Texaphyrin compounds having improved functionalization

Texaphyrin metal complexes having improved functionalization include the addition of electron-donating groups to positions 2, 7, 12, 15, 18 and/or 21 and/or the addition of electron-withdrawing groups to positions 15 and/or 18 of the macrocycle. Electron-donating groups at positions 2, 7, 12, 15, 18 and/or 21 contribute electrons to the aromatic .pi. system of the macrocycle which stabilizes the metal complex to demetallation and the imine bonds to hydrolysis. The addition of substituents to the 12 and 21 positions of the macrocycle offer steric protection for the imine bonds against possible in vivo enzyme hydrolysis. Electron-withdrawing groups at positions 15 and/or 18 render the macrocycle more readily reduced, i.e. the redox potential is lower and the macrocycle more readily gains an electron to form a radical. Such texaphyrins having a low redox potential and imine bond stabilization are useful in a variety of applications.

FIELD OF THE INVENTION 
The present invention relates to the field of expanded porphyrins, in 
particular, to texaphyrins having improved functionalization. 
BACKGROUND OF THE INVENTION 
Certain texaphyrin compounds are described in U.S. Pat. Nos. 4,935,498, 
5,162,509, 5,252,720, 5,272,142 and 5,256,399, each of which is 
incorporated by reference herein. "Texaphyrin" refers to a particular 
"expanded porphyrin" pentadentate macrocyclic ligand. The compound is 
capable of existing in both its free-base form and of supporting the 
formation of a 1:1 complex with a variety of metal cations, such as 
Cd.sup.2+, Hg.sup.2+, In.sup.3+, Y.sup.3+, Nd.sup.3+, Eu.sup.3+, 
Sm.sup.3+, La.sup.3+, Lu.sup.3+, Gd.sup.3+, and other cations of the 
lanthanide series that are too large to be accommodated in a stable 
fashion within the 20% smaller tetradentate binding core of the 
well-studied porphyrins. 
Large, or "expanded" porphyrin-like systems are of interest for several 
reasons: They could serve as aromatic analogues of the better studied 
porphyrins or serve as biomimetic models for these or other naturally 
occurring pyrrole-containing systems. In addition, large 
pyrrole-containing systems offer possibilities as novel metal binding 
macrocycles. For instance, suitably designed systems could act as 
versatile ligands capable of binding larger metal cations and/or 
stabilizing higher coordination geometries than those routinely 
accommodated within the normally tetradentate ca. 2.0 .ANG. radius 
porphyrin core. The resulting complexes could have important application 
in the area of heavy metal chelation therapy, serve as contrast agents for 
magnetic resonance imaging (MRI) applications, act as vehicles for 
radioimmunological labeling work, or serve as new systems for extending 
the range and scope of coordination chemistry. 
The desirable properties of texaphyrins are: 
1) appreciable solubility, particularly in aqueous media; 
2) biolocalization in desired target tissue; 
3) low intrinsic toxicity; 
4) the ability to attach to solid matrices; 
5) the ability to be attached to biomolecules; 
6) efficient chelation of divalent and trivalent metal cations; 
7) absorption of light in the physiologically important region of 690-880 
nm; 
8) high chemical stability; 
9) ability to stabilize diamagnetic complexes that form long-lived triplet 
states in high yield and that act as efficient photosensitizers for the 
formation of singlet oxygen; 
10) ability to chelate Gd(III) for magnetic resonance imaging; 
11) a redox potential lower than that of oxygen for use as a 
radiosensitizer. 
One of the disadvantages of the texaphyrin metal complexes of prior patents 
is their short half-life. The Y.sup.3+ and In.sup.3+ complexes of the 
basic texaphyrin have half-lives for decomplexation and/or ligand 
decomposition of about 3 weeks in 1:1 methanol-water mixtures. While such 
stability is adequate for some in vitro or in vivo applications, a greater 
degree of stability in aqueous solution is desirable. For example, a 
desired solution-phase shelf life of 2-3 years would facilitate the 
formulation of texaphyrin metal complexes as pharmaceutical products. The 
new molecules of the present invention address the problems of 
demetallation of the texaphyrin metal complex and the susceptibility of 
the imine bonds of the macrocycle to hydrolysis. The solution to these 
problems is expected to provide a texaphyrin which has a more desirable 
shelf life. 
SUMMARY OF THE INVENTION 
The present invention seeks to solve the above problems by providing 
texaphyrin metal complexes having improved functionalization compared to 
those previously described. The improved functionalization is two-fold: 
firstly, addition of electron-donating groups to positions 2, 7, 12, 15, 
18 and/or 21 of the macrocycle contributes electrons to the aromatic .pi. 
system of the macrocycle which stabilizes the metal complex to 
demetallation and stabilizes the imine bonds to hydrolysis; and secondly, 
the addition of electron-withdrawing groups to positions 15 and/or 18 
renders the macrocycle more readily reduced, i.e. the redox potential will 
be lower and the macrocycle will more readily gain an electron to form a 
radical. The addition of substituents to the 12 and 21 positions of the 
macrocycle also offers steric protection for the imine bonds against 
possible in vivo enzyme hydrolysis. Thus, the macrocycles of the present 
invention represent molecules where an attempt has been made to optimize 
their structure and properties in terms of imine bond stabilization and 
low redox potential, properties that are expected to be important for 
radiosensitization as well as other applications. 
Exemplary electron-donating groups that may be employed in the practice of 
the invention include, among others, amino, alkylamino, hydroxyl, 
acylamino, alkoxy, acyloxy, alkyl, aryl, and alkenyl. Electron-withdrawing 
groups include halide other than iodide, haloalkyl other than iodoalkyl, 
formyl, acyl, carboxylic acid, ester, acyl chloride, sulfonic acid, and 
nitro, among others. Other potential electron-donating or withdrawing 
groups will be apparent to one of skill in the art in light of the present 
disclosure. 
In certain embodiments, the present invention provides a texaphyrin having 
the structure: 
##STR1## 
M is H, a divalent metal cation, or a trivalent metal cation. 
R.sub.1 -R.sub.4, R.sub.7 and R.sub.8 are independently hydrogen, halide, 
hydroxyl, alkyl, aryl, haloalkyl, nitro, formyl, acyl, hydroxyalkyl, 
oxyalkyl, oxyhydroxyalkyl, saccharide, carboxy, carboxyalkyl, 
carboxyamidealkyl, a site-directing molecule, a catalytic group, or a 
couple to a site-directing molecule or to a catalytic group. 
R.sub.6 and R.sub.9 are independently selected from the groups of R.sub.1 
-R.sub.4, R.sub.7 and R.sub.8, with the proviso that the halide is other 
than iodide and the haloalkyl is other than iodoalkyl. 
R.sub.5 and R.sub.10 -R.sub.12 are independently hydrogen, alkyl, aryl, 
hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, carboxyalkyl, carboxyamidealkyl 
or a couple to a saccharide, to a site-directing molecule or to a 
catalytic group. 
For this invention, at least one of R.sub.5, R.sub.6, R.sub.9, R.sub.10, 
R.sub.11 and R.sub.12 is other than hydrogen. 
The charge, Z, is an integer value less than or equal to 5. Here, as would 
be apparent to one skilled in the art, the charge Z would be adjusted so 
as to account for the choice of metal, M, the pH under consideration, and 
the substituents R.sub.1 -R.sub.12. For instance, if R.sub.1 =carboxyl and 
R.sub.2 -R.sub.12 =alkyl and the metal M=Gd.sup.3+, and the solution is 
pH=7 (so that R.sub.1 .dbd.CO.sub.2 --), the charge Z would be zero. The 
charge would be negative when substituents have a sufficient number of 
negative charges, for example, when a substituent is an oligonucleotide. 
The charge would be +5, for example, when the M is Gd.sup.3+ and the net 
charge of a substituent(s) is three positive charges. 
An aspect of the present invention is an embodiment where a substituent may 
be an electron-donating group. In this case, R.sub.1 -R.sub.4 and R.sub.6 
-R.sub.9 are independently hydrogen, hydroxyl, alkyl, aryl, hydroxyalkyl, 
oxyalkyl, oxyhydroxyalkyl, saccharide, carboxyalkyl, carboxyamidealkyl, a 
site-directing molecule, a catalytic group, or a couple to a 
site-directing molecule or to a catalytic group. R.sub.5 and R.sub.10 
-R.sub.12 are independently hydrogen, alkyl, aryl, hydroxyalkyl, oxyalkyl, 
oxyhydroxyalkyl, carboxyalkyl, carboxyamidealkyl or a couple to a 
saccharide, to a site-directing molecule or to a catalytic group. At least 
one of R.sub.5, R.sub.6, R.sub.9, R.sub.10, R.sub.11 and R.sub.12 is other 
than hydrogen and Z is an integer less than or equal to 5. 
In another embodiment of the present invention, R.sub.6 or R.sub.9 may have 
an electron-withdrawing group. In that case, R.sub.1 -R.sub.4, R.sub.7 and 
R.sub.8 are independently hydrogen, halide, hydroxyl, alkyl, aryl, 
haloalkyl, nitro, formyl, acyl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, 
saccharide, carboxy, carboxyalkyl, carboxyamidealkyl, a site-directing 
molecule, a catalytic group, or a couple to a site-directing molecule or 
to a catalytic group. R.sub.5 and R.sub.10 -R.sub.12 are independently 
hydrogen, alkyl, aryl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, 
carboxyalkyl, carboxyamidealkyl or a couple to a saccharide, to a 
site-directing molecule or to a catalytic group. R.sub.6 and R.sub.9 are 
independently hydrogen, halide other than iodide, formyl, acyl, carboxy, 
or nitro, where at least one of R.sub.6 and R.sub.9 is other than hydrogen 
and Z is an integer less than or equal to 5. 
A couple may be an amide, disulfide, thioether, or ether covalent bond. A 
site-directing molecule may have binding specificity for localization to a 
treatment site. 
It is contemplated that the texaphyrins of the present invention are useful 
in a variety of applications including use as a photodynamic therapy 
agent, as a magnetic resonance imaging agent, as a radiation sensitizer, 
for RNA hydrolysis, and for DNA photocleavage. The use of a texaphyrin 
diamagnetic-metal complex having a substituent at the 2, 7, 12, 15, 18 
and/or 21 position and an absorption range from about 730 to about 770 
nanometers includes the following methods which take advantage of the 
ability of these compounds to produce singlet oxygen: i) a method of 
deactivating a retrovirus or enveloped virus in an aqueous fluid, the 
method comprising the steps of adding said texaphyrin metal complex to 
said aqueous fluid and exposing the mixture to light to effect the 
formation of singlet oxygen; ii) a method of producing light-induced 
singlet oxygen comprising subjecting said texaphyrin metal complex to 
light in the presence of oxygen; iii) a method of photosensitization 
comprising photoirradiating said texaphyrin; iv) a method of DNA 
light-induced photocleavage comprising placing said texaphyrin in contact 
with the RNA or DNA and photoirradiating said texaphyrin; and v) a method 
of treating a host harboring atheroma or neoplastic tissue comprising 
administering to the host an effective amount of said texaphyrin complex, 
the complex exhibiting selective biolocalization in the atheroma or 
neoplastic tissue relative to surrounding tissues, and photoirradiating 
the texaphyrin complex in proximity to the atheroma or neoplastic tissue. 
Further aspects of the present invention include the use of a texaphyrin 
paramagnetic-metal complex having a substituent at the 2, 7, 12, 15, 18 
and/or 21 position in the following methods which take advantage of the 
high relaxivity of these compounds: i) a method of enhancement of 
relaxivity comprising the administration of said texaphyrin; ii) a method 
of magnetic resonance image enhancement comprising administering to a 
subject an effective amount of said texaphyrin followed by MR imaging of 
the subject; iii) a method of detection of atheroma or neoplastic tissue 
in a subject comprising administering to the subject said texaphyrin in an 
amount effective to enhance a magnetic resonance image and detecting the 
atheroma or neoplastic tissue by MR imaging of said subject; iv) a method 
of imaging an organ in a subject comprising administering to the subject 
said texaphyrin in an amount effective to enhance a magnetic resonance 
image of the organ and detecting the organ by MR imaging of said subject; 
v) a method of imaging an atheroma in a subject comprising administering 
to the subject said texaphyrin in an amount effective to enhance a 
magnetic resonance image of the atheroma and detecting the atheroma by MR 
imaging of said subject; and vi) a method of RNA hydrolysis comprising 
placing said texaphyrin in contact with the RNA. 
A method of treating a host harboring atheroma or neoplastic tissue is also 
an aspect of the present invention, such method comprising administering 
to the host as a first agent a texaphyrin detectable-metal complex of the 
present invention, said complex exhibiting selective biolocalization in 
the atheroma or neoplastic tissue relative to surrounding tissue; 
determining localization sites in the host by reference to such 
texaphyrin-detectable metal complex; administering to the host as a second 
agent a texaphyrin diamagnetic-metal complex having a substituent at the 
2, 7, 12, 15, 18 and/or 21 position and having essentially identical 
biolocalization property and exhibiting the ability to generate singlet 
oxygen upon exposure to light; and photoirradiating the second agent in 
proximity to said atheroma or neoplastic tissue. 
