CA2792724A1 - Chelating compounds and immobilized tethered chelators - Google Patents

Abstract

Novel compounds useful as chelators, intermediates for their production and methods for removing trivalent and tetravalent metal ions from solution are presented.

Description

CHELATING COMPOUNDS AND IMMOBILIZED lETHERED CHELATORS
This application is a continuation-in-part of U.S. Patent Application Serial No.
13/052,477, filed 21 March 2011 which is a divisional of U.S. Patent Application Serial No.
12/104,066, filed 16 April 2008 which is now issued U.S. Patent 7,932,326, the entire disclosures of which are incorporated herein by reference.

TECHNICAL FIELD AND INDUSTRIAL APPLICABILITY OF THE INVENTION
[0001] The present invention relates generally to the chemical field and, more particularly, to novel chelating agents, useful intermediates for synthesizing those chelating agents, the immobilization of those agents on a solid support resin, and the use of those chelating resins to remove metal ions from aqueous solutions.

BACKGROUND OF THE INVENTION

[0002] A chelator or chelating agent is a polydentate ligand that bonds to more than one coordination site of a metal ion. Chelating agents have long been known in the art to be useful in chemical analysis, in environmental remediation and in medicine. In chelation therapy, a chelating agent is employed to bind a poisonous metal agent such as mercury, arsenic, iron, lead or aluminum in order to displace the ion from biological ligands such as proteins and convert the metal ion into a less toxic form that can be excreted without further interaction with the body.

[0003] The present invention relates to (1) novel chelating agents or compounds, (2) novel immobilized, tethered chelators comprising the novel chelating compounds linked to immobilized supports and (3) methods of employing the novel compounds and chelators to remove trivalent and tetravalent metal ion such as Al3+ and Pu4+ from aqueous systems in situ, in vivo and in vitro.

[0004] There have been previous studies of tripodal, trihydroxamic acids.
Most of these ligands are based on tripodal platforms of tris(2-aminethyl)amine (tren) (Matsumoto et al., Chem. Commun. 2001, 978-979; Matsumoto et al., Inorg. Chem., 2001, 40:
190-191;

Matsumoto et al., Inorg. Chem. 2004, 43: 8538-8546; Ng et al., Inorg. Chem.
1989, 28:
2062-2066), tris(3-aminopropyl)amine (Matsumoto et al., Eur. J. Inorg. Chem.
2001, 2481-2484); or nitrilotriacetic acid (nta) (Lee et al, J. Med. Chem. 1985, 28: 317-323; Hara et al., Inorg. Chem. 2000, 39: 5074-5082). These studies teach that such ligands form Fe3+
complexes with binding constants in the range of 1028 to 1033, so long as there are five or six atoms connecting the bridgehead atom of the platform and the first atom of the hydroxamate functional group on the sidearm (Matsumoto et al., Eur. J. Inorg. Chem. 2001, 2481-2484;
Matsumoto et al., Inorg. Chem. 2001, 40: 190-191; Ng et al., Inorg. Chem.
1989, 28: 2062-2066). These ligands include amide functional groups in the sidearms, and the iron complexes appear to be stabilized by intramolecular hydrogen bonding between the amide functional groups (Matsumoto et al., Inorg. Chem. 2001, 40:190-191).

[0005] The common feature of all the above ligands is that the bridgehead atom is a tertiary nitrogen. To attach these ligands to a solid support via this nitrogen would require the formation of a quaternary ammonium group. This is expected to have an adverse effect on the chelating ability of the ligand. It will introduce a permanent positive charge on the ligand, resulting in electrostatic repulsion of the target metal ion. In some cases, it will also require a change in the conformation of the metal complex.

[0006] A few tripodal tris(hydroxamate) ligands have been prepared in which the bridgehead atom is a carbon, rather than a nitrogen. These ligands are built on tripodal bases of either 1,1,1-tris(hydroxymethypethane (Motekaitis et al., Inorg. Chem.
1991, 30: 1554-1556) or 1,1,1-tris(hydroxymethyl)propane (D ayan et al., Inorg. Chem. 1993, 32: 1467-1475). Hydroxamate groups were added to these tripodal bases through ether linkages.
These studies teach that one needs 4 or 5 atoms between the bridgehead carbon and the first atom of the hydroxamate functional group for strong metal binding. The Fe3+
complexes of these ligands have binding constants of 1026 to 1028. However, it is not possible to link these ligands to a polymeric support through the quaternary carbon bridgehead atom.

[0007] The current invention is based in the use of hydroxyalkylaminomethanes, especially the common buffer tris (1,1,1-tris(hydroxymethyl)aminomethane), as the tripodal base. The use of hydroxylalkylaminomethanes allows us to construct tripodal chelating functional groups that will mimic the high metal binding affinities of the ligands already in the literature, but it also provides a free amine group that can be used to easily attach the ligands to a variety of solid supports.

[0008] In issued U.S. Patent 7,932,326, all of the hydroxamate ligands are derived from tris[tris(hydroxymethyl)aminomethane]. The oxygen atoms of tris are alkylated with alkyl groups of various lengths terminating in hydroxamic acids. The amine of the tris is linked to a polymer support via sulfonamide, carboxamide or urea groups amongst others, to give the resin supported ligands. This document relates to an extension of that work.

SUMMARY OF THE INVENTION

[0009] In accordance with the purposes of the present invention as described herein, novel di- and tripodal compounds are disclosed for use as chelating agents.
Such compounds include, but are not limited to, novel tripodal trihydroxamate chelating agents having a tris(hydroxylalky)aminomethane platform, such chelating agents bonded to a polymeric resin, useful intermediates for making such chelating agents and to a method of removing a trivalent metal such as aluminum from a solution using such chelating agents.

[0010] In the following description there is shown and described several different embodiments of the invention, simply by way of illustration of some of the modes best suited to carry out the invention. As it will be realized, the invention is capable of other different embodiments and its several details are capable of modification in various, obvious aspects all without departing from the invention. Accordingly, the drawings and descriptions will be regarded as illustrative in nature and not as restrictive.

BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The accompanying drawings, incorporated herein and forming a part of the specification, illustrate several aspects of the present invention and together with the description serve to explain certain principles of the invention. In the drawings:

[0012] Figure 1 is a linear free energy relationship showing the correlation between the binding affinities of Fe3+ and Al3+ with hydroxamate ligands. Each data point represents a ligand, with the log f3 value for Fe3+ as the x-coordinate and the log r3 value for Al3+ as the y-coordinate. The open symbols represent reference compounds described in the literature.

The data points are: 1-3 represent the 1:1, 1:2, and 1:3 complexes with acetohydroxamic acid. Points 4 - 7 represent a series of linear dihydroxamates, in which the hydroxamate groups are separated by 4, 5, 6, or 7 methylene groups. Points 8 and 9 are the binding constants of the desferrioxamine (DFO) complex and the protonated complex of DFO. Point is mesitylenetrihydroxamic acid. The filled triangles represent compounds from the current invention. Point 11 represents the complexes of Ligand 1, point 12 represents the protonated complexes of Ligand 1, and point 13 represents the complexes of Ligand 7.

[0013] Figure 2 is a graph demonstrating the binding of A13' to 50 mg Resin 1 in which the concentration of free A13+ remaining in solution after the addition of 50 mcg Al at time 0 to either 100 ml or 5 ml of 4-morpholineethanesulfonic acid (MES) buffer at pH 5 has been determined by electrothermal atomic absorption spectroscopy (ETAAS).

[0014] Figure 3 is a spectrophotometric assay showing the binding of Al3+ to Resin 1 following addition of 22.8 mg of Resin 1 to 3 ml of 0.15 mM Al-ferron at pH 5.
Spectra show the decrease in the absorbance of Al-ferron at 364 nm and the increase in the absorbance of free ferron at 434 nm. Spectrum 10 shows the reference spectrum for 0.15 mM ferron.

[0015] Figure 4 is a graph illustrating the binding of A134 to Resin 1 following the sequential addition of six aliquots of 100 mcg of Al to 50 mg of Resin 1 suspended in 100 ml of pH 5 MES buffer. The first aliquot of Al was added at time = 0. Five subsequent additions were made at 12 hr intervals at the time indicated by the arrows on the graph. The free Al concentration was determined by ETAAS; and [0016] Figure 5 is a graph illustrating the binding of A134 to Resin 1 following the addition of 250 mg of Resin 1 to 0.5 ml of 0.23M calcium gluconate containing ¨ 9000 ng Al/ml. The free Al concentration was determined by ETAAS.

[0017] Figure 6 is a plot of the fraction of Al remaining in solution as a function of time during the extraction of A134 by resin 9. Three solutions were extracted:
a 0.1 M MES
buffer at pH 6.06, which had been spiked with 6.3 ppm Al; a solution of 0.46 M
gluconate containing 3.4 ppm Al, which had been adjusted to pH 6.4 by the addition of tetramethylammonium hydroxide; and a commercial sample of 0.23 M
calcium(gluconate)2, which contained 5.9 ppm Al and had a pH of 6.07. In each experiment, approximately 240 mg of resin 9 was added to 10 ml of solution. The mixtures were stirred by a magnetic overhead stirrer during the extraction. At periodic times, a 100 uL aliquot was removed from the sample and analyzed by inductively coupled plasma-mass spectrometry to determine the Al concentration.

