U.S. patent application number 11/405359 was filed with the patent office on 2006-10-19 for biocompatible materials and probes.
Invention is credited to Manssur Yalpani.
Application Number | 20060235193 11/405359 |
Document ID | / |
Family ID | 29250865 |
Filed Date | 2006-10-19 |
United States Patent
Application |
20060235193 |
Kind Code |
A1 |
Yalpani; Manssur |
October 19, 2006 |
Biocompatible materials and probes
Abstract
The present invention relates to fluorinated biopolymer and
polymer derivatives useful as imaging probes, diagnostic agents and
contrast agents and to imaging methods employing the fluorinated
biopolymers and polymers.
Inventors: |
Yalpani; Manssur; (Rancho
Sante Fe, CA) |
Correspondence
Address: |
SALIWANCHIK LLOYD & SALIWANCHIK;A PROFESSIONAL ASSOCIATION
PO BOX 142950
GAINESVILLE
FL
32614-2950
US
|
Family ID: |
29250865 |
Appl. No.: |
11/405359 |
Filed: |
April 17, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10411972 |
Apr 11, 2003 |
7030208 |
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11405359 |
Apr 17, 2006 |
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60372500 |
Apr 11, 2002 |
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Current U.S.
Class: |
528/328 ;
525/430; 528/310 |
Current CPC
Class: |
C08B 31/12 20130101;
C08B 37/0072 20130101; C08B 11/193 20130101; C08B 37/0021 20130101;
C08H 1/06 20130101; C08H 1/00 20130101; C08B 13/00 20130101; C08B
11/06 20130101; C08B 37/00 20130101; C08B 11/12 20130101 |
Class at
Publication: |
528/328 ;
528/310; 525/430 |
International
Class: |
C08G 69/10 20060101
C08G069/10 |
Claims
1. A fluorinated biopolymer of the Formula XIII ##STR19## where for
Formula XIII: K represents H, OH, X, OX, OZ, (Y).sub.f; L
represents H, OH, X, OX, OZ, (Y).sub.f; W represents H,
(CH.sub.2).sub.d, CO.sub.2H, CH, CX, X; T represents H, OH, X, OX,
OZ; V represents (CH.sub.2).sub.d, CH.sub.2OX, CH.sub.2OZ,
CH.sub.2X, CH.sub.2NHX; S represents H, O, X, NHX, (Y).sub.f; T
represents H, O, X, NHX, (Y).sub.f; R.sub.1 represents H, X and d
represents 1-3 inclusive; f, and n represent 1-1,500 inclusive and
for all of the above Formulas: X represents a fluorine containing
moiety, a luminescent residue, a fluorescent residue, a fluorinated
luminescent residue or a fluorinated fluorescent residue; Y
represents an amino acid residue or a fluorinated amino acid
residue; and Z represents acyl, alkyl.
2. The fluorinated biopolymer of any of claim 1, wherein X is
fluoroalkyl, fluoroaryl, fluoroacyl, perfluoroalkyl, perfluoroaryl,
perfluoroacyl, perfluoropolymer, fluoroamine, fluorocarbamate,
fluorotriazine, fluorosulfonylalkyl derivatives, CF.sub.2Cl,
SO.sub.2[CF.sub.2].sub.xCF.sub.3, F, CF.sub.3, COC.sub.xF.sub.y,
C.sub.xF.sub.yH.sub.z,
([CH.sub.2].sub.mO).sub.x(CH.sub.2CF.sub.2O).sub.y(CF.sub.2CF.sub.2O).sub-
.z(CF.sub.2).sub.2CF.sub.2CH.sub.2O(CH.sub.2).sub.pOH,
CH.sub.2C(OH)C.sub.xF.sub.yH.sub.z, C.sub.xF.sub.yH.sub.zO.sub.p,
COC.sub.xF.sub.yH.sub.z,
OCH.sub.2C.sub.xF.sub.z[C.sub.xF.sub.zO].sub.mF,
CH.sub.2C(CH.sub.3)CO.sub.2C.sub.xH.sub.z(CF.sub.2).sub.mCF.sub.3,
CH.sub.2(CF.sub.2O).sub.x(CF.sub.2CF.sub.2O).sub.y(CF.sub.2O).sub.zCF.sub-
.2CH.sub.2OH, COCF(CF.sub.3)--[CF(CF.sub.3)CF.sub.2O].sub.mF,
NHC.sub.xF.sub.yH.sub.zO.sub.p,
CH.sub.2CF.sub.2O[CF.sub.2CF.sub.2O].sub.m(CF.sub.2OCF.sub.2CH.sub.2OH,
COC.sub.xH.sub.z(CF.sub.2).sub.mCF.sub.3,
COCF.sub.2O[CF.sub.2CF.sub.2O].sub.nCF.sub.2OCF.sub.2CO.sub.2H,
([CH.sub.2].sub.mO).sub.x(CH.sub.2CF.sub.2O).sub.y(CF.sub.2CF.sub.2O).sub-
.zCF.sub.2CH.sub.2O(CH.sub.2).sub.pOH,
N[C.sub.xF.sub.yH.sub.z].sub.p,
C.sub.xH.sub.zCO.sub.2C.sub.xH.sub.z(CF.sub.2).sub.mCF.sub.3,
COC.sub.xF.sub.y[C.sub.pF.sub.zO].sub.mF, and m, x, p, y, z are
integers from 1 to 150 inclusive.
3. A method of improving the effectiveness of magnetic resonance
imaging (MRI) which comprises: a. administering an effective amount
of one or more fluorinated polymers of claim 1, to a patient; b.
subjecting the patient to an MRI of a tissue/organ where the
administered polymer is expected to accumulate; and c. evaluating
the tissue/organ from the MRI images obtained.
Description
CROSS-REFERENCE TO A RELATED APPLICATION
[0001] This application claims the benefit of provisional patent
application Ser. No. 60/372,500, filed Apr. 11, 2002, which is
hereby incorporated by reference in its entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to fluorinated biopolymer and
polymer derivatives useful as imaging probes, diagnostic agents and
contrast agents and to imaging methods employing the fluorinated
biopolymers and polymers.
BACKGROUND
[0003] Hyaluronic acid (or hyaluronan, HA) is a high molecular
weight copolymer of 1.fwdarw.3-.beta.-linked
N-acetyl-D-glucosamine-1.fwdarw.4-.beta.-D-glucuronic acid from the
glycosaminoglycans family of biopolymers with unusual rheological
properties. Its physiological functions include the lubrication and
protection of cells, maintenance of tissue structural integrity,
and transport of molecules to and within cells. HA is found in the
extracellular matrix (ECM) and plays an integral role in its
organization and structure. Hyaluronan influences cellular
proliferation and migration in developing, regenerating and
remodeling tissues and in tissues undergoing malignant tumor-cell
invasion (see, e.g., B. P. Toole S. D. Banerjee, Oligosaccharides
reactive with hyaluronan-binding protein, monoclonal antibodies
recognizing hyaluronan-binding protein, and use in cancer therapy,
U.S. Pat. No. 5,902,795, 1999; S. Kumar, D. West, D. B. Rifkin, M.
Klagsburn (eds.) Hyaluronic acid and its degradation products
modulate angiogenesis in vivo and in vitro. In Current
Communications in Molecular Biology; Angiogenesis: Mechanism and
Pathobiology, Cold Spring Harbor Laboratory, Cod Spring Harbor,
N.Y., pp. 90-94, 1987.).
[0004] HA binds specifically to proteins in the ECM, within the
cytosol and on cell-surface receptors. The prevalence of
hyaluronan-binding proteins indicates the importance of HA
recognition in tissue organization, proliferation and
differentiation, growth factor activities, and the control of
cellular adhesion and motility. HA's role extends to embryonic
development, modulation of inflammation, stimulation of
angiogenesis and wound healing, and morphogenesis.
[0005] A number of extracellular matrix and cellular proteins, the
hyaladherins, have specific affinities to HA within the
extracellular matrix. These include aggrecan, cartilage
link-protein, hyaluronectin, neurocan and versican. Cellular
hyaluronan receptors such as CD44 (CD="cluster of differentiation")
and RHAMM (receptor for hyaluronate-mediated motility) are also
known. Recent evidence implicates the CD44-HA interaction in cancer
metastasis (for reviews, see Entwistle, J.; Hall, C. L.; Turley, E.
A. J. Cell. Biochem., 61, 569-577, 1996; Bajorath, J. Proteins:
Struct. Funct. Genet., 39, 103-111, 2000.). Melanoma cells
expressing high CD44 levels show increased cell motility and
metastatic potential compared to the same cell types that expressed
low receptor levels (see e.g., Birch, M.; Mitchell, S.; Hart, I.
Cancer Res., 51, 6660-6667, 1991.). The presence of specific HA
cell receptors provides therefore potential uses in cancer
diagnosis and therapy. Other biomedical uses include cataract
surgery, osteoarthritis, and prevention of post-surgical adhesions.
HA also displays useful wetting and moisture-preserving functions
that are of interest in cosmetic and topical medical areas. HA
sources include rooster combs, umbilical cords, shark skin, bull's
eye and fermentation.
[0006] The integrin receptor family binds to ECM receptors (S. M.
Abelda, Role of integrins and other cell adhesion molecules in
tumor progression and metastasis, Lab Invest., 68, 4-17, 1993.).
Integrins are heterodimeric glycoproteins with two subunits
(.alpha. and .beta.). A given .beta.-subunit can pair with a number
of .alpha.-subunits, resulting in various integrins with unique
binding properties. Thus, .alpha.2.beta.1 constitutes a collagen
receptor that does not interact with laminin on platelets (C. J.
Anderson, Bioconjugate Chem. 12, 1057-65, 2001.)