The present invention provides a method of radiation therapy for a host 
harboring atheroma or neoplastic tissue, the method comprising 
administering to the host a texaphyrin of the present invention, and 
administering ionizing radiation to the host in proximity to the atheroma 
or neoplastic tissue. The radiation may be administered either before or 
after administration of the texaphyrin. The texaphyrin exhibits greater 
biolocalization in the atheroma or neoplastic tissue relative to 
surrounding tissues and has radiosensitization properties. An additional 
step may be included, the step being the determination of localization 
sites of the atheroma or neoplastic tissue in the host by monitoring 
texaphyrin concentrations. 
One skilled in the art would recognize in light of the present disclosure 
that sapphyrin-conjugated texaphyrin metal complexes may be used in 
methods for generating singlet oxygen. Sapphyrin compounds are disclosed 
in U.S. Pat. Nos. 5,159,065 and 5,120,411 which are incorporated by 
reference herein. 
Texaphyrin metal complexes having increased solution phase stability are 
expected to be more stable in vivo. Increased stability achieved via 
specific, designed modifications of the texaphyrin skeleton could give 
rise to products with modified biolocalization properties. Selective 
targeting would improve the efficacy and utility of texaphyrins as 
diagnostic or therapeutic agents for the range of applications discussed 
herein.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS 
The present invention involves metal complexes of texaphyrins having a 
substituent(s) at the 2, 7, 12, 15, 18 and/or 21 position(s) of the 
texaphyrin macrocycle and the synthesis and uses thereof. The nomenclature 
as used herein defines a substituent R.sub.11 attached to position 2, 
R.sub.12 attached to position 7, R.sub.5 attached to position 12, R.sub.6 
attached to position 15, R.sub.9 attached to position 18 and R.sub.10 
attached to position 21 of the macrocycle. The following structure shows a 
correlation of the IU nomenclature for the positions of the atoms 
around the periphery of the macrocycle with the positions of the R groups 
of the present invention. 
##STR2## 
Substituents at the R.sub.6 and R.sub.9 positions on the B (benzene ring) 
portion of the macrocycle are incorporated into the macrocycle by their 
attachment to ortho-phenylenediamine in the 3 and 6 positions of the 
molecule. Substituents at the R.sub.5 and R.sub.10 positions on the T 
(tripyrrane) portion of the macrocycle are incorporated by appropriate 
functionalization of carboxyl groups in the 5 positions of the tripyrrane 
at a synthetic step prior to condensation with a substituted 
ortho-phenylenediamine. 
In a method for synthesizing a texaphyrin metal complex having a 
substituent at the 2, 7, 12, 15, 18 or 21 position, the method comprises 
the steps of: i) mixing, in an organic solvent, a nonaromatic texaphyrin 
having a substituent at the 2, 7, 12, 15, 18 and/or 21 position, a 
trivalent metal salt, a Br.o slashed.nsted base and an oxidant; and ii) 
allowing the mixture to react to form an aromatic texaphyrin metal complex 
having a substituent at the 2, 7, 12, 15, 18, and/or 21 position. A 
preferred means is to stir at ambient temperature or heat the mixture at 
reflux for at least two hours. 
The corresponding nonaromatic texaphyrin having a substituent at the 2, 7, 
12, 15, 18, and/or 21 position is conveniently produced by condensation of 
a tripyrrane aldehyde or ketone having structure A and a substituted 
ortho-phenylenediamine having structure B: 
##STR3## 
In this embodiment, R.sub.1 -R.sub.4, R.sub.7 and R.sub.8 are independently 
hydrogen, halide, hydroxyl, alkyl, aryl, haloalkyl, nitro, formyl, acyl, 
hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, saccharide, carboxy, 
carboxyalkyl, carboxyamidealkyl, a site-directing molecule, or a couple to 
a site-directing molecule. 
R.sub.6 and R.sub.9 are independently selected from the groups of R.sub.1 
-R.sub.4, R.sub.7 and R.sub.8, with the proviso that the halide is other 
than iodide and the haloalkyl is other than iodoalkyl. 
R.sub.5, R.sub.10, R.sub.11 and R.sub.12 are independently hydrogen, alkyl, 
aryl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, carboxyalkyl, 
carboxyamidealkyl or a couple to a saccharide or to a site-directing 
molecule. At least one of R.sub.5, R.sub.6, R.sub.9, R.sub.10, R.sub.11 
and R.sub.12 is other than hydrogen. 
In a preferred method of synthesis, the Br.o slashed.nsted base is 
triethylamine or N,N,N',N'-tetramethyl-1,8-diaminonaphthalene ("proton 
sponge"), and the oxidant is air saturating the organic solvent, oxygen, 
platinum oxide, o-chloranil or 2,3-dichloro-5,6-dicyano-1,4-benzoquinone. 
The stirring or heating at reflux step may comprise stirring or heating at 
reflux the mixture for at least 24 hours. The organic solvent may comprise 
methanol, or methanol and chloroform, or methanol and benzene, or methanol 
and dimethylformamide. 
In the texaphyrins of the present invention, M is hydrogen, a divalent 
metal cation, or a trivalent metal cation. The divalent metal cation may 
be selected from, but is not limited to, the group consisting of Ca(II), 
Mn(II), Co(II), Ni(II), Zn(II), Cd(II), Hg(II), Fe(II), Sm(II) and 
UO.sub.2 (II). The trivalent metal cation may be selected from, but is not 
limited to, the group consisting of Mn(III), Co(III), Ni(III), Fe(III), 
Ho(III), Ce(III), Y(III), In(III), Pr(III), Nd(IZI), Sm(III), Eu(III), 
Gd(III), Tb(III), Dy(III), Er(III), Tm(III), Yb(III), Lu(III), La(III), 
and U(III). 
The alkyl, aryl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, saccharide, 
carboxyalkyl, carboxyamidealkyl, site-directing molecule, or molecule 
couple is covalently bonded to the texaphyrin via a carbon-carbon, a 
carbon-nitrogen or a carbon-oxygen bond. 
The aryl may be a phenyl group, unsubstituted or substituted with a nitro, 
carboxy, sulfonic acid, hydroxy, oxyalkyl or halide other than iodide 
substituent. In this case, the substituent on the phenyl group may be 
added in a synthetic step after the condensation step which forms the 
macrocycle. 
Representative examples of alkanes useful as alkyl group substituents of 
the present invention include methane, ethane, straight-chain, branched or 
cyclic isomers of propane, butane, pentane, hexane, heptane, octane, 
nonane and decane, with methane, ethane and propane being preferred. 
Representative examples of alkenes useful as alkenyl group substituents 
include ethene, straight-chain, branched or cyclic isomers of propene, 
butene, pentene, hexene, heptene, octene, nonene and decene, with ethene 
and propene being preferred. Representative examples of alkynes useful as 
alkynyl group substituents include ethyne, straight-chain, branched or 
cyclic isomers of propyne, butyne, pentyne, hexyne, heptyne, octyne, 
nonyne and decyne, with ethyne and propyne being preferred. Representative 
examples of substituted alkyls include alkyls substituted by two or more 
functional groups as described herein. 
Among the halide substituents, chloride, bromide, fluoride and iodide are 
contemplated in the practice of this invention with the exception that 
R.sub.6 and R.sub.9 are not iodide. R.sub.6 and R.sub.9 may have chloride, 
bromide or fluoride substituents. Representative examples of haloalkyls 
used in this invention include halides of methane, ethane, propane, 
butane, pentane, hexane, heptane, octane, nonane and decane, with halides, 
preferably chlorides or bromides, of methane, ethane and propane being 
preferred. 
Representative examples of hydroxyalkyls include alcohols of methane, 
ethane, straight-chain, branched or cyclic isomers of propane, butane, 
pentane, hexane, heptane, octane, nonane and decane, with alcohols of 
methane, ethane or propane being preferred. "Hydroxyalkyl" is meant to 
include glycols and polyglycols having hydroxyl groups; diols of ethane, 
straight-chain, branched or cyclic isomers of propane, butane, pentane, 
hexane, heptane, octane, nonane and decane, with diols of ethane or 
propane being preferred; polyethylene glycol, polypropylene glycol and 
polybutylene glycol as well as polyalkylene glycols containing 
combinations of ethylene, propylene and butylene. 
Representative examples of oxyalkyls include the alkyl groups as herein 
described having ether linkages. The number of repeating oxyalkyls within 
a substituent may be up to 100, preferably is from 1-10, and more 
preferably, is 2-3. A preferred oxyalkyl is O(CH.sub.2 CH.sub.2 O).sub.x 
CH.sub.3 where x=1-100, preferably 1-10, and more preferably, 2-3. 
Representative examples of thioalkyls include thiols of ethane, thiols of 
straight-chain, branched or cyclic isomers of propane, butane, pentane, 
hexane, heptane, octane, nonane and decane, with thiols of ethane 
(ethanethiol, C.sub.2 H.sub.5 SH) or propane (propanethiol, C.sub.3 
H.sub.7 SH) being preferred. Sulfate-substituted alkyls include alkyls as 
described above substituted by one or more sulfate groups, a 
representative example of which is diethyl sulfate ((C.sub.2 
H.sub.5).sub.2 SO.sub.4). 
Representative examples of phosphates include phosphate or polyphosphate 
groups. Representative examples of phosphate-substituted alkyls include 
alkyls as described above substituted by one or more phosphate or 
polyphosphate groups. Representative examples of phosphonate-substituted 
alkyls include alkyls as described above substituted by one or more 
phosphonate groups. 
Representative examples of carboxy groups include carboxylic acids of the 
alkyls described above as well as aryl carboxylic acids such as benzoic 
acid. Representative examples of carboxyamides include primary 
carboxyamides (CONH.sub.2), secondary (CONHR') and tertiary (CONR'R") 
carboxyamides where each of R' and R" is a functional group as described 
herein. 
Representative examples of useful amines include a primary, secondary or 
tertiary amine of an alkyl as described hereinabove. 
Hydroxyalkyl means alkyl groups having hydroxyl groups attached. Oxyalkyl 
means alkyl groups attached to an oxygen. Oxyhydroxyalkyl means alkyl 
groups having ether or ester linkages, hydroxyl groups, substituted 
hydroxyl groups, carboxyl groups, substituted carboxyl groups or the like. 
Saccharide includes oxidized, reduced or substituted saccharide; hexoses 
such as D-glucose, D-mannose or D-galactose; pentoses such as D-ribulose 
or D-fructose; disaccharides such as sucrose, lactose, or maltose; 
derivatives such as acetals, amines, and phosphorylated sugars; 
oligosaccharides, as well as open chain forms of various sugars, and the 
like. Examples of amine-derivatized sugars are galactosamine, glucosamine, 
sialic acid and D-glucamine derivatives such as 1-amino-1-deoxysorbitol. 
Carboxyamidealkyl means alkyl groups containing any number of functional 
groups, one of which is a secondary or tertiary amide. Carboxyalkyl means 
alkyl groups containing any number of functional groups, one of which is a 
carboxyl group. 
For the above-described texaphyrins, oxyhydroxyalkyl may be alkyl having 
independently hydroxy substituents and ether branches or may be 
C.sub.(n-x) H.sub.((2n+1)-2x) O.sub.x O.sub.y or OC.sub.(n-x) 
H.sub.((2n+1)-2x) O.sub.x O.sub.y where n is a positive integer from 1 to 
10, x is zero or a positive integer less than or equal to n, and y is zero 
or a positive integer less than or equal to ((2n+1)-2x). 
The oxyhydroxyalkyl or saccharide may be C.sub.n H.sub.((n+1)-q) O.sub.y 
R.sub.q.sup.a, OC.sub.n H.sub.((2n+1)-q) O.sub.y R.sub.a.sup.1 or 
(CH.sub.2).sub.n CO.sub.2 R.sup.a where n is a positive integer from 1 to 
10, y is zero or a positive integer less than ((2n+1)-q), q is zero or a 
positive integer less than or equal to 2n+1, R.sup.a is independently H, 
alkyl, hydroxyalkyl, saccharide, C.sub.(m-w) H.sub.((2m+1)-2w) O.sub.w 
O.sub.z, O.sub.2 CC.sub.(m-w) H.sub.((2m+1)-2w) O.sub.w O.sub.z or 
N(R)OCC.sub.(m-w) H.sub.((2m+1)-2w) O.sub.w O.sub.z, where m is a positive 
integer from 1 to 10, w is zero or a positive integer less than or equal 
to m, z is zero or a positive integer less than or equal to ((2m+1)-2w), R 
is H, alkyl, hydroxyalkyl, or C.sub.m H.sub.((2m+1)-r) O.sub.z 
R.sub.r.sup.b where m is a positive integer from 1 to 10, z is zero or a 
positive integer less than ((2m+1)-r), r is zero or a positive integer 
less than or equal to 2m+1, and R.sup.b is independently H, alkyl, 
hydroxyalkyl, or saccharide. 