[0018] Figure 7 is a plot of the fraction of Al remaining in a solution of commercial calcium(gluconate)2 during the extraction of Al3+ by resin 9 (filled and open circles) and by the commercial chelating resin Chelex (filled triangles). In each experiment, approximately 240 mg of resin was added to 10 ml of solution. The mixtures were stirred by a magnetic overhead stirrer during the extraction. At periodic times, a 100 uL aliquot was removed from the sample and analyzed by inductively coupled plasma-mass spectrometry to determine the Al concentration.

[0019] Figures 8a-8c illustrate three different embodiments of cartridges that may be filled with the immobilized chelating agents described herein.

[0020] Figure 8d is a partially cross sectional view of the cartridge illustrated in Figure 8a.

[0021] Figures 9a-9c illustrate three different embodiments, the first two of a vessel holding a flow-through packet containing the immobilized chelating agents described herein, and the third holding free immobilized chelating agents as described herein.

[0022] Figure 10 is a plot of percent removal of Al versus flow rate using the device illustrated in Figure 8a filled with resin 9.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS OF THE INVENTION

[0023] The present invention relates generally to novel chelating compounds having a general formula of Ri¨N¨R3 [0024] wherein It' = hydrogen or tosylate, R2 = hydrogen, methyl, ethyl, n-propyl or isopropyl and 5 , , and R3=

a.) 0, pH

µC-N, ( \CH2/) x (Rc4H2):ji C-N
0õ d y =R4 OH

(CH2) z =R4 wherein x, y, and z vary independently from 2 to 4, and R4 = hydrogen or CI-C)0 straight or branched alkyl;

b.) HO

N-CI, (CH2)x / R4 OH 0 µ ii 1 ,(CH2)-N-C\
0õ 0 y R4 H

cCH2); N¨C=R4 wherein x, y, and z vary independently from 2 to 4 and R4 = hydrogen or C1-C10 straight or branched alkyl;

C) \C0N(R4)0H

\ \
0) c0 / CON(R4)0H

0 CON(R4)0H

wherein R4 = hydrogen or C1-C10 straight or branched alkyl;

d.) --CON(R4)0H

CON(R4)0H
o)C0 0---/

ON /OTh CON(R4)0H

wherein R4 = hydrogen or C1-C10 straight or branched alkyl;

e.) 0 (CR52)-1---N/ OH
/ NH / n R4 0 0 (CR52)_1/ n ,OH0 0 \ 5(CR 2) n R4 wherein n = 2 or 3, R5 = hydrogen or methyl, and R4 = hydrogen or C1-C10 straight or branched alkyl;

f.) C¨N OH
sR4 C¨N
sR4 wherein R4 = hydrogen or C1-C10 straight or branched alkyl;
g) 0 OOH 0 OH /A

'R4 wherein R4 = hydrogen or C1-C10 straight or branched alkyl; and OH
h.) CLN 0 \R4 0\ )1 N 0 OH

wherein R5 = hydrogen or methyl and R4 = hydrogen or C1-C10 straight or branched alkyl.

[0025] The novel compounds of the present invention are particularly useful as chelators or chelating agents. One preferred use of the free ligands would be in vivo chelation therapy to remove metal ions such as Fe3+ and Al3+ from the body.

[0026] The compounds include an amine functional group that allows the ligands to be easily linked to an insoluble matrix via a sulfonamide linkage, an amide linkage or a urea linkage to provide immobilized, tethered chelators. Typically, the insoluble matrix comprises a resin support. The resin support may take the form of a macro-porous polystyrene such as commercially available under the trademark XAD-4 sold by Rohm and Haas. Other polymer resins useful in the present invention include but are not limited to, polyacrylate, sepharose and silica gel.
10027] The overall process of adding a chelating compound of the present invention to a polystyrene resin via a sulfonamide bond is shown in Scheme 1, where NR2H-Ligand in , this and subsequent schemes refers to, the free amine form (R1 = H; R2 =
hydrogen, methyl, ethyl, n-propyl or isopropyl) of any of the free ligands represented by R3 = a through h.

, o CI ES-Lagani \\___14H,Ligand INSOLUBLE ,/=\ CO INSOLUBLE __(--=-\4. a - INSOLUBLE

- MATRIX ,,__J '' IRE Ts? MATRIX \ /7 II
MATRIX n__/
0 . 0 macroporous modified resin resin linker ligand polystyrene resin, e.g. XAD-4 [0028] Scheme 1 [0029] The overall process of adding a chelating compound of the present invention to a resin support by means of an amide linkage is shown in Scheme 2.

Et3N, THT, DCC

INSOLUBLE ¨7-.= INSOLUBLE i 1 C /
/P. --- \

MATRIX MATRIX \ ITH2-Dgard \ / \ 0H
\ NH=ligand \¨

[0030] Scheme 2 [0031] The overall process of adding a chelating compound of the present invention to a resin support by means of a urea linkage is shown in Scheme 3.

Insoluble Matrix 410 Insoluble Matrix, 4.

I, NH2 NH)0¨N

7 0 \

N-0-'-' 2 Et3N, THF 0 r I

Insoluble Matrix, .
)- , HR2N¨Ligand N NR--Ligand H

[0032] Scheme 3 [0033] For certain applications it may be desirable to elongate the linker by adding polyethylene glycol units between the resin support and the ligand in order to increase the rate of metal binding to the resin-hound ligand. These elongated linkers are added using commercially available amine capped polyethylene glycols of variable length, with the use of a urea functional group to covalently bind the ligand and linker moieties.

[0034] The elongation process is illustrated in Scheme 4 using the linker 3-oxa-1,5-pentanediamine as a specific example.

II 11 Et3N, THEa-Insoluble Matrix 11 Insoluble Matrix/

0 H2Nas-'-'N H2 0 Et3N, THF
Insoluble Matrix; 11 0 0 H HR2N¨Ligand Insoluble Matrix] Ito H 0 NR¨Ligand [0035] Scheme 4 [0036] Other commercially available amine-capped polyethyleneglycols include the compounds 0 .1"-\\ 0 NH2 ) 2 n= 10-12 n which give chelating resins with the structures shown below Insoluble Matrix, = S¨NH 0 0 NH NR2¨Ligand Insoluble Matrix, 4. 0 H CH3 0 0 CH3 n=10-12 NH NR2¨Ligand [0037] The immobilized, tethered chelators of the present invention comprise the chelating compounds identified above bound to a resin support through an appropriate linkage. The immobilized, tethered chelators of the present invention may be generally described as having the following formula:

wherein R6 =

\ 0 Insoluble 9 Insoluble Matrix 0 Matrix Insoluble Insoluble CH2¨NH¨
Matrix ¨CH2¨NH Matrix Insoluble 0 Matrix !I
Insoluble Matrix 0 HNH

Insoluble \ 9 H CH3 0 Matrix 0 µ.., r,u 1 13 fl n = 10-12 R2 = hydrogen, methyl, ethyl; n-propyl or isopropyl and R3 =

a.) ¨Nõ 4 N
/C orc1-1H2),._ OH
(\CH2)x ¨

Y sR4 ?I pH

(CH2)z wherein x, y, and z vary independently from 2 to 4 and R4 = hydrogen or Cl-C10 straight or branched alkyl;

b.) HO ;0 (CH2)x OHO
r(CH2)-N-C
0 y R4 OH n /;-CH2)--N¨Cs z R-wherein x, y, and z vary independently from 2 to 4, and R4 = hydrogen or C1-C10 straight or branched alkyl;

c.) õO/ \CON(R4)0H

\CON(R4)0H
OD( 0/

\ /CON(R4)0H

wherein R4 = hydrogen or C1-C10 straight or branched alkyl;

d.) --CON(R4)0H

CON(R4)0H

0\
/0-\CON(R4)0H

wherein R4 = hydrogen or C1-C10 straight or branched alkyl;

, ' e.) 5 0 0 /(CR 2)__ILn NPH\
fiLNIFI R4 0 0 i(CIR-2)n-..fiN,OH c 0 ,C0............õõ)---NH
\

\ ,NH OH
II \ 51L N:
0 (CR-2)n se wherein n = 2 or 3, R5 = hydrogen or methyl, and R4 = hydrogen or CI-C10 straight or branched alkyl;

0 pH
f.) 0 8-N , FrA

9¨N
---E-30 * 0 OH

841, wherein R4 = hydrogen or C1-C10 straight or branched alkyl;

g) OOH
. C-14, 0H4 ogirf ,ip OOH

%R4 wherein R4 = hydrogen or C1-C10 straight or branched alkyl; and h.) O OH
rN \R4 0\ ) N

wherein R5 = hydrogen or methyl and R4 = hydrogen or C1-C10 straight or branched alkyl.