[0007] Normal human tissue cells express various integrins such as
.alpha.1.beta.1, .alpha.2.beta.1, .alpha.3.beta.1, and
.alpha.6.beta.1 that are required for adhesion to collagen and
laminin (J. L. Lauer, C. M. Gendron, G. B. Fields, Effect of ligand
conformation on melanoma cell alpha3beta1 integrin mediated signal
translocation event Implication for a collagen structural
modulation mechanism of tumor cell invasion, Biochemistry, 37,
5279-87, 1998.). Radiolabeled ECM fragments are useful imaging
agents since their integrins are upregulated in certain tumors and
can be targeted for diagnosis and therapy. Integrins promote
adhesion, signal transduction and linkage between intracellular
proteins and ligands. ECM fragments are used as imaging agents as
their integrins are upregulated in certain tumor types and can be
targeted for diagnostic or therapeutic use.
[0008] The ubiquitous nature of HA in biological systems, coupled
with its antitumor and diverse range of other medical activities
make diagnostic probe-carrying HA derivatives attractive for
diagnostic and therapeutic uses. There is furthermore growing
evidence that oligosaccharides derived from hyaluronan also bind to
CD44. Thus, if an antagonist could be found for the CD44 receptor
that would prevent HA binding, it would be possible consequently to
limit metastasis. Such small molecules would have advantages over
HA itself in that they would possibly be water soluble, membrane
penetrating, and easy to administrate. Minimally, a 6-mer
(hexasaccharide) is required for binding to CD44 and the 10-mer
(decasaccharide) is required to displace HA from the HA-CD44
complex.
[0009] Other acidic polysaccharides, such as alginate and pectin
are possibly also biologically active, as some evidence indicates
in the literature (A. Kawada, N. Hiura, S. Tajima, H. Takahara,
Alginate oligosaccharides stimulate VEGF-mediated growth and
migration of human endothelial cells, Arch. Dermatol. Res., 291,
542-7, 1999; M. Sakurai, H. T. Matsumoto, H. Kiyohara, H. Yamada,
B-cell proliferation activity of pectic polysaccharides from a
medicinal herb, Immunology, 97, 540-7, 1999; H. Yamada,
Contribution of pectins on health care, in J. Visser, A. G. J.
Voragen eds., Pectins and Pectinases, Elsevier, Amsterdam, 173-190,
1996; H. Yamada, H. Kiyohara, Complement-activating polysaccharides
from medicinal herbs, in H. Wagner ed., Immunomodulatory Agents
from Plants, Birkhauser Verlag, Basel, 1999.). The preparation of
alginate oligosaccharides (A, Martinsen, G. Skjak-Braek, O.
Smidsrod, Carbohydr. Polym., 15, 171-173, 1991. Ikeda, H-F, A. A
Takemura, H Ono, Carbohydr. Polym., 42, 421-425, 2000.) and pectic
oligosaccharides (N. O. Maness, A. J. Mort, Anal. Biochem., 178,
248-254, 1989.) has been reported.
[0010] Hyaluronan has attracted considerable interest as
biocompatible, resorbable material for tissue engineering and a
wide range of other biomedical applications (for reviews, see D.
Campoccia, P. Doherty, M. Radice, P. Brun, G. Abatangelo, D. F.
Williams, Semisynthetic resorbable materials from hyaluronan
esterification, Biomaterials, 19, 2101-2127, 1998; E. Milella, E.
Brescia, C. Massaro, P. A. Ramires, M. R. Miglietta, V. Fiori, P.
Aversa, Physico-chemical properties and degradability of non-woven
hyaluronan benzylic esters as tissue engineering scaffolds,
Biomaterials, 23, 1053-1063, 2002.) A considerable number of
hyaluronan derivatives have been reported (see, e.g., K. P.
Vercruysee, G. D. Prestwich, Hyaluronate derivatives in drug
delivery, Crit. Rev. Therapeut. Carrier Syst., 15, 514-555, 1998;
Y. Luo, G. D. Prestwich, Hyaluronic acid-N-hydroxysuccinimide: a
useful intermediate for bioconjugation, Bioconjugate Chem. 12,
1085-88, 2001.). Collagen, the other major component of the
extracellular matrix, constitutes over 30% of the human protein
content and is associated with a number of diseases. Collagen has
therefore been similarly widely employed as biocompatible matrix,
as have hybrid materials derived from collagen and hyaluronan
(S.-N. Park, J-C. Park, H. O. Kim, M. J. Song, H. Suh,
Characterization of porous collagen/hyaluronic acid scaffold
modified by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide
cross-linking, Biomaterials, 23, 1205-1212, 2002). Collagen
features an unusual amino acid composition: glycine constitutes
over 30%, proline and hydroxyproline about 20%, whilst it lacks
tryptophan and cysteine (i.e., no disulfide bonds).
[0011] Poly(glutamic acids), and in particular
poly(.gamma.-glutamic acid) (.gamma.-PGA) are new biodegradable
materials with many potential biomedical uses (I.-L. Shih, Y.-T.
Van, The production of poly(.gamma.-glutamic acid) from
microorganisms and its various applications, Bioresource Techn.,
79, 207-225, 2001). .gamma.-PGA, elaborated by various Bacillus
species (e.g., B. licheniformis), is an unusual polypeptide with
its glutamic acid residues linked linearly through the
.gamma.-carboxyl function. .gamma.-PGA assumes an .alpha.-helix
conformation in solution, and, unlike the synthetic .alpha.-PGA
analog, is a well-defined, high molecular weight homopolymer.
[0012] .gamma.-PGA's polyanionic nature renders it highly water
soluble and permits modulation of its solution conformation by
co-solutes. PGAs ability to undergo conformational changes in
response to different pH values offers the potential to affect
targeted delivery. .gamma.-PGA features a high molecular weight
range and different solution conformations, is biocompatible,
biodegradable (it biodegrades to glutamic acid monomers),
non-toxic, and non-immunogenic nature. .gamma.-PGA is also highly
mucoadhesive, a key feature for localizing it site-specifically as
a drug delivery vehicle in the small intestinal or colonic
mucosa.
[0013] Radiolabeled peptide hormone analogues are of interest as
diagnostic and therapeutic vehicles for treating cancer (Cutler C.
S. Lewis J. S. Anderson C. J. Adv. Drug Deliv. Res., 37, 189-211,
1999. Anderson C. J. Welch M. J., Chem. Rev., 99, 2219-2234, 1999;
Anderson C. J. Dehdashti F Cutler P. D. Schwarz S W. Laforet R.
Bass L. R. Lewis J. S. McCarthy D. W., J. Nucl. Med., 42, 213-2334,
2001.). These radiolabeled peptide receptor ligands can target
upregulated cell surface receptors on tumors. For example,
.sup.111In-DTPA-octreotide is employed for imaging of
neuroendocrine tumors that overexpress the somastatin receptor (E.
P. Krenning, D. J. Kwekboom, W. H. Bakker, W. A. P. Breeman, P. P.
M. Kooji, H. Y. Oei, M. van Hagen, P. T. E. Postema, M. de Jong, J.
C. Reubi, T. J. Visser, A. E. M. Reji, L. L. J. Holland, J. W.
Kuuper, S. W. J. Lamberts, Somatostatin receptor scintography with
[.sup.111In-DTPA-D-Phe] and [.sup.111In-Tyr]octreotide, Eur. J.
Nucl. Med., 20, 716-731, 1993.).
[0014] Primary human tumors from colon, ovary, skin and stomach and
their metastatic sites show high levels of .alpha.3.beta.1, and
similarly cultured human cell lines (e.g., breast, ovarian
carcinoma) express .alpha.3.beta.1. Non-invasive means of
monitoring .alpha.3.beta.1 expression could be useful as a
diagnostic tool for assessing metastasis prior to surgery. Since
natural collagens are integrin ligands radiolabeled collagen
fragments can serve as imaging agents.
[0015] There is a considerable demand for versatile non-invasive
diagnostic probes, and fluorine's diagnostic value is of particular
interest in non-invasive imaging applications. Apolar oxygen
imparts paramagnetic relaxation effects on .sup.19F nuclei
associated with spin-lattice relaxation rates (R.sub.1) and
chemical shifts. This effect is proportional to the partial
pressure of O.sub.2 (pO.sub.2). .sup.19F NMR can therefore probe
the oxygen environment of specific fluorinated species in cells and
other biological structures.
[0016] Noth et al. (U. Noth, P. Grohn, A. Jork, U. Zimmermann, A.
Haase, J. Lutz, .sup.19F-MRI in vivo determination of the partial
oxygen pressure in perfluorocarbon-loaded alginate capsules
implanted into the peritoneal cavity and different tissues, Magn.
Reson. Med., 42(6), 1039-47, 1999) employed perfluorocarbon-loaded
alginate capsules in MRI experiments to assess the viability and
metabolic activity of the encapsulated materials. Quantitative
.sup.19F-MRI was performed on perfluorocarbon-loaded alginate
capsules implanted into rats, in order to determine in vivo the
pO.sub.2 inside the capsules at these implantation sites. Fraker et
al. reported recently a related method with perfluorotributylamine
(C. Fraker, L. Invaeradi, M. Mares-Guia, C. Ricordi, PCT WO
00/40252, 2000).
[0017] Although a large range of fluorinated products is available
commercially, most PFCs suffer from a number of shortcomings. Many
commercial PFCs currently employed for diagnostic purposes were
originally selected for blood substitution. Their physicochemical
properties [J. G. Reiss et al., Biomat. Artif. Cells Artif. Organs,
16, 421-430, 1988.] are therefore not targeted towards specific
diagnostic or other biomedical uses, particularly for MRI. The
molecular features of these PFCs are not optimized for
high-sensitivity .sup.19F-MRI studies. Their T.sub.1 relaxation
times are relatively long, T.sub.2 relaxation times are short, and
severe J-modulation effects and chemical shift artifacts can
profoundly limit their MRI utility. Whilst their immiscibility in
water offers benefits in some respects, it necessitates the use of
emulsifiers. Thus, for PFC-in-water emulsions, such as F-44E,
perfluorohexyl bromide (PFHB), perfluorooctyl bromide (PFOB,
Perflubron.TM.), perfluoromethyldecalin (PMD), perfluorooctyl
ethane (PFOE), perfluorotripropylamine (FTPA), and the blood
substitutes Fluosol.TM. and Oxygent.TM., lecithins or poloxamers
are employed to disperse the PFCs and stabilize the emulsion.
