993, 32, 4193- 4199). Among these, ProHance and
Dotarem are tetraaza-based (e.g. DO3A and DOTA) macrocyclic
ligands. In the case of prodrug-procontrast, macrocyclic
ligand-metal complexes can deliver prodrugs into the targeted
cancer cells, thus reducing the toxic dose of active drugs.
Simultaneously, they also act as contrast agents allowing the
measurement of drug activity in the body. The efficiency of the
whole process is dependent on the metal complexes and ultimately on
the ligands employed (Frullano, L. et al., Inorg. Chem. 2006, 45,
8489-8491).
[0003] Accurate diagnosis needs effective imaging outcome, which is
dependent on the characteristics of contrast agents and ultimately
is dependent on the characteristics of ligands. Therefore, there
remains a need in developing novel ligands for better imaging in
MRI studies to avoid confusing situations in identification,
investigation and effective treatment of diseases. In artificial
metallonuclease or fluorescence probe areas, it is also essential
to develop novel ligands to effectively cleave nucleic acid at a
desirable site or to function as efficient fluorescence probes.
SUMMARY
[0004] In one aspect, macrocyclic compounds are provided. The
compounds of the present disclosure have the following formula
(I):
##STR00001##
or a enantiomer, a tautomer, a pharmaceutically acceptable salt, or
a prodrug thereof;
[0005] wherein:
[0006] Ar is an aryl or a heterocyclic ring optionally substituted
with --R.sub.1;
[0007] G is N, O, or S;
[0008] J is --CH.sub.2--, or --CH.sub.2CH.sub.2--;
[0009] L.sub.1 is --CH.sub.2--, or --CH.sub.2CH.sub.2--;
[0010] K is H, --CH.sub.2--, or --CH.sub.2CH.sub.2--;
[0011] L is --C--, --S(O)--, or --P(OH)--, when K is not H;
[0012] each R.sub.1 is independently --(CH.sub.2).sub.nOR.sub.4,
--(CH.sub.2).sub.nSR.sub.4, --(CH.sub.2).sub.nN(R.sub.4)R.sub.5,
--(CH.sub.2).sub.nNHN(R.sub.4)R.sub.5,
--(CH.sub.2).sub.nC(O)OR.sub.4, --(CH.sub.2).sub.n-alkyl,
--(CH.sub.2).sub.n-aryl, --(CH.sub.2).sub.n-alkenyl,
--(CH.sub.2).sub.n-alkynyl, --(CH.sub.2).sub.n--N.sub.3,
--(CH.sub.2).sub.n--C(O)N(R.sub.4)R.sub.5,
--(CH.sub.2).sub.n--S(O).sub.2N(R.sub.4)R.sub.5, or
--(CH.sub.2).sub.n--P(O)(OR.sub.4).sub.2;
[0013] each R.sub.2 is absent when G is O or S, or is independently
H, optionally substituted alkyl, optionally substituted cycloalkyl,
or --CH.sub.2R.sub.7;
[0014] each R.sub.3 is independently --OH, or --NHR.sub.6,
[0015] each R.sub.4 and R.sub.5 are same or different and
independently H, alkyl, or aryl;
[0016] each R.sub.6 is independently H, optionally substituted
alkyl, optionally substituted alkenyl, optionally substituted
alkynyl, optionally substituted aryl, or optionally substituted
heterocyclic;
[0017] each R.sub.7 is independently cycloalkyl, aryl, or
heterocyclic;
[0018] m is 2, 3, or 4; and
[0019] each n is 0, 1, 2, or 3, and
[0020] with the proviso that if m is 2, Ar is phenyl, K is H, and
R.sub.2 is H or absent, then G is not N or O.
[0021] In one embodiment, the compound of formula (I) may be a
compound of formula (II):
##STR00002##
[0022] In one embodiment, the compound of formula (I) may be a
compound of formula (III):
##STR00003##
[0023] wherein R.sub.2 is methyl, p-methoxybenzyl, or
6-methyl-pyridien-2-carboxylic acid.
[0024] In some embodiments, the compound of formula (I) may further
comprise at least one group of formula (IV):
--W--(X.sub.s--Y.sub.t).sub.p-- (IV)
that bounds the compound of formula (I) to a biomolecule.
[0025] wherein: W is --O--, --S--, --CH.sub.2--, -alkynyl-, --SO--,
S(O).sub.2--, --C(O)--, --NHNH--, --C(O)NH--, --N(H)CO, --NHCONH--,
CONHCO--, NHC(O)N(H)O, --C(O)O--, --C--O--CH.sub.2--,
--CH.sub.2C(O)--, --N(H)O--, --ONH--, --CH.dbd.CH--,
--CH.sub.2NH--, --NHCH.sub.2--, --NH--, --NR.sub.8--,
--NH(C.dbd.NH)NH--, --CH.dbd.N--, --CH.dbd.N--O--, --N.dbd.CH--,
--S(O).sub.2NH--, --NHS(O).sub.2--, --NH--C(S)--, --NHC(S)NH--,
NH(C.dbd.NH)NH--O--, --OC(O)--, --S--S--, --N.dbd.N--, methylene
carbamate, methylene thiocarbamate, methylene isourea, methylene
isothiourea, methylene guanidine, --C.dbd.C(R)-COOR.sub.8,
--C.dbd.C(R)-COR.sub.8, --C.dbd.NH, --C.dbd.NOH, --C.dbd.NR.sub.8,
--C.dbd.NOR.sub.8, allyl-OCOR.sub.8, allyl ester, phenylene,
alkylene, ethynydiyl, ethylenediyl, a polymerizable group, a boron,
a silicon, phosphorous, selenium, haloacetamido, isothiocyanate,
maleimido, dichlorotriazinylamino, dichlorotriazinyl, thioester,
pyridyldithio, aminooxy, amino, hydrazide, carboxylic acid, or acid
halide;
[0026] R.sub.8 is alkyl, cycloalkyl, or aryl;
[0027] X and Y same or different and independently alkyl,
cycloalkyl, aryl, or heterocylic, wherein any two carbon atoms are
optionally interrupted with one or more N, O, S, Si, P, Se or B;
and
[0028] s, t, and p are an integer from 0 to 100.
[0029] In some embodiments, the X and Y may further comprise
positively charged carboxylate or negatively charged ammonium
moieties.
[0030] In some embodiments, the biomolecule may be proteins,
protein-proteins, oligopeptides, oligonucleotides, DNA, RNA,
haptens, antibodies, bleomycins, modified oligopeptides or
oligonucleotides, sugar- or base modified protein, oligonucleotide
or polynucleotide, oligo-saccharide, polysaccharide, PNA, LNA,
drug, phosphoslipide, lectine, receptor binding ligand, polymers,
proteoglycans, liposomes, aerogels, steroids, microorganisms,
hormones dendrimers, nanparticles, microparticles, or amyloid
binding moieties.
[0031] In some embodiments, the modified oligopeptides or
oligonucleotides may be phosphoroamidate, phosphoromonothioate, or
a phosphorodithioate, and the polymers may be polyethyleneglycol or
polylysine.
[0032] In another aspect, metal complexes of compounds of formula
(I) are provided. The metal complexes may comprise metal ions and a
compound of formula (I) as set forth above.
[0033] In some embodiments, the metal ions may comprise transition
metal ions, lanthanide metal ions, Al, Ga, Y, In, Sn, TI, Pb, or Bi
ions.
[0034] In some embodiments, the metal ions may be radioactive
isotopes comprising Cr-51, Ga-66, Ga-67, Ga-68, Y-90, In-111,
Ce-134, Nd-140, Eu-157, Sm-153, Tb-161, Dy-165, Ho-166, Ho-166,
Er-169, Lu-177, Er-165, or Ho-161.