Carboxyamidealkyl may be alkyl having secondary or tertiary amide linkages 
or (CH.sub.2).sub.n CONHR.sup.a, O(CH.sub.2).sub.n CONHR.sup.a, 
(CH.sub.2).sub.n CON(R.sup.a).sub.2, or O(CH.sub.2).sub.n 
CON(R.sup.a).sub.2 where n is a positive integer from 1 to 10, R.sup.a is 
independently H, alkyl, hydroxyalkyl, saccharide, C.sub.(m-w) 
H.sub.((2m+1)-2w) O.sub.w O.sub.z, O.sub.2 CC.sub.(m-w) H.sub.((2m+1)-2w) 
O.sub.w O.sub.z or N(R)OCC.sub.(m-w) H.sub.((2m+1)-2w) O.sub.w O.sub.z, 
where m is a positive integer from 1 to 10, w is zero or a positive 
integer less than or equal to m, z is zero or a positive integer less than 
or equal to ((2m+1)-2w), R is H, alkyl, hydroxyalkyl, or C.sub.m 
H.sub.((2m+1)-r) O.sub.z R.sub.r.sup.b where m is a positive integer from 
1 to 10, z is zero or a positive integer less than ((2m+1)-r), r is zero 
or a positive integer less than or equal to 2m+1, and R.sup.b is 
independently H, alkyl, hydroxyalkyl, or saccharide. 
The carboxyalkyl may be alkyl having a carboxyl substituted ether, an amide 
substituted ether or a tertiary amide removed from an ether or C.sub.n 
H.sub.((2n+1)-q) O.sub.y R.sub.q.sup.c or OC.sub.n H.sub.((2n+1)-q) 
O.sub.y R.sub.q.sup.c where n is a positive integer from 1 to 10; y is 
zero or a positive integer less than ((2n+1)-q), q is zero or a positive 
integer less than or equal to 2n+1, R.sup.c is (CH.sub.2).sub.n CO.sub.2 
R.sup.d, (CH.sub.2).sub.n CONHR.sup.d or (CH.sub.2).sub.n 
CON(R.sup.d).sub.2 where n is a positive integer from 1 to 10; R.sup.d is 
independently H, alkyl, hydroxyalkyl, saccharide, C.sub.(m-w) 
H.sub.((2m+1)-2w) O.sub.w O.sub.z, O.sub.2 CC.sub.(m-w) H.sub.((2m+1)-2w) 
O.sub.w O.sub.z or N(R)OCC.sub.(m-w) H.sub.((2m+1)-2w) O.sub.w O.sub.z, 
where m is a positive integer from 1 to 10, w is zero or a positive 
integer less than or equal to m, z is zero or a positive integer less than 
or equal to ((2m+1)-2w), R is H, alkyl, hydroxyalkyl, or C.sub.m 
H.sub.((2m+1)-r) O.sub.z R.sub.r.sup.b where m is a positive integer from 
1 to 10, z is zero or a positive integer less than ((2m+1)-r), r is zero 
or a positive integer less than or equal to 2m+1, and R.sup.b is 
independently H, alkyl, hydroxyalkyl, or saccharide. 
A couple may be described as a linker, i.e., a reactive group for attaching 
another molecule at a distance from the texaphyrin macrocycle. An 
exemplary linker or couple is an amide, disulfide, thioether or ether 
covalent bond as described in the examples for attachment of 
oligonucleotides and antibodies. 
Certain reactions utilizing the texaphyrin complexes of the present 
invention, such as hydrolytic cleavage of phosphate ester bonds for 
example, by may be enhanced by additional catalytic groups appended to the 
texaphyrin metal complex or to a texaphyrin complex-site directing 
molecule conjugate. The term "catalytic group" means a chemical functional 
group that assists catalysis by acting as a general acid, Br.o 
slashed.nsted acid, general base, Br.o slashed.nsted base, nucleophile, or 
any other means by which the activation barrier to reaction is lowered or 
the ground state energy of the substrate is increased. Exemplary catalytic 
groups contemplated include, but are not limited to, imidazole; guanidine; 
substituted saccharides such as D-glucosamine, D-mannosamine, 
D-galactosamine, D-glucamine, and the like; amino acids such as 
L-histidine and L-arginine; derivatives of amino acids such as histamine; 
polymers of amino acids such as poly-L-lysine, (LysAla).sub.n or 
(LysLeuAla).sub.n where n is from 1-30 or preferably 1-10 or more 
preferably 2-7, and the like; derivatives thereof; and texaphyrin metal 
complexes. The term "appended to the texaphyrin-site directing molecule 
conjugate" means that the catalytic groups are attached either directly to 
the texaphyrin metal complex or to the texaphyrin complex via a linker or 
couple of variable length, or are attached to the ligand portion of a 
texaphyrin complex-ligand conjugate either with or without a linker or 
couple of variable length. 
In one embodiment of the present invention, the texaphyrin is coupled to 
site-directing molecules to form conjugates for targeted in vivo delivery. 
"Specificity for targeted sites" means that upon contacting the texaphyrin 
conjugate with the targeted site, for example under physiological 
conditions of ionic strength, temperature, pH and the like, specific 
binding will occur. The interaction may occur due to specific 
electrostatic, hydrophobic, entropic or other interaction of certain 
residues of the conjugate with specific residues of the target to form a 
stable complex under conditions effective to promote the interaction. 
Exemplary site-directing molecules contemplated in the present invention 
include, but are not limited to: oligonucleotides, including 
oligodeoxyribonucleotides and oligoribonucleotide analogs; polyamides 
including peptides having affinity for a biological receptor and proteins 
such as antibodies, low density lipoproteins, the APO protein of 
lipoprotein; steroids and steroid derivatives; hormones such as estradiol, 
or histamine; hormone mimics such as morphine; and further macrocycles 
such as sapphyrins and rubyrins. 
Representative examples of useful oligonucleotides include nucleotides, 
oligonucleotides and polynucleotides primarily composed of adenine, 
cytosine, guanine, thymine or uracil bases. An oligonucleotide may be 
derivatized at the base, the sugar, the ends of the chain, or at the 
phosphate groups of the backbone to promote in vivo stability. 
Modification of the phosphate groups is preferred in one embodiment of the 
invention since phosphate linkages are sensitive to nuclease activity. 
Preferred derivatives are methylphosphonates, phosphotriesters, 
phosphorothioates, and phosphoramidates, and the like. Additionally, 
phosphate linkages may be completely substituted with non-phosphate 
linkages such as amide linkages. Appendages to the ends of the 
oligonucleotide chain also provide exonuclease resistance. Sugar 
modifications may include alkyl groups attached to an oxygen of a ribose 
moiety in a ribonucleotide. In particular, the alkyl group generally has 1 
to 4 carbon atoms, and preferably is a methyl group and the methyl group 
is attached to the 2' oxygen of the ribose. Other alkyl groups may be 
ethyl or propyl. It is understood that the terms "nucleotide" and 
"oligonucleotide", as used herein, refer to both naturally-occurring and 
synthetic nucleotides, poly- and oligonucleotides and to analogs and 
derivatives thereof. 
The term "texaphyrin-oligonucleotide conjugate" means that an 
oligonucleotide is attached to the texaphyrin in a 5' or a 3' linkage, or 
to both types of linkages to allow the texaphyrin to be an internal 
residue in the conjugate. The oligonucleotide or other site-directing 
molecule may be attached either directly to the texaphyrin via a linker, 
or a couple of variable length. During treatment, for example, the 
texaphyrin portion of a texaphyrin metal complex-oligonucleotide conjugate 
of the present invention is envisioned as being placed in the vicinity of 
the targeted tissue upon binding of the oligonucleotide to its 
complementary DNA or RNA. 
Representative examples of useful steroids include any of the steroid 
hormones of the following five categories: progestins (e.g. progesterone), 
glucocorticoids (e.g., cortisol), mineralocorticoids (e.g., aldosterone), 
androgens (e.g., testosterone) and estrogens (e. g., estradiol). 
Representative examples of useful amino acids of peptides or polypeptides 
include amino acids with simple aliphatic side chains (e.g., glycine, 
alanine, valine, leucine, and isoleucine), amino acids with aromatic side 
chains (e.g., phenylalanine, tryptophan, tyrosine, and histidine), amino 
acids with oxygen and sulfur-containing side chains (e.g., serine, 
threonine, methionine, and cysteine), amino acids with side chains 
containing carboxylic acid or amide groups (e.g., aspartic acid, glutamic 
acid, asparagine, and glutamine), and amino acids with side chains 
containing strongly basic groups (e.g., lysine and arginine), and proline. 
Representative examples of useful peptides include any of both naturally 
occurring and synthetic di-, tri-, tetra-, pentapeptides or longer 
peptides derived from any of the above described amino acids (e.g., 
endorphin, enkephalin, epidermal growth factor, poly-L-lysine, or a 
hormone). Representative examples of useful polypeptides include both 
naturally occurring and synthetic polypeptides (e.g., insulin, 
ribonuclease, and endorphins) derived from the above described amino acids 
and peptides. 
The term "a peptide having affinity for a biological receptor" means that 
upon contacting the peptide with the biological receptor, for example 
under appropriate conditions of ionic strength, temperature, pH and the 
like, specific binding will occur. The interaction may occur due to 
specific electrostatic, hydrophobic, entropic or other interaction of 
certain amino acid or glycolytic residues of the peptide with specific 
amino acid or glycolytic residues of the receptor to form a stable complex 
under the conditions effective to promote the interaction. The interaction 
may alter the three-dimensional conformation and the function or activity 
of either or both the peptide and the receptor involved in the 
interaction. A peptide having affinity for a biological receptor may 
include an endorphin, an enkephalin, a growth factor, e.g. epidermal 
growth factor, poly-L-lysine, a hormone, a peptide region of a protein and 
the like. A hormone may be estradiol, for example. 
For the above-described texaphyrins, the couple may be an amide, disulfide, 
thioether or ether covalent bond; the oligonucleotide, the antibody, the 
hormone or the sapphyrin may have binding specificity for localization to 
a treatment site; and the biological receptor may be localized to a 
treatment site. 
Generally, water-soluble texaphyrins are preferred for the applications 
described herein. "Water-soluble" means soluble in aqueous fluids to about 
1 mM or better. Such characteristics allow these texaphyrins to be useful 
in a biological environment. Improved water solubility can be achieved by, 
for example, substituents chosen from saccharides or hydroxylated 
substituents. 
A preferred embodiment of the present invention is a texaphyrin wherein 
when either R.sub.5 or R.sub.10 is other than hydrogen, then R.sub.6 or 
R.sub.9, respectively, is hydrogen, halide other than iodide (preferably 
fluoro), or hydroxyl. 
A further preferred embodiment of the present invention is a then R.sub.5 
or R.sub.10, respectively, is hydrogen or methyl. 
Other preferred functionalizations are where R.sub.6 and R.sub.9 are 
hydrogen, then R.sub.5, R.sub.10, R.sub.11 and R.sub.12 are aryl, lower 
alkyl or lower hydroxyalkyl. The lower alkyl is preferably methyl or 
ethyl, more preferably methyl. The lower hydroxyalkyl is preferably of 1 
to 6 carbons and 1 to 4 hydroxy groups, more preferably 3-hydroxypropyl. 
The aryl is preferably phenyl, either unsubstituted or substituted, 
preferably unsubstituted. 
Further preferred embodiments of the present invention are where R.sub.2 
and R.sub.3 are CH.sub.2 CH.sub.3 and R.sub.4 is CH.sub.3, where R.sub.5 
and R.sub.10 are methyl, or where R.sub.5 and R.sub.10 are 
(CH.sub.2).sub.n CH.sub.3 where n is 0, 1, 2, 3 or 4. Furthermore, R.sub.5 
and R.sub.10 may be aryl having an R.sub.13 substituent where R.sub.13 is 
hydrogen, nitro, carboxy, sulfonic acid, hydroxy, oxyalkyl or halide. A 
presently preferred aryl is phenyl. The derivatization of the R.sub.13 
group may occur after the condensation of the macrocycle. Preferred 
substituents for R.sub.6 include carboxy, alkyl or carboxyamidealkyl 
having a tertiary amide linkage. Preferred substituents for R.sub.7, 
R.sub.8 and R.sub.9 are oxyalkyl or hydroxyalkyl. 
Further preferred texaphyrins are wherein each of R.sub.1 -R.sub.12 is any 
one of the substituents of Tables A and B described herein below; more 
preferred texaphyrins are texaphyrins A1-A56 of Tables A and B described 
herein below. Preferred metals are Mn(II), Mn(III), Y(III), Lu(III), 
La(III), In(III), Gd(III), Eu(III), and Dy(III). 
Electron-donating substituents at the 2, 7, 12, 15, 18 and/or 21 positions 
of the macrocycle stabilize the molecule against decomposition processes 
involving hydrolysis of the imine bonds. Such substituents also stabilize 
the resulting complex against demetallation by contributing electrons to 
the aromatic .pi. system. demetallation by contributing electrons to the 
aromatic .pi. system. Such electron-donating groups include hydroxyl, 
alkyl, haloalkyl other than iodoalkyl, aryl, hydroxyalkyl, oxyalkyl, 
oxyhydroxyalkyl, saccharide, carboxyalkyl, carboxyamidealkyl, a 
site-directing molecule, or a couple to any of these molecules. 
Hydrolysis-resistant texaphyrin metal complexes are useful for 
localization, magnetic resonance imaging, radiosensitization, radiation 
therapy, fluorescence imaging, photodynamic therapy and applications 
requiring singlet oxygen production for cytotoxicity. 