[0038] PREPARATION OF COMPOUNDS OF THIS INVENTION
[0039] Selected chelating agents and chelating resins from this invention are listed in Table 1.
Table 1. Partial list of chelating agents and chelating resins included in this invention Ri-N-R3 , . . .
Free Ligands (R1 = Tosyl, 12-=H) Ligand IC x y z R2 R5 Ligand 1 a 2 2 2 H
Ligand 2 a, 3 3 3 H
Ligand".,. a 4 4 4 H
Ligand 4 '.: 4 4 2 H
Ligand 5 c H
Ligand 6 d H
Ligand 7 b Methyl Ligand a 3 3 2 H
Ligand 9 a Resins (R6 = polystyrenesulfonate, R4=H) Resin 1 a 2 2 2 H
Resin 2 h Methyl [0040] Example 1 [0041] Synthesis of Ligand 1 [0042] The overall synthesis of Ligand 1 is shown in Scheme 5. The aminotriol (1, Tris buffer) was reacted with acrylonitrile in the presence of a catalytic amount of base to give the trinitrile (Intermediate 1) (Newkome, G.R. and X. Lin, Symmetrical, four-directional, poly(ether-amide) cascade polymers. Macromolecules, 1991, 24(6):
1443-1444).
Reaction of Intermediate in refluxing methanolic HC1 gave the tris(methyl ester) (Intermediate 2). Reaction of Intermediate 2 with tosyl chloride gave the sulfonamide tris ester (Intermediate 3). This ester was converted to the trihydroxamic acid (Ligand 1) by reaction with 0-trimethylsilyi hydroxylamine (NH,OTMS) in methanol.

dioxane, KOH (cat), H2N_e00 CNI oN Me0H, HCI, reflux 33% 1.25 eq per OH
\ /C N
55%

Intermediate 1 //CO2Me /CO2Me Et3N, THF

H2NOCOMe g¨IF\i¨e ---¨0O2Me =0 so201 CO2Me Intermediate 3 CO2Me Intermediate 2 ,CONHOH
NH2OTMS Me0H KOH
AcOH Me0H
s-N
\-0 'CONHOH

Ligand 1 CONHOH

[0043] Scheme 5 [0044] Synthesis of Intermediate [0045] To a stirred solution of tris(hydroxymethyl)aminomethane (50.0 g, 412.0 mmol) and KOH (2.3 g, 4.5% of the weight of alcohol) in 1,4-dioxane (150 mL) was added acrylonitrile (71.17 g, 1342.4 mmol) drop wise over a period of lh, after which a clear solution was obtained. After stirring at room temperature for 24 h, the mixture was made acidic (¨ pH = 2) by the addition of dil. HC1. After extraction with CH2C12 (3 x 100 mL) the combined organic layers were dried over sodium sulfate and evaporated to give tris[(cyanoethoxy)methyl]aminomethane (Intermediate 1), 43.2 g (33.5%). IR
(neat) 3588, 3368, 2251 cm-3, IHNMR (CDC13) 6 3.68 (t, J=6.0 Hz, 6H), 3.44 (s, 61-I), 2.61 (t, J=6.0 Hz), 1.68 (br s, 2H); 13C NMR (CDC13) 6 118.2, 72.7, 65.9, 56.3, 19.0; HRMS (El, MH') calcd for C13H211\1403: 281.16147, found: 281.16138. (Newkome, G.R. and X. Lin, Symmetrical, four-directional, poly(ether-amide) cascade polymers. Macromolecules, 1991.
24(6): p.

1443-1444.

[0046] Synthesis of Intermediate [0047] Dry HC1 gas was passed through a solution of intermediate 1 (52.6 g, 187.0 mmol) in dry methanol (150 mL) until the solution was saturated with HC1. The mixture was refluxed overnight. After the solution was cooled, NH4C1 was removed by filtration, and the filtrate was concentrated to give a gum. The gum was taken up in THF, filtered, and the filtrate was concentrated to get the tris ester (Intermediate 2) 37.0 g (55.0%). IR (neat) 3394, 1735 cm-1; 11-1 NMR (CDC13) 63.69 (t,2Jh_h=6.31 Hz, 6H), 13C NMR (CDC13) 5 172.4, 69.0, 67.1, 59.6, 52.0, 34.7; HRMS (El, MH+) calcd for Cl6H30N09: 380.19217, found:
380.19205. (Nierengarten, J.F.; Habicher, T.; Kessinger, R.; Cardullo, F., Diuederich, F.;
Gramlich, V.; Gisselbrecht, J.P.; Boudon, D.; Gross, M., Macrocylization on the fullerene core. Direct regio- and diasterioselective multi-functionalization of [60]fullerene, and synthesis offullerene-dendrimer derivatives. Helv. Chim. Acta, 1997, 80: 2238-2276).
[0048] Synthesis of Intermediate 3 100491 To a stirred solution of tosyl chloride (10.0 g, 52.4 mmol) and the tris ester (Intermediate 2) (19.90 g, 52.4 mmol) in CH2C12 was added NEt3 (6.37 g, 62.9 mmol) and the mixture was heated at reflux overnight. The solvent was removed in vacuo, and the residue was redissolved in CH2C12 (200 mL) and washed with water (3 x 100 mL).
The organic layer was dried over Na2SO4 and concentrated to give a gum. Column chromatography using silica gel with 50% ethyl acetate in hexane yielded a gummy solid of Intermediate 3 (20.4 g, 73%), which later crystallized on storing at room temperature.
Finally it was characterized by X-ray crystallography. IR (neat) 3610, 3287, 1736 cm-1; 114 NMR (CDC13) 5 7.78, 7.76, 7.27, 7.24 (s each, 4H), 3.68 (s, 9H), 3.51 (s, 6H), 3.51 (t, 24-h=
6.5 Hz, 6H), 2.41 (t, 23h_h= 6.5 Hz, 6H) 2.41 (s, 3H); 13C NMR (CDC13) 5 172.1, 142.8, 140.5, 129.2, 127.0, 69.9, 66.7, 62.4, 51.9, 34.7, 21.6; BERMS (El, MH+) calcd for C23H36N0NS:
534.20095, found: 534.20093.
[0050] Synthesis of Ligand 1 [0051] To a stirred solution of the tris(ester) (Intermediate 3) (8.23 g, 15.4 mmol) in methanol (100 mL) was added NH2OTMS (9.74 g, 92.5 mmol) followed by KOH (2.60 g, 46.0 mmol). After 6 h at room temperature, the reaction mixture was treated with 20 g of prewashed Amberlyst-15 and swirled for 1 h. The resin was filtered off and the filtrate was evaporated to give a gum. Recrystallization from acetone:hexane (1:1) yielded the tris hydroxamate (Ligand 1), 5.02 g, (61%) which was characterized by X-ray crystallography.

IR (neat) 3184, 1631 cm-1; 1H NMR (CDC13) ö 7.76, 7.73, 7.42, 7.39 (s each, 4H), 3.46 (t, J =

5.8 Hz, 6H), 3.40 (s, 6H), 3.31 (s, 3H, Me0H), 2.40 (s, 3H), 2.30 (t, J = 5.8 Hz, 6H); 13C

NMR (CDC13) 8 171.1, 145.0, 139.0, 130.1, 127.0, 69.2, 67.0, 63.0, 49.2 (CH3OH), 33.3, 21.0 HRMS (El, MH ) calcd for C20H33N4O1S: 537.18677, found: 537.18665.

[0052] Example 2.

[0053] Synthesis of Ligand 2 [0054] The overall synthesis of Ligand 2 is shown in Scheme 6. The trimethyl orthoester of 4-iodo-1-butyric acid (3), in which the vulnerable sp2 carbon has been protected, is known to alkylate alkoxides (Srivastava, R. P., Hajda, J.
Stereospecific synthesis of ether phospholipids. Preparation of 1-0-(3'-carboxypropyl)-glycero-3-phosphoserine from glyceric acid. Tetrahedron Lett. 1991, 32, 6525-6528) (Method A). Thus treatment of the BOC-protected trio! (2) with sodium hydride and the trimethyl ortho ester (3) in DMF, followed by deprotection with anhydrous methanolic HC1 gives the triester (Intermediate 4).

Alternatively, reductive alkylation of the trimethylsilylated trio! (BSA, reflux) with 3-cyanopropionaldehyde (Iwanami, K., Kentaro Y., Takeshi, 0. An Efficient and Convenient Method for the Direct Conversion of Alkyl Silyl Ethers into Corresponding Alkyl Ethers Catalyzed by Iron (III) Chloride. Synthesis 2005, 2669-2672) (Method B), followed by treatment with anhydrous HC1 in refluxing methanol should also yield intermediate 4.

Intermediate 4 is tosylated to give Intermediate 5, which is then converted to the corresponding hydroxamic acid (Ligand 2) by treatment with 0-(trimethylsilyphydroxylamine).

CO2Me 1, R = H HO¨KOH OMeOMe 3 /
11?.flec(319 1 NaH, OMe t Method A 0¨v--0 CO2Me 2, R = Boc RHN OH 2. HC1, Me0H
H2NI \-0_TCO2Me 1 FeC13, Et3S1H, H(0=)CCH2CH2CN Intermediate 4 Method B

2 Me0H, HO

r CO2Me (- --1"CONHOH
'¨

Et3N, THE / NH2OTMS Me0H KOH

= 0CO2Me AcOH Me0H =g4 11 0 0\ F-CONHOH
0 0\ ,-002Me O Intermediate 5 Ligand 2 [0055] Scheme 6 [0056] Example 3.

[0057] Synthesis of Ligand 3.

[0058] The overall synthesis of Ligand 3 is shown in Scheme 7. Treatment of the BOC-protected triol (2) with sodium hydride and the trimethyl ortho ester or 5-iodo-1-pentanoic acid (4) in DMF, followed by deprotection with anhydrous methanolic HCI gives the triester (Intermediate 6). Alternatively, reductive alkylation of the trimethylsilylated triol (Bis silylacetamide, reflux) with 4-cyanobutryoaldehyde (Iwanami, K., Kentaro Y., Takeshi, 0.. An Efficient and Convenient Method for the Direct Conversion of Alkyl Silyl Ethers into Corresponding Alkyl Ethers Catalyzed by Iron (III) Chloride. Synthesis 2005, 2669-2672) (Method B), followed by treatment with anhydrous HC1 in refluxing methanol should also yield intermediate 6. Intermediate 6 is tosylated to give Intermediate 7, which is then converted to the corresponding hydroxamic acid (Ligand 3) by treatment with 0-(trimethylsilyl)hydroxylamine).