Fluosol.TM. was a 20% w/v mixture of 14% perfluorodecalin and 6%
perfluorotripropylamine emulsified primarily with Pluronic
F-68.TM.. Oxygent.TM. is a 60% emulsion consisting mostly of PFOB
and perfluoro-decylbromide, water, salts, and a lecithin. However,
surfactants are problematic in that their use adds processing
requirements and some of them can be unstable, chemically
ill-defined or polydisperse, or cause potential undesirable side
effects. Thus, Pluronic F-68, the surfactant in Fluosol.TM., caused
a transitory anaphylactic reaction in certain patients. Further,
the stability of Pluronic F-68-based emulsions was limited;
requiring frozen storage and mixing with two annex solutions prior
to administration. The use of emulsions poses the additional
disadvantage that the PFCs' fluorine content is effectively diluted
(often by 50% or more), diminishing their spectral and imaging
signal intensities and, hence diagnostic benefit. The impact of
such dilutions is particularly evident in tumor oxygenation studies
where only .about.10% of the injected PFC emulsion dose reaches the
tumor, necessitating time consuming T.sub.1 measurements. This
dilution effect is even more pronounced, when only a portion of the
available PFCs' fluorine resonances is of diagnostic value. This is
often the case, as severe chemical shift artifacts need to be
circumvented by selectively exciting only a narrow chemical shift
range containing one resonance (or a closely spaced group of
resonances). Although F-44E, for instance, has a high fluorine
content (74%) with largely acceptable spectral features, many MRI
studies have selectively excited its trifluoromethyl resonance,
representing only one third of the total F-content, which on
emulsification (at 90%) is further diluted to .about.22%.
Similarly, for MRI with perfluorononane the choice is between the
selective acquisition of the single trifluoromethyl resonance (6
fluorines with a spectral width of 50 kHz at 7 Tesla) or multiple
difluoromethylene resonances (14 fluorines with a 1300 kHz spectral
dispersion) (see, e.g., S. L. Fossheim; K A Il'yasov, J. Hennig, A.
Bjornerud, Acad. Radiol., 7(12), 1107-15, 2000.).
[0018] Ideally, PFC imaging agents should combine the following
features: non-toxic, biocompatible, chemically pure and stable, low
vapor pressure, high fluorine content, reasonable cost and
commercial availability. Additionally, they should meet several
.sup.19F-NMR criteria, including a maximum number of chemically
equivalent fluorines resonating at one or only few frequencies,
preferably from trifluoromethyl functions. Some of the other
spectral criteria have been discussed in detail elsewhere (C. H.
Sotak, P. S. Hees, H. N. Huang, M. H. Hung, C. G. Krespan, S.
Raynolds, Magn. Reson. Med., 29, 188-195, 1993.). For MRI, it would
furthermore be desirable to have control over the amount of
magnetically responsive material for specific uses, and to employ
temperature-responsive and pH-dependent imaging agents for special
uses. These could have applications in MRI-based temperature
monitoring for use in general hyperthermia treatment (see, e.g., S.
L. Fossheim; K. A. Il'yasov, J. Hennig, A. Bjornerud, Acad.
Radiol., 7(12), 1107-15, 2000.) of tumors and for monitoring the
efficacy of chemotherapy, respectively (see, e.g., N. Rhagunand, R.
Martinez-Zagulan, S. H. Wright, R. J. Gilles, Biochem. Pharmacol.,
57, 1047-1058, 1999; I. F Tannock, D. Rotin, Cancer Res., 49,
4373-4383, 1989.). Furthermore, water solubility would enhance the
PFC functionality in many biomedical settings, as it would obviate
the need for emulsifiers.
[0019] Although selected efforts have been directed at developing
new fluorinated MRI probes, none are water soluble compounds [e.g.,
perfluoro-[15]-crown-5 ether)], and some are commercially
unavailable [e.g.,
perfluoro-2,2,2',2'-tetramethyl-4,4'-bis(1,3-dioxalane)-PTBD]. It
appears no attempts have so far focused on screening available PFCs
from the thousands of commercial fluorinated products in order to
identify potentially more suitable MRI probes for biomedical uses.
It seems furthermore that no studies have attempted to establish
structure activity relations (SARs) of related PFCs for MRI
purposes. Noteworthy is also the fact that all PFCs examined to
date have molecular weights under 1,000, typically between 400-600
Da. This is partly a reflection of the specific requirements for
blood substitution agents, but also due to the widely held belief
that higher molecular weight or polymeric fluorinated agents would
not be detectable by .sup.19F-NMR due to anticipated excessive line
broadening, and would therefore be unsuitable. Thus, with the
exception of the polymer-encapsulated PFCs noted above, this
important class of materials had so far been excluded from
consideration.
[0020] Paramagnetic ions, such as gadolinium (Gd.sup.3+) decrease
the T.sub.1 of water protons in their vicinity, thereby providing
enhanced contrast. Gadolinium's long electron relaxation time and
high magnetic moment make it a highly efficient T.sub.1 perturbant.
Since uncomplexed gadolinium is very toxic, gadolinium chelate
probes, such as gadolinium diethylenetriamine pentaacetic acid
(GdDTPA M.sub.w 570 Da), albumin-GdDTPA (Gadomer-17, M.sub.w, 35 or
65 kDa), have been employed extensively in MRI of tumors and other
diseased organs and tissues. Several other developmental chelators
have also been reported, including dual-labeled agents,
oligonucleotide-derived, dextran-derived GdDTPA, and TAT and other
peptide-derived chelators. However, presently approved MRI contrast
agents are either not tissue specific, e.g., GdDTPA, or target only
normal tissue, which limits their utility in diagnosis of
metastases or neoplasia. MRI studies with GdDTPA, for instance, do
not correlate with the angiogenic factor or the vascular
endothelial growth factor (VEGF). Attempts have also been made to
overcome the low relaxivities of small Gd-DTPA chelates by
preparing polymer conjugates of Gd(DTPA).sup.(2-) [see e.g., M. R.
A. Duarte, M. G. Gil, M. H. Peters, J. A. Colet, J. M. Elst, L.
Vander; R. N. Muller, C. F. G. C. Geraldes, Bioconjug. Chem., 21,
170-177, 2001.]. However, the relaxivity of these polymer
conjugates was only slightly improved and they were also cleared
very quickly from the blood of rats, indicating that they are of
limited value as blood pool contrast agents for MRI.
[0021] Whilst much can be achieved with currently available imaging
and contrast agents, there are still unmet needs for novel
diagnostic agents, particularly for those exploiting biological
specificity. Imaging agents suitable for targeting metastases or
neoplasia would substantially enhance the MRI sensitivity and
utility for tumor detection and prevention. Although selected
efforts have been directed at developing such new probes, a broader
investigation of these agents is urgently needed. Similarly, new
imaging probes are needed as noninvasive means to detect and image
cells, tissues and organs undergoing apoptosis. An even greater
demand exists for biocompatible materials in tissue engineering and
various other biomedical applications.
BRIEF DESCRIPTION OF THE DRAWING
[0022] FIG. 1 shows a NMR spectrum of the perfluoroalkyl hyaluronan
of Example 2.
SUMMARY OF THE INVENTION
[0023] The present invention relates to fluorinated biopolymer and
polymer derivatives (Formulas I-XX) useful as imaging probes,
diagnostic agents and contrast agents. Additionally, the present
invention relates to imaging methods employing the present
compounds of Formulas I-XX
[0024] Novel compositions comprising modified biopolymers of the
present invention include the compounds of general formula I to
VIII and their use as new biomaterials, imaging probes, diagnostic
tools and contrast agents: ##STR1## Where For Formula I: R.sub.1=H,
X; R.sub.2=H, X; R.sub.3=H, OH, OY, OX, NHX For Formula II:
R.sub.1=H, X; R.sub.2=H, X; R.sub.3=H, OY, OX, NHX For Formula III:
R.sub.1=H, X; R.sub.2=H, X; R.sub.3=H, Y, X For Formula IV:
R.sub.1=H, X; R.sub.2=H, X, R.sub.3=CO.sub.2H, CO.sub.2X,
CH.sub.2X, CH.sub.2NHX; R.sub.4=H, X; R.sub.5=H, X; R.sub.6=H, X;
R.sub.7=X, COCH.sub.3, COX For Formula V: R.sub.1=H, X; R.sub.2=H,
X; R.sub.3=CO.sub.2H, CO.sub.2X, CH.sub.2X, CH.sub.2NHX; R.sub.4=H,
SO.sub.3H, X; R.sub.5H, SO.sub.3H, X; R.sub.6=H, X;
R.sub.7=COCH.sub.3, COX, X For Formula VI: R.sub.1=H, X; R.sub.2=H,
X; R.sub.3=CO.sub.2H, CO.sub.2X, CH.sub.2X, CH.sub.2NHX; R.sub.4=H,
X; R.sub.5=SO.sub.3H, X; R.sub.6=H, X; R.sub.7=COCH.sub.3, COX, X
For Formula VIII: R.sub.1=H, X; R.sub.2=H, X; R.sub.3=H, X;
R.sub.4=SO.sub.3H, X; R.sub.5=SO.sub.3H, X; R.sub.6=H, X;
R.sub.6=H, X; R.sub.7=COCH.sub.3, COX, X For Formula VIII:
R.sub.1=H, X; R.sub.2=H, X; R.sub.3=CO.sub.2H, CO.sub.2X,
CH.sub.2X, CH.sub.2NHX; R.sub.4=H, X; R.sub.5=H, X; R.sub.6=H, X;
R.sub.7=H, X; R.sub.8=COCH.sub.3, COX, X Wherein for all of the
above Formulas X=fluoroalkyl, fluoroaryl, fluoroacyl,
perfluoroalkyl, perfluoroaryl, perfluoroacyl, perfluoropolymer,
fluoroamine, fluorocarbamate, fluorotriazine, fluorosulfonylalkyl
derivatives, CF.sub.2C.sub.1, SO.sub.2[CF.sub.2].sub.xCF.sub.3, F,
CF.sub.3, COC.sub.xF.sub.y, C.sub.xF.sub.yH.sub.2,
([CH.sub.2].sub.mO).sub.x(CH.sub.2CF.sub.2O).sub.y(CF.sub.2CF.sub.2O).sub-
.z(CF.sub.2).sub.2CF.sub.2CH.sub.2O(CH.sub.2).sub.pOH,
CH.sub.2C(OH)C.sub.xF.sub.yH.sub.z, C.sub.xF.sub.yH.sub.zO.sub.p,
COC.sub.xF.sub.yH.sub.z,
OCH.sub.2C.sub.xF.sub.z[C.sub.xF.sub.zO].sub.mF,
CH.sub.2C(CH.sub.3)CO.sub.2C.sub.xH.sub.z(CF.sub.2).sub.mCF.sub.3,
CH.sub.2(CF.sub.2O).sub.x(CF.sub.2CF.sub.2O).sub.y(CF.sub.2O).sub.zCF.sub-
.2CH.sub.2OH, COCF(CF.sub.3)--[CF(CF.sub.3)CF.sub.2O].sub.mF,
NHC.sub.xF.sub.yH.sub.zO.sub.p,
CH.sub.2CF.sub.2O[CF.sub.2CF.sub.2O].sub.m(CF.sub.2OCF.sub.2CH.sub.2OH,
COC.sub.xH.sub.z(CF.sub.2).sub.mCF.sub.3,
COCF.sub.2O[CF.sub.2CF.sub.2O].sub.nCF.sub.2OCF.sub.2CO.sub.2H,
([CH.sub.2].sub.mO).sub.x(CH.sub.2CF.sub.2O).sub.y(CF.sub.2CF.sub.2O).sub-
.zCF.sub.2CH.sub.2O(CH.sub.2).sub.pOH,
N[C.sub.xF.sub.yH.sub.z].sub.p,
C.sub.xH.sub.zCO.sub.2C.sub.xH.sub.z(CF.sub.2).sub.mCF.sub.3,
COC.sub.xF.sub.y[C.sub.pF.sub.zO].sub.mF, a luminescent residue, a
fluorescent residue, a fluorinated luminescent residue or a
fluorinated fluorescent residue and m, x, p, y, z are integers from
1 to 150, and where m is more preferably 10-100, and most
preferably 10-50, and where x, p, y, z are more preferably 10-75,
even more preferably 10-50, and most preferably 10-20. Acyl and
alkyl residues in the above formulas comprise lipophilic moieties,
including saturated and unsaturated aliphatic residues with C.sub.k
chains, where k is 2 to 100, more preferably 2-50, and most
preferably 2-20, and aryl residues comprise aromatic moieties,
including benzyl, biphenyl, phenyl polycyclic aromatics, and
heteroatom-containing aromatics; and Y=saccharide branch residue
comprised of mono-, di-, oligo- or polysaccharide, fluorinated
saccharide branch residue comprised of mono-, di-, oligo- or
polysaccharide.