[0035] In another aspect, pharmaceutical compositions comprising
pharmaceutically acceptable carriers and compounds of formula (I)
or metal complexes that comprise metal ions and compounds of
formula (I) are provided.
[0036] In another aspect, a method of making compound of formula
(I) is provided. The method may comprise: reacting a compound of
formula (V):
##STR00004##
[0037] wherein J is --CH.sub.2--, or --CH.sub.2CH.sub.2--; G is N,
O or S;
[0038] or a compound of formula (VI):
##STR00005##
[0039] wherein R.sub.2 is independently H, optionally substituted
alkyl, optionally substituted cycloalkyl, or --CH.sub.2R.sub.7;
R.sub.7 is independently cycloalkyl, aryl, or heterocyclic ; and X
is fluoro, chloro, or bromo;
[0040] with Ar dibromide,
[0041] wherein Ar is an aryl or a heterocyclic ring optionally
substituted with --R.sub.1; R.sub.1 is independently
--(CH.sub.2).sub.nOR.sub.4, --(CH.sub.2).sub.nSR.sub.4,
--(CH.sub.2).sub.nN(R.sub.4)R.sub.5,
--(CH.sub.2).sub.nNHN(R.sub.4)R.sub.5,
--(CH.sub.2).sub.nC(O)OR.sub.4, --(CH.sub.2).sub.n-alkyl,
--(CH.sub.2).sub.n-aryl, --(CH.sub.2).sub.n-alkenyl,
--(CH.sub.2).sub.n-alkynyl, --(CH.sub.2).sub.n--N.sub.3,
--(CH.sub.2).sub.n--C(O)N(R.sub.4)R.sub.5,
--(CH.sub.2).sub.n--S(O).sub.2N(R.sub.4)R.sub.5, or
--(CH.sub.2).sub.n--P(O)(OR.sub.4).sub.2; R.sub.4 and R.sub.5 are
same or different and independently H, alkyl, or aryl; and each n
is 0, 1, 2, or 3; and
[0042] reducing the resulting salt under suitable conditions to
form a compound of formula (VII):
##STR00006##
[0043] wherein:
[0044] Ar is an aryl or a heterocyclic ring optionally substituted
with --R.sub.1;
[0045] G is N, O, or S;
[0046] J is --CH.sub.2--, or --CH.sub.2CH.sub.2--;
[0047] L.sub.1 is --CH.sub.2--, or --CH.sub.2CH.sub.2--;
[0048] each R.sub.1 is independently --(CH.sub.2).sub.nOR.sub.4,
--(CH.sub.2).sub.nSR.sub.4, --(CH.sub.2).sub.nN(R.sub.4)R.sub.5,
--(CH.sub.2).sub.nNHN(R.sub.4)R.sub.5,
--(CH.sub.2).sub.nC(O)OR.sub.4, --(CH.sub.2).sub.n-alkyl,
--(CH.sub.2).sub.n-aryl, --(CH.sub.2).sub.n-alkenyl,
--(CH.sub.2).sub.n-alkynyl, --(CH.sub.2).sub.n--N.sub.3,
--(CH.sub.2).sub.n--C(O)N(R.sub.4)R.sub.5,
--(CH.sub.2).sub.n--S(O).sub.2N(R.sub.4)R.sub.5, or
--(CH.sub.2).sub.n--P(O)(OR.sub.4).sub.2;
[0049] each R.sub.2 is absent when G is O or S, or is independently
H, optionally substituted alkyl, optionally substituted cycloalkyl,
or --CH.sub.2R.sub.7;
[0050] each R.sub.4 and R.sub.5 are same or different and
independently H, alkyl, or aryl;
[0051] each R.sub.7 is independently cycloalkyl, aryl, or
heterocyclic;
[0052] m is 2, 3, or 4; and
[0053] each n is 0, 1, 2, or 3.
[0054] In another aspect, a method of acquiring an image is
provided. The method may comprises: administrating to an issue,
cell, or mammal an effective amount of a metal complex comprising
metal ions and a compound of formula (I); and acquiring a magnetic
image of the cell, tissue or mammal.
[0055] In yet another aspect, a method of cleaving a nucleic acid
is provided. The method may comprise using a metal complex of
comprising metal ions and a compound of formula (I).
BRIEF DESCRIPTION OF DRAWINGS
[0056] The foregoing aspects and many of the attendant advantages
of the embodiments of present disclosure will become more readily
appreciated as the same become better understood by reference to
the following detailed description, when taken in conjunction with
the accompanying drawings, wherein:
[0057] FIG. 1A is a graph showing the titration curve of
Dimer-12N.sub.3O with Cu.sup.2+ ion by standard base.
[0058] FIG. 1B is a graph showing titration curves of
Dimer-12N.sub.3O with Cu.sup.2+ and Zn.sup.2+ ions.
[0059] FIG. 2A is a graph showing titration curves of Dimer-ODO2A
with Ca.sup.2+ and Zn.sup.2+ ions by standard base.
[0060] FIG. 2B is a graph showing titration curves of Dimer-ODO2A
with Lanthanide metal (Eu and Yb) ions.
[0061] FIG. 3 is a graph showing titration curves of
methooxybenxyl-DO2A-Dimer with Eu.sup.3+ and Yb.sup.3+ ions.
DETAILED DESCRIPTION OF VARIOUS EMBODIMENTS
[0062] The present disclosure provides mono- or bi-functional
macrocyclic compounds and their metal complexes that are useful in
various biomedical fields, including contrast agents in MRI
studies, artificial nuclease to cleave phosphodiester bond of DNA
and RNA molecules, fluorescence probes, and nuclear medicines.
[0063] As used herein, "R" refers to a substituent on an atom.
Unless otherwise specifically assigned, R represents any single
atom or any one of the substituent groups defined below. When there
is more than one R in a molecule, the "R" may independently at each
occurrence refer to a single atom or any one of the substituent
groups defined below.
[0064] "Alkyl" refers to CI-C20 straight chain and branched alkyl
groups. Representative straight chain alkyl groups include methyl,
ethyl, n-propyl, n-butyl, n-pentyl, and n-hexyl groups. Unless
stated otherwise in the specification, an alkenyl group may be
optionally substituted by one of the following groups:
--(CH.sub.2).sub.nOR.sub.5, --(CH.sub.2).sub.nSR.sub.5,
--(CH.sub.2).sub.nN(R.sub.5)R.sub.6,
--(CH.sub.2).sub.nNHN(R.sub.5)R.sub.6,
--(CH.sub.2).sub.nC(O)OR.sub.5, --(CH.sub.2).sub.n-alkyl,
--(CH.sub.2).sub.n-aryl, --(CH.sub.2).sub.n-alkenyl,
--(CH.sub.2).sub.n-alkynyl, --(CH.sub.2).sub.n--N.sub.3,
--(CH.sub.2).sub.n--C(O)N(R.sub.5)R.sub.6,
--(CH.sub.2).sub.n--S(O).sub.2N(R.sub.5)R.sub.6, or
--(CH.sub.2).sub.n--P(O)(OR.sub.5).sub.2, where n is 0, 1, 2, or 3
and R.sub.5 and R.sub.6 are and same or different and independently
alkyl, or aryl.
[0065] "Alkenyl" refers to a straight or branched hydrocarbon chain
containing at least one double bond and having 2-10 carbon atoms.
Representative alkenyl groups include ethenyl, propylenyl,
1-butenyl, 2-butenyl, isobutylenyl, and 2,3-dimethlyl-2-butenyl
groups. The alkenyl group may be optionally substituted as defined
above for the alkyl group.