CO2Me OMe /\/\/ CO2Me HO¨OH 4 1. NaH, I OMe Ve6c1?1 r Method A

2, R = Boc RHNI \¨OH 2. HCI, Me0H

CO2Me 1. FeCI3, Et3S1H, H(0=)CCH2CH2CH2CN
Method B Intermediate 6 2. Me0H, HCI

CO Me CONHOH

(;_0/\/.\/
CONHOH
Et3N, THF 0 9/\/\/co2Me NH2OTMS, Me0H, KOH ik 0 S N
,-N
AcOH Me0H " H

SO2CI oo\/v\CO2Me \¨ \,/\/\CONHOH

Intermediate 7 Ligand 3 [0059] Scheme 7 10060] Example 4 [0061] Synthesis of Ligand 4. The binding constants for Ligand 1 (see below) indicate that two arms of the ligand bind to metal ions very strongly, but that steric hindrance weakens the binding of the third arm. In the heteropodal Ligand 4, the length of two of the ligand arms have been extended to relieve this internal strain. The synthesis of ligand 4 is shown in Scheme 8. To prepare heteropodal trihydroxamic ligands, two of the hydroxyls on the aminotriol (tris) are first blocked by a protecting group. The aminotriol (1) is converted to the known cyclic acetal (5) using a published 2 step, 1 pot procedure (Ooi, H., Ishibashi, N., Iwabuchi, U., Ishihara, J., Hatakeyama, S.. A concise Route to (+)-Lactacystin. J. Org.
Chem. 2004, 69, 7765-7768). Alternatively, the diol can be protected as the benzylidene (6a) (Balakumar, V., A highly regio- and chemoselective reductive cleavage of benzylidene acetals with EtA1C12-Et3SiH, Synlet, 2004, 647-650; Low, J.N., B.F. Milne, J.-N. Ross, and J.L. Wardell, Derivatives of 1V,N'-bis[2-Hydroxy-1,1-bis(hydroxymethyl)ethyl]ethanediamide. Journal of the Brazilian Chemical Society, 2002, 13: 207-217) using similar reaction conditions, which results in additional options for deprotection later in the synthetic sequence. Addition of the remaining free alcohol to acrylonitrile yields the mononitrile (7) (Newkome, G. R., Lin, X. Symmetrical, four-directional, poly(ether-amide) cascade polymers. Macromolecules. 1991, 24, 1443-1444).
Simultaneous deprotection of the acetal and methanolysis of the nitrile with a refluxing solution of methanolic HC1 yields the monoester-diol (8). The diol is then alkylated by the addition of the trimethyl ortho ester of 5-iodopentanoic acid to form the triester (Intermediate 8). An additional complication in the alkylation of the monoester diol is base-catalyzed beta elimination of the alkoxy group beta to the ester. Intermediate 8 is tosylated to give Intermediate 9, which is converted to the corresponding trihydroxamate (Ligand 4) by the addition of 0-(trimethylsilyphydroxylamine.

, /ON

HOThr-OH
B20, DMF then HO-v-Ov NaOH, DMSO

¨KO

Me2C(OMe)2, pTSA
CN õ...-...-::-.' HNI \¨Of\
H,N 0K
HO-v-0 ''_ R

13o c 5 H"-0 BOG
6a, R = H
6b. R = OMe OMe iCO2Me HCI, Me0H
r CO2Me 1 NaH, ,.....,..)<OMe r--\_102Me _ \O--v¨OH
I
OMe OThr _,.
2. HCI, Me0H
H2N7\¨OH
H2 NI \--0\ _...
02Me Intermediate 8 (CO2Me rCONHOH

\CD

Et3N,THF

/¨\_/CO2Me p Fici----\_ JCONHOH
I

t.
40.
NH2OTMS, Me0H, KOH
, = S¨N

H
AcOH Me0H

0\ j--\

\___I-7LONHOH
CO2Me Intermediate 9 Ligand 4 [0062]
Scheme 8 [0063]
Example 5.
[0064]
Synthesis of Ligand 5. The synthesis of the heteropodal Ligand 5 is described in Scheme 9. The extension of one arm of the ligand is achieved by the reaction of the acetal protected aminotriol (5) with chloroacetic acid, followed by selective reduction of the carboxylic acid and deprotection of the cyclic acetal to give the unsymmetric triol (9).
Adding the triol to acrylonitrile gives the trinitrile Intermediate 10, and methanolysis of the nitrile gives the tris(ester) (Intermediate 11).
This compound is tosylated to give Intermediate 12. The addition of 0-(trimethylsilyl)hydroxylamine to Intermediate 12 gives the heteropodal Ligand 5.

=

HO
HO

HO¨v¨Oy NaH, CICH2CO2H BH3 THF

HN/ \-0/ \
Eoc 5 HN1 \-0/\
Boc Boc HO

HCI, H20, Me0H 'o OHNaOH, DMSO
OO CN

H2N/ \--OH CN

/ \ Intermediate 10 0 CO2Me HCI, Me0H, reflux - Ox0 CO2Me /¨\ Et3N, THF

SO2C' H2N 0 CO2Me Intermediate 11 /\ / \
/\./.0 CO2Me /\,0 CONHOH

II 2e 0/ \CO M NH2OTMS, Me0H, KOH II
0/ \CONHOH
S ¨N =S¨N
8 H AcOH Me0H 8 H
0 CO2Me 0 CONHOH
\
Ligand 5 Intermediate 12 = [0065] Scheme 9 [0066] Example 6.

[0067] Synthesis of Ligand 6 [0068] The synthesis of Ligand 6 is shown in Scheme 10. The diol (8) from Scheme 8 is reprotected at the amine with a Boc group to give (10). The remaining hydroxyls are reductively alkylated with aldehyde (11) followed by methanolysis to yield the triester (Intermediate 13). The aldehyde (11) is easily prepared in two steps from glycol. Although synthesis of the ester (12) would provide a more direct approach, the reaction would be complicated by competing, rapid lactonization to lactone (13). Intermediate 13 is tosylated to give intermediate 14. This compound is treated with 0-(trimethylsilyl)hydroxylamine to give the heteropodal trihydroxamic acid Ligand 6.

, .

(CO2Me (CO2Me 0 0---\ 11 /---\
CO2Me 1, FeCI3, Et3SiH \\--/ CN . 0¨v-0 0--/

0¨v¨OH 2. HCI, Me0H, reflux H2N/ \--0 0¨\ \--/ CO2Me RC OH

8, R = H Intermediate 10, R = Boc 1. Glycol (10 equiv.), NaH HO 0¨\ PCC, CH2Cl2 0 0¨\
HO 0¨\ CD) \¨/ CO2Me CN --,-' \-1 ON

('CO2Me KCO2Me /---\ CO2Me 0¨v¨O 0¨/ Et3N, THF ip ___.(c_ /¨\
ICO2Me NH2OTMS, Me0H. KOH
. = s-N 0 0_, ).--ll H AcOH Me0H
H2N/ \-0 0--\ * SO2CI 0 0 0¨\

\¨/ CO2Me \¨/ CO2Me Intermediate 13 Intermediate 14 (CONHOH

o______ /--\ CONHOH

11 i¨N 0 0¨/

\--/ CONHOH

Ligand 6 [0069] Scheme 10 [0070] Example 7.

[0071] Synthesis of Ligand 7. The overall synthesis of Ligand 7 is shown in Scheme 11.

/ \ /CO2Me OH dioxane, KOH (cat) 0 CN Me0H, HCI, reflux 0/

H2N¨& H2N -CCH3 OH --;"---CN 1.25 eq per OH 0 CN 55 A 0 \ /
33%

Intermediate 15 CO2Me Intermediate 16 /CO2Me Et3N, THF 0= 0/

g1CH3 SO2C! I I \-0 Intermediate 17 CO2Me ,CONHOH
NH2OTMS Me0H KOH 0 CH3 /
= 4_,1 \co AcOH Me0H

Ligand 7 CONHOH

[0072] Scheme 11 [0073] Synthesis of Intermediate 15 [0074] To a stirred solution of 2-amino-2-methy1-1.3-propanediol (50.0 g, 475.5 mmol) and KOH (1.0 g, 2% of the weight of diol) in 1,4-dioxane (100 mL) was added acrylonitrile (56.7 g, 1070.0 mmol) dropwise over a period of 1 h, after which a clear solution was obtained. After stirring at room temperature for 24 h, 200 mL of CH2Cl2 was added to the mixture. The mixture was extracted with water and the organic layer was dried over sodium sulfate. The solvent was evaporated to yield a thick oil.
Distillation under reduced pressure (160f 'C/ 10 mm Hg) yielded Intermediate 15, 39.5 g (39%). IR
(neat) 3517, 3360, 2250 cm-1; 1H NMR (CDC13) 8 3.68 (t, J=6.1 Hz, 4H), 3.33 (Abq, Av = 14.2 Hz, J=8.5 Hz, 4H), 2.60 (t, J=6.1 Hz, 4H), 1.44 (br s, 2H), 1.06(s, 3H; 13C NMR
(CDC13) 6 118.2, 76.5, 65.8, 52.8, 22.6, 19.0 HR.MS (El, MI-1 ) calcd for C10H18N302:
212.14002, found:

212.13989.