[0025] The novel compositions are comprised of modified
biopolymers, wherein said biopolymers include biopolymers that are
selected from the group consisting of amylose, cellulose, dextran,
dextrins, galactan, .beta.-glucans, glycosaminoglycans, including
chondroitin sulfate, dermatan sulfate, heparan sulfate, heparin,
hyaluronate, and keratin sulfate, maltodextrins, mannans, pustulan,
starch, xylans and their copolymers, linear or cyclic oligomers,
hybrids, salts and derivatives.
[0026] Novel compositions comprising modified biopolymers of the
present invention include the compounds of general formula IX to
XII and are disclosed herein as new biomaterials, imaging probes,
diagnostic tools and contrast agents: ##STR2## wherein For Formula
IX: R.sub.1=H, X, Z; R.sub.2=H, X, Z; R.sub.3=H, X; R.sub.4=H, X,
Z; R.sub.5H, X; R.sub.6=H, X; R.sub.7=H, X, Z For Formula X:
R.sub.1=H, X, CH.sub.2OGOCO.sub.2H, CH.sub.2OGCO.sub.2X,
CH.sub.2OGCONX, CH.sub.2OGCH.sub.2NX; R.sub.2=H, X, Z; R.sub.3=H,
X, CH.sub.2OGOCO.sub.2H, CH.sub.2OGCO.sub.2X, CH.sub.2OGCONX,
CH.sub.2OGCH.sub.2NX; R.sub.4=H, COCH.sub.3, COX, X,
CH.sub.2OGOCO.sub.2H, CH.sub.2OGCO.sub.2X, CH.sub.2OGCONX,
CH.sub.2OGCH.sub.2NX For Formula XI: R.sub.1=H, OH, X, OX, OZ,
CH.sub.2OGOCO.sub.2H, CH.sub.2OGCO.sub.2X, CH.sub.2OGCONX,
CH.sub.2OGCH.sub.2NX; R.sub.2=H, OH, X, OX, OZ,
CH.sub.2OGOCO.sub.2H, CH.sub.2OGCO.sub.2X, CH.sub.2OGCONX,
CH.sub.2OGCH.sub.2NX; R.sub.3=CH.sub.2OH, CH.sub.2OX, CH.sub.2OZ,
CH.sub.2X, CH.sub.2NHX, CO.sub.2H, CO.sub.2X, CONX, CH.sub.2OGOH,
CH.sub.2OGOX, CH.sub.2OGOCO.sub.2H, CH.sub.2OGCO.sub.2X,
CH.sub.2OGCONX, CH.sub.2OGCH.sub.2NX; R.sub.4=OH, Z, OX, X;
G=alkyl, hydroxyalkyl For Formula XII: K=H, OH, X, OX, OZ; L=H, OH,
X, OX, OZ; W=H, OH, X, OX, OZ; T=H, OH, X, OX, OZ;
V=anhydrofuranosyl, anhydropyranosyl, and m, n, p, q, r=1-500
inclusive; n=1-8 inclusive. Wherein for all of the above Formulas
X=fluoroalkyl, fluoroaryl, fluoroacyl, perfluoroalkyl,
perfluoroaryl, perfluoroacyl, perfluoropolymer, pfluoroamine,
fluorocarbamate, fluorotriazine, fluorosulfonylalkyl derivatives,
F, CF.sub.2Cl, SO.sub.2[CF.sub.2].sub.xCF.sub.3, CF.sub.3,
COC.sub.xF.sub.y, C.sub.xF.sub.yH.sub.z,
([CH.sub.2].sub.mO).sub.x(CH.sub.2CF.sub.2O).sub.y(CF.sub.2CF.sub.2O).sub-
.z(CF.sub.2).sub.2CF.sub.2CH.sub.2O(CH.sub.2).sub.pOH,
CH.sub.2C(OH)C.sub.xF.sub.yH.sub.z, C.sub.xF.sub.yH.sub.zO.sub.p,
COC.sub.xF.sub.yH.sub.z,
OCH.sub.2C.sub.xF.sub.z[C.sub.xF.sub.zO].sub.mF,
CH.sub.2C(CH.sub.3)CO.sub.2C.sub.xH.sub.z(CF.sub.2).sub.mCF.sub.3,
CH.sub.2(CF.sub.2O).sub.x(CF.sub.2CF.sub.2O).sub.y(CF.sub.2O).sub.zCF.sub-
.2CH.sub.2OH, COCF(CF.sub.3)--[CF(CF.sub.3)CF.sub.2O].sub.mF,
NHC.sub.xF.sub.yH.sub.zO.sub.p,
CH.sub.2CF.sub.2O[CF.sub.2CF.sub.2O].sub.m(CF.sub.2OCF.sub.2CH.sub.2OH,
COC.sub.xH.sub.z(CF.sub.2).sub.mCF.sub.3,
COCF.sub.2O[CF.sub.2CF.sub.2O].sub.nCF.sub.2OCF.sub.2CO.sub.2H,
([CH.sub.2].sub.mO).sub.x(CH.sub.2CF.sub.2O).sub.y(CF.sub.2CF.sub.2O).sub-
.zCF.sub.2CH.sub.2O(CH.sub.2).sub.pOH,
N[C.sub.xF.sub.yH.sub.z].sub.p,
C.sub.xH.sub.zCO.sub.2C.sub.xH.sub.z(CF.sub.2).sub.mCF.sub.3,
COC.sub.xF.sub.y[C.sub.pF.sub.zO].sub.mF, a luminescent residue, a
fluorescent residue, a fluorinated luminescent residue or a
fluorinated fluorescent residue and m, x, p, y, z are integers from
1 to 150, and where m is more preferably 10-100, and most
preferably 10-50, and where x, p, y, z are more preferably 10-75,
even more preferably 10-50, and most preferably 10-20. Acyl and
alkyl residues in the above formulas comprise lipophilic moieties,
including saturated and unsaturated aliphatic residues with C.sub.k
chains, where k is 2 to 100, more preferably 2-50, and most
preferably 2-20, and aryl residues comprise aromatic moieties,
including benzyl, biphenyl, phenyl polycyclic aromatics, and
heteroatom-containing aromatics. Y=saccharide branch residue
comprised of mono-, di-, oligo- or polysaccharide, fluorinated
saccharide branch residue comprised of mono-, di-, oligo- or
polysaccharide. Z=acyl, alkyl
[0027] The present compositions are comprised of modified
biopolymer that include biopolymers that are selected from the
group consisting of linear, branched, cyclic, ionic or neutral
glycans, such as acacia, agar, alginate, arabinogalactans,
arabinoxylans, carageenans, cyclodextrins, fructooligosaccharides,
fucoidan, gellan, galactomannans, glucomannans, inulins, pectin,
pullulan, tragacanth, xanthan, and xyloglucans, carboxyalkyl
glycans, such as carboxymethyl cellulose, carboxymethyl chitosan,
and carboxymethyl dextran, glycan esters, such as cellulose
acetate, cellulose phosphate, cellulose sulphate and starch
acetate, aminoglycans, such as chitin, chitosan, emulsan, and
poly(galactosamine), hydroxyalkyl glycans, such as hydroxyethyl
cellulose, hydroxypropyl cellulose, alkylglycans such as
ethylcellulose and methylcellulose, polysialic acid, and their
oligomers, hybrids, salts and derivatives.