[0066] "Alkynyl" refers to a straight or branched hydrocarbon chain
having containing at least one triple bond and having 2-10 carbon
atoms. Representative alkynyl groups include ethynyl, propynyl, and
1-butynyl groups. The alkynyl group may be optionally substituted
as defined above for the alkyl group.
[0067] "Aryl" refers to aromatic monocyclic or multicyclic
hydrocarbon ring system consisting only of hydrogen and carbon and
containing from 6 to 19 carbon atoms in the ring portion.
Representative aryl groups include phenyl, biphenyl, naphthyl, and
anthracenyl groups. The aryl group may be optionally substituted as
defined above for the alkyl group.
[0068] "Cycloalkyl" refers to a non-aromatic, monocyclic or
polycyclic ring comprising carbon and hydrogen atoms.
Representative cycloarkyl groups include cyclopropyl, cyclopentyl
groups. The cycloalkyl group may be optionally substituted as
defined above for the alkyl group.
[0069] "Complex" and "metal complex" refer to a compound of the
present disclosure, e.g. Formula (I), complexed or coordinated with
a metal.
[0070] "Enantiomers" refers to two stereoisomers whose molecules
are nonsuperimposeable mirror images of one another. A
"stereoisomer" refers to a compound made up of the same atoms
bonded by the same bonds but having different three-dimensional
structures, which are not interchangeable.
[0071] "Heterocyclic" means aromatic groups that include one or
more heteroatoms (e.g., oxygen, nitrogen, sulfur, or phosphorus) in
the aromatic ring. Representative heteroaromatic groups that
include a single heteroatom in the aromatic ring include furanyl,
pyridinyl, pyrrolyl, and thiophenyl groups. Representative
heteroaromatic groups that include more than one heteroatom in the
aromatic ring include C.sub.4N.sub.2, C.sub.3N.sub.3,
C.sub.3N.sub.2, C.sub.3NO, C.sub.3NS, and C.sub.2N.sub.3 groups.
The heterocyclic group may be optionally substituted as defined
above for the alkyl group.
[0072] "Halogen" or "halo" refers to bromo, choloro, fluoro, or
iodo.
[0073] "Mammal" includes humans and domestic animals, such as cats,
dogs, cattle, rabbits, mice, rats, and the like.
[0074] "Optional" or "optionally" means that the subsequently
described event of circumstances may or may not occur, and that the
description includes instances where the event of circumstance
occurs and instances in which it does not. For example, "optionally
substituted aryl" means that the aryl radical may or may not be
substituted and that the description includes both substituted aryl
radicals and aryl radicals having no substitution.
[0075] "Pharmaceutically acceptable salt" includes both acid and
base addition salts.
[0076] A "pharmaceutical composition" refers to a formulation of a
compound of the disclosure and a medium generally accepted in the
art for the delivery of the biologically active compound to
mammals, e.g., humans. Such a medium includes all pharmaceutically
acceptable carriers, diluents or excipients therefor.
[0077] "Prodrugs" is meant to indicate a compound that may be
converted under physiological conditions or by solvolysis to a
biologically active compound of the invention. Thus, the term
"prodrug" refers to a metabolic precursor of a compound that is
pharmaceutically acceptable. A prodrug may be inactive when
administered to a subject in need thereof, but is converted in vivo
to an active compound. Prodrugs are typically rapidly transformed
in vivo to yield the parent compound of the invention, for example,
by hydrolysis in blood. The prodrug compound often offers
advantages of solubility, tissue compatibility or delayed release
in a mammalian organism (see, Bundgard, H., Design of Prodrugs
(1985), pp. 7-9, 21-24 (Elsevier, Amsterdam).
[0078] A "tautomer" refers to a proton shift from one atom of a
molecule to another atom of the same molecule. The present
disclosure includes tautomers of any said compound.
[0079] In one aspect, macrocyclic compounds are provided. The
compounds of the present disclosure have the following formula
(I):
##STR00007##
or a enantiomer, a tautomer, a pharmaceutically acceptable salt, or
a prodrug thereof.
[0080] In formula (I), Ar is an aryl or a heterocyclic ring
optionally substituted with --R.sub.1; G is N, O, or S; J is
--CH.sub.2--, or --CH.sub.2CH.sub.2--; L.sub.1 is --CH.sub.2--, or
--CH.sub.2CH.sub.2--; K is H, --CH.sub.2--, or
--CH.sub.2CH.sub.2--; L is --C--, --S(O)--, or --P(OH)--; each
R.sub.1 is independently --(CH.sub.2).sub.nOR.sub.4,
--(CH.sub.2).sub.nSR.sub.4, --(CH.sub.2).sub.nN(R.sub.4)R.sub.5,
--(CH.sub.2).sub.nNHN(R.sub.4)R.sub.5,
--(CH.sub.2).sub.nC(O)OR.sub.4, --(CH.sub.2).sub.n-alkyl,
--(CH.sub.2).sub.n-aryl, --(CH.sub.2).sub.n-alkenyl,
--(CH.sub.2).sub.n-alkynyl, --(CH.sub.2).sub.n--N.sub.3,
--(CH.sub.2).sub.n--C(O)N(R.sub.4)R.sub.5,
--(CH.sub.2).sub.n--S(O).sub.2N(R.sub.4)R.sub.5, or
--(CH.sub.2).sub.n--P(O)(OR.sub.4).sub.2; each R.sub.2 is absent
when G is O or S, or is independently H, optionally substituted
alkyl, optionally substituted cycloalkyl, or --CH.sub.2R.sub.7;
each R.sub.3 is independently --OH, or --NHR.sub.6, each R.sub.4
and R.sub.5 are same or different and independently H, alkyl, or
aryl; each R.sub.6 is independently H, optionally substituted
alkyl, optionally substituted alkenyl, optionally substituted
alkynyl, optionally substituted aryl, or optionally substituted
heterocyclic; each R.sub.7 is independently cycloalkyl, aryl, or
heterocyclic; m is 2, 3, or 4; and each n is 0, 1, 2, or 3; with
the proviso that if m is 2, Ar is phenyl, K is H, and R.sub.2 is H
or absent, then G is not N or O.
[0081] In one embodiment, the compound of formula (I) is a compound
of formula (II)
##STR00008##
[0082] In one embodiment, the compound of formula (I) is a compound
of formula (III)
##STR00009##
[0083] Preferably, R.sub.2 is methyl, p-methoxybenzyl, or
6-methyl-pyridien-2-carboxylic acid.
[0084] In some embodiments, the compound of formula (I) may
comprise at least one group of formula (IV):
--W--(X.sub.s--Y.sub.t).sub.p-- (IV)
that bounds the compound of formula (I) to a biomolecule.
[0085] In formula (IV), W is --O--, --S--, --CH.sub.2--, -alkynyl-,
--SO--, S(O).sub.2--, --C(O)--, --NHNH--, --C(O)NH--, N(H)CO--,
--NHCONH--, CONHCO--, NHC(O)N(H)O, --C(O)O--, --C--O--CH.sub.2--,
--CH.sub.2C(O)--, --N(H)O--, --ONH--, --CH.dbd.CH--,
--CH.sub.2NH--, --NHCH.sub.2--, --NH--, --NR.sub.8--,
NH(C.dbd.NH)NH--, --CH.dbd.N--, --CH.dbd.N--O--, --N.dbd.CH--,
--S(O).sub.2NH--, --NHS(O).sub.2--, NH--CS--, NHC(S)NH--,
NH(C.dbd.NH)NH--O, --OC(O)--, --S--S--, --N.dbd.N--, methylene
carbamate, methylene thiocarbamate, methylene isourea, methylene
isothiourea, methylene guanidine, --C.dbd.C(R)--COOR.sub.8,
--C.dbd.C(R)--COR.sub.8, --C.dbd.NH, --C.dbd.NOH, --C.dbd.NR.sub.8,
--C.dbd.NOR.sub.8, allyl-OCOR.sub.8, allyl ester, phenylene,
alkylene, ethynydiyl, ethylenediyl, a polymerizable group, boron,
silicon, phosphorous, selenium, haloacetamido, isothiocyanate,
maleimido, dichlorotriazinylamino, dichlorotriazinyl, thioester,
pyridyldithio, aminooxy, amino, hydrazide, carboxylic acid, or acid
halide; R.sub.8 is alkyl, cycloalkyl, or aryl; X and Y are same or
different and independently alkyl, cycloalkyl, aryl, or
heterocylic, wherein any two carbon atoms are optionally
interrupted with one or more N, O, S, Si, P, Se or B; and s, p, and
t are an integer from 0 to 100.