[0075] Synthesis of Intermediate 16 [0076] Dry HCI gas was passed through a solution of Intermediate 16 (39.48 g, 186.1 mmol) in dry methanol (150 mL) until the solution was saturated with HC1. The mixture was refluxed overnight. After cooling, NH4C1 was removed by filtration, and the filtrate was concentrated to give a gum. The gum was redissolved in THF, filtered, and the filtrate was concentrated to get the diester (Intermediate 16) 30.0 g (57.0%). IR (neat) 3409, 1727 cm-1;
'H NMR (CDC13) 8 3.79 (t, J=6.1 Hz, 2H), 3.70 (2, 6H), 3.62 (s, 4H). 2.64 (t, J=6.1 Hz, 411), 1.42 (s, 3H); 13C NMR (CDC13) 8 172.6, 71.7, 67.2, 58.0, 52.1, 34.8, 18.4;
HRMS (EL mir) calcd for Cl2H24N06: 278.16047, found: 278.16037.
[0077] Synthesis of Intermediate 17 [0078] To a stirred solution of tosyl chloride (10.0 g, 52.4 mmol) and Intermediate 16 (14.54 g, 52.4 mmol) in CH2C12 was added NEt3 (6.37 g, 62.9 mmol) and the mixture was heated at reflux overnight. The solvent was removed in vacuo, and the residue was redissolved in CH2C12 (200 mL) and washed with water (3 x 100 mL). The organic layer was dried over Na2SO4 and concentrated to give a gum. Column chromatography using silica gel with 40% ethyl acetate in hexane yielded a gummy solid of Intermediate 17, (16.9 g, 74%). IR (neat) 3604, 1736, 1735 cm-1; IFINMR (CDC13) 8 7.82, 7.55, 7.29, 7.26 (s each, 4H), 3.70 (s, 6H), 3.65 (t, J=6.2 Hz, 4H), 3.33 (Abq, Av = 47.0 Hz, J = 9.1Hz, 4H), 2.53 (t, J
= 6.2 Hz, 411), 2.41 (s, 311), 1.10 (s, 3H); 13C NMR (CDCI3) 8 172.2, 143.1, 140.7, 129.6, 127.0, 73.7, 66.8, 58.9, 51.9, 34.9, 21.7, 18.2; HRMS (El, MH+) calcd for Ci9H30N08S:
432.16931, found: 432.16922.
[0079] Synthesis of Ligand 7 [0080] To a stirred solution of the ester (Intermediate 17) (3.61 g, 8.3 mmol) in methanol (50 mL) was added NH2OTMs (3.52 g, 33.4 mmol) followed by KOH (0.94 g, 16.7 mmol). After 6 h at room temperature, the mixture was treated with 7.0 g of prewashed Amberlyst-15 and swirled for 1 h. The resin was filtered off and the filtrate was evaporated to give a gum. Recrystallization from CH2C12:ether (1:1) yielded Ligand 7, 2.46 g, (68%).
IR (neat) 3233, 1633 cm-1; NMR (CDC13) 8 7.76, 7.68, 7.42, 7.39 (s each, 411), 3.54 (m, 411), 3.35 (s, 411), 3.30 (Abq, Av 22.6 Hz, J = 10.0 Hz, 4H), 2.40 (s, 311), 2.34 (t, J 5.8 Hz, 611), 1.08 (s, 3H); 13C NMR (CDC13) 6 171.1, 145.0, 139.0, 130.2, 127.0, 73.5, 67.0, 59.5, 49.3, 21.0, 18.8; HRMS (El, MH ) calcd for C17H28N308S: 434.15983, found:
434.15970.
[0081] Example 8 [0082] Synthesis of Resin 1. Resin 1 was prepared by synthesizing the chelating functional group (R3= a) of Ligand 1 on the surface of a polystyrene resin.
The synthesis of Resin 1 is shown in Scheme 12. Macro-porous polystyrene beads (14, Amberlite XAD-4) were reacted with chlorosulfonic acid to give the polymeric sulfonyl chloride (15) (Emerson, D. W., Emerson, R.R., Joshi, S.C., Sorensen, E. M., Turek, J.E. Polymer-bound sulfonylhydrazine functionality. Preparation, characterization, and reactions of copoly(styrene-divinylbenzenesulfonylhydrazine). J. Org. Chem. 1979, 44: 4634-4640; Hu, J.
-B., Zhao, G., Ding, Z. -D. Enantioselective reduction of ketones catalyzed by polymer-supported sulfonamide using NaBH4/Me3SiC1 (or BF3*OEt2) as reducing agent.
Angewandte Chemie, International Edition 2001, 40: 1109-1111). The procedures in Scheme 5 were used to prepare the methyl ester of the free amine form of ligand 1 (Intermediate 2). Addition of Intermediate 2 to the sulfonyl chloride form of the resin (15) gave the sulfonamide triester (Intermediate 18). The ester functional groups were converted to hydroxamic acids by reaction with 0-trimethylsily1 hydroxylamine in methanol to give Resin 1. The successful conversion of the esters to hydroxamic acids was judged from the IR spectra. The number of ligand molecules on the surface of the resin was calculated from the S and N combustion analysis of the resin to be 0.3 mmoles ligand per gram of resin.

41 Cii= ISO3H 4<¨¨S02C1 Et3N, THF \ 0 H r---CO2Me 14 H2N-0/¨0O2Me Intermediate 18 Intermediate 2 f"-CONHOH
NH2OTMS MeOH, KOH 0 H _C ZCONHOH¨
AcOH, Me0H 8 _-CONHOH
Resin 1 [0083] Scheme 12 [0084] Synthesis of Sulfonyl Chloride Resin:

[0085] To 35 g of macroporous styrene-divinylbenzene copolymer (20-60 mesh, avg.

pore diameter: 40A, Amberlite XAD-4) in 100 mL of 1,2-dichloroethane was added 160 g (1.37 mol) of technical grade chlorosulfonic acid with occasional swirling.
The mixture was kept at room temperature for 12 h. The product was filtered using a glass frit and was washed successively with two portions of dichloromethane (DCM), two portions of DCM-THF mixture, two portions of THF, and a final wash with DCM. The vacuum dried polymer was ready to use and was stored under argon at low temperature. IR (neat) 3521, 1369, 1171 cm ; Anal. Found: C, 57.17; H, 5.50; S, 10.23; Cl, 8.49; calculated loading S, 3.22 mmol/g, Cl, 2.39 mmol/g.

[0086] Synthesis of Intermediate 18 [0087] To polymeric sulfonyl chloride (15) (2.0 g, 5.0 mmol) in THF (50 mL) was added a solution of the tris ester of the free ligand (Intermediate 2) (7.58 g, 20.0 mmol) in THF (30 mL) followed by triethylamine (2.0 g, 20.0 mmol) and the mixture was swirled for four days at room temperature. The polymer was then filtered off and washed successively with THF, water, THF, DCM and dried in vacuo. IR (neat) 3494, 1732, 1169 cm-1;
Anal.

Found: C, 58.96; H, 6.81; S, 8.22; N, 2.66; calculated loading: S, 2.57 mmol/g, N, 1.90 mmol/g.

[0088] Synthesis of Resin 1 [0089] To the resin-bound triester (Intermediate 18) (1.7 g, 4.25 mmol) in methanol (40 mL) was added NH2OTMs (4.02 g, 38.2 mmol) dropwise with stirring at room temperature. KOH (2.14 g, 38.2 mmol) was added and the mixture was swirled for 12 h.
The product, resin 1, was filtered off and washed successively with methanol, water, and methanol. The resin was then swirled with dil. acetic acid for an hour and washed successively with methanol, water, THF, and dried over pump. IR (neat) 3479, 1644, 1173 cm-1; Anal. Found: C, 54.65 H, 5.57; S, 9.51; N, 1.59; Loading: S, 2.97 mmol/g, N, 1.14 mmol/g.
[0090] Example 9 [0091] Synthesis of Resin 2. The overall synthesis of Resin 2 is shown in Scheme 13.

* CISO3H
so2c1 Et3N, THF
\ 0 H

15 H2N-4---CH3 /O r¨N-0O2Me 0002Me ¨"\---0O2Me Intermediate 19 Intermediate 16 NH2OTMS Me0H, KOH
H_C
r---CONHOH
AcOH, Me0H
/

L'¨\.--CONHOH
Resin 2 [0092] Scheme 13.
[0093] Synthesis of Intermediate 19.
The synthesis of the chlorosulfonated polystyrene resin (15) is described in Scheme 12. To this polymeric sulfonyl chloride (5.2 g, 13.0 mmol) in THF (80 mL) was added a solution containing Intermediate 16 from Scheme 11, (14.4 g, 52.0 mmol) in THF (50 mL) followed by triethylamine (5.26 g, 52.0 mmol). The suspension was swirled for four days at room temperature. The product, Intermediate 19, was then filtered off and washed successively with THF, water, THF, DCM and dried over pump. IR (neat) 3492, 1735, 1170 cm-1; Anal. Found: C, 60.28; H, 7.04; S, 8.42; N, 2.61;
calculated loading: S, 2.63 mmol/g, N, 1.86 mmol/g.