[0028] Novel compounds of this invention include compositions
comprising modified biopolymers of general formula XIII as new
biomaterials, imaging probes, diagnostic tools and contrast agents:
##STR3## Where For Formula XIII: K=H, OH, X, OX, OZ, (Y).sub.f;
L=H, OH, X, OX, OZ, (Y).sub.f; W=H, (CH.sub.2).sub.d, CO.sub.2H,
CH, CX, X; T=H, OH, X, OX, OZ; V=(CH.sub.2).sub.d, CH.sub.2OX,
CH.sub.2OZ, CH.sub.2X, CH.sub.2NHX; S=H, O, X, NHX, (Y).sub.f; T=H,
O, X, NHX, (Y).sub.f; R.sub.1=H, X and d=1-3; f, n=1-1,500;
preferably f, n=100-1,000 Wherein Wherein for all of the above
Formulas X=fluoroalkyl, fluoroaryl, fluoroacyl, perfluoroalkyl,
perfluoroaryl, perfluoroacyl, perfluoropolymer, fluoroamine,
fluorocarbamate, fluorotriazine, fluorosulfonylalkyl derivatives,
F, CF.sub.3, CF.sub.2Cl, SO.sub.2[CF.sub.2].sub.xCF.sub.3,
COC.sub.xF.sub.y, C.sub.xF.sub.yH.sub.z,
([CH.sub.2].sub.mO).sub.x(CH.sub.2CF.sub.2O).sub.y(CF.sub.2CF.sub.2O).sub-
.z(CF.sub.2).sub.2CF.sub.2CH.sub.2O(CH.sub.2).sub.pOH,
CH.sub.2C(OH)C.sub.xF.sub.yH.sub.z, C.sub.xF.sub.yH.sub.zO.sub.p,
COC.sub.xF.sub.yH.sub.z,
OCH.sub.2C.sub.xF.sub.z[C.sub.xF.sub.zO].sub.mF,
CH.sub.2C(CH.sub.3)CO.sub.2C.sub.xH.sub.z(CF.sub.2).sub.mCF.sub.3,
CH.sub.2(CF.sub.2O).sub.x(CF.sub.2CF.sub.2O).sub.y(CF.sub.2O).sub.zCF.sub-
.2CH.sub.2OH, NHC.sub.xF.sub.yH.sub.zO.sub.p,
CH.sub.2CF.sub.2O[CF.sub.2CF.sub.2O].sub.m(CF.sub.2OCF.sub.2CH.sub.2OH,
COC.sub.xH.sub.z(CF.sub.2).sub.mCF.sub.3,
CO--CF.sub.2O[CF.sub.2CF.sub.2O].sub.nCF.sub.2OCF.sub.2CO.sub.2H,
COCF(CF.sub.3)--[CF(CF.sub.3)CF.sub.2O].sub.mF,
([CH.sub.2].sub.mO).sub.x(CH.sub.2CF.sub.2O).sub.y(CF.sub.2CF.sub.2O).sub-
.zCF.sub.2CH.sub.2O(CH.sub.2).sub.pOH,
N[C.sub.xF.sub.yH.sub.z].sub.p,
C.sub.xH.sub.zCO.sub.2C.sub.xH.sub.z(CF.sub.2).sub.mCF.sub.3,
COC.sub.xF.sub.y[C.sub.pF.sub.zO].sub.mF, a luminescent residue, a
fluorescent residue, a fluorinated luminescent residue or a
fluorinated fluorescent residue and m, x, p, y, z are integers from
1 to 150, and where m is more preferably 10-100, and most
preferably 10-50, and where x, p, y, z are more preferably 10-75,
even more preferably 10-50, and most preferably 10-20. Acyl and
alkyl residues in the above formulas comprise lipophilic moieties,
including saturated and unsaturated aliphatic residues with Ck
chains, where k is 2 to 100, more preferably 2-50, and most
preferably 2-20, and aryl residues comprise aromatic moieties,
including benzyl, biphenyl, phenyl polycyclic aromatics, and
heteroatom-containing aromatics. Y=amino acid residue, fluorinated
amino acid residue. Z=acyl, alkyl.
[0029] The novel compositions are comprised of modified biopolymers
that include biopolymers that are selected from the group
consisting of collagens, elastins, gelatins, poly(amino acids),
including poly(aspartic acid), poly(glutamic acid), and
poly(lysine), biopolyesters, including poly(glycolic acid,
poly(hydroxy alkanoates), and poly(lactic acid), their copolymers,
oligomers, hybrids, salts and derivatives.
[0030] Novel compositions of this invention also include compounds
comprising modified polymers of general formula XIV to XX and their
use as new biomaterials, imaging probes, diagnostic tools and
contrast agents: ##STR4## Where For Formula XIV: R.sub.1=H, X;
R.sub.2=H, Z, X. For Formula XV: R.sub.1=H, CH.sub.3,
(CH.sub.2).sub.mCH.sub.3, CF.sub.3, (CF.sub.2).sub.mCF.sub.3;
R.sub.2=H, CH.sub.3, (CH.sub.2).sub.mCH.sub.3, CF.sub.3,
(CF.sub.2).sub.mCF.sub.3; R.sub.3=H, CH.sub.3,
(CH.sub.2).sub.mCH.sub.3, CF.sub.3, (CF.sub.2).sub.mCF.sub.3;
R.sub.4=H, CH.sub.3, (CH.sub.2).sub.mCH.sub.3, CF.sub.3,
(CF.sub.2).sub.mCF.sub.3; R.sub.5=H, CH.sub.3,
(CH.sub.2).sub.mCH.sub.3, CF.sub.3, (CF.sub.2).sub.mCF.sub.3,
CH.sub.2X, CH.sub.2NHX; R.sub.6=H, CH.sub.3,
(CH.sub.2).sub.mCH.sub.3, CF.sub.3, (CF.sub.2).sub.mCF.sub.3,
CH.sub.2X, CH.sub.2NHX; m=1-10; n=1-3,000. For Formula XVI:
R.sub.1=OH, X, OX, OZ; R.sub.2=OH, X, OX, OZ; m, n=1-15,000,
preferably m, n=500-2,000. For Formula XVI: R.sub.1=O, CH.sub.2X,
CH.sub.2NHX, OZ; R.sub.2=CH.sub.3, (CH.sub.2).sub.mCH.sub.3;
R.sub.3=CH.sub.3, (CH.sub.2).sub.mCH.sub.3, CF.sub.3,
(CF.sub.2).sub.mCF.sub.3; R.sub.4=(CH.sub.2).sub.n;
R.sub.5=(CH.sub.2).sub.q; Rr=H, OH, OX, X; R.sub.7=H, OH, OX, X; m,
n=1-30; p, q=0-3,000; preferably p, q=50-500. For Formula XVII:
R.sub.1=O, OX, CF.sub.3, (CF.sub.2).sub.mCF.sub.3;
R.sub.2=CH.sub.3, (CH.sub.2).sub.nCH.sub.3, CF.sub.3,
(CF.sub.2).sub.nCF.sub.3; R.sub.3=CH.sub.3,
(CH.sub.2).sub.nCH.sub.3, CF.sub.3, (CF.sub.2).sub.nCF.sub.3;
R.sub.4=(CH.sub.2).sub.m; R.sub.5=(CH.sub.2).sub.q; R.sub.6=OH, OX,
X; R.sub.7=H, X; m=1-2; n=1-10; p, q=1-3,000, preferably p,
q=100-1,000. For Formula XVIII: R.sub.1=H, CH.sub.3,
(CH.sub.2).sub.mCH.sub.3, CF.sub.3, (CF.sub.2).sub.mCF.sub.3;
R.sub.2=H, OH, OX, X; R.sub.3=H, OH, OX, X; m=1-30; n=1-3,000,
preferably n=500-1,000. For Formula XIX: R.sub.1=CH.sub.2,
CH.sub.2CH.sub.2, CF.sub.2, CF.sub.2CF.sub.2; R.sub.2=CH.sub.2,
CF.sub.2; R.sub.3=OH, OX, OZ; R.sub.4=H, X, Z; where
Z=Y[(OCH.sub.2CH.sub.2).sub.m].sub.q; Y=multidentate core, such as
trivalent or tetravalent residues; m, n=1-80,000, preferably m,
n=1,000-20,000; q=1-10. For Formula XX: R.sub.1=H, X; R.sub.2=H, X,
(CH.sub.2CH.sub.2N).sub.m, (CH.sub.2CH.sub.2NX).sub.m; R.sub.3=H,
X, (CH.sub.2CH.sub.2N).sub.m, (CH.sub.2CH.sub.2NX).sub.m;
R.sub.4=H, X, (CH.sub.2CH.sub.2N).sub.mCH.sub.2CH.sub.2NH.sub.2,
(CH.sub.2CH.sub.2N).sub.mCH.sub.2CH.sub.2NHX; m=1-3,000;
n=5-80,000, preferably n=500-15,000. Wherein for all of the above
Formulas [0031] X=fluoroalkyl, fluoroaryl, fluoroacyl,
perfluoroalkyl, perfluoroaryl, perfluoroacyl, perfluoropolymer,
fluoroamine, fluorocarbamate, fluorotriazine, fluorosulfonylalkyl
derivatives, F, CF.sub.3, COC.sub.xF.sub.y, C.sub.xF.sub.yH.sub.z,
([CH.sub.2].sub.gO).sub.x(CH.sub.2CF.sub.2O).sub.y(CF.sub.2CF.sub.2O).sub-
.z(CF.sub.2).sub.2CF.sub.2CH.sub.2O(CH.sub.2).sub.pOH,
CH.sub.2C(OH)C.sub.xF.sub.yH.sub.z, C.sub.xF.sub.yH.sub.zO.sub.p,
COC.sub.xF.sub.yH.sub.z,
OCH.sub.2C.sub.xF.sub.z[C.sub.xF.sub.zO].sub.gF,
CH.sub.2C(CH.sub.3)CO.sub.2C.sub.xH.sub.z(CF.sub.2).sub.gCF.sub.3,
CH.sub.2(CF.sub.2O).sub.x(CF.sub.2CF.sub.2O).sub.y(CF.sub.2O).sub.zCF.sub-
.2CH.sub.2OH, CF.sub.2Cl, SO.sub.2[CF.sub.2].sub.xCF.sub.3,
NHC.sub.xF.sub.yH.sub.zO.sub.p,
CH.sub.2CF.sub.2O[CF.sub.2CF.sub.2O].sub.g(CF.sub.2OCF.sub.2CH.sub.2OH,
COC.sub.xH.sub.z(CF.sub.2).sub.gCF.sub.3,
CO--CF.sub.2O[CF.sub.2CF.sub.2O].sub.nCF.sub.2OCF.sub.2CO.sub.2H,
CO--CF(CF.sub.3)--[CF(CF.sub.3)CF.sub.2O].sub.gF
([CH.sub.2].sub.gO).sub.x(CH.sub.2CF.sub.2O).sub.y(CF.sub.2CF.sub.2O).sub-
.zCF.sub.2CH.sub.2O(CH.sub.2).sub.pOH,
N[C.sub.xF.sub.yH.sub.z].sub.p,
C.sub.xH.sub.zCO.sub.2C.sub.xH.sub.z(CF.sub.2).sub.gCF.sub.3,
COC.sub.xF.sub.y[C.sub.pF.sub.zO].sub.mF, a luminescent residue, a
fluorescent residue, a fluorinated luminescent residue or a
fluorinated fluorescent residue and g, x, p, y, z are integers from
1 to 150, and where g is more preferably 10-100, and most
preferably 10-50, and where x, p, y, z are more preferably 10-75,
even more preferably 10-50, and most preferably 10-20. Acyl and
alkyl residues in the above formulas comprise lipophilic moieties,
including saturated and unsaturated aliphatic residues with C.sub.k
chains, where k is 2 to 100, more preferably 2-50, and most
preferably 2-20, and aryl residues comprise aromatic moieties,
including benzyl, biphenyl, phenyl polycyclic aromatics, and
heteroatom-containing aromatics. [0032] Z acyl, alkyl.