[0086] In some embodiment, the biomolecule may comprise proteins,
protein-proteins, oligopeptides, oligonucleotides, DNA, RNA,
haptens, antibodies, bleomycins, modified oligopeptides or
oligonucleotides, sugar- or base modified protein, oligonucleotide
or polynucleotide, oligo-saccharide, polysaccharide, PNA, LNA,
drug, phosphoslipide, lectine, receptor binding ligand, polymers,
proteoglycans, liposomes, aerogels, steroids, microorganisms,
hormones dendrimers, nanparticles, microparticles, or amyloid
binding moieties.
[0087] Preferably, the modified oligopeptides or oligonucleotides
are phosphoroamidate, phosphoromonothioate, or phosphorodithioate.
Preferably, the polymers are polyethyleneglycol or polylysine.
[0088] In another aspect, metal complexes are provides. The metal
complexes of the present disclosure may comprise metal ions and
compounds of formula (I) as set forth above.
[0089] Preferably, the metal ions are transition metal ions,
lanthanide metal ions, Al, Ga, Y, In, Sn, TI, Pb, or Bi ions.
[0090] The metal complexes of compounds of formula (I) with various
radioactive isotopes are useful in positron emission tomography
(PET) to diagnose disease. Preferably, the radioactive isotopes
comprising Cr-51, Ga-66, Ga-67, Ga-68, Y-90, In-111, Ce-134,
Nd-140, Eu-157, Sm-153, Tb-161, Dy-165, Ho-166, Ho-166, Er-169,
Lu-177, Er-165, or Ho-161.
[0091] Compounds of formula (I) and their metal complexes as set
forth above can be applied in the various fields such as, as a
contrast agent or molecular probe in imaging studies (MRI, optical,
etc.), as artificial nuclease/hydrolase/peptidase to cleave
phosphodiester bond of DNA and RNA molecules or peptide bond, as a
nuclear medicine, or applied in other biomedical applications
(therapeutics and diagnosis).
[0092] In another aspect, the present disclosure provides
pharmaceutical compositions comprising pharmaceutically acceptable
carriers and compounds of formula (I) or metal complexes of
compounds of formula (I) as set forth above.
[0093] The pharmaceutically acceptable carrier, also known in the
art as an excipient, vehicle, auxiliary, adjuvant, or diluent, is
typically a substance which is pharmaceutically inert, confers a
suitable consistency or form to the composition, and does not
diminish the therapeutic or diagnostic efficacy of the conjugate.
The carrier is generally considered to be "pharmaceutically
acceptable" if it does not produce an unacceptably adverse,
allergic or other untoward reaction when administered to a mammal,
especially a human.
[0094] The selection of a pharmaceutically acceptable carrier will
also, in part, be a function of the route of administration. In
general, the metal complexes of the present disclosure can be
formulated with conventional pharmaceutically acceptable carriers
for any route of administration so long as the target tissue is
available via that route. For example, suitable routes of
administration include, but are not limited to, oral, parenteral
(e.g., intravenous, intraarterial, subcutaneous, subcutaneous,
intramuscular, intracapsular, intraspinal, or intraperitoneal),
intravesical, intrathecal, enteral, pulmonary, intralymphatic,
intracavital, transurethral, intradermal, intramammary, buccal,
orthotopic, intralesional, percutaneous, endoscopical,
transmucosal, and intestinal administration.
[0095] Pharmaceutically acceptable carriers for use in the
compositions of the present disclosure are well known to those of
ordinary skill in the art and are selected based upon a number of
factors: the particular complex used, and its concentration,
stability and intended bioavailability; the disease, disorder or
condition being diagnosed with the composition; the subject, its
age, size and general condition; and the route of administration.
Suitable pharmaceutically acceptable carriers include those that
are suitable for injection such as aqueous buffer solutions; e.g.,
tris(hydroxymethyl)amino methane (and its salts), phosphate,
citrate, bicarbonate, etc., sterile water for injection,
physiological saline, and balanced ionic solutions containing
chloride and/or bicarbonate salts of normal blood plasma cations
such as Ca, Na, K and Mg, and other halides, carbonates, sulphates,
phosphates of Na, K, Mg, Ca. The vehicles may advantageously
contain a small amount (e.g., from about 0.01 to about 15.0 mole %)
of a chelating agent such as ethylenediamine tetraacetic acid
(EDTA), calcium disodium EDTA, or other pharmaceutically acceptable
chelating agents such as calcium monosodium DTPA-BMEA (Versetamide;
Mallinckrodt Inc.). The composition may further comprise
non-radiographic additives selected from the group consisting of
excipients, such as, for example, glycerol, polyethylene glycol or
dextran, and anticlotting agents, such as, for example, heparin or
hirudin. A thorough discussion of pharmaceutically acceptable
carriers, diluents, and other excipients is presented in
REMINGTON'S PHAEPHACEUTICAL SCIENCES (Mack Pub. Co., N.J. current
edition).
[0096] In another aspect, the present disclose provides a method of
acquiring an image that may comprises administrating to a tissue,
cell, or mammal an effective amount of a compound of formula (I) or
a metal complex of compound f formula (I) as set forth above and
acquiring a magnetic image of the cell, tissue or patient.
[0097] In another aspect, the present disclose provide a method of
cleaving a nucleic acid using a compound of formula (I) or a metal
complex of compound of formula (I) as set forth above.
[0098] In particular, compounds of formula (I) of the present
disclosure may be prepared by the following Schemes. It is
understood that other compounds of the present disclosure may be
prepared in a similar manner as described below or by methods known
to one of ordinary skill in the art.
[0099] Compound 9 (12N.sub.3O) and Compound 10 (Cyclen) are
intermediates in the preparation of compounds of formula (I). They
are prepared as described blew in Scheme 1. First, Compounds
diethyleneglycol (1), diethylenetriamine (2), and
bis-(2-Hydroxy-ethyl)-amine (3) are protected and converted into
their corresponding tosylated derivatives, Compound 4
(1,5-ditosyloxy-3-oxapentane), Compound 5
(N,N',N''-tritosyldiethylenetriamine), and Compound 6
(bis-(2-Tosyloxy-ethyl)-methylamine)), respectively, using tosyl
chloride. Compound 5 is then reacted with sodium hydride to obtain
corresponding salt. Reacting the resulting salt of Compound 5 with
Compound 4 or Compound 6 affords cyclized Compound 7 (12N3Ts3O) or
Compound 8 (12N4Ts4), respectively. Compound 7 and Compound 8 are
deprotected using concentrated sulphuric acid to get corresponding
Compound 9 (12N.sub.3O) and Compound 10 (cyclen).
##STR00010##
[0100] Referring to the following Scheme 2, further protection of
Compound 9 with dimethyl acetal yields Compound 11
(12N.sub.3O-tricyclic form), which is then reacted with m-xylene
dibromide to afford Compound 12 (Dimer-12N.sub.3O-tricyclic form).