[0094] Synthesis of Resin 2.
[0095] To the resin-bound diester (Intermediate 19) (5.27 g, 13.17 mmol) in methanol (60 mL) was added NH2OTMs (11.08g, 105.3 mmol) dropwise with stirring at room temperature to give Resin 2. KOH (2.95 g, 52.6 mmol) was added and the mixture was swirled for 12 h. The resin was filtered off and washed successively with methanol, water, and methanol. The resin was then swirled with dilute acetic acid for an hour and washed successively with methanol, water, THF, and dried over pump. IR (neat) 3468, 1643, 1176 cm-I; Anal. Found: C, 53.09 H, 5.59; S, 8.82; N, 1.95; Loading: S, 2.75 mmol/g, N, 1.39 mmol/g.
[0096] Example 10 [0097] Ligands immobilized via an amide linkage [0098] The triester intermediate of each ligand containing a free amine group (R1 = H) is coupled to a resin bearing a carboxylic acid using a coupling reagent such as dicyclohexylcarbodiimide (DCC) or N-ethyl-N"-(3-dimethylaminopropyl) carbodiimide (EDC) with a tertiary amine base in TI-IF solution. The esters are then converted to the hydroxamic acid as described above for the sulfonamide linked system. This process is described in Scheme 14 using Intermediate 2 as an example.

z_./CO2Me _/CO2Me Et3N, THF, DCC

H2N-"e0--¨0O2Me Matrix Insoluble =cu 0 o Insoluble C ¨OH
CO2Me Matrix CO2Me Intermediate 20 Intermediate 2 /...._/CONHOH

NH2OTMS, Me0H, KOH
Insoluble 4/lik9 1-1Ã.9 matrix 0--¨CONHOH

AcOH, Me0H C¨N_ '-0 CONHOH
Resin 3 [0099] Scheme 14 [00100] Example 11 [00101] Ligands immobilized via a urea linker [00102] The amine of a triester intermediate is coupled to a resin bearing an amine via a urethane linkage using a reagent such as N,N-disuccinimidyl carbonate (16), carbonyl diimidazole or triphosgene with a tertiary amine base in THF solution. The esters are then converted to the hydroxamic acid as described above for the sulfonamide linked system.

This process is shown in Scheme 15 using Intermediate 2 as an example.

, , . .

Insoluble Matrix 0I
' Insoluble Matrix, O S

NH).0-N

N- 0--r-'-' /CO2Me Et3N, THE

Insoluble Matrix, it CO Me 2 NAN-6 / H H
0 CO2Me \

H2N_e?0,, Intermediate 21 \
CO2Me CO2Me \

\
CO2Me Intermediate 2 /CONHOH
NH2OTMS Me0H KOH0 /

> Insoluble MatrixiI =
AcOH Me0H0-\___CONHOH

\
Resin 4 \
CONHOH

[00103] Scheme 15 [00104] Example 12 [00105] Extension of the Linker Group. In Paragraph [0032]
and [0036] we showed three options for longer linkers that might be used to connect the chelating agent to the polymer resin. These linkers insert polyethylene glycol units between the aromatic ring of the resin and the amine group attached to the bridgehead carbon of the chelating agent.

[00106] The invention includes the use of three amine capped polyethylene glycol (PEG) based linkers, 3-oxa-1,5-diaminopentane (17), 4,7,10-trioxa-1,13-tridecanediamine (18), and the 2-aminopropane capped polyethylene glycol with 10-12 PEG units (19), all of which are commercially available in bulk. A representative attachment scheme using Intermediate 2 and the 2 PEG unit diamine (17) is shown in Scheme 16, along with the structures of the two other PEG linkers (18,19).

, . , Et3N THF0 Insoluble Matrix' = II
Insoluble Matrix 4. SO2CI . 1 S_ N'C)NH

15 H2N7-..õ..Ø.,...---.,NH2 20 , Insoluble Matrix' 4.
"

\ 2 /CO2Me Et3N, THE 0 0 Insoluble Matrix' =
' /CO2Me 1 8 H H H 0 CO2Me / \
\
Intermediate 22 CO2Me CO2Me \\

Intermediate 2 CO2Me CONHOH
//

NH2OTMS Me0H KOH 0 0 ' Insoluble Matrix; 0 ¨ N '---'''' N j---- N-C-0-\_CONHOH

AcOH Me0H 0 H \

\CONHOH
Resin 5 Alternative Linkers ..(-0 NH2 )2H2N )J,0c),N H2 0 ' n 18 n = 10-12CH3 [00107] Scheme 16 [00108] Each amine capped polyethylene glycol linker is attached to the activated resin (15) by using an excess of the diamine to ensure complete capping. The sulfonamide linked tether (20) is activated as the N-hydroxysuccinimide (NHS) with N,N'-disuccinimidyl carbonate (16) (Takeda, K., Y. Akagi, A. Saiki, T. Tsukahara, and H. Ogura, Studies on activating methods of functional groups. Part X. Convenient methods for syntheses of active carbamates, ureas, and nitroureas using N,N'-disuccinimido carbonate (DSC).
Tetrahedron Letters, 1983, 24: 4569-4572.), followed by washing to remove the excess carbonate and the N-hydroxysuccinimide byproduct from the resin to produce (21). Reaction of the activated urethane with the amino-triester (Intermediate 2) provides the resin capped product Inteimediate 22 which is expected to be stable to hydroxylamine and aqueous conditions.
Final conversion to the trihydroxamate (Resin 5) is accomplished with 0-trimethylsily1 hydroxylamine in methanol.
[00109] Example 13. Binding of Metal Ions by the Free Ligands [00110] The acid dissociation constants for the trihydroxamate Ligand 1 and the dihydroxamate Ligand 7 have been determined by potentiometric titration of the free ligands in 0.1 M KNO3 at 25 C. The overall ligand protonation constants for Ligand 1 are log 13011 =
10.26, log 13012 = 19.68, and log 13013 = 28.15. The overall ligand protonation constants for Ligand 7 are log Pon = 9.80 and log 13m: = 18.49. These protonation constants have been used in the calculations of the metal chelate stability constants described below.
[00111] The binding of A134-, Fe3+, an a series of divalent metal ions to Ligand 1 has been evaluated by potentiometric titration in 0.1 M KNO3 at 25 C. For most of the metal ions, two complexes were detected. In one complex, all three of the hydroxamate groups were coordinated to the central metal one. The stability of these complexes is described by the overall binding constant [ML] (3) A = {M][L]

where L refers to the fully deprotonated. "nonionic form of ligand 1, and charges on the species have been omitted for clarity.
[00112] The potentiometric analysis also detected a protonated metal chelate, designated as MHL. The position of the 112 and-to-metal charge transfer band in the visible spectrum of the MI-IL complex of Fe3-1 indicated that in the MHL complexes, two of the hydroxamate groups are coordinated to the metal ion, while the third hydroxamate group is protonated and not bound to the metal ion. The stability of the MHL complexes is described by the overall binding constant =

[MHL]
All = [M1{1-1-11] (4) [00113] The calculated binding constants for the complexes of Ligand 1 are listed in Table 2. The binding constant for Al3+ is log 1311 0= 21.44. Ligands with only two hydroxamates have binding constants of about log f3110 ¨ 15 (Evers, A., Hancock, R.D., Martell, A.E., Motekaitis, R.J.., Metal ion recognition in ligands with negatively charged oxygen donor groups. Complexation of Fe(111), Ga(II1), In (III), Al(II1), and other highly charged metal ions, Inorg. Chem. 1989, 28: 2189-2195). The larger value of log p,,0 for Ligand 1 confirms that all three hydroxamate groups of the ligand are bound to the Ar'.

Table 2: Binding constants for metal complexes of the trihydroxamate ligand, Ligand 1.

Fe3+ Al3+ Cu2+ Ni24 Zr124 Mn2+ Ca2' Log 1311, 27.60 26.27 23.61 19.10 19.13 17.06 13.34 Log 13,10 23.78 21.44 10.73 10.13 8.95 3.71 [00114] The data in Table 2 confirm that Ligand 1 shows very high selectivity for the binding of trivalent metal ions such as Al3+ and Fe3+ in preference to the binding of Ca'''.

This is a critical property, as it allows this ligand to bind trivalent metal ions in the presence of very high concentrations of Ca2+.

[00115] Ligand 1 showed good selectivity for Al3+ and Fe3' in comparison to the divalent transition metal ions Cu2+, Ni2+, Zn2+, and Mn2 . However, the binding affinities for these metal ions were still appreciable, especially for the binding of Cu2+.
Thus it is not claimed that the invention can remove Al' and/or Fe3+ from pharmaceutical solutions without also removing significant amounts of Cu2+ and Zn2+. The proposed process for reducing Al3+ in total parenteral nutrition (TPN) solutions involves the removal of Al'' from the calcium gluconate and sodium phosphate component solutions, rather than treating the final TPN solution. Treating the final TPN solution with the invention is likely to remove a large percentage of the essential ions Cu2+ and Zn2"-.

[001161 Example 14. Metal Binding by Ligand 7 [001171 The dihydroxamate Ligand 7 forms 1:1 complexes with all the metal ions studied in which both hydroxamate groups are coordinated to the metal ion. The stability of these complexes is characterized by the values of log 13110 shown in Table 3.
The 1:1 complex of Al3+, Zn2+ and Mn2+ hydrolyze to form the mixed-ligand hydroxo complexes ML(OH). characterized by the overall binding constant [ML(OH)][H] (5) /811-1[M][L]

Speciation calculations based on the stability constants in Table 3 indicated that the Al complex of Ligand 7 existed as a mixture of the ML and ML(OH) complexes over the pH

range of 3 to 7. If an immobilized form of Ligand 7 (Resin 2) is used to remove Al3+ from solutions within this pH range, the formation of the ML(OH) complex will stabilize the immobilized Al and facilitate removal of Al3+ from the solution.