[0033] The novel compositions are comprised of modified polymers
that include polymers that are selected from the group consisting
of biocompatible polymers, including poly(acrylates),
poly(acrylamides), poly(alkylene glycols), including poly(ethylene
glycols), poly(ethylene oxides) and poly(propylene glycols),
poly(allylamines), poly(butadienes), poly(caprolactones),
poly(ethylene imines), poly(methacrylates), poly(orthoesters),
poly(tetrahydrofurans), poly(vinyl pyrrolidones), poly(vinyl
acetates), and poly(vinyl alcohols), their copolymers, oligomers,
hybrids, salts and derivatives and where acyl and alkyl residues of
this disclosure comprise lipophilic moieties, including saturated
and unsaturated aliphatic residues with Ck chains, where k is 2 to
100, more preferably 2-50, and most preferably 2-20, and where aryl
residues comprise aromatic moieties, including benzyl, biphenyl,
phenyl, polycyclic aromatics, and heteroatom-containing
aromatics.
[0034] The fluorinated and/or paramagnetic polymers of the present
invention are used to improve the imaging available in an MRI
examination procedure. The method of improving the effectiveness of
magnetic resonance imaging (MRI) comprises: [0035] a. administering
an effective amount of one or more fluorinated and/or paramagnetic
polymers of claims 14 to a patient; [0036] b. subjecting the
patient to an MRI of a tissue/organ where the administered polymer
is expected to accumulate; and [0037] c. evaluating the
tissue/organ from the MRI images obtained.
[0038] One of ordinary skill in the art would readily be able to
evaluate the results of the MRI by observation and/or comparisons
to a patient's prior MRI results or standard MRI results used for
diagnosis purposes.
DETAILED DESCRIPTION OF THE INVENTION
[0039] Compositions of this invention comprised of carbohydrate,
polymer and protein residues are obtained by treating the
respective starting materials (backbone or substrate moiety) with
fluorine moieties employing routine fluorination chemistry such as
those described below.
[0040] Three general approaches can be employed to prepare the new
fluorinated biopolymers of the instant invention: (1) low molecular
weight fluorinated substituents can be employed (as illustrated in
the Examples); (2) high molecular weight polyfluorinated residues
can be employed, such as with functional perfluoropolymers; and (3)
fluorinated monomers can be incorporated into polymeric materials
by chemical or enzymatic processes. The above approaches permit the
preparation of fluorinated biopolymers with a broad range of
fluorine substituent types and incorporation levels (5-40% or more
as illustrated in the following Examples) that can be tailored to
either diagnostic or therapeutic uses. The optimum fluorine content
will be determined in each case by the diagnostic requirements for
sensitivity on one hand and the extent to which the maximum
fluorine substitution does not interfere with the probe's
biological or physicochemical properties, e.g., its solubility or
receptor binding ability. An important parameter in these
considerations will be the type of fluorine substitution and its
position on the probe substrate. Generally preferable F levels are
1040%, and more preferable 20-40%.
[0041] Linking of fluorinated residues to biopolymer and polymer
starting materials of this invention can be accomplished by a
number of reactions, many of which have been described generally in
conjugate chemistry (for reviews see, for instance: G. T.
Hermanson, Bioconjugate Chemistry, Academic Press, New York, 1996;
S. S. Wong, Chemistry of protein conjugation and cross-linking, CRC
Press, Boca Raton, 1993; R. L. Lundblad, Techniques in Protein
Modification, CRC Press, Boca Raton, 1994; C. F. Meares (ed.),
Perspectives in Bioconjugate Chemistry, American Chemical Society,
Washington, 1993).
[0042] A terminal hydroxyl group on the biopolymers and polymers of
this disclosure can be allowed to react with bromoacetyl chloride
to form a bromoacetyl ester that in turn is allowed to react with
an amine precursor to form the --NH--CH.sub.2--C(O)-- linkage. A
terminal hydroxyl group also can be allowed to react with
1,1'-carbonyl-bisimidazole and this intermediate in turn allowed to
react with an amino precursor to form a --NH--C(O)O-- linkage (see
Bartling et al., Nature, 243, 342, 1973). A terminal hydroxyl also
can be allowed to react with a cyclic anhydride such as succinic
anhydride to yield a half-ester which, in turn, is allowed to react
with a precursor of the formula C.sub.xF.sub.yH, --NH.sub.2 using
conventional peptide condensation techniques such as
dicyclohexylcarbodiimide, diphenylchlorophosphonate, or
2-chloro-4,6-dimethoxy-1,3,5-triazine (see e.g., Means et al.,
Chemical Modification of Proteins, Holden-Day, 1971). A terminal
hydroxyl group can also be allowed to react with 1,4-butanediol
diglycidyl ether to form an intermediate having a terminal epoxide
function linked to the polymer through an ether bond. The terminal
epoxide function, in turn, is allowed to react with an amino or
hydroxylprecursor (Pitha et al., Eur. J. Biochem., 94, 11, 1979;
Elling and Kula, Biotech. Appl. Biochem., 13 354, 1991; Stark and
Holmberg, Biotech. Bioeng., 34, 942, 1989).
[0043] Halogenation of a hydroxyl group permits subsequent reaction
with an alkanediamine such as 1,6-hexanediamine. The resulting
product then is allowed to react with carbon disulfide in the
presence of potassium hydroxide, followed by the addition of
proprionyl chloride to generate a isothiocyanate that in turn is
allowed to react with an amino precursor to yield a
--N--C(S)--N--(CH.sub.2).sub.6--NH-- linkage (see e.g., Means et
al., Chemical Modification of Proteins, Holden-Day, 1971).
[0044] A carboxylic acid group of the biopolymers and polymers can
be activated with N,N'-dicyclohexylcarbodiimide and then allowed to
react with an amino or hydroxyl group to form an amide or ether
respectively. Anhydrides and acid chlorides will produce the same
links with amines and alcohols. Alcohols can be activated by
carbonyldiimidazole and then linked to amines to produce urethane
linkages. Alkyl halides can be converted to an amine or allowed to
react with an amine, diamines, alcohols, or diols. A hydroxy group
can be oxidized to form the corresponding aldehyde or ketone. This
aldehyde or ketone then is allowed to react with a precursor
carrying a terminal amino group to form an imine that, in turn, is
reduced with sodium borohydride or sodium cyanoborohydride to form
the secondary amine (see Kabanov et al., J. Controlled Release, 22,
141, 1992; Methods Enzymology, XLVII, Hirs & Timasheff, eds.,
Academic Press, 1977). The precursor terminating in an amino group
can also be allowed to react with an alkanoic acid or fluorinated
alkanoic acid, preferably an activated derivative thereof, such as
an acid chloride or anhydride, to form a linking group --CONH--.
Alternatively, an amino precursor can be treated with an
.alpha.-.omega.-diisocyanoalkane to produce a
--NC(O)NH(CH.sub.2).sub.6NHC(O)--N-- linkage (see Means, Chemical
Modification of Proteins, Holden-Day, 1971). Furthermore, linkages
that are unsymmetrical, such as --CONH-- or --NHCOO--, can be
present in the reverse orientation; e.g., --NHCO-- and --OCONH--,
respectively. Examples of an activated carbonyl group include
anhydride, ketone, p-nitrophenylester, N-hydroxysuccinimide ester,
pentafluorophenyl ester and acid chloride.