Followed by deprotection, Compound 13 (Dimer-12N.sub.3O) is
alkylated to give Compound 14 (dimer-ODO2A 14).
##STR00011##
[0101] General procedures A and B are employed to synthesize
cyclen-containing compounds of formula (I). Referring to the
following Scheme 3, Compounds 19 and 24 are prepared according to
the general procedure A. Initially, Compound 10 (cyclen) is
protected with glyoxal to give Compound 15. Compound 15 is then
reacted with alkyaryl or alkyl halide (RX) to yield Compound 16 and
Compound 21, respectively. Compound 16 or Compound 21 is reacted
with m-xylene dibromide to obtain Compound 17 or Compound 22,
respectively. Compound 17 or Compound 22 is then deprotected with
hydrazine to yield Compound 18 or Compound 23, respectively.
Finally, Compound 18 or Compound 23 is alkylated to yield Compound
19 or Compound 24, respectively. R may be methyl or p-methoxy
benzyl substituent.
##STR00012##
[0102] Another approach to synthesize Compound 24 by procedure B is
illustrated in the following Scheme 4. First, m-xylene dibromide is
reacted with protected Compound 15 to obtain Compound 20. Reacting
Compound 20 with methyl iodide affords Compound 22. After
deprotection and alkylation, Compound 24 is obtained through
Compound 23.
##STR00013##
##STR00014##
[0103] Using general procedure B, Compound 28 of formula (I) is
prepared as described in Scheme 5. Compound 22 is reacted with
Compound 32 first to obtain Compound 25. After deprotection and
hydrolysis, Compound 28 is obtained by alkylation of Compound 27
using bromoacetic acid under basic condition.
[0104] The synthesis of intermediate Compound 32 is shown in the
following Scheme 6. Pyridine-2,6-dicarboxylic acid is treated with
thionyl chloride in methanol to obtain its diester compound 30,
which is partially reduced with sodium borohydride to get Compound
31. Compound 31 is then reacted with phosphorous tribromide to
yield compound 32.
##STR00015##
[0105] Compound 37 of formula (I) is prepared according to a
general procedure C as described below in Scheme 7. First, Compound
20 is converted to Compound 35 via the protection-deprotection
transformations through compound 33 and 34, then it is alkylated to
obtain compound 36 and finally formyl protection is removed to
obtain Compound 37.
##STR00016##
[0106] An example of preparing a bioconjutage of a compound of
formula (I) is described below in Scheme 8.
##STR00017##
[0107] Metal complexes of compound 13, 14, or 19 are prepared as
described below in Scheme 9. A zinc complex of Compound 13 is
obtained by reacting zinc ion with Compound 13 under basic
condition. The pH of the resulting reaction mixture is then
adjusted to about pH 6.5 using a tetramethyl ammonium hydroxide
solution to form zinc complex of compound 13. Metal complexes of
compound 14 and 19 are prepared under the same condition.
##STR00018## ##STR00019##
[0108] The methods provided in the present disclosure are simple
and efficient to synthesize macrocyclic compounds of formula (I).
The salts of compounds of formula (I) are stable and can be stored
in inert atmosphere as these salts are hygroscopic in nature. In
addition, the aromatic ring of compounds of formula (I) are
connected directly to ring nitrogen of macrocyclic rings through
the Linker L.sub.1, L.sub.2 or L.sub.3. After chelating with the
metal ion, this would allow a free rotation of the metal complex
and reduced steric hindrance and strain-stress in the overall
molecule. As a result, energy transfer from one metal ion to
another metal ion via polar groups will become more efficient, thus
enhancing the physicochemical properties of metal complexes. This
will certainly improve biomedical applications of these metal
complexes.
[0109] In the present disclosure, all technical and scientific
terms herein used are having the same meaning as understood by
ordinary skill person in the art, otherwise defined. The
nomenclatures and procedures described herein are commonly employed
in the laboratory practice. Standard techniques are used in
synthesis and for chemical analysis ["Advanced Organic Chemistry:
Reactions, Mechanisms and Structure" by March, 3.sup.rd ed. (1985)
John Willey & Sons, New York, N.Y.]
[0110] The synthesis of compounds of the present disclosure are
illustrated by, but not limited to the following examples.
EXAMPLES
Example 1
Synthesis of Compound 4 (1,5-ditosyloxy-3-oxapentane)
[0111] Diethyleneglycol (10.61 g, 0.1 mol) was added into
triethylamine (100 ml) and stirred with an overhead stirrer. To
this mixture, tosylchloride (0.25 mol, 47.7 g) in ether (250 ml)
was added slowly in 30 min at 0.degree. C. The reaction mixture was
stirred overnight at ambient temperature. The white solid
precipitated in the reaction mixture was filtered and washed with
water and then with a small amount of ether. The white solid was
recrystallized from ethanol to afford compound 4 in 93% yield. mp:
88-89.degree. C.; .sup.13C NMR (CDCl.sub.3, 300MHz): .delta. 21.5,
68.6, 69.0, 127.8, 129.8, 132.6, 144.9.
Synthesis of Compound 5 (N,N',N''-tritdyldiethylenetriamine)
[0112] Diethylenetriamine (0.1 mol, 10.32 g) was added into a
sodium hydroxide solution (0.33 mol, 100 mL) in a flask and stirred
with an overhead stirrer. To this solution, tosylchloride (0.35
mol, 66.74 g) in ether (350 ml) was added dropwise using an
addition funnel over 30 min at 0.degree. C. The reaction mixture
was stirred for 14-16 h at ambient temperature. The white
precipitate was filtered and washed with water and then with ether.
The crude product was recrystallized from ethanol to yield compound
5 in 88%. mp: 175; .sup.13C NMR (CDCl.sub.3, 300MHz) .delta. 21.2,
41.8, 48.6, 126.8, 130.0, 135.4, 137.5, 143.9.
Synthesis of Compound 7 (12N.sub.3Ts.sub.3O)
[0113] Compound 4 (0.03 mol, 12.53 g) was dissolved in DMF and
heated to 70.degree. C. in a flask. Sodium hydride (4 eq, 0.12 mol)
was added slowly into a solution of compound 5 (0.03 mol, 17.00 g)
in DMF in a beaker over 30 min at 0.degree. C. to form sodium salt
of compound 5. After the complete addition of sodium hydride, the
reaction mixture was stirred for 15-20 min and then filtered under
vacuum. The filtrate was added to the solution of compound 4
dropwise over a 30 min at 70.degree. C. The reaction mixture was
heated to 120.degree. C. and stirred for .about.16 h at this
temperature. The pale yellow reaction mixture was concentrated. The
resulting sticky residue was poured into water with strong
stirring. The precipitated crude compound was filtered and then
recrystallized from chloroform/ethanol to obtain compound as a
white solid in 86% yield. mp 199-201.degree. C.; .sup.13C NMR
(CDCl.sub.3, 300 MHz): .delta. 21.5, 48.0, 50.5, 51.0, 72.0, 127.3,
129.8, 134.9, 143.7.
Synthesis of Compound 9 (12N30)
[0114] Concentrated sulphuric acid (100 ml) was added into compound
7 (0.04 mol, 25.55 g) at ambient temperature and the reaction
mixture was heated to 140.degree. C. for 1 day. Diethyl ether (300
ml) was added to the reaction mixture at 0.degree. C. to wash the
black solution twice or thrice to obtain a black sticky solid. The
black sold was dissolved in water (100 ml) and was discolored by
charcoal. After filtration, a colorless filtrate was obtained. The
pH of the solution was adjusted to 12 or above 12 with a sodium
hydroxide solution. This solution was extracted with chloroform 5
times, dried over magnesium sulfate, and then concentrated to
obtain compound 9 as a white solid in 87% yield. mp 78-79.degree.