Electron-withdrawing substituents at the 15, 16, 17 and/or 18 positions of 
the macrocycle destabilize the aromatic .pi. system and render the 
macrocycle more readily reduced, i.e. more easily able to gain an electron 
to form a radical. Such electron-withdrawing groups include halide other 
than iodide, formyl, acyl, carboxy, or nitro substituents. Readily 
reducible texaphyrin metal complexes are useful for radiosensitization 
where the extent of radiation damage is dependent on the generation of 
hydroxyl and texaphyrin radicals. 
The photophysical properties of prior texaphyrin metal complexes are 
reported in U.S. Pat. No. 5,252,720 and include strong low energy optical 
absorptions in the 690-880 nm spectral range, a high triplet quantum yield 
and efficient production of singlet oxygen. Texaphyrin metal complexes of 
grandparent application Ser. No. 08/135,118, incorporated by reference 
herein, demonstrate enhanced cytotoxicity from radiation and enhanced 
nucleic acid strand scission in the presence of a gadolinium(III) 
metallotexaphyrin complex. U.S. Pat. No. 5,252,720 describes 
photosensitized inactivation of enveloped viruses and magnetic resonance 
imaging (MRI) of atheroma, liver, kidney and tumor using various 
substituted texaphyrin metal complexes. Altering the polarity and 
electrical charges of side groups of the texaphyrin macrocycles alters the 
degree, rate, and site(s) of binding to free enveloped viruses such as 
HIV-1 and to virally-infected peripheral mononuclear cells, thus 
modulating photosensitizer take-up and photosensitization of leukemia or 
lymphoma cells contaminating bone-marrow. Powerful techniques include the 
use of these texaphyrins in magnetic resonance imaging followed by 
photodynamic therapy in the treatment of atheroma and benign and malignant 
tumors, or followed by sensitized X-ray treatment. 
It is contemplated that the texaphyrins of the present invention will prove 
useful in a variety of applications. One example is in a method of 
deactivating a retrovirus or enveloped virus in an aqueous fluid. Such a 
method comprises the step of adding a texaphyrin metal complex having a 
substituent at the 2, 7, 12, 15, 18 and/or 21 position to said aqueous 
fluid and exposing the mixture to light to effect the formation of singlet 
oxygen. The aqueous fluid may be a biological fluid, blood, plasma, edema 
tissue fluid, ex vivo fluid for injection into body cavities, cell culture 
media, or a supernatant solution from cell culture and the like. 
In blood, an exemplary viral deactivating method would include: i) mixing 
with blood in vitro or ex vivo a texaphyrin metal complex having a 
substituent at the 2, 7, 12, 15, 18 and/or 21 position capable of 
producing singlet oxygen when irradiated in the presence of oxygen; and 
ii) photoirradiating the mixture in vitro or ex vivo to produce singlet 
oxygen in a quantity cytotoxic to said retrovirus or enveloped virus. 
Exemplary retroviruses or enveloped viruses include herpes simplex virus 
I, cytomegalovirus, measles virus, or human immunodeficiency virus HIV-1. 
However, it is contemplated that the utility of the invention is not 
limited to these viruses. Preferred metal cations are diamagnetic metal 
cations and a preferred metal complex is the Lu(III), La(III) or In(III) 
complex of said texaphyrin. 
A further application of the present invention is a method of light-induced 
singlet oxygen production comprising subjecting a texaphyrin metal complex 
having a substituent at the 2, 7, 12, 15, 18 and/or 21 position to light 
in the presence of oxygen. A method of photosensitization comprising the 
production of singlet oxygen by irradiating a texaphyrin metal complex 
having a substituent at the 2, 7, 12, 15, 18 and/or 21 position and an 
absorption range from about 730 to about 770 nanometers to form long-lived 
triplet states in high yield is another embodiment of the present 
invention. A texaphyrin metal complex having a substituent at the 2, 7, 
12, 15, 18 and/or 21 position has the structure as described previously 
herein; however, for these applications, M is a diamagnetic metal cation, 
for example, In(III), Zn(II), Cd(II), Lu(III) or La(III). "Intrinsic 
biolocalization selectivity" means having an inherently greater affinity 
for certain tissues relative to surrounding tissues. 
Further aspects of the present invention include the use of a texaphyrin 
paramagnetic-metal complex having a substituent at the 2, 7, 12, 15, 18 
and/or 21 position in the following methods which take advantage of the 
high relaxivity of these compounds: i) a method of enhancement of 
relaxivity comprising the administration of said texaphyrin; ii) a method 
of magnetic resonance image enhancement comprising administering to a 
subject an effective amount of said texaphyrin; iii) a method of detection 
of neoplastic tissue in a patient comprising the steps of administering to 
a patient said texaphyrin in an amount effective to enhance a magnetic 
resonance image and detecting neoplastic tissue by magnetic resonance 
imaging of said patient; iv) a method of imaging an organ in a patient 
comprising administering to a patient said texaphyrin in an amount 
effective to enhance a magnetic resonance image of the organ and detecting 
the organ by magnetic resonance imaging of said patient (the organ may be 
liver, kidney or the upper GI tract); v) a method of imaging atheroma in a 
patient comprising administering to a patient said texaphyrin in an amount 
effective to enhance a magnetic resonance image of atheroma and detecting 
atheroma by magnetic resonance imaging of said patient. 
For use in these imaging applications, the texaphyrin paramagnetic-metal 
complex has the structure as described herein; however, M is a 
paramagnetic metal cation, such as a trivalent lanthanide metal other than 
La(III), Lu(III) and Pm(III). In particular, M may be Mn(II), Mn(III), 
Fe(III) or Gd(III) and is preferably Gd(III). 
A method of treating a host harboring atheroma or benign or malignant tumor 
cells is also an aspect of the invention. An exemplary preferred method 
includes administering to the host as a first agent a texaphyrin 
detectable-metal complex having a substituent at the 2, 7, 12, 15, 18 
and/or 21 position, said complex exhibiting selective biolocalization in 
such atheroma or tumor cells relative to surrounding tissue; determining 
localization sites in the host by reference to such detectable metal; 
administering to the host as a second agent a texaphyrin diamagnetic-metal 
complex having a substituent at the 2, 7, 12, 15, 18 and/or 21 position 
and having essentially identical biolocalization property and exhibiting 
the ability to generate singlet oxygen upon exposure to light; and 
photoirradiating the second agent in proximity to said atheroma or tumor 
cells. 
In the above-described method, the first agent is further defined as being 
a texaphyrin paramagnetic-metal complex, the paramagnetic metal serving as 
the detectable metal. In this case, determination of localization sites 
occurs by magnetic resonance imaging; and the second agent is a texaphyrin 
diamagnetic-metal complex. The paramagnetic metal is most preferably 
Gd(III) and the diamagnetic metal is most preferably La(III), Lu(III) or 
In(III). A variation of this method uses as a first agent a 
texaphyrin-gamma emitting metal complex that serves as a detectable metal, 
determination of localization sites occurs by gamma body scanning and the 
second agent is a texaphyrin-diamagnetic metal complex. A further 
variation uses as a first agent a texaphyrin which exhibits fluorescence, 
e.g., a texaphyrin that is non-metallated (M.dbd.H) or is complexed with a 
diamagnetic metal. Localization means is then by fluorescent spectroscopy. 
where M is hydrogen or a detectable metal, preferably detectable by 
magnetic resonance imaging, by gamma scanning or fluorescence 
spectroscopy. "Detectable" as used herein means that the location may be 
foundby localization means such as magnetic resonance imaging if the metal 
is paramagnetic, gamma ray detection if the metal is gamma emitting or 
using monochromatic X-ray photon sources or by fluorescence. "Selective 
biolocalization" means having an inherently greater affinity for certain 
tissues relative to surrounding tissues. "Essentially identical 
biolocalization property" means the second agent is a texaphyrin 
derivative having about the same selective targeting characteristics in 
tissue as demonstrated by the first agent. 
A method of treating a host harboring tumor cells comprises the steps of: 
i) administering to the host an effective amount of a texaphyrin 
diamagnetic-metal complex having a substituent at the 2, 7, 12, 15, 18 
and/or 21 position, the complex exhibiting selective biolocalization in 
the tumor cells relative to surrounding tissue; and ii) photoirradiating 
the texaphyrin-diamagnetic metal complex in proximity to the tumor cells. 
The photoirradiating is generally at a wavelength of about 730 to 770 
nanometers or may be from laser light. In these embodiments, the 
diamagnetic metal will typically be In(III), La(III) or Lu(III). 
The present invention provides a method of radiation therapy for a host 
harboring a tumor. The method includes the steps of administering to the 
host a texaphyrin having a substituent in the 2, 7, 12, 15, 18 and/or 21 
position(s), and administering ionizing radiation to the host in proximity 
to the tumor either before or after administration of the texaphyrin, 
following procedures as described in U.S. Ser. No. 08/135,118, 
incorporated herein by reference. The texaphyrin exhibits greater 
biolocalization in the tumor relative to non-tumor tissue and has 
radiosensitization properties. A tumor may be a benign or malignant tumor 
or may be atheroma. A texaphyrin having radiosensitization properties 
enhances cytotoxicity from ionizing radiation as compared to control 
experiments without the texaphyrin. Ionizing radiation includes but is not 
limited to x-rays, and internal and external gamma emitting radioisotopes. 
The texaphyrin may be complexed with a metal; however, a metal is not 
necessary for radiosensitization. The metal is important to the stability 
of the texaphyrin complex. The ionizing radiation may be from an external 
source or the metal may be a radioactive metal. In the latter case, the 
ionizing radiation is from the radioactive metal in combination with 
radiation from an external source. 
An improved method of treating a host harboring a tumor comprises the 
further step of determining localization sites in the host by monitoring 
texaphyrin concentrations. "Monitoring texaphyrin concentrations" means 
measuring fluorescence of an administered free base texaphyrin or by 
reference to the metal of an administered texaphyrin metal complex. If the 
metal is paramagnetic, then magnetic resonance imaging is used for 
measurement; if the metal is a gamma emitting radioactive metal, then 
gamma emission is used for measurement. 
A further improved method of treating a host harboring a tumor comprises 
the additional steps of administering to the host as a second agent a 
texaphyrin-diamagnetic metal complex having a substituent at the 2, 7, 12, 
15, 18 and/or 21 position and having essentially identical biolocalization 
property and administering ionizing radiation and photoirradiation in 
proximity to the tumor. 
In these methods, determining localization sites may occur by observing 
fluorescence from the texaphyrin. When the first agent is complexed with a 
metal, the metal may be a gamma-emitting metal and determining 
localization sites would occur by gamma body imaging, or the metal may be 
a paramagnetic metal and determining localization sites would occur by 
magnetic resonance imaging. 
"Exhibiting greater biolocalization in the tumor relative to non-tumor 
tissue" means having an inherently greater affinity for tumor tissue 
relative to non-tumor tissue. The second agent has essentially identical 
biolocalization property as the first agent and exhibits the ability to 
generate singlet oxygen upon exposure to light. The photodynamic effect 
may be derived from anaerobic electron transfer processes. A preferred 
diamagnetic metal texaphyrin complex is the Lu(III), La(III) or In(III) 
complex of a texaphyrin. "Essentially identical biolocalization property" 
means the second agent is a texaphyrin derivative having about the same 
selective targeting characteristics in tissue as demonstrated by the first 
agent. The first agent and the second agent may be the same texaphyrin. 
A preferred embodiment of the present invention is a method of radiation 
therapy for a host harboring a tumor comprising the steps of i) 
administering to the host a pharmaceutically effective amount of the Gd 
complex of a texaphyrin having a substituent at the 2, 7, 12, 15, 18 
and/or 21 position(s); and ii) administering ionizing radiation to the 
host in proximity to the tumor, either before or after administration of 
the texaphyrin metal complex. 
Another aspect of this invention is a method of imaging atheroma in a host 
comprising the administration to the host as an agent a detectable-metal 
texaphyrin complex having a substituent at the 2, 7, 12, 15, 18 and/or 21 
position(s), said complex exhibiting selective biolocalization in such 
atheroma; and imaging the atheroma in the host by reference to such 
detectable metal. The agent is preferably a detectable-metal texaphyrin 
complex having a paramagnetic metal serving as said detectable metal, and 
imaging of the atheroma occurs by magnetic resonance imaging. The 
paramagnetic metal is preferably Gd(III). 
For the above-described uses, texaphyrins are provided as pharmaceutical 
preparations. A pharmaceutical preparation of a texaphyrin may be 
administered alone or in combination with pharmaceutically acceptable 
carriers, in either single or multiple doses. Suitable pharmaceutical 
carriers include inert solid diluents or fillers, sterile aqueous solution 
and various organic solvents. The pharmaceutical compositions formed by 
combining a texaphyrin of the present invention and the pharmaceutically 
acceptable carriers are then easily administered in a variety of dosage 
forms such as injectable solutions. 
For parenteral administration, solutions of the texaphyrin in sesame or 
peanut oil, aqueous propylene glycol, or in sterile aqueous solution may 
be employed. Such aqueous solutions should be suitably buffered if 
necessary and the liquid diluent first rendered isotonic using, for 
example, saline or glucose. These particular aqueous solutions are 
especially suitable for intravenous, intramuscular, subcutaneous and 
intraperitoneal administration. In this connection, sterile aqueous media 
which can be employed will be known to those of skill in the art in light 
of the present disclosure. 