[0045] Suitable fluorinated starting materials for making the novel
compositions of the present invention include, but are not limited
to inorganic fluorinating agents, such as
trifluoromethylhypofluorite, sulfur tetrafluoride or potassium
fluoride, organic fluorinating agents, such as Selecffluor.TM.,
fluoroalkylcarboxylic acids, fluoroalkylaldehydes, anhydrides,
esters, ketones, acid chlorides of fluoroalkylcarboxylic acids,
such as monofluoroacetic acid, difluoroacetic acid, trifluoroacetic
acid, pentafluoro-propionic acid, heptafluorobutyric acid,
heptafluorobutyric anhydride, heptafluorobutyrylchloride,
nonafluoropentanoic acid, tridecafluoroheptanoic acid,
pentadecafluorooctanoic acid, heptadecafluorononanbic acid,
nonadecafluorodecanoic acid, perfluorododecanoic acid,
perfluorotetradecanoic acid; fluoroalkanols, such as
2,2,3,3,4,4,4-heptafluoro-1-butanol,
2,2,3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,11,11,11-heneicosafluoro-1-undecano-
l, 2,2,3,3,4,4,5,5,6,6,7,7,8,9,9,9-hepta-decafluoro-1-nonanol,
2,2,3,3,4,4,5,5,6,6,7,7,8,8,8-penta-decafluoro-1-octanol,
2,2,3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-nonadeca-fluoro-1-decanol,
Krytox and Zonyl derivatives, fluoroarylesters, fluoroalkylamines,
fluoroarylamines, fluorinated polymers containing reactive terminal
groups, fluoroalkyl halides, such as perfluoroethyl iodide,
perfluoropropyl iodide, perfluorohexyl bromide, perfluoroheptyl
bromide, perfluorooctyl bromide, perfluorodecyl iodide,
perfluorooctyl iodide,
1,1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8-heptadecafluoro-10-iododecane,
1,1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8-heptadecafluoro-10-iododecane,
polytetrafluoroethyleneoxide-co-difluoromethyleneoxide-co-bis(methylcarbo-
xylate), dihydroxy-propanoxymethyl derivatives of
perfluoropolyoxyalkane, hydroxypolyethylenoxy derivatives of
perfluoropolyoxyalkane and the like. Suitable modification
procedures have been described in several monographs (J. J. Clark,
D. Walls. T. W. Bastock, Aromatic Fluorination, CRC Press, Boca
Raton, Fla., 1996; M. Hudlicky, A. E. Pavlath, Chemistry of Organic
Fluorine Compounds, ACS, Washington, D.C. 1995; M. Howe-Grant ed.,
Fluorine Chemistry, A Comprehensive Treatment, Wiley, New York,
1995; G. A. Olah, G. K. Sarya Prakash, R. D. Chambers, eds.
Synthetic Fluorine Chemistry, Wiley, New York, 1992).
[0046] The present invention also contemplates paramagnetic
polymers for those polymers capable of forming salts or conjugates
with paramagnetic ions. Suitable paramagnetic ions include any
paramagnetic ion of the transition metal or lanthanide series,
including gadolinium (III), iron (III), manganese (II and III),
chromium (III), copper (II), dysprosium (III), terbium (III),
holmium (III), erbium (III), and europium (III); most preferred are
gadolinium (III), dysprosium (III), iron (III), and manganese (II).
The magnetic materials of this invention can be used as
contrast-enhancing agents for in vivo MR imaging and magnetic
resonance angiography.
[0047] Specific compounds of Formulas I-XX may require the use of
protecting or blocking groups to enable their successful
elaboration into the desired structure. Protecting groups may be
chosen with reference to Greene, T. W., et al., Protective Groups
in Organic Synthesis, John Wiley & Sons, Inc., 1991. The
blocking groups are readily removable, i.e., they can be removed,
if needed, by procedures that will not cause cleavage or other
disruption of the remaining portions of the molecule. Such
procedures include chemical and enzymatic hydrolysis, treatment
with chemical reducing or oxidizing agents under mild conditions,
treatment with fluoride ion, treatment with a transition metal
catalyst and a nucleophile, and catalytic hydrogenation.
[0048] Examples of suitable hydroxylprotecting groups are:
trimethylsilyl, triethylsilyl, o-nitrobenzyloxycarbonyl,
p-nitrobenzyloxycarbonyl, t-butyldiphenylsilyl,
t-butyldimethylsilyl, benzyloxycarbonyl, t-butyloxycarbonyl,
2,2,2-trichloroethyloxycarbonyl, and allyloxycarbonyl. Examples of
suitable carboxylprotecting groups are benzhydryl, o-nitrobenzyl,
p-nitrobenzyl, 2-naphthylmethyl, allyl, 2-chloroallyl, benzyl,
2,2,2-trichloroethyl, trimethylsilyl, t-butyidimethylsilyl,
t-butyldiphenylsilyl, 2-(trimethylsilyl) ethyl, phenacyl,
p-methoxybenzyl, acetonyl, p-methoxyphenyl, 4-pyridylmethyl and
t-butyl.
[0049] The compounds used in the method of the invention can be
prepared readily according in the following detailed examples using
readily available starting materials, reagents and conventional
synthetic procedures. Additional variants are also possible that
are known to those of ordinary skill in this art, but which are not
mentioned in greater detail. The following examples illustrate the
practice of the present invention but should not be construed as
limiting its scope.
EXAMPLE 1
N-[3-[2-(Perfluorohexyl)-2-ethoxy]-2-hydroxypronyl].gamma.-Polyplutamic
Acid
[0050] A solution of
3-[2-(perfluorohexyl)-2-ethoxy]-1,2-epoxypropane in methylene
chloride (0.6 equivalents) was added to .gamma.-polyglutamic acid
and stirred at ambient temperature for 6 hours. The suspension was
filtered, washed with methylene chloride and acetone, dialyzed and
dried, yielding 3-[2-(perfluorohexyl)-2-ethoxy]-2-hydroxypropyl
.gamma.-polyglutamic acid with F 13.38%. ##STR5##
EXAMPLE 2
3-[2-(Perfluorohexyl)-2-ethoxy]-2-hydroxypropyl Hyaluronic Acid
[0051] An aqueous solution of hyaluronic acid was treated with
3-[2-(perfluorohexyl)-2-ethoxy]-1,2-epoxypropane (0.6 equivalents)
and the resulting viscous paste was stirred at ambient temperature
for 6 hours. The reaction mixture was precipitated with acetone,
washed with acetone, filtered, dialyzed and dried, yielding
3-[2-(perfluorohexyl)-2-ethoxy]-2-hydroxypropyl hyaluronate with F
33.12%. ##STR6##
EXAMPLE 3
3-[2-(Perfluorohexyl)-2-ethoxy]-2-hydroxypropyl Maltodextrin
[0052] An aqueous solution of maltodextrin was treated with NaOH
(1.3 equivalents) and then with
3-[2-(perfluorohexyl)-2-ethoxy]-1,2-epoxypropane in DMSO (0.6
equivalents) and the resulting viscous paste was stirred at ambient
temperature for 6 hours. The reaction mixture was precipitated with
acetone, washed with acetone, filtered, dialyzed and dried,
yielding 3-[2-(perfluorohexyl)-2-ethoxy]-2-hydroxypropyl
maltodextrin with F 21.52%. ##STR7##
EXAMPLE 4
Perfluorophenylhydrazone Carboxymethyl Cellulose
[0053] A solution of perfluorophenylhydrazine in DMSO (0.6
equivalents) was added to an aqueous solution of carboxymethyl
cellulose and stirred at ambient temperature for 6 hours. The
suspension was filtered, washed with methylene chloride and
acetone, dialyzed and dried, yielding perfluorophenylhydrazone CMC
with F 18.94%. ##STR8##
EXAMPLE 5
Di-.alpha.,.omega.-(heptafluorobutyryl) Polyethylene Glycol
[0054] A solution of heptafluorobutyryl chloride in dioxane (0.6
equivalents) was added to polyethylene glycol (Mw 1,000) in dioxane
containing triethylamine (0.6 equivalents) and stirred at ambient
temperature for 6 hours. The reaction mixture was precipitated in
ether, and the crude fluorinated PEG product was chromatographed on
silica gel, yielding heptafluorobutyryl PEG with F 19.95%.
CH.sub.2F(CF.sub.2).sub.3[OCH.sub.2CH.sub.2].sub.nO(CF.sub.2).sub.3CH.sub-
.2F n=1-50,000
EXAMPLE 6
Perfluoroaniline Hyaluronate
[0055] A solution of perfluoroaniline (1.6 equivalents) in DMSO was
added to an aqueous solution of sodium hyaluronate and stirred at
40.degree. C. for 4 hours. The reaction mixture was cooled, treated
with sodium cyanoborohydride (10 equivalents) for 9 hours,
precipitated with acetone, washed with acetone, filtered, dialyzed
and dried, yielding perfluoroaniline hyaluronate with F 17.75%.
##STR9##
EXAMPLE 7
Perfluorohexanoate Hyaluronate
[0056] Sodium hyaluronate was dissolved in water and the pH of the
solution was adjusted to pH 4.75 by addition of 0.1 N HCl. Then EDC
(1.5 equivalents) was added followed by methylperfluorohexanoate
methyl ester (1.05 equivalents). The pH of the reaction mixture
then rises to 6.2 over two hours. The reaction mixture was kept at
room temperature for five hours, after which it forms a thick
insoluble hydrogel. This hydrogel is dialyzed with a 1 M NaCl
solution and lyophilized to yield perfluorohexanoate hyaluronate F
29.52%. ##STR10##
EXAMPLE 8
Trifluoroacetate Hydroxypropyl Cellulose
[0057] A solution of ethyl trifluoroacetate in pyridine (1.6
equivalents) was added to a pyridine solution of hydroxypropyl
cellulose and stirred at ambient temperature for 9 hours. The
solution was precipitated in ice water, filtered, washed with
methanol and acetone, dialyzed and dried, yielding trifluoroacetate
hydroxypropyl cellulose with F 34.56%. ##STR11##
EXAMPLE 9
3-(Perfluoro-n-octyl)-2-hydroxypropyl Hyaluronic Acid
[0058] An aqueous solution of hyaluronic acid was treated with
3-(perfluoro-n-octyl)-1,2-epoxypropane (1.2 equivalents) and the
resulting viscous paste was stirred at ambient temperature for 6
hours. The reaction mixture was precipitated with acetone, washed
with acetone, filtered, dialyzed and dried, yielding
3-(perfluoro-n-octyl)-2-hydroxypropyl hyaluronate with F 31.52%.