C.; .sup.1H NMR (CDCl.sub.3, 300 MHz): .delta. 2.53-2.56 and
2.66-2.75 (m, 12H, NCH.sub.2), 3.50-3.54 (t, 4H, OCH.sub.2);
.sup.13C NMR (CDCl.sub.3, 300 MHz): .delta. 46.1, 46.3, 66.3.
Synthesis of Compound 11 (12N.sub.3O-tricylci form)
[0115] Compound 9 (0.03 mol, 5.22 g) was dissolved in toluene (90
ml) and heated to 60.degree. C. under nitrogen. To this solution,
dimethylformamide dimethyl acetal (0.03 mol, 3.57 g) was added
slowly and the reaction mixture was stirred for 6 h at 60.degree.
C. The reaction mixture was concentrated and dried to get yellowish
oily residue in 84% yield. .sup.1H NMR (CDCl.sub.3, 300 MHz):
.delta. 2.6-2.8 (m, 12H, NCH.sub.2), 3.3-3.4 (m, 4H, OCH.sub.2),
4.7 (s, 1H, CH); .sup.13C NMR (CDCl.sub.3, 300 MHz) .delta. 51.7,
53.3, 53.8, 71.5, 100.8.
Synthesis of Compound 12 (Dimer-12N3O-tricyclic form)
[0116] Compound 11, 12N.sub.3O-tricyclic form (0.031 mol, 5.67 g)
was dissolved in dry acetonitrile and heated to 50.degree. C. under
nitrogen. To above solution, the solution of m-dibromo xylene
(0.0125 mol, 3.3 g) in dry acetonitrile (15 ml) was added dropwise.
The reaction mixture was stirred for 6 h and the resulting white
solid dibromide salt, compound 12 (12N.sub.3O-tricyclic-dimer
bromide salt) was obtained after filtration.
Synthesis of Compound 13 (Dimer-12N.sub.3O)
[0117] Compound 12, dibromide salt, was refluxed in 80 ml sodium
hydroxide (2 N) solution for 12 h. After cooling, the reaction
mixture was extracted with chloroform for 5 times. The combined
organic layer was dried over magnesium sulfate and concentrated to
obtain a white solid of compound 12 in 94% yield. .sup.1H
NMR(CDCl.sub.3, 300 MHz): .delta. 2.58, 2.63, 2.73 (three broad s,
24H, NCH.sub.2), 3.53 (s, 4H, benzylic), 3.58 (s, 8H, OCH.sub.2),
7.19-7.31 (m, 4H, aromatic); .sup.13C NMR(CDCl.sub.3, 300 MHz)
.delta. 44.1, 46.2, 51.4, 59.2, 66.3, 128.1, 128.5, 130.1,
138.8.
Synthesis of Zinc Complex of Compound 13 (Dimer-12N.sub.3O)
[0118] Compound 13 (Dimer-12N.sub.3O) was chelating with copper and
zinc ion to form respective metal ion complexes which was observed
from the thermodynamics titration experiments.
[0119] Zinc perchlorate was added into aqueous solution of ligand
and the pH of the solution was adjusted to 6.5 with tetraammonium
hydroxide to obtain zinc complex. Standardization of ligand stock
solution was carried out by complexing with Cu.sup.2+ ion. Besides,
protonation constants will also be known by titration raw data with
calculation software--"Hyperquad 2008".
[0120] The formation of the complexes is shown in FIGS. 1A and
1B.
[0121] Experiment condition: [L']=1 mM, [M]=2 mM (for M.sub.2L'),
I=0.1 M with TMACI, at 25.degree. C.
Example 2
Synthesis of Compound 14 (Dimer-ODO2A)
[0122] Compound 13 (Dimer-12N.sub.3O) (5.7 mmol, 2.56 g) was
dissolved in 40 ml of water and the pH of the solution was adjusted
to 11 (or above 11) with sodium hydroxide solution. Bromoacetic
acid (5 equiv.) was added to the above solution and the reaction
mixture was stirred at 70.degree. C. for 3 days. During the course
of reaction, the pH of the reaction mixture was maintained above
11. After cooling the reaction mixture to room temperature, the pH
was adjusted to 3.5 with hydrochloric acid and the solution was
concentrated by rotavapor to obtain a pale yellow solid. The solid
was dissolved in a minimum amount of water and purified by cation
exchange resin to yield a pure white solid in 85% yield. .sup.1H
NMR (CDCl.sub.3, 200 MHz): .delta. 3.06 (m), 3.43 (m), 3.51 (m)
(three m, 24H, NCH.sub.2), 3.78-3.83 (m, 20H), 7.28 (m, 4H,
aromatic); .sup.13C NMR (CDCl.sub.3, 200 MHz): .delta. 45.9, 50.2,
54.5, 55.6, 57.2, 64.7, 130.1, 131.5, 132.7, 132.8, 168.0;
Molecular ion peak [M+1].sup.+=681.
Synthesis of Lanthanide ion complex of Compound 14
(Dimer-ODO2A)
[0123] Compound 14 (Dimer-ODO2A) and lanthanide ion forms complexes
in aqueous solution at a pH 6.5 which was adjusted by tetraammonium
hydroxide. Standardization of ligand stock solution was carried out
by complexing with Ca.sup.2+ and Zn.sup.2+ ions. Besides,
protonation constants will also be known by titration raw data with
calculation software--"Hyperquad 2008".
[0124] The formation of the complexes is shown in FIGS. 2A and
2B.
[0125] For calculation of stability constants of lanthanide metal
(Eu, Yb) complexes: Experiment condition: [L]=1 mM, [M]=1 mM (for
ML'), 2 mM (for M.sub.2L'), I=0.1 M with TMACI, at 25.degree.
C.
Example 3
Synthesis of Compound 16 (Mono-N.sub.1 alkylated cyclen)
[0126] Compound 15, cyclen glyoxal (1.58 g, 8.13 mmol) was
dissolved in dry tetrahydrofuran (20 ml) under nitrogen atmosphere.
To this solution, 4-methoxy benzyl chloride (1.18 g, 7.39 mmol)
solution in THF (6 ml) was added dropwise. Then the reaction
mixture was heated to 40.degree. C. and stirred for 2 days. The
white precipitated salt was washed several times with anhydrous THF
and the resulting solid was dried under high vacuum to get compound
16 as a white solid (1.98 g) in 76% yield. .sup.1H NMR (200 MHz,
D.sub.2O): .delta. 7.42, 7.37, 7.01, 6.96 (4H, q, C.sub.6H.sub.6),
4.76, 4.72, 4.58, 4.49 (2H, d, --CHCH--), 3.87, 3.62 (2H, d,
--CH.sub.2-Aromatic), 3.75 (3H, s, --OCH.sub.3), 3.48.about.2.37
(16H, m, --NCH.sub.2CH.sub.2N--); .sup.13C NMR (200 MHz, D.sub.2O):
.delta. 160.72, 134.11, 119.13, 114.91 (C.sub.aromatic), 82.32,
71.68 (C.sub.animal), 61.10, 56.93, 55.52, 51.28, 48.32, 48.18,
47.60, 43.80 (--NCH.sub.2).
Synthesis of Compound 17
[0127] Compound 16 (1.8 g, 5.13 mmol) was dissolved in dimethyl
formamide (70 ml) at ambient temperature under argon atmosphere.
The solution of m-xylene dibromide (0.658 g, 2.44 mmol) in DMF (25
ml) was added to above solution dropwise. After addition, the
reaction mixture was heated at 45.degree. C. for 4 days. Then the
reaction mixture was centrifuged and the solid was collected and
washed with a small amount of DMF and then with acetonitrile. The
resulting solid was dried under vacuum to obtain a white solid in
50% yield. .sup.1H NMR (200 MHz, D.sub.2O): .delta.