The pharmaceutical forms suitable for injectable use include sterile 
aqueous solutions or dispersions and sterile powders for the 
extemporaneous preparation of sterile injectable solutions or dispersions. 
In all cases the form must be sterile and must be fluid to the extent that 
easy use with a syringe exists. It must be stable under the conditions of 
manufacture and storage and must be preserved against the contaminating 
action of microorganisms, such as bacteria and fungi. The carrier can be a 
solvent or dispersion medium containing, for example, water, ethanol, 
polyol (for example, glycerol, propylene glycol, and liquid polyethylene 
glycol, and the like), suitable mixtures thereof, and vegetable oils. The 
proper fluidity can be maintained, for example, by the use of a coating, 
such as lecithin, by the maintenance of the required particle size in the 
case of dispersion andby the use of surfactants. The prevention of the 
action of microorganisms can be brought about by various antibacterial and 
antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic 
acid, thimerosal, and the like. In many cases, it will be preferable to 
include isotonic agents, for example, sugars such as mannitol or dextrose 
or sodium chloride. A more preferable isotonic agent is a mannitol 
solution of about 2-8% concentration, and, most preferably, of about 5% 
concentration. Prolonged absorption of the injectable compositions can be 
brought about by the use in the compositions of agents delaying 
absorption, for example, aluminum monostearate and gelatin. 
Sterile injectable solutions are prepared by incorporating the active 
compounds in the required amount in the appropriate solvent with various 
of the other ingredients enumerated above, as required, followed by 
filtered sterilization. Generally, dispersions are prepared by 
incorporating the various sterilized active ingredients into a sterile 
vehicle which contains the basic dispersion medium and the required other 
ingredients from those enumerated above. In the case of sterile powders 
for the preparation of sterile injectable solutions, the preferred methods 
of preparation are vacuum-drying and freeze-drying techniques which yield 
a powder of the active ingredient plus any additional desired ingredient 
from a previously sterile-filtered solution thereof. 
As used herein, "pharmaceutically acceptable carrier" includes any and all 
solvents, dispersion media, coatings, antibacterial and antifungal agents, 
isotonic and absorption delaying agents and the like. The use of such 
media and agents for pharmaceutically active substances is well known in 
the art. Except insofar as any conventional media or agent is incompatible 
with the active ingredient, its use in the therapeutic compositions is 
contemplated. Supplementary active ingredients can also be incorporated 
into the compositions. 
The following examples describe the synthesis of texaphyrin metal complexes 
having a substituent(s) at the 2, 7, 12, 15, 18 and/or 21 position(s) of 
the macrocycle. Unless defined otherwise, all technical and scientific 
terms used herein have the same meaning as commonly understood by one of 
ordinary skill in the art to which this invention belongs. Although any 
methods and materials similar or equivalent to those described herein can 
be used in the practice or testing of the present invention, the preferred 
methods and materials are now described. Unless mentioned otherwise, the 
techniques employed herein are standard methodologies well known to one of 
ordinary skill in the art. 
EXAMPLE 1 
Synthesis of Compounds A3, A5, A6 and A7 
This example describes the synthesis of a texaphyrin metal complex having 
substituents at the 12 (R.sub.5), 15 (R.sub.6), 18 (R.sub.9) and 21 
(R.sub.10) positions of the macrocycle as depicted in Scheme A, parts 1 
and 2; a tripyrrane ketone A5, a substituted ortho-phenylenediamine A3, a 
nonaromatic texaphyrin A6, and a metal complex of aromatic texaphyrin A7. 
All solvents and reagents are of reagent grade quality, available 
commercially, and are used without further purification. Sigma lipophilic 
Sephadex (LH-20-100) and Merck type 60 (230-400 mesh) silica gel are used 
for column chromatography. 
.sup.1 H and .sup.13 C NMR spectra are obtained on a General Electric 
QE-300 (300 MHz.) spectrometer. Electronic spectra are recorded on a 
Beckman DU-7 spectrophotometer in CHCl.sub.3. Infrared spectra are 
recorded, as KBr pellets, from 4000 to 600 cm.sup.-1 on a Nicolet 510P 
FT-IR spectrophotometer. Chemical ionization mass spectrometric analyses 
(CI MS) are made using a Finnigan MAT 4023. Low resolution and high 
resolution fast atom bombardment mass spectrometry (FAB MS) are performed 
with a Finnigan-MAT TSQ-70 and VG ZAB-2E instruments, respectively. A 
nitrobenzyl alcohol (NBA) matrix is utilized with CHCl.sub.3 as the 
co-solvent. Elemental analyses are performed by Atlantic Microlab, Inc. 
Melting points are measured on a Mel-temp apparatus and are uncorrected. 
##STR4## 
Tripyrrane ketone A5: An example of the synthesis of a precursor to a 
tripyrrane ketone, the 
2,5-bis[(3-(3-hydroxypropyl)-5-carboxyl-4-methylpyrrol-2-yl)methyl]-3,4-di 
ethylpyrrole F5, Scheme F, was presented in prior application, U.S. Ser. 
No. 08/135,118, incorporated by reference herein. In this example, R.sub.1 
is 3-hydroxypropyl, R.sub.2 and R.sub.3 are ethyl and R.sub.4 is methyl. 
The synthesis of compound F5 provides teachings for the synthesis of A4, 
precursor to tripyrrane ketone A5 as shown in Scheme F and described 
herein. 
##STR5## 
2,5-Bis[(5-benzyloxycarbonyl-4-methyl-3-methoxycarbonylethylpyrrol-2-yl)met 
hyl]-3,4-diethylpyrrole. F3, Scheme F. In a 500 mL round bottom flask was 
placed 250 mL of ethanol from an unopened bottle which is purged with dry 
nitrogen for ten minutes. 3,4-Diethylpyrrole F2 (1.29 g, 0.01 mol) and 
2-acetoxymethyl-5-benzyloxycarbonyl-4-methyl-3-methoxycarbonylethylpyrrole 
F1 (7.83 g, 0.02 mol) were added and the mixture heated until all of the 
pyrroles dissolved. p-Toluenesulfonic acid (65 mg) was added and the 
reaction temperature maintained at 60.degree. C. The reaction slowly 
changed color from a clear yellow to a dark red with the product 
precipitating out of the solution as the reaction progressed. After ten 
hours the reaction was cooled to room temperature, the volume reduced to 
one half on a rotary evaporator, and then placed in the freezer for 
several hours. The product was collected by filtration, washed with a 
small amount of cold ethanol to afford 4.61 g of an off-white fine powder 
(61%): .sup.1 H NMR (CDCl.sub.3, 250 MHz): .delta.1.14 (6H, t, CH.sub.2 
CH.sub.3), 2.23 (6H, s, pyrrole-CH.sub.3), 2.31 (4H, t, CH.sub.2 CH.sub.2 
CO.sub.2 CH.sub.3), 2.50 (4H, q, CH.sub.2 CH.sub.3), 2.64 (4H, t, CH.sub.2 
CH.sub.2 CO.sub.2 CH.sub.3), 3.60 (10H, br s, CH.sub.3 CO.sub.2 -- and 
(pyrrole).sub.2 --CH.sub.2), 4.44 (4H, br s, C.sub.6 H.sub.5 CH.sub.2), 
6.99-7.02 (4H, m, aromatic), 7.22-7.26 (6H, m, aromatic), 8.72 (1H, s, 
NH), 10.88 (2H, br s, NH); .sup.13 C NMR (CDCl.sub.3, 250 MHz): 
.delta.10.97, 16.78, 17.71, 19.40, 22.07, 35.09, 51.46, 65.32, 117.37, 
119.34, 122.14, 126.58, 126.79, 127.36, 128.19, 133.55, 136.62, 162.35, 
173.49; CI MS (M+H).sup.+ 750; HRMS 749.3676 (calc. for C.sub.44 H.sub.51 
N.sub.3 O.sub.8 : 749.3676). 
A synthetic scheme is presented in Scheme G for the attachment of an ester, 
a carboxyl and a tertiary amide as R.sub.2 and R.sub.3 substituents. The 
synthesis of compound G1 is described in Kaesler et al. (1983). 
##STR6## 
2,5-Bis[(5-benzyloxycarbonyl-3-(3-hydroxypropyl)-4-methylpyrrol-2-yl)methyl 
]-3,4-diethylpyrrole- F4, Scheme F. 
2,5-Bis[(5-benzyloxycarbonyl-4-methyl-3-methoxycarbonylethylpyrrol-2-yl)me 
thyl]-3,4-diethylpyrrole F3 (5.00 g, 0,007 mol) was placed in a three 
necked 100 mL round bottom flask and vacuum dried for at least 30 minutes. 
The flask was equipped with a thermometer, an addition funnel, a nitrogen 
inlet tube, and a magnetic stir bar. After the tripyrrane was partially 
dissolved into 10 mL of dry THF, 29 mL of borane (1M BH.sub.3 in THF) was 
added dropwise with stirring. The reaction became mildly exothermic and 
was cooled with a cool water bath. The tripyrrane slowly dissolved to form 
a homogeneous orange solution which turned to a bright fluorescent orange 
color as the reaction went to completion. After stirring the reaction for 
one hour at room temperature, the reaction was quenched by adding methanol 
dropwise until the vigorous effervescence ceased. The solvents were 
removed under reduced pressure and the resulting white solid redissolved 
into CH.sub.2 Cl.sub.2. The tripyrrane was washed three times with 0.5M 
HCl (200 mL total), dried over anhydrous K.sub.2 CO.sub.3, filtered, and 
the CH.sub.2 Cl.sub.2 removed under reduced pressure until crystals of the 
tripyrrane just started to form. Hexanes (50 Ml) was added and the 
tripyrrane allowed to crystallize in the freezer for several hours. The 
product was filtered and again recrystallized from CH.sub.2 Cl.sub.2 
/ethanol. The product was collected by filtration and vacuum dried to 
yield 3.69 g of an orangish white solid (76%): mp 172.degree.-173.degree. 
C.; .sup.1 H NMR (CDCl.sub.3, 300 MHz): .delta.1.11 (6H, t, CH.sub.2 
CH.sub.3), 1.57 (4H, p, CH.sub.2 CH.sub.2 CH.sub.2 OH), 2.23 (6H, s, 
pyrrole--CH.sub.3), 2.39-2.49 (8H, m, CH.sub.2 CH.sub.3 and CH.sub.2 
CH.sub.2 CH.sub.2 OH), 3.50 (4H, t, CH.sub.2 CH.sub.2 CH.sub.2 OH), 3.66 
(4H, s, (pyrrole).sub.2 --CH.sub.2), 4.83 (4H, s, C.sub.6 H.sub.5 
--CH.sub.2), 7.17-7.20 (4H, m, aromatic), 7.25-7.30 (6H, m, aromatic), 
8.64 (1H, s, NH), 9.92 (2H, s, NH); .sup.13 C NMR (CDCl.sub.3, 300 MHz): 
.delta.10.97, 16.72, 17.68, 20.00, 22.38, 33.22, 62.01, 65.43, 117.20, 
119.75, 120.72, 122.24, 127.23, 127.62, 128.30, 132.95, 136.60, 162.13; 
FAB MS (M.sup.+) 693. 
2,5-Bis[(3-(3-hydroxypropyl)-5-carboxyl-4-methyl pyrrol-2-yl) 
methyl]-3,4-diethylpyrrole F5, Scheme F. 2,5 
-Bis[(3-(3-hydroxypropyl)-5-benzyloxycarbonyl-4-methylpyrrol-2-)methyl]-3, 
4 -diethylpyrrole F4 (15.0 g, 0.02 mol) was placed in a 1 L round bottom 
flask and dried in vacuo for ca. 30 min. The tripyrrane was dissolved in 
dry THF (600 mL) with triethylamine (10 drops) and 10% Pd on carbon (600 
mg) and the reaction was stirred at room temperature under one atmosphere 
of H.sub.2. After 15 h, the suspension was filtered through celite to 
remove the catalyst and the resulting clear solution was concentrated 
under reduced pressure to yield a light pink solid. This material, 
obtained in near quantitative yield, was taken on to the next step without 
further purification. 
A carboxyl tripyrrane A4 (a specific example presented as F5 in Scheme F) 
(0.02 mol) is placed in a 250 mL round bottom flask and dried in vacuo for 
ca. 1 h. At room temperature under nitrogen, trifluoroacetic acid (31 mL, 
0.40 mol) is added dropwise via syringe. The tripyrrane dissolves with 
visible evolution of CO.sub.2 to form a homogeneous yellow solution. The 
reaction is stirred at room temperature for ca. 15 min, then cooled to 
0.degree. C. using a water/ice bath. A triethyl-ortho-ester (or 
trimethyl-ortho-ester, ca. 18 eq) is added to the reaction mixture 
dropwise with stirring after which the reaction is stirred for an 
additional 15 minutes at 0.degree. C. If the ester is acetate, then a 
methyl group would be attached, propionate would attach an ethyl group, 
for example. The reaction is warmed to room temperature and 100 mL of 
water added dropwise. After stirring the resulting two phase mixture for 
ca. 30 minutes, the reaction mixture is extracted three times with 
CH.sub.2 Cl.sub.2. The CH.sub.2 Cl.sub.2 extracts are combined and washed 
three times with 1M aq. NaHCO.sub.3, once with water, dried over anhydrous 
sodium sulfate, filtered, and the solvent removed under reduced pressure. 