##STR12##
EXAMPLE 10
Perfluoro-2,5,8,11-tetramethyl-3,6,9,12-tetraoxopentadecanoate
Hyaluronic Acid
[0059] An aqueous solution of hyaluronic acid was treated with
perfluoro-2,5,8,11-tetramethyl-3,6,9,12-tetraoxopentadecanoic acid
methyl ester (1.2 equivalents) and catalytic amounts of sulfuric
acid and the resulting viscous paste was stirred at ambient
temperature for 14 hours. The reaction mixture was washed with
acetone, filtered, dialyzed and dried, yielding
perfluoro-2,5,8,11-tetramethyl-3,6,9,12-tetraoxopentadecanoate
hyaluronate with F 34.65%. ##STR13##
EXAMPLE 11
4,4,4-Trifluoro-3-hydroxy-3-(trifluoromethyl)butanoate
Hydroxypropyl Cellulose
[0060] An aqueous solution of
4,4,4-trifluoro-3-hydroxy-3-(trifluoromethyl)butyric acid (3.2
equivalents) was acidified to pH 4.75 and treated with EDC (3.5
equivalents) followed by hydroxypropyl cellulose in water and the
resulting viscous paste was stirred at ambient temperature for 14
hours. The reaction mixture was dialyzed and lyophilized, yielding
4,4,4-trifluoro-3-hydroxy-3-(trifluoromethyl)butanoate
hydroxypropyl cellulose with F 28.76%. ##STR14##
EXAMPLE 12
4,4,4-Trifluoro-3-hydroxy-3-(trifluoromethyl)butanoate Dextran
[0061] An aqueous solution of
4,4,4-trifluoro-3-hydroxy-3-(trifluoromethyl)butyric acid (4.2
equivalents) was acidified to pH 4.75 and treated with EDC (4.5
equivalents) followed by dextran (Mw 500,000) in water and the
resulting paste was stirred at ambient temperature for 14 hours.
The reaction mixture was precipitated with acetone, filtered,
redissolved in water, dialyzed and lyophilized, yielding
4,4,4-trifluoro-3-hydroxy-3-(trifluoromethyl)butanoate dextran with
F 24.57%. ##STR15##
EXAMPLE 13
Trifluoroacetate Collagen
[0062] A solution of ethyl trifluoroacetate in pyridine (1.6
equivalents) was added to a pyridine solution of collagen and
stirred at ambient temperature for 6 hours. The solution was
dialyzed and lyophilized, yielding trifluoroacetate collagen with F
23.16%. ##STR16## wherein m+n+p=.about.0.5
EXAMPLE 14
Trifluoroacetate Poly(vinyl alcohol)
[0063] A solution of trifluoroacetic anhydride in pyridine (1.2
equivalents) was added to a pyridine solution of poly(vinyl
alcohol) (Mw 30,000) and stirred at ambient temperature for 6
hours. The solution was precipitated with ice water, dialyzed and
lyophilized, yielding trifluoroacetate PVA with F 39.98%.
##STR17##
EXAMPLE 15
Heptafluorobutyryl Polyethylene Imine
[0064] A solution of heptafluorobutyryl chloride in DMSO (2.6
equivalents) was added to a DMSO solution of polyethylene imine (Mw
70,000) containing triethylamine (2.6 equivalents) and stirred at
ambient temperature for 6 hours. The reaction mixture was dialyzed,
yielding heptafluorobutyryl PEI with F 45.35%. ##STR18## wherein
m+n+p+v=1
EXAMPLE 16
Superparamagnetic Iron Oxide Hyaluronate Particles
[0065] To a stirred dispersion of superfine iron oxide particles (3
nanometer, 0.5 g) in water (50 mL) was added an aqueous solution of
sodium hyaluronate (50 mg, 5 mL). The dispersion was sonicated,
centrifuged and the supernatant filtered through 0.22..mu.m filter.
A magnetization curve revealed that the hyaluronate particles were
superparamagnetic.
EXAMPLE 17
Paramagnetic Gadolinium Hyaluronate Beads
[0066] To a rapidly stirred, aqueous solution of gadolinium (III)
acetate (1.1 equivalents) was added dropwise an aqueous solution of
sodium hyaluronate through a syringe. The resulting gel beads were
centrifuged, dialyzed and lyophilized. A magnetization curve
revealed that the hyaluronate particles were paramagnetic.
Use of New Biocompatible Materials and Probes
[0067] The fluorine-modified biopolymers of the instant invention
are useful as diagnostic tools (MRI, NMR and the like). As
illustrated by the Examples, the methods of the instant invention
permit the preparation of diagnostic agents with dual
functionalities. Thus, the simultaneous incorporation of .sup.19F
or superparamagnetic residues and fluorescent moieties into
biopolymers and polymers affords diagnostic probes that can be
employed for both MRI and fluorescent studies. Examples of such
dual function diagnostic probes are those biopolymers and polymers
that contain both a fluorine moiety as described herein and a
fluorescent moiety or a fluorinated fluorescent moiety such as:
4-trifluoromethyl-7-aminocoumarin, 4-trifluoromethyl-umbelliferone
(or its acetate or butyrate derivatives),
4-fluoro-7-sulfamyl-benzofurazam, certain BODIPY dyes, e.g.,
N-(4,4'-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)-methyl
iodoacetamide,
N-(4,4'-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-2-yl)--
iodoacetamide and
4,4'-difluoro-5-phenyl-4-bora-3a,4a-diaza-s-indacene-3-propionic
acid,
3-chloro-1-(3-chloro-5-(trifluoromethyl)-2-pyridimyl)-5-(trifluoromethyl)-
-2[1H]-pyridinone,
6-carboxymethylthio-2',4,'5,7'-tetrabromo-4,5,7-trifluorofluorescein
(Eosin F3S), and Oregon Green carboxylic acid.
[0068] The fluorinated polymers of the present invention display
sensitivity in their T.sub.1 relaxation times to different oxygen
partial pressures (pO.sub.2), producing linear correlation over a
range of pO.sub.2. This demonstrates their potential utility as
oxygen sensitive imaging probes. The fluorinated polymers also
display chemical shift and temperature sensitivity, indicating
their utility as temperature sensitive imaging probes. These novel
agents of this invention are suitable for many diagnostic uses, and
provide the ability to image in vivo or non-invasively monitor
tissues, organs and cellular implants, for example, pancreatic
islet .beta.-cells that are encapsulated with the present
fluorinated polymers, and measure their mass, function, viability
or evidence of inflammation. Additionally, engraftment of
transplanted isolated pancreatic islets can be monitored, using,
for example, islets labeled with .beta.-cell specific
oxygen-sensitive fluorinated probes. .sup.19F-MRI with these novel
agents permits monitoring of other disorders, such as cancer, the
comparison of normal or diseased cells, organs or tissues, the
viability of transplanted cells or other tissues when those
fluorinated agents have specificity for target tissues. This new
methodology is instrumental in the development of clinical
examinations for monitoring disease progress and response to
therapy in diabetics and in people strongly at risk for diabetes
and other patient populations.
[0069] The paramagnetic polymers of the present invention are used
as contrast agents and are administered orally (or elsewhere in the
gastrointestinal tract), intravascularly or intraperitoneally in
physiological buffer or other physiologically acceptable carriers.
The dosage depends on the sensitivity of the NMR imaging
instrumentation and on the composition of the contrast agent. Thus,
a contrast agent containing a highly paramagnetic substance, e.g.,
gadolinium (III), generally requires a lower dosage than a contrast
agent containing a paramagnetic substance with a lower magnetic
moment, e.g., iron (III). In general, dosage will be in the range
of about 0.001-1 mmol/kg, more preferably about 0.01-0.1 mmol/kg.
In one embodiment, the products are dispersed in a suitable
injection medium, such as distilled water or normal saline, to form
a dispersoid that is introduced into the subject's vascular system
by intravenous injection. The particles are then carried through
the vascular system to the target organ where they are taken
up.
[0070] When intravascularly administered, the paramagnetic
compounds will be preferentially taken up by organs which
ordinarily function to cleanse the blood of impurities, notably the
liver, spleen, and lymph nodes, and the other organs which tend to
accumulate such impurities, notably bone and neural tissue and to
some extent, lung. In each of these organs and tissues, the uptake
into the reticuloendothelial cells will occur by phagocytosis,
wherein the paramagnetic compounds enter the individual cells in
membrane-bound vesicles; this permits a longer half-life in the
cells, as such membrane-bound paramagnetic compounds will not tend
to clump or aggregate (aggregates are rapidly metabolized and
cleared from the organ/tissue). Other uptake mechanisms are
possible, e.g., pinocytosis. Also, it is possible that the other
cells of the liver (hepatocytes) may absorb the paramagnetic
componds. Because cancerous tumor cells can lack the ability of
phagocytic uptake, the intravascularly administered particles can
serve as valuable tools in the diagnosis of cancer in the
above-mentioned organs, as tumors will be immediately
distinguishable on any image obtained. In another embodiment, the
paramagnetic compounds are administered as dispersoids into the
gastrointestinal tract, which includes the esophagus, stomach,
large and small intestine, either orally, by intubation, or by
enema, in a suitable medium such as distilled water. The
paramagnetic compounds are preferentially absorbed by the cells of
the tract, especially those of the intestine and, like the
intravascularly introduced paramagnetic compounds, will exert an
effect on T.sub.2 of the organ or tissue. In this manner, cancers
and other debilitating diseases of the digestive system such as
ulcers can be diagnosed and affected areas pinpointed.
[0071] The new compositions of this disclosure also display unusual
surfactant and emulsification properties.
[0072] All patents, patent applications, provisional applications,
and publications referred to or cited herein are incorporated by
reference in their entirety, including all figures and tables, to
the extent they are not inconsistent with the explicit teachings of
this specification.
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Pat. No. 6,054,492 dated Apr. 25, 2000 Fluorinated copolymeric
pharmaceutical adjuncts [0107] INVENTOR(S)-- Kabanov, Alexander V.;
Vinogradov, Serguei V. [0108] PATENT ASSIGNEE(S)--Supratek Pharma
Inc.
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