7.79.about.7.67, 7.48, 7.44, 7.04, 7.00 (12H, s, H.sub.aromatic),
5.04.about.4.84 (4H, d, H.sub.animal), 4.81, 4.75 (4H, s,
CH.sub.2-Aromatic) 3.76 (6H, s, OCH.sub.3) 3.50.about.2.88 (32H, m,
NCH.sub.2CH.sub.2N); .sup.13C NMR (200 MHz, D.sub.2O): .delta.
164.86, 135.84, 135.12, 133.98, 131.11, 127.94, 118.41, 115.03
(C.sub.aromatic) 78.29, 77.21 (C.sub.animal) 61.04, 60.69, 60.08,
55.49, 54.45, 46.04, 42.70 (NCH.sub.2).
Synthesis of Compound 18
[0128] Compound 17 (600 mg, 0.58 mmol) was dissolved in 4 ml
hydrazine monohydrate and stirred under argon at 110.degree. C. for
12 h. Then the reaction mixture was cooled to ambient temperature
and filtered to remove excess hydrazine. The solid was collected
and dissolved in 20 ml ethanol and concentrated by rotavapour. This
process was repeated twice or thrice and then the solid was dried
under high vacuum to obtain compound 18 in 95.3% (0.38 g). .sup.1H
NMR (200 MHz, CDCl.sub.3): 7.24.about.7.46, 6.88, 6.92 (Benzene,
12H), 3.79 (--OCH.sub.3, 6H), 3.59, 3.57 (benzylic CH.sub.2, 8H),
2.63 (--NCH.sub.2, 32H); .sup.13C NMR (200 MHz, CD.sub.3OD) 158.97,
139.19, 130.72, 130.11, 129.93, 126.73, 127.77, 123.48
(C.sub.aromatic), 59.69, 59.06 (NCH.sub.2-Aromatic,) 54.26
(OCH.sub.3), 51.00, 50.83, 44.70 (NCH.sub.2).
Synthesis of Compound 19 (Methoxybenzyl-DO2A-Dimer)
[0129] Compound 18, methoxybenzyl dimer12N4 (0.38 g, 0.55 mmol) was
dissolved in dd water (30 ml) and pH of solution was adjusted to
11-12 with sodium hydroxide solution. To above solution,
bromoacetic acid (0.46 g, 3.31 mmol) in 20 ml dd water was added
slowly at 60.degree.. The reaction mixture was maintained at pH
11-12 for 3 days at 60.degree. C. When the pH of the reaction
mixture was not decreased (approximately after 3 days), the pH of
the reaction mixture was adjusted to 4 with 6N hydrochloric acid
solution. The reaction mixture was concentrated by rotavapor to get
a solid residue, to which dry methanol (40 ml) was added. Sodium
chloride was eliminated by filtration. The filtrate was
concentrated and the residue was purified on cation exchange resin
to obtain compound 19 (MB-DO2A-Dimer) as white solid in 70% yield.
.sup.1H NMR (200 MHz, D.sub.2O) .delta.: 7.50.about.7.60,
7.26.about.7.30, 6.83.about.6.87 (12H, s, Aromatic), 4.38, 4.22(8H,
s, Aromatic-CH.sub.2N), 3.62 (6H, s, OCH.sub.3) 2.88 (8H, s,
CH.sub.2COOH), 3.23, 2.80, 2.77 (32H, s, NCH.sub.2CH.sub.2N);
.sup.13C NMR (200 MHz, D.sub.2O) .delta. 174.03 (--COOH), 160.40,
134.91, 133.33, 132.78, 131.01, 129.47, 120.53, 115.05(C.sub.Ar),
57.58 (NCH.sub.2C.sub.6H.sub.6), 55.40 (CH.sub.2COOH), 53.50
(OCH.sub.3), 50.14, 49.87, 48.35 (NCH.sub.2CH.sub.2N).
[0130] Synthesis of Lanthanide Complexes of Compound 19
(Methoxybenzyl-DO2A-Dimer)
[0131] Compound 19 (Methoxybenzyl-DO2A-Dimer) and lanthanide ion
forms complexes in aqueous solution at pH 6.5 which was adjusted by
tetraammonium hydroxide. Standardization of ligand stock solution
was carried out by complexing with Ca.sup.2+ and Zn.sup.2+ ions.
Besides, protonation constants will also be known by titration raw
data with calculation software--"Hyperquad 2008".
[0132] FIG. 3 is a graph showing titration curves of
methooxybenxyl-DO2A-Dimer with Eu.sup.3+ and Yb.sup.3+ ions.
[0133] For calculation of stability constants of lanthanide (Eu,
Yb) complexes: Experiment condition: [L]=1 mM, [M]=1 mM (for ML'),
2 mM (for M.sub.2L'), I=0.1M with TMACI, at 25.degree. C.
Example 4
Synthesis of Compound 20
[0134] Solution of m-xylene dibromide (0.618 g, 2.34 mmol) in dry
acetonitrile (10 ml) was added dropwise to the solution of compound
15 (1 g, 5.15 mmol, 2.2 eq.) in acetonitrile (20 ml) at ambient
temperature under argon. The reaction mixture was stirred for 2.5
days. The white salt precipitate was washed several times with
acetonitrile till washing was colorless. The white salt was dried
under high vacuum to get 1.07 g of compound 20(.about.100% yield).
.sup.1H NMR (200 MHz, D.sub.2O) .delta.2.35-2.6 (m, 2H), 2.6-3.0
(m, 8H), 3.0-3.35 (m, 10H), 3.35-3.6 (m, 8H), 3.73 (s, 2H), 4.0 (s,
2H), 4.15 (t, 2H), 4.66-4.96 (dd, 4H), 7.6-7.8 (m, 4H); .sup.13C
NMR (200 MHz, D.sub.2O) .delta. 43.67, 47.52, 48.13, 48.24, 51.25,
56.97, 60.72, 61.39, 71.56, 8308, 128.4, 130.92, 134.99,
135.98.
Synthesis of Compound 21
[0135] Compound 15, cyclen glyoxal (3.35 g, 17.24mmol) was
dissolved in 42 ml anhydrous tetrahydrofuran under nitrogen
atmosphere. To this solution, methyl iodide (1.07 ml, 16.89 mmol)
solution in tetrahydrofuran (14.5 ml) was added dropwise at ambient
temperature. The reaction mixture was stirred at same temperature
for 2 days. The mono-salt precipitate was filtered and washed with
dry tetrahydrofuran, dried under high vacuum to obtain compound 21
as a white solid in 72.59% yield (4.12 g). .sup.1H NMR (200 MHz,
D.sub.2O) .delta. 3.43.about.3.85, 3.07.about.3.24,
2.67.about.2.86, 2.37.about.2.46 (18H, mCHCH and NCH.sub.2) 3.27
(3H, s, CH.sub.3); .sup.13C NMR (200 MHz, D.sub.2O) .delta. 83.44,
71.51 (--CHCH--) 65.43, 61.07, 51.22, 48.26, 48.08, 47.79, 44.04
(--NCH.sub.2) 47.60 (CH.sub.3).
Synthesis of Compound 22 (by method A, SCHEME 3)
[0136] Compound 20 (70 mg, 0.107 mmol) was dissolved in dimethyl
formamide (2.5 ml) under argon. To this solution, methyl iodide
(0.1 ml, in excess) was added at ambient temperature and the
reaction mixture was stirred at ambient temperature for 4 days.