The resulting solid is recrystallized from CH.sub.2 Cl.sub.2 / hexanes. 
Substituted ortho-phenylenediamine: The synthesis of an 
ortho-phenylenediamine substituted at the 4 and 5 positions is described 
in U.S. Pat. No. 5,252,720 and application Ser. No. 08/135,118. 
Texaphyrin macrocycles having a free carboxyl or a free amino group for 
further derivatization on the benzene ring portion of the molecule may be 
synthesized by replacing ortho-phenylenediamine with 3,4-diaminobenzoic 
acid or 3,4-diaminoaniline. One skilled in the art of organic synthesis 
would realize in light of the present disclosure that other substituted 
1,2-o-phenylenediamines may be used as a precursor, e.g., a 
1-2-o-phenylenediamine that is differentially substituted in the 4 and 5 
positions. This substitution may be the result of different 
functionalities being present or specific protection and standard organic 
and/or biochemical transformations having been carried out. Such 
macrocycles can be further functionalized to derivatives having an 
antibody, oligonucleotide, protein, peptide, sapphyrin and the like on one 
position of the B portion of the molecule. 
Synthesis of A3, Scheme A: Compound A1 of Scheme A (a 
1,2-dialkyl-4,5-dinitrobenzene) is reacted with an alkyl halide where the 
halide is chloride, bromide or iodide in the presence of a Lewis acid such 
as AlCl.sub.3, for example. The 3 and 6 positions of the phenyl ring are 
derivatized with the alkyl group to form compound A2. A mixture of 
reactants having a single halide and different alkyl groups may be used to 
generate different alkyl derivatives at the 3 and 6 positions. The yield 
of a particular product would be lower in this case. 
A diemine A3 (Scheme A) is obtained by reduction of the corresponding 
substituted dinitrobenzene (A2, Scheme A) with hydrazine hydrate (1 mL) 
and 10% palladium on carbon (50 mg) in 40 mL refluxing absolute methanol. 
The resulting suspension may bubble for approximately 15-20 minutes and 
then turn colorless after 1 hour. At this point the reduction is complete 
as verified by TLC. The reaction solution is hot filtered through celite 
into a dry flask, covered with aluminum foil, and then concentrated to an 
oil. The diamine is taken to the next step without further purification. 
Ammonium formate in the presence of palladium (10% on carbon) catalyst may 
act as a mild, inexpensive and safe alternative to hydrazine hydrate in 
the above reaction and would be used, for example, when sensitive groups 
such as amide are present at other positions of the molecule. 
Condensation of a tripyrrane ketone and a substituted 
ortho-phenylenediamine to form a nonaromatic texaphyrin having 
substituents at the 2, 7, 12, 15, 18 and/or 21 position(s): A tripyrrane 
ketone and a substituted ortho-phenylenediamine having substituents at the 
3 and/or 6 position(s) are placed in a 2 L round bottom flask with 1000 mL 
of toluene and 200 mL of methanol. The solvents are purged with nitrogen 
prior to use. Concentrated HCl (0.5 mL) is added and the reaction heated 
to reflux under nitrogen. After 5 h the reaction is cooled to room 
temperature and the solvents removed under reduced pressure until the 
product precipitates out of solution. The remainder of the solvent is 
decanted off and the macrocycle is dried in vacuo. The product is 
recrystallized from methanol/diethylether and characterized by .sup.1 H 
NMR and .sup.13 C NMR. 
Condensation of a diformyltripyrrole and a substituted 
ortho-phenylenediamine yields a nonaromatic texaphyrin having substituents 
in the 15, 16, 17 or 18 positions. 
General procedure for the synthesis of a metal complex of texaphyrin (A7, 
Scheme A). One equivalent of the hydrochloride salt of the macrocycle A6, 
1.5 equivalents of the M(OAc.sub.3).sub.3.XH.sub.2 O metal salt (where 
M=metal ion), and triethylamine (ca. 1 mL) are mixed together in methanol 
and heated to reflux under air. After completion of the reaction (as 
judged by the UV/vis spectrum of the reaction mixture), the solution is 
cooled to room temperature, the solvent is removed under reduced pressure 
and the crude complex dried in vacuo for several hours. A solution of 
dichloromethane/methanol (99:1 v/v) is added to the crude complex and the 
suspension is sonicated a few min. The suspension is filtered in order to 
remove impurities in the filtrate (incomplete oxidation products and 
excess triethylamine). The resulting solid is dissolved in methanol and 
then chloroform is added to reduce the polarity of the mixture (1:2 v/v). 
This solution is filtered through celite and loaded on a 
(pre-treated/pre-washed 1M NaNO.sub.3) neutral alumina column (10 cm). The 
column is first eluted with a 1:10 (v/v) methanol/chloroform solution by 
gravity to remove any impurity. The metal complex is then obtained by 
eluting the column with chloroform containing increasing amounts of 
methanol (20-50%). The purified lanthanide(III) texaphyrin complex is 
recrystallized by dissolving the complex in methanol/chloroform and 
carefully layering the solution with a small amount of methanol, then with 
diethylether. The layered solution is kept at room temperature in the dark 
for a few days. The texaphyrin metal complex is recrystallized twice for 
analytically pure measurements and characterizations. 
Alternatively, the crude metal complex may be isolated by mixing the 
complex with an aqueous solution of a salt at a temperature above the 
freezing point of the resulting mixture, and then recovering the 
precipitated texaphyrin from the mixture. The salt can be any salt that is 
soluble in water or a water/organic solvent mixture and does not cause 
transmetallation of the texaphyrin metal complex. 
The texaphyrin metal complex may alternatively be purified by dissolving 
the complex in water and methanol, and acid-washed zeolites (such as 
LZY-54 zeolite) are then added to the solution. The mixture is agitated 
for a period of time and is then filtered to remove the zeolites. This 
procedure may be repeated 2 or more times until significantly all of the 
free metal ion is removed. 
Lanthanum(III), Cerium(III), Praseodymium(III), Neodymium(III), 
Samarium(III), Europium(III), Gadolinium(III), Terbium(III), 
Dysprosium(III), Holmium(III), Erbium(III), Thulium(III), Ytterbium(III), 
Lutetium(III) complexes of texaphyrin: The hydrochloride salt of 
macrocycle A6 (0.407 mmol), and one of the following lanthanide salts: 
La(OAc.sub.3).sub.3.6H.sub.2 O (0.814 mmol), Ce(OAc.sub.3).sub.3.6H.sub.2 
O (0.611 mmol), Pr(OAc.sub.3).sub.3.5H.sub.2 O (0.611 mmol), 
Nd(OAc.sub.3).sub.3.6H.sub.2 O (0.611 mmol), Sm(OAc.sub.3).sub.3.5H.sub.2 
O (0.611 mmol), Eu(OAc.sub.3).sub.3.5H.sub.2 O (0.65 mmol), 
Gd(OAc.sub.3).sub.3.5H.sub.2 O (1.5 mmol), Tb(OAc.sub.3).sub.3.6H.sub.2 O 
(0.611 mmol), Dy(OAc.sub.3).sub.3.5H.sub.2 O (0.611 mmol), 
Ho(OAc.sub.3).sub.3.5H.sub.2 O (0.611 mmol), Er(OAc.sub.3).sub.3.5H.sub.2 
O (0.611 mmol), Tm(OAc.sub.3).sub.3.5H.sub.2 O (0.611 mmol), 
Yb(OAc.sub.3).sub.3.5H.sub.2 O (0.611 mmol), or 
Lu(OAc.sub.3).sub.3.H.sub.2 O (0.611 mmol), together with TBANO.sub.3 (1.0 
mmol) and triethylamine (ca. 0.5 mL) in 350 mL methanol are heated to 
reflux under air for 5-24 h. The workup uses the general procedure 
outlined above. The thulium and lutetium complexes may be more difficult 
to purify due to their lower solubility in methanol/chloroform solutions, 
which leads to a lower yield. 
EXAMPLE 2 
Synthesis of Compounds B4, C5 and D5 
Ortho-phenylenediamine compounds having substituents bound to the phenyl 
ring via an oxygen are prepared as indicated in Schemes B and C. 
##STR7## 
2,3,4-Trihydroxybenzoic acid B1, is reacted with an alkyl halide where the 
halide is chloride, bromide, or iodide in the presence of potassium 
carbonate and acetonitrile to form a trialkoxy derivative B2. The alkyl 
group of the halide may be a primary or secondary alkyl having one or more 
hydroxy, alkoxy, carboxy, ester, amine, amide or protected amine 
substituents at positions at least one carbon removed from the site of 
halide attachment. These alkyl groups may be unsubstituted, singly or 
multiply functionalized. They may be branched or unbranched. Preferred 
alkyl groups are methyl, hydroxypropyl or methoxy(ethoxy).sub.n ethoxy 
(n=1-100; a polyethylene glycol substituent). Compound B2 is reacted with 
90% nitric acid to form the dinitro derivative B3 which is then reacted 
with either hydrazine hydrate or ammonium formate and 10% palladium on 
carbon in methanol to form compound B4. 
In a similar synthesis, starting with 2,3,4-trihydroxybenzaldehyde C1 
(Scheme C), reduction of the trialkoxy derivative C2 with hydrazine in KOH 
results in a methyl derivative at the R.sub.6 position to form 
1,2,3-trialkoxy-4-methylbenzene C3. The diamine is formed as depicted in 
Scheme B and described above. 
Scheme D shows the formation of a tertiary amine at the R.sub.6 position. 
The starting material is 2,3,4-trihydroxybenzoic acid (D1). Compound D3 
(B3) is treated with an amine component in 1,3-dicyclohexylcarbodiimide 
and dimethylformamide to form D4 having an amide linkage. Alternative 
coupling reagents include 1,1'-carbonyldiimidazole (CDI) or ECC. Reduction 
as described above yields the diamine for condensation with a tripyrrane 
ketone. 
##STR8## 
EXAMPLE 3 
Synthesis of a T2B4 Texaphyrin 
Scheme E, parts 1 and 2, shows the synthesis of a lanthanide metal complex 
of a T2B4 texaphyrin. A diformyltripyrrole E5 is condensed with a 
substituted ortho-phenylenediamine E4 to form the nonaromatic precursor 
E6. The synthesis of the substituted ortho-phenylenediamine E4 was 
described in example 2 and the diformyltripyrrole was described in U.S. 
Pat. No. 5,252,720. In this example, R' may be polyethylene glycol (PEG) 
where the number of repeating ethoxy units may be as many as 200, a 
saccharide, a polyhydroxy substituent or the like. R may be methoxy, 
methyl or hydrogen. 
##STR9## 
EXAMPLE 4 
Synthesis of a Tripyrrane Having Meso-substituents 
Scheme A, parts 1 and 2, refers to the structure of a metallotexaphyrin 
with substituents in the 2 and 7 positions (meso-positions). Texaphyrin 
macrocycles having meso-substitution on the periphery of the aromatic 
macrocycle may be synthesized by first preparing new 
methylene-functionalized tyripyrrane dialdehydes described in Scheme I, 
parts 1 and 2. One skilled in the art of organic synthesis would realize 
in light of the present disclosure that a variety of 
1,2-o-phenylenediamines may be used to react with these new functionalized 
tripyrranes. The organic synthesis required for the various 
transformations illustrated in Scheme I is derived from classic 
pyrrole/porphyrin chemistry. 
Synthesis of I3, Scheme I, part 1: Pyrrole I1 (readily available from 
Aldrich Chemical Co., Milwaukee, Wis.) of Scheme I is reacted with 
sulfuryl chloride in dichloromethane, followed by hydrolysis with sodium 
acetate, and acidification to afford the acid pyrrole, I2 (see A. R. 
Battersby et al., J. C. S. Perkin I, 1976, 1008). Decarboxylation via 
trifluoroacetic acid yields I3 (see M. J. Cyr, Ph.D. Dissertation, 
University of Texas at Austin, 1992). 
##STR10## 
Synthesis of I5. The acid-catalyzed condensation between compound I3 and 
the t-butylester derived pyrrole I4 (pyrrole I4 is described in D. H. R. 
Barton and S. Zard, J.C.S. Chem. Commun., 1985, 1098-1100), in the 
presence of an aldehyde (R.sub.12 =alkyl, aryl, etc.) will afford a 
mixture of three dipyrromethanes. The desired mixed-ester derived 
dipyrromethane I5 is obtained by column chromatography. The preparation of 
dipyrromethanes is well-documented in the literature (see, Sessler et al., 
J. Org. Chem., 1986, 51, 2838). 
Synthesis of I7. The t-butylester of compound I5 is selectively deprotected 
and decarboxylated via trifluoroacetic acid and subsequently condensed via 
acid-catalysis with pyrrole I3 in the presence of an aldehyde (R.sub.11 
=alkyl, aryl, etc.) to afford the desired tripyrrane I7. 
Synthesis of the diformyl tripyrrane I9. With compound I7 in hand, the 
tripyrrane is transformed to the desired diformyl tripyrrane I9 (R.sub.5 
=H) by standard organic synthesis reported earlier (U.S. Pat. No. 