Table 3. Binding Constants for Metal Complexes of Ligand 7 Al3+ Cu2+ Ni2+ Zn2+ Mn2+

Pilo 16.07 13.97 9.02 9.18 7.15 P11-1 11.06 0.35 -0.1 [00118]

[00119] Example 15. Binding of Al to Resin 1 [00120] The compounds and compositions of the present invention are useful in a method of removing a trivalent metal ion such as A13+ from an aqueous solution. This is performed by treating the aqueous solution with an effective amount of the compound or composition of the present invention. In the most preferred embodiment, the invention consists of a resin to which the chelating agent is attached by a covalent bond to form a chelating resin.
1001211 In one method of use, the resin is stirred in a solution. After the metal ions from the solution bind to the resin, the metal-depleted solution and the metal-laden resin are separated by filtration or decantation.
[00122] In a second method of use, the resin is packed in a column, and the metal-containing solution is passed through the column. The metal ions are retained on the column, while the metal-depleted solution exits from the outlet of the column.
1001231 In one possible application, the invention would be used to reduce the amount of Al3+ contained in total parenteral nutrition solutions, particularly for TPN solutions given to neonates. The binding constants shown in Tables 1 and 2 indicate that treatment of the final TPN solution with the invention is likely to remove essential metal ions such as Fe3+, Cu2+ and Zn2+ in addition to A13+. Thus the strongly preferred process is to use the invention to remove the Al3+ from small volume parenteral (SVP) solutions that are used in the preparation of TPN solutions.
1001241 The primary "culprit" SVP solutions, which are contaminated with aluminum thereby contributing aluminum to the final TPN admixture and therefore to the patient, are calcium gluconate and sodium phosphate (Driscoll, M. and D.F. Driscoll, Am. J.
Health-Syst. Pharm. 2005, 62: 312-315). It should be appreciated that removal of Al3+
from these solutions is difficult because the anions of these salts, gluconate and phosphate, respectively, are themselves strong Al-binding agents (R.J. Motekaitis and A.E. Martell, Inorg. Chem.
1984, 23: 18-23; K. Atkari, T. Kiss, R. Bertani, and R.B. Martin, Inorg. Chem.
1996, 35:
7089-7094). Thus the invention must compete against high concentrations of these anions in order to remove Al3+ from the solution.
[00125] In one possible application, the compositions of the present invention are loaded into a flow-through filter device such as illustrated in Figures 8a-8d and described in greater detail below. As the SVP solution flows through the device, the aluminum is extracted from the solution. The device is provided in-line between the container of the SVP
culprit solution and the TPN bag being prepared by the automated TPN
compounder. The device has on its outlet side a membrane filter with a pore size small enough to sterilize the solution by filtration, retain the resin in the device and block release of large particles from the device. A screen on the inlet side contains the resin. Leur lock or similar connectors on the inlet and outlet sides enable easy connection to standard i.v. fluid administration sets.
[00126] In another medical application, the compounds and compositions of the present invention are utilized to ensure that aluminum is not inadvertently included in the dialysis solution used in peritoneal dialysis or hemodialysis. Another example is home peritoneal dialysis, where tap water is used to prepare the dialysate. If the tap water contains significant aluminum, which might have been introduced during the water treatment process, or might enter in the raw water, and which is not adequately removed during the water treatment process, the aluminum could enter the patient. In addition the compounds and compositions of the present invention could be used on a bulk scale in industry to remove aluminum from solutions such as the solutions that go into SVP containers or any material or process that is contaminated with aluminum, such as the guanine nucleotide-binding regulatory component (G/F) of adenylate cyclase, with which aluminum binds to activate adenylate cyclase. The following experimental data support the utility of the claimed compounds and compositions.
[00127] To demonstrate the ability of Resin I to bind Al, 50 mg of the resin was suspended in 100 ml of a buffered (0.10 M 4-morpholineethanesulfonic acid) aqueous solution at pH 5, and 25 mcg Al was added, as an acidic solution of aluminum chloride. The free Al3+ concentration in the sample solution was measured as a function of time by ETAAS. The results are shown in Figure 2. The 50 mg of resin removed 94% of the Al3+
from the solution after 94 hours and 97.4% of the Al'' from the solution after 287 hours (see Fig. 2 and Table 5). The removal of the A13+ followed first order kinetics, with a half-life of 10.5 hrs.
[00128] The Al3+ removal experiment was repeated by adding 25 mcg of Ar and 50 mg of Resin 1 to a smaller volume of only 5 ml of MES buffer at pH 5. The results are shown in Figure 2. Under these conditions, 98% of the Al3+ was removed from the solution within 12 hr and 99.9% after 24 hours.
[00129] The removal of A131- from MES buffer was also followed by a spectrophotometric assay in which the weaker chelating agent 7-iodo-8-hydroxyquinoline-5-sulfonic acid (ferron) was used as an indicator for free A13. The data are shown in Figure 3.
The starting solution contains a 1:1 ratio of 150 microMolar A13" and ferron in a total volume of 3.0 ml, and the initial spectrum shows the peak at 360 nm indicative of the Al-ferron complex. A total of 25 mg of Resin 1 was added to the solution, and the removal of A13"
from the solution was monitored by the loss of the absorbance of the Al-ferron complex at 360 nm and the corresponding increase in the absorbance of free ferron at 440 nm. Based on a comparison to the final absorbance to that of a standard solution of free ferron, it is estimated that the resin removed approximately 80% of the A134. The rate of Al removal corresponds to a half-life of approximately 90 min. The smaller percentage of Al removed reflected the competition for A13 from the ferron. These data were used to estimate an equilibrium constant for the binding of Al3+ to the Resin 1 as described below.
[00130] To determine the capacity of Resin 1 to bind Al, sequential aliquots of 100 mcg of A134 were added at 8 hr intervals to 50 mg of the resin suspended in 100 ml of MES
buffer at pH 5. The total amount of Al added to the solution was 6,000 ng/ml.
The concentration of free Al remaining in the solution was followed by ETAAS. The results are shown in Figure 4. Because of the slow rate of Al removal in dilute solutions, the concentration of A134- accumulates to a total of approximately 4,000 ng/ml after the addition of the final aliquot of A134. However, after 200 hrs the resin removed about 85% of the added Al3+, reducing the free Al concentration to about 1,000 ng/ml. This indicates that the binding capacity of Resin 1 is at least 10,000 mcg A134 per gram of resin.
1001311 The binding affinity of Resin 1 has been evaluated from four different types of experiments and the results are summarized in Table 5. Binding constants for the immobilized ligand were calculated using the speciation program HySS. A
multicomponent equilibrium model was constructed for each reaction solution, in which the binding constant of Resin 1 was the only unknown binding constant. This constant was then adjusted manually until the HySS speciation model results matched the experimentally determined value for the percentage of Al bound to the resin. In all the calculations of the binding constants for the resin, the protonation constants for the immobilized ligand are assumed to be the same as those of the free ligand so that the resulting equilibrium constant can be expressed as a value of log13110, rather than a pH-dependent effective binding constant.

[00132] The binding affinity of Resin 1 was determined from the final Al concentrations shown in Figure 2 for the removal of A13 from MES buffer. In these calculations, the only competitive binding agent was hydroxide ion. The calculations used hydrolysis constants for the Al3+ for 0.1 M ionic strength taken from Mesmer and Baes (The Hydrolysis of Cation, Wiley, New York, 1976). The values of 13110 for the immobilized ligand of Resin 1 are listed in Table 5.
[00133] The aluminum binding constant for Resin 1 has been calculated from the spectrophotometric data shown in Figure 3. In addition to A13" hydrolysis constants, these calculations included the Al binding constants of ferron from Martell and Smith (Critical Stability Constants, Vol 3, Plenum, New York, 1979). The binding constant for Resin 1 is listed in Table 5.
[00134] The Al binding constant for Resin 1 has been determined by competition against the well-known hexadentate chelating agent 1,10- diaza-4,7-di oxadecane-1,1,10,10-tetraacetic acid (EGTA). A known amount of Resin 1 was allowed to equilibrate in a pH 5.7 solution containing both EGTA and A13 . After equilibration, the concentration of Al bound in the solution to EGTA was determined by ETAAS. Protonation constants and the Al-binding constant for EGTA were taken from Martell and Smith (Critical Stability Constants, vol 1, Plenum, New York, 1974). The binding constant for Resin 1 is listed in Table 5.
[00135] Aluminum binding constants for the Resin 1 were also measured by competition against gluconic acid. A 50 mg aliquot of the resin was added to 1 ml of a commercial solution of 0.23 M Ca(gluconate)2. Analysis by ETAAS showed that the untreated solution contained 115 mcMolar Al3+ as a contaminant. No other Al was added to the solution. Resin 1 removed approximately 10% of the Al from this solution.
Based on the known binding constants for Al-gluconate, HySS was used to calculate the binding constant for the immobilized ligand on Resin 1. The binding constant for Resin 1 is listed in Table 5.
[00136] Competition experiments versus gluconate were repeated using samples in which the commercial Ca(gluconate)2 solution was diluted with pH 5 MES buffer.
These solutions contained gluconate concentrations of 0.215 M, 0.1 M, and 0.046 M
gluconic acid.
The results are listed in Table 5.