Then acetonitrile (20 ml) was added to the stirred reaction mixture
slowly. The precipitate was washed with acetonitrile several times
till washing was colorless. Tetra-salt product was dried under high
vacuum to obtain 50 mg of compound 22 in 58% yield.
[0137] Synthesis of Compound 22 (by method B, SCHEME 4)
[0138] Compound 21 (4.07 g, 12.1 mmol) was dissolved in 40 ml DMF
under nitrogen atmosphere, to which m-xylene dibromide (1.56 g,
5.764 mmol) in DMF (13 ml) was added dropwise at ambient
temperature. The reaction mixture was stirred at room temperature
for 2 days. The tetra-salt precipitate was centrifuged and washed
once with a small amount of DMF and then washed with acetonitrile
several times. The white solid salt (compound 22) was dried under
high vacuum to obtain compound 22 in 54% yield (3.49 g). .sup.1H
NMR(200 MHz, D.sub.2O) .delta. 7.70.about.7.86 (4H,
H.sub.Aromatic), 4.75.about.5.06 (4H, CHCH), 4.68 (4H,
Aromatic-CH.sub.2), 3.38 (6H, --CH.sub.3), 4.2.about.4.36,
3.80.about.4.05, 3.31.about.3.63, 2.93.about.3.20(--NCH.sub.2);
.sup.13C NMR (200 MHz, D.sub.2O) .delta. 134.83, 134.14, 130.16,
126.86 (C.sub.aromatic), 76.97 (C.sub.animal), 45.08 (--CH.sub.3),
63.86, 60.11, 59.21, 58.01, 53.53, 45.55, 45.29, 41.92, 41.59
(--NCH.sub.2).
Synthesis of Compound 23
[0139] Compound 22 (3.35 g, 2.82 mmol) was heated with hydrazine
monohydrate (20 ml) at 110.degree. C. in argon. After cooling, the
precipitate was filtered and remaining hydrazine was removed by
rotatory evaporator as an azotropic mixture using ethanol as the
solvent. The same procedure was repeated twice. The resulting
product was dried under high vacuum to obtain white oil (1.25 g,
93%). The white oil was further treated with hydrochloric acid (6
N) at 0.degree. C. to convert it to HCl salt form, compound 23.
.sup.1H NMR (HCl salt form, 200 MHz, D.sub.2O) .delta. 7.39, 7.36,
7.27, 7.23 (4H, m, Aromatic H), 3.78 (4H, s, Benzylic-CH.sub.2),
2.77.about.3.15 (32H, m, NCH.sub.2), 2.26 (6H, s, CH.sub.3);
.sup.13C NMR (HCl form, 200 MHz, D.sub.2O) .delta. 133.13, 132.73,
131.01, 129.53 (C.sub.Aromatic), 57.52 (C.sub.Benzlic-C), 51.43,
48.30, 42.41, 42.31 (--NCH.sub.2) 42.64 (CH.sub.3).
Synthesis of Compound 24
[0140] Compound 23 (1.93 g, 2.62 mmol) was dissolved in 55 ml dd
water and basified with an aqueous sodium hydroxide solution until
the pH reached 11-12. To this solution, an aqueous solution of
bromoacetic acid (2.2 g, 15.86 mmol) in 30 mL dd H.sub.2O was added
slowly. The reaction mixture was heated at 60.degree. C., and the
pH of the reaction mixture was maintained at 11-12 by using the
sodium hydroxide solution. After 3 days, the reaction mixture was
acidified using hydrochloride (6 N) till the pH of the mixture
reached 4. The solvent was removed by rotatory evaporator. The
residue was dissolved in anhydrous methanol (40 ml) and the mixture
was filtered to remove sodium chloride salt. The filtrate was
evaporated and the resulting solid was purified on cation exchange
resin to obtain compound 24 as a white solid in 70% yield (1.75 g).
.sup.1H NMR (400 MHz, D.sub.2O) .delta. 7.49, 7.42 (4H, s,
Aromatic), 4.42 (4H, s, benzulic CH.sub.2), 2.97.about.3.40 (40H,
m, NCH.sub.2 CH.sub.2COOH), 2.83 (6H, s, CH.sub.3), .sup.13C NMR
(400 MHz, D.sub.2O) .delta. 174.66 (--COOH), 133.90, 132.97,
131.51, 130.17 (C.sub.aromatic), 56.90 (benzene--CH.sub.2), 52.98
(CH.sub.2COOH), 54.50, 50.61, 49.08, 48.69 (NCH.sub.2), 43.14
(CH.sub.3).
Synthesis of Compound 30
[0141] In a solution of pyridine -2,6-dicarboxylic acid (compound
29, 10 g, 59.8 mmol) in 100 ml methanol (ACS grade) under argon,
thionyl chloride (8 ml, 109 mmol) was added slowly at 0-5.degree.
C. The reaction mixture was refluxed for 18 -20 hrs, and then
cooled to ambient temperature and then to 0.degree. C. Compound 30
was crystallized out from the solution. The solid was filtered
under vacuum and washed with cold methanol (30 ml). The resulting
solid was dried to get 9.3 g of compound 30 (80%). MP=122.degree.
C.; R.sub.f=0.61 in ethyl acetate; .sup.1H NMR (200, CDCl.sub.3)
.delta. 3.98 (s, 6H, ester CH.sub.3), 7.98 (t, 1H, .gamma. proton
of pyridine ring), 8.26 (d, 2H, .beta. protons of pyridine
ring).
Synthesis of Compound 31
[0142] In a solution of compound 30 (9.3 g, 47.7 mmol) in methanol
(200 ml) at 0.degree. C. under argon, sodium borohydride (6.32 g,
166.3 mmol) was slowly. The reaction mixture was stirred at
0.degree. C. for 3 hrs. Reaction was monitored by TLC. Saturated
sodium bicarbonate was added to decompose sodium borohydride. The
reaction mixture was concentrated and the solution was extracted
with chloroform (3.times.200 ml). The organic layers were combined
and dried over magnesium sulphate. After removing solvent, the
white residue was purified on silica gel column (60 g) using 30-40%
ethyl acetate/chloroform as eluent to obtain compound 31 (4.5 g,
56%). MP=83.degree. C.; R.sub.f=0.38 in ethyl acetate; .sup.1H NMR
(200, CDCl.sub.3) .delta. 3.90 (s, 3H, OCH.sub.3), 4.36 (br s, 1H,
OH), 4.79 (s, 2H, CH.sub.2), 7.5 (d, 1H, Ar--H), 7.76 (t, 1H,
.gamma. proton of pyridine ring), 7.93 (d, 1H, Ar--H).
Synthesis of Compound 32
[0143] Into a solution of compound 31 (6.4 g, 38.32 mmol) in
chloroform (50 ml) at 0.degree. C. under argon, phosphorous
tribromide (6 ml, 63 mmol) was added slowly. Then the reaction
mixture was stirred at ambient temperature for 14-16 hrs. The
reaction was quenched with a 20% potassium carbonate solution at
0.degree. C. till pH=8-9. The reaction mixture was extracted with
chloroform. The organic layer was dried over magnesium sulphate.
After removing the solvent, the residue was purified on silica gel
column using chloroform as eluent. Compound 32 was obtained in 6.5
g (70%).R.sub.f=0.60 (in 3:1 hexane:ethyl acetate). .sup.1H NMR
(200, CDCl.sub.3) .delta. 4.01 (s, 3H, OCH.sub.3), 4.65 (s, 2H,
benzylic protons), 7.71 (d, 1H, ArH), 7.87 (t, 1H, ArH), 8.04 (d,
1H, ArH); .sup.13H NMR (200, CDCl.sub.3) .delta. 33.01, 52.87,
124.25, 126.93, 138.02, 147.45, 157.23, 165.15.
* * * * *