5,252,720). Compound I7 is reduced by borane/THF, followedby acetylation 
via acetic anhydride or acetyl chloride to afford tripyrrane I8. At this 
point, debenzylation of I8, followed by subsequent Clezy formylation of 
the intermediate, and basic hydrolysis with lithium hydroxide, provides 
tripyrrane I9. 
Tripyrrane I9 may then be condensed with an ortho-phenylenediamine to 
construct a texaphyrin macrocycle as depicted in Scheme A. Substituents in 
these meso-positions are expected to further stabilize the macrocycle. 
EXAMPLE 5 
2,5-Bis 
[(3-acetoxypropyl-5-benzoyl-4-methylpyrrol-2-yl)methyl]-3,4-diethylpyrrole 
(J2). 
2,5-Bis[(3-acetoxypropyl-5-carboxyl-4-methylpyrrol-2-yl)methyl]-3,4-diethy 
lpyrrole J1 (1.00 g, 1.67 mmol) was placed in a 100 mL three-neck 
round-bottom flask and dried under high vacuum for ca. 1 hr. The 
round-bottom flask was equipped with an argon inlet line and for magnetic 
stirring. At room temperature under argon, CH.sub.2 Cl.sub.2 (10 mL) was 
added to the flask and the resulting mixture stirred to form a suspension. 
Trifluoroacetic acid (2.7 mL) was then added all at once to the 
suspension. The tripyrrane dissolved to form a light orange solution. The 
reaction was stirred at room temperature under argon for ca. 45 min, after 
which it was cooled to 0.degree. C. using an ice/water bath. 
Triethylorthobenzoate (3.8 mL) was added dropwise to the reaction with 
stirring over a two minute period under a flow of argon. The reaction was 
stirred for 40 min at 0.degree. C. then allowed to warm to room 
temperature over 20 min. Water (20 mL) was added to the reaction and 
stirring continued for another 2 hr. Transferred reaction to a separatory 
funnel, separated and discarded the upper aqueous phase, and basified the 
lower organic layer with sat. aqueous NaHCO.sub.3 (Caution: gas evolution 
and frothing occurs). Separated the two layers and washed the organic 
phase once with sat. aqueous NaHCO.sub.3 and once with water. Dried 
organic phase over anhydrous MgSO.sub.4, filtered off the drying agent, 
removed the solvent under reduced pressure, and dried the resulting 
orange-red oil under high vacuum overnight. The oil was dissolved into a 
minimum amount of CH.sub.2 Cl.sub.2 (5-10 mL), the solution layered with 
hexanes (ca. 50 mL), and the tripyrrane allowed to crystallize at 
-20.degree. C. The product was collected by filtration and dried under 
high vacuum to yield 1.06 grams of a tan solid (J2) (88%). .sup.1 H NMR 
(CDCl.sub.3, 300 MHz): .delta.1.06 (6H, t, CH.sub.2 CH.sub.3), 1.67 (4H, 
p, CH.sub.2 CH.sub.2 CH.sub.2 OAc), 1.81 (6H, s, CH.sub.3 CO.sub.2 --), 
2.02 (6H, s, pyrr-CH.sub.3), 2.37-2.44 (8H, m, CH.sub.2 CH.sub.3 and 
CH.sub.2 CH.sub.2 CH.sub.2 OAc), 3.71 (4H, s, (pyrr).sub.2 --CH.sub.2), 
3.99 (4H, t, CH.sub.2 CH.sub.2 CH.sub.2 OAc), 7.29-7.48 (10H, m, 
aromatic), 9.16 (1H, s, NH), 9.66 (2H, s, NH); .sup.13 C NMR (CDCl.sub.3): 
.delta.11.8, 16.4, 17.7, 20.1, 20.9, 22.7, 29.1, 63.9, 120.9, 121.5, 
127.3, 128.1, 128.2, 129.1, 130.8, 135.6, 140.1, 171.3, 185.7; FAB MS, 
M.sup.+ : m/e 717. 
4,5-Diethyl-9,24-bis 
(3-hydroxypropyl)-16,17-dimethoxy-10,23-dimethyl-12,21-diphenyl-13,20,25,2 
6,27-pentaazapentacyclo-[20.2.1.1.sup.3,6.1.sup.8,11.0.sup.14,19 
]heptacosa-3,5,8,10,12,14,16,18,20,22,24-undecaene (J6). 
2,5-Bis[(3-acetoxypropyl-5-benzoyl-4-methylpyrrol-2-yl)methyl]-3,4-diethyl 
pyrrole J2 (100 mg, 0.14 mmol) and 4,5-dimethoxy-1,2-phenylenediamine J5 
(23 mg, 0.14 mmol) were dissolved into 100 mL of absolute methanol under 
argon. Concentrated HCl (5 drops) was added and the reaction heated at 
reflux under argon. After heating for 2 days, the reaction was cooled to 
room temperature and the solvent removed under reduced pressure. The 
resulting red solid was dissolved into CH.sub.2 Cl.sub.2 (5 mL), filtered, 
and the CH.sub.2 Cl.sub.2 solution layered with hexanes (20 mL). The 
product was allowed to slowly precipitate out of solution at room 
temperature overnight. The mother liquor was decanted off and the 
remaining solid washed with hexanes. After drying the solid under high 
vacuum, 39 mg of dark red product (J6) was obtained. FAB MS, (M+H).sup.+ : 
m/e 766. 
Cadmium(II) complex of 4,5-diethyl-9,24-bis (3-hydroxypropyl)-16, 
17-dimethoxy-10,23-dimethyl-12,21-diphenyl-13,20,25,26,27-pentaazapentacyc 
lo [20.2.1.1.sup.3,6.1.sup.8,11.0.sup.14,19 ]-heptacosa-1,3,5,7,9,11(27), 
12,14 (19), 15, 17,20,22(25),23-tridecaene (J8). The protonated form of 
the macrocycle J6 (11 mg, 0.014 mmol), cadmium(II) chloride (11 mg, 0.06 
mmol) and triethylamine (20 mL) in 20 mL of methanol were heated at reflux 
under air for 2 days. The reaction was cooled to room temperature, the 
solvent removed under reduced pressure, and the complex dried in vacuo 
overnight to give the final texaphyrin-Cd(II) metal complex (J8). UV/vis 
(CH.sub.3 OH) [.lambda..sub.max, nm]: 472.0, 756.0; FAB MS, (M+H).sup.+ : 
m/e 875. 
##STR11## 
EXAMPLE 6 
2,5-bis[(5-benzoyl-3-ethyl-4-methylpyrrol-2-yl)methyl]-3,4-diethylpyrrole 
(J4). 
2,5-Bis[(5-carboxyl-3-ethyl-4-methylpyrrol-2-yl)methyl]-3,4-diethylpyrrole 
J3 (1.00 g, 2.20 mmol) was placed in a 100 mL three-neck round-bottom 
flask and dried under high vacuum for 1 hr. The round-bottom flask was 
equipped with an argon inlet line and for magnetic stirring. At room 
temperature under argon, CH.sub.2 Cl.sub.2 (10 mL) was added to the 
reaction flask and the resulting mixture stirred to form a suspension. 
Trifluoroacetic acid (3.5 mL) was then added all at once to the 
suspension. The tripyrrane dissolved to form a yellowish orange solution. 
The reaction was stirred at room temperature under argon for ca. 35 min, 
after which it was cooled to 0.degree. C. using an ice/water bath. 
Triethylorthobenzoate (5.0 mL) was added dropwise to the reaction with 
stirring over a two minute period under a flow of argon. The reaction was 
stirred for 40 min at 0.degree. C. then allowed to warm to room 
temperature over 20 min. Water (20 mL) was added to the reaction and 
stirring continued for another 1 hr. Transferred reaction to a separatory 
funnel, separated and discarded the upper aqueous phase, and basified the 
lower organic layer with sat. aqueous NaHCO.sub.3 (30 mL) (Caution: gas 
evolution and frothing occurs). Separated the two layers and washed the 
organic phase once with sat. aqueous NaHCO.sub.3 and once with water. 
Dried organic phase over anhydrous MgSO.sub.4, filtered off the drying 
agent, and removed the solvent under reduced pressure to yield a dark oil 
with some precipitate. The oil and solid were dissolved into a minimum 
amount of CH.sub.2 Cl.sub.2 (5 mL), the solution layered with hexanes (ca. 
50 mL), and the product allowed to crystallize at -20.degree. C. The 
product was collected by filtration, washed with a small amount of 
hexanes, and dried under high vacuum to yield 0.88 grams of a tan solid 
(J4) (70%). .sup.1 H NMR (CDCl.sub.3, 300 MHz): .delta.0.95 (6H, t, 
CH.sub.2 CH.sub.3), 1.05 (6H, t, CH.sub.2 CH.sub.3), 1.80 (6H, s, 
pyrr--CH.sub.3), 2.32-2.40 (8H, m, CH.sub.2 CH.sub.3), 3.67 (4H, s, 
(pyrr).sub.2 --CH.sub.2), 7.27-7.48 (10H, m, aromatic), 9.27 (1H, s, NH), 
9.66 (2H, s, NH); .sup.13 C NMR (CDCl.sub.3): .delta.11.7, 15.1, 16.3, 
17.1, 17.7, 22.8, 120.8, 121.4, 124.8, 127.0, 128.0(6), 128.1(4), 129.2, 
130.6, 135.5, 140.2, 185.7; FAB MS, (M+H).sup.+ : m/e 574. 
4,5,9,24-Tetraethyl-16,17-dimethoxy-10,23-dimethyl-12,21-diphenyl-13,20,25, 
26,27-pentaazapentacyclo-[20.2.1.1.sup.3,6.1.sup.8,11.0.sup.14,19 
]heptacosa-3,5, 8,10,12,14,16,18,20, 22,24-undecaene (J7). 
2,5-Bis[(5-benzoyl-3-ethyl-4-methylpyrrol-2-yl)methyl]-3,4-diethylpyrrole 
J4 (101 mg, 0.18 mmol) and 4,5-dimethoxy-1,2-phenylenediamine J5 (30 mg, 
0.18 mmol) were dissolved into 200 mL of toluene and 100 mL of absolute 
methanol. The solvents were sparged with argon for approximately 5 min 
before the reaction was started. Concentrated HCl (3 drops) was then added 
and the reaction heated at reflux under an atmosphere of argon. After 
heating for ca. 2.75 days, the reaction was cooled to room temperature, 
the solvent removed under reduced pressure and the remaining solid dried 
in vacuo. The macrocycle was dissolved into CH.sub.2 Cl.sub.2 (10 mL), 
filtered, and the CH.sub.2 Cl.sub.2 solution layered with hexanes (80 mL). 
The product was allowed to slowly precipitate out of solution at 
-20.degree. C. overnight. The macrocycle was collected by filtration, 
dissolved into a minimum amount of ethanol, and the solution layered with 
hexanes. The macrocycle was allowed to slowly precipitate out of solution 
at -20.degree. C. for several days. The macrocycle was collected by 
filtration, washed with a small amount of hexanes, and dried under high 
vacuum to yield 28 mg of dark red product (J7). FAB MS, (M+H).sup.+ : m/e 
707. 
Macrocycle J7 can be oxidized and metallated to give the corresponding 
texaphyrin metal complex following the procedures previously described 
herein. 
##STR12## 
EXAMPLE 7 
R.sub.5, R.sub.6, R.sub.9 and/or R.sub.10 Substituents 
Scheme H, parts 1 and 2, shows a synthetic scheme for attaching a nitro 
group at position R.sub.6 or R.sub.9. 
##STR13## 
A 1,2-dialkyl-4,5-dinitrobenzene (H1, also A1) is reduced with ammonium 
formate to the diamino derivative and an amine protecting group is 
attached before the nitration step. Amine protecting groups include amides 
such as N-acetyl, and carbamates such as CBZ, for example. An acetyl 
protecting group is later removed by refluxing in HCl. Protection and 
deprotection procedures are well known to those of skill in the art in 
light of the present disclosure (Greene et al. 1991). The deprotected 
nitro derivative H5 is condensed with a diformyltripyrrane H6 to form a 
nonaromatic texaphyrin having a nitro group at the 15 position. 
A bromine is introduced at the R.sub.6 and R.sub.9 positions of the 
macrocycle by reacting 1,2-dialkyl-4,5-dinitrobenzene with bromine in the 
presence of FeBr.sub.3 or AlBr.sub.3. The 3 and 6 positions of the phenyl 
ring are derivatized with bromide and reduction to the amine as described 
in example 2 prepares the precursor for condensation with a 
diformyltripyrrole or a tripyrrane ketone. 
Preferred texaphyrins having a substituent on the 2, 7, 12, 15, 18 and/or 
21 position of the macrocycle are listed in Tables A and B. Substituents 
R.sub.1 -R.sub.6 are provided in Table A and R.sub.7 -R.sub.12 are 
provided in Table B for a given texaphyrin ("TXP"). 
TABLE A 
__________________________________________________________________________ 
Representative Substituents for Texaphyrin Macrocycles A1-A50 of the 
Present Invention. 
Substituents for R.sub.1 -R.sub.6 are provided in TABLE A and for R.sub.7 
-R.sub.12 in TABLE B. 
TXP 
R.sub.1 R.sub.2 R.sub.3 R.sub.4 
R.sub.5 R.sub.6 
__________________________________________________________________________ 
A1 CH.sub.2 (CH.sub.2).sub.2 OH 
CH.sub.2 CH.sub.3 
CH.sub.2 CH.sub.3 
CH.sub.3 
H COOH 
A2 " " " " " COOH 
A3 " " " " " CONHCH--(CH.sub.2 OH).sub.2 
A4 " " " " " " 
A5 " " " " " H 
A6 " " " " " OCH.sub.3 
A7 " " " " " " 
A8 " " " " " " 
A9 " " " " " " 
A10 
" " " " " " 
A11 
" " " " " " 
A12 
" " " " " " 
A13 
" " " " " CH.sub.3 
A14 
" " " " " " 
A15 
" " " " " " 
A16 
" " " " " " 
A17 
" " " " CH.sub.3 H 
A18 
" " " " " " 
A19 
" " " " " " 
A20 
CH.sub.2 (CH.sub.2).sub.2 OH 
CH.sub.2 CH.sub.3 
CH.sub.2 CH.sub.3 
CH.sub.3 
CH.sub.3 H 
A21 
" " " " " " 
A22 
" " " " " " 
A23 
" " " " " " 
A24 
" " " " " " 
A25 
" " " " " " 
A26 
" " " " " OH 
A27 
" " " " " F 
A28 
" " " " CH.sub.2 (CH.sub.2).sub.6 OH 
H 
A29 
" " " " H Br 
A30 
" " " " " NO.sub.2 
A31 
" " " " " COOH 
A32 
" " " " " CH.sub.3 
A33 
" " " " C.sub.6 H.sub.5 
H 
A34 
" COOH COOH " CH.sub.2 CH.sub.3 
" 
A35 
" COOCH.sub.2 CH.sub.3 
COOCH.sub.2 CH.sub.3 
" CH.sub.3 " 
A36 
CH.sub.2 CH.sub.2 CON(CH.sub.2 CH.sub.2 OH).sub.2 
CH.sub.2 CH.sub.3 
CH.sub.2 CH.sub.3 
" " " 
A37 
CH.sub.2 CH.sub.2 ON(CH.sub.3)CH.sub.2 -- 
" " " " " 
(CHOH).sub.4)CH.sub.2 OH 
A38 
CH.sub.2 CH.sub.3 
" " " CH.sub.2 (CH.sub.2).sub.6 OH 
" 
A39 
CH.sub.2 (CH.sub.2).sub.2 OH 
CH.sub.2 CH.sub.3 
CH.sub.2 CH.sub.3 
CH.sub.3 
CH.sub.3 or CH.sub.2 CH.sub.3 
H 
A40 
" " " " " " 
A41 
" " " " " " 
A42 
" " " " " " 
A43 
" " " " " " 
A44 
" " " " " " 
A45 
" " " " " " 
A46 
" " " " " " 
A47 
" " " " " " 
A48 
" " " " " " 
A49 
" " " " " " 
A50 
" " " " " " 
A51 
" " " " H " 
A52 
" " " " " " 
A53 
" " " " " " 
A54 
" " " " " " 
A55 
" " " " CH.sub.3 or CH.sub.2 CH.sub.3 
" 
A56 
" " " " " " 
__________________________________________________________________________ 
TABLE B 
__________________________________________________________________________ 
Representative Substituents for Texaphyrin Macrocycles A1-A50 of the 
Present Invention. 
Substituents for R.sub.1 -R.sub.6 Are Provided in TABLE A and for R.sub.7 
-R.sub.12 in TABLE B. 
TXP 
R.sub.7 R.sub.8 R.sub.9 R.sub.10 R.sub.11 R.sub.12 
__________________________________________________________________________ 
A1 O(CH.sub.2).sub.3 OH 
O(CH.sub.2).sub.3 OH 
O(CH.sub.2).sub.3 OH 
H H H 
A2 O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
COOH " " " 
A3 O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
O-saccharide 
" " " 
A4 " " O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
" " " 
A5 " O(CH.sub.2).sub.3 CON-linker-oligo 
" " " " 
A6 H OCH.sub.2 CON-linker-oligo 
OCH.sub.3 " " " 
A7 " OCH.sub.2 CO-poly-L-lysine 
" " " " 
A8 " OCH.sub.2 CO-estradiol 
" " " " 
A9 " O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
" " " " 
A10 
O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
" " " " " 
A11 
" OCH.sub.2 CON-linker-oligo 
" " " " 
A12 
" OCH.sub.2 CO-estradiol 
" " " " 
A13 
" O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
" " " 
A14 
" OCH.sub.2 CO-estradiol 
" " " " 
A15 
O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
O(CH.sub.2 CH.sub.2 O).sub.120 CH.sub.3 
OCH.sub.3 " " " 
A16 
H saccharide " " " " 
A17 
O(CH.sub.2).sub.3 OH 
O(CH.sub.2).sub.3 OH 
H CH.sub.3 " " 
A18 
H O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
" " " " 
A19 
O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
" " " " " 
A20 
H OCH.sub.2 CON-linker-oligo 
H CH.sub.3 " " 
A21 
" OCH.sub.2 CO-estradiol 
" " " " 
A22 
" OCH.sub.2 CON(CH.sub.2 CH.sub.2 OH).sub.2 
" " " " 
A23 
O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
O(CH.sub.2 CH.sub.2 O).sub.120 CH.sub.3 
" " " " 
A24 
" OCH.sub.2 CON-linker-oligo 
" " " " 
A25 
H CH.sub.2 CON(CH.sub.3)CH.sub.2 -- 
" " " " 
(CHOH).sub.4 CH.sub.2 OH 
A26 
O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
OH " " " 
A27 
" " F " " " 
A28 
" " H CH.sub.2 (CH.sub.2).sub.6 OH 
" " 
A29 
" " Br H " " 
A30 
" " NO.sub.2 " " " 
A31 
" " COOH " " " 
A32 
" " CH.sub.3 " " " 
A33 
" " H C.sub.6 H.sub.5 
" " 
A34 
" " " CH.sub.2 CH.sub.3 
" " 
A35 
" " " CH.sub.3 " " 
A36 
" " " " " " 
A37 
OCH.sub.3 OCH.sub.3 " " " " 
A38 
H OCH.sub.2 CO.sub.2 -glucosamine 
" CH.sub.2 (CH.sub.2).sub.6 OH 
" " 
A39 
O(CH.sub.2).sub.3 OH 
O(CH.sub.2).sub.3 OH 
H CH.sub.3 or CH.sub.2 CH.sub.3 
CH.sub.3 or CH.sub.2 
CH.sub.3 CH.sub.3 or 
CH.sub.2 CH.sub.3 
A40 
O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
" " " " 
A41 
O(CH.sub.2).sub.3 OH 
O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
" " " " 
A42 
H O(CH.sub.2).sub.n CON-linker-oligo, 
" " " " 
n = 1,2,3 
A43 
H O(CH.sub.2).sub.n CO-estradiol, 
" " " " 
n = 1,2,3 
A44 
H saccharide " " " " 
A45 
O(CH.sub.2).sub.3 OH 
O(CH.sub.2).sub.n CON-linker-oligo, 
" " " " 
n = 1,2,3 
A46 
" O(CH.sub.2).sub.n CO-estradiol, 
" " " " 
n = 1,2,3 
A47 
" saccharide " " " " 
A48 
O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
O(CH.sub.2).sub.n CON-linker-oligo, 
" " " " 
n = 1,2,3 
A49 
" O(CH.sub.2).sub.n CO-estradiol, 
" " " " 
n = 1,2,3 
A50 
" saccharide " " " " 
A51 
" O(CH.sub.2).sub.n CON-linker-oligo 
" H " " 
n = 1,2,3 
A52 
" O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
" " " " 
A53 
" " " " CH.sub.2 (CH.sub.2).sub.2 
CH.sub.2 (CH.sub.2). 
sub.2 OH 
A54 
" O(CH.sub.2).sub.n CON-linker-oligo 
" " " " 
n = 1,2,3 
A55 
" " " CH.sub.3 or CH.sub.2 CH.sub.3 
" " 
A56 
" O(CH.sub.2 CH.sub.2 O).sub.3 CH.sub.3 
" " " " 
__________________________________________________________________________ 
A substituent on the R.sub.5, R.sub.10, R.sub.11 or R.sub.12 position of 
the macrocycle may be derivatized after condensation of the macrocycle. 
Substituents may include an alkyl group having up to 5 carbon atoms or a 
phenyl group which may be further derivatized with a nitro, carboxyl, 
sulfonic acid, hydroxyl, halide or alkoxy where the alkyl of the alkoxy 
may be hydroxyalkyl and like, as described in U.S. Pat. No. 5,252,720 and 
application Ser. No. 08/135,118. 
EXAMPLE 8 
Further Derivatives of Texaphyrin 
One skilled in the art of organic synthesis in light of the present 
disclosure could extend and refine the basic synthetic chemistry outlined 
in this application, in U.S. Pat. No. 5,252,720 and in application Ser. 
No. 08/135,118 so as to produce texaphyrins having various substituents, 
yet having basic utility to those specifically detailed in the present 
examples. For example, polyether-linked polyhydroxylated groups, catechol 
(i.e. benzene diol) derivatives bearing further hydroxyalkyl substituents 
off the tripyrrane-derived portion of the macrocycle, saccharide 
substitutions in which the saccharide is appended via an acetal-like 
glycosidic linkage, an oligosaccharide or a polysaccharide may be 
similarly linked to a texaphyrin. A doubly carboxylated texaphyrin in 
which the carboxyl groups are linked to the texaphyrin core via aryl 
ethers or functionalized alkyl substituents could be converted to various 
esterified products wherein the ester linkages serve to append further 
hydroxyl-containing substituents. Polyhydroxylated texaphyrin derivatives 
may be synthesized via the use of secondary amide linkages. Saccharide 
moieties may be appended via amide bonds. Polyhydroxylated texaphyrin 
derivatives containing branched polyhydroxyl (polyol) subunits may be 
appended to the texaphyrin core via aryl ethers or ester linkages. 
Treatment of carboxylated texaphyrins with thionyl chloride or 
p-nitrophenol acetate would generate activated acyl species suitable for 
attachment to monoclonal antibodies or other biomolecules of interest. 
Standard in situ coupling methods (e.g. 1,1'-carbonyldiimidazole (CDI)) 
could be used to effect the conjugation. 
The selectivity of the texaphyrins may be enhanced by covalently linking 
oligonucleotides onto the periphery of the macrocycle. Amides, ethers and 
thioethers are representative of linkages which may be used for this 
purpose. Oligonucleotides functionalized with amines at the 5'-end, the 
3'-end, or internally at sugar or base residues may be modified 
post-synthetically with an activated carboxylic ester derivative of the 
texaphyrin complex. Alternatively, oligonucleotide analogs containing one 
or more thiophosphate or thiol groups may be selectively alkylated at the 
sulfur atom(s) with an alkyl halide derivative of the texaphyrin complex. 
The resultant oligodeoxynucleotide-texaphyrin complex conjugates may be 
designed so as to provide optimal catalytic interaction between a target 
nucleic acid and the bound texaphyrin. The oligonucleotide may be large 
enough to bind probably at least 15 nucleotides of complementary nucleic 
acid. 
A general method for preparing oligonucleotides of various lengths and 
sequences is described by Caracciolo et al. (1989). Preferred 
oligonucleotides resistant to in vivo hydrolysis may contain a 
phosphorothioate substitution at each base (J. Org. Chem., 55:4693-4699, 
1990). Oligodeoxynucleotides or their phosphorothioate analogues may be 
synthesized using an Applied Biosystem 380B DNA synthesizer (Applied 
Biosystems, Inc., Foster City, Calif.). Specific methods for preparing 
texaphyrin-oligonucleotide conjugates are disclosed in PCT publication WO 
94/29316, the disclosure of which is incorporated herein by reference. 
Another means of gaining selectivity may be to covalently link the 
texaphyrin complex to a sapphyrin (sap) molecule, (U.S. Pat. No. 
5,159,065; U.S. Pat. No. 5,120,411; U.S. Pat. No. 5,041,078, all 
incorporated by reference herein.) Since sapphyrins bind DNA, 
K.about.10.sup.6 M.sup.-1, (U.S. Ser. No. 07/964,607, incorporated by 
reference herein) the linked texaphyrin-sapphyrin complex (txph-sap) could 
effectively increase the texaphyrin concentration at locations adjacent to 
the sapphyrin binding sites. Sapphyrins have a higher fluorescent quantum 
yield than texaphyrins, allowing greater fluorescence detection. A laser 
system may be employed where the molecules are optimized to the laser 
wavelength; an excited sapphyrin may transfer its energy to the conjugated 
texaphyrin for detection. The texaphyrin molecule may further be designed 
to pass through cell membranes for selective radiosensitization. 
New texaphyrin derivatives are characterized fully using normal 
spectroscopic and analytical means, including, X-ray diffraction methods. 
It is understood that the examples and embodiments described herein are for 
illustrative purposes only and that various modifications in light thereof 
will be suggested to persons skilled in the art and are to be included 
within the spirit and purview of this application and scope of the 
appended claims.