[00137] To determine the ability of Resin 1 to bind Al at ratios of mg resin per ml of Ca(gluconate) that more closely model conditions one expects in a filtration device, another competition experiment versus gluconate was conducted using samples in which 250 mg of Resin 1 was added to 0.50 ml of the commercial Ca(gluconate)2 solution. The initial Al concentration was 9130 ng/ml. The concentration of free Al remaining in the solution was followed by ETAAS. The results are shown in Figure 5.
Table 5: Summary of Experiments to Calculate the Al binding constant (f3i10) for Resin 1 Solution Total Volume pH mg resin Total Al % Al Log plio (m1) (microMolar)removed 0.1 MMES 100 5.0 50 9.26 97.4 18.9 0.1 MMES 5 5.0 50 185 99.9 18.2 0.15 mM ferron 3 5 25 150 78 19.2 mM EGTA 1 5.7 50 14.9 58 18.8 0.23 M Ca(glu)2 1 5.3 50 115 10 18.5 0.108 M Ca(glu)2 1 5.2 50 53.8 18 18.5 0.05 M Ca(glu)2 1 5.4 50 25.0 44 18.7 0.023 M Ca(glu)2 1 5.3 50 11.5 72 18.9 0.23 M Ca(glu)2 0.5 4.3 250 338 40 19.0 [00138] The overall average binding constant for the immobilized ligand of Resin 1 from the data in Table 5 is log 13110 = 18.7 + 0.3. This value is about 2.7 log units less than the binding constant for free Ligand 1. Unfavorable steric interactions with the resin may be a factor, particularly when large, bulky ligands are bonded to the resin (M.
Feng, L. van der Does, and A. Bantjes, J. Appl. Polymer Sci., 1995, 56: 1231-1237). The invention includes resins with longer groups linking the ligand to the polymer. It is anticipated this elongation of the linker will result in better agreement between the binding constants of the free and immobilized ligands.
[00139] It is significant that the binding constants calculated from competition with the gluconate solutions are in good agreement with the other values in Table 5.
This confirms that the presence of high concentrations of Ca2+ in the gluconate solutions has essentially no impact on the ability of the resin to remove Al from gluconate solutions.

[00140] EXPANDED DETAILED
DESCRIPTION OF THE PREFERRED EMBODIMENTS OF
THE INVENTION
[00141] The present invention relates to generally novel chelating agents having general formula of [00142] wherein R1 =
hydrogen, sulfonamide, urea, carbamate, carboxamide, aryl or alkyl, R2 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent [00143] and R3 =
a) (CH2)x-0 0 R4 / R4 N-OH
(CH2)z-0 0(CH2)y-OM'NLOH
o HO N-R4 [00144] wherein x, y, and z vary independently from 1 to 4, X = CH2 and 0, and R4 ¨
hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent.

[00145] [Note: when x=y=z=1, this is the TRIS platform]

b) 0 pH
>\¨N
µR4 (CH2)x-0 0 (CH2)y-01-- N'R4 (CH2)z-0 0H

[00146] wherein x, y, and z vary independently from 1 to 4, and R4 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent.

[00147] Note: when x=y=z=1, this is the TRIS platform]

c) Ho, 0 (CH2)x-0 0 (CH2)y-ON--\
(CH2)z-0 HO

0 \

[00148] wherein x and y vary independently from 1 to 4, X = CH2 and 0, and R4 =

hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent.

d) pH
(CH2)x-0 0R4 (CH2)y ON N
(CH2)z-0 HO ) pH
0 \
[00149] wherein x and y vary independently from 1 to 4, X CH, and 0, and R4 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent.

e) (CH2)x-0¨(CH2)y¨CON(OH)¨CH2(CF12)zCONR4OH
c R5 (CH2)x-0¨(CH2)y¨CON(OH)¨CH2(CH2)zCONR4OH

[00150] wherein x varies from 1-4, y varies from 1-2, and z varies independently from 2 to 8, R4 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent, and R5 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent.

(CH2)x-0¨(CH2)y¨CON(OH)¨CH2(CH2)zCH2N(COR6)0H
c R5 (CH2)x-0¨(CH2)y¨CON(OH)¨CH2(CH2)zCH2N(COR6)0H

[00151] wherein x varies from 1-4, y varies from 1-2, and z varies independently from 2 to 8, R5 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent, and R6 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent,or Ph or similar aryl substituent [00152] The present invention also includes generally novel tripodal triesters as precursors or useful intermediates to chelating ligands having general formula of [00153] wherein R' = hydrogen, sulfonamide, urea, carboxamide or benzyl, R2 --hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent and R7 =

g) (CH2)x--o (CH2)y-00R8 (CH2)z-0 0 [00154] wherein x, y, and z vary independently from 1 to 4, X = CH2 and 0, and R8 ¨
hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent and Ph or similar aryl substituent.
h) (CH2)x-0 0 (CH2)z-0(CE12)y¨OLOR8 [00155] wherein x, y, and z vary independently from 1 to 4, X = CH2 and 0, and R8 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent and Ph or similar aryl substituent.

[00156] The novel compounds of the present invention are particularly useful as chelators or chelating agents. One preferred use of the free ligands would be in vivo chelation therapy to remove metal ions such as Fe3+ and A13+ from the body.

[00157] The compounds include an amine functional group that allows the ligands to be easily linked to an insoluble matrix via a sulfonamide linkage, an amine linkage, an amide linkage, or a urea linkage to provide immobilized, tethered chelators.
Typically, the insoluble matrix comprises a resin support. The resin support may take the form of a macro-porous polystyrene such as commercially available under the trademark XAD-4 sold by Rohm and Haas. Other polymer resins useful in the present invention include but are not limited to, polyacrylate, sepharose and silica gel.

[00158] The overall process of adding a chelating compound of the present invention to a polystyrene resin via a sulfonamide bond, where NR2H-Ligand in this and subsequent schemes refers to the free amine form (le = H; R2 = hydrogen, methyl, ethyl, n-propyl or isopropyl) of any of the free ligands represented by R3 = a through f is shown in scheme 17.

Scheme 17 CISO3H 0 Et3N, THF
Insoluble Matrix Insoluble Matrix , g CI
8 RHN-ligand macroporous polystyren resin modified resin e.g. XAD-4 Insoluble Matrix, 411 g¨N¨Ligand linker ligand resin [00159] An alternative process of adding a chelating compound of the present invention to a polystyrene resin via a sulfonamide bond, where NR2H-Ligand-prcursor in this and subsequent schemes refers to the free amine form (1t1 = H; R2 = hydrogen, methyl, ethyl, n-propyl or isopropyl) of any of the triesters represented by R3 = g through h is shown in Scheme 18.

[00160] Scheme 18 O CISO3H 0 Et3N, THF
Insoluble Matrixl S¨CI ' Insoluble Matrix;
411 g CI

ligand precursor macroporous polystyren resin modified resin e.g. XAD-4 O H NH2OR', Me0H, Me0Na Insoluble Matrix; g¨N¨Ligand precursor ii O R' = H, TMS

Insoluble Matrixi 4r ¨N¨Ligand linker ligand resin CO2Me (CH2)x-0 RHN (CH2)y (CH2)z-0 0 CO2Me S-CI

CO2Me a it 0, ri (cH2)y-0,CO2Me (CH2)x-0 CONHOH
8 (cH2)z¨o 0R (CH2)x-0 CO2Me N (CH2)y_cy----v=CONHOH

6 (cH2)z--0\

CONHOH

[00161] The overall process of adding a chelating compound of the present invention to a resin support by means of a urea linkage is shown in Scheme 19.

[00162] Scheme 19 ' , , , , ' Et3N, THF

Insoluble Matrix! .
NH2 0=0=N-ligand precursor k Insoluble Matrix! ii N A NH-ligand precursor H
modified resin NI-120R', Me0H, Me0Na , Insoluble Matrix l . 1 NANH-ligand R' = H, TMS

H

Insoluble (-1 Matrix = New /1CO2Me /CO2Me itif lik (CH2)x-0 COCl2, CH2Cl2 (CH2)x-0 /

0CO2Me CO2Me , H2N ( (CH2)y (CH2)z-0 aq NaHCO ' 3 0/
/0, ,N (CH2)z-0(CH2)y-0 Et3N, THF
\
\
\

\
CO2Me CO2Me / /CO2Me (?' (CH2)x-0 NH2OR', Me0H, Me0Na NH--"N c (CH2)y-0 CO,Me H
R' = H, TMS
(CH2)z-0 \

CONHOH
\
CO2Me Giv<¨
//
ICI' (CH2)x-0 NI-1--I''-N c (CH2)y-0--- CONHOH
H
(CH2)z-0 \
\
CONHOH

1001631 An alternative process of adding a chelating compound of the present invention to a resin support by means of a urea linkage is shown in Scheme 20.

1001641 Scheme 20 , .
, COCl2, CH2C1 2 1 Insoluble Matrix . , Insoluble Matrix, =

NH2 aq NaHCO3 N=C=0 modified resin Et3N, THF i 0 NH2OR', Me0H, Me0Na - Insoluble Matrix, 411 A
,, RHN-ligand precursor N NR-ligand precursor H
R' = H, TMS

Insoluble Matrix; .

NANR-ligand H

CO2Me Insoluble ri (c H2)x-o Matrix = '40.- RHN c (CH2)y _cy\-0O2Me (CH2)z-0 \--\
e= II COCl2, CH2Cl2 . it 11 CO2Me ) NH2 aq NaHCO3 NCO Et3N, THF

CO2Me /
/
(cH2)x-0 CO2Me NH2OR', Me0H, Me0Na it .0 N HN c (CH2)y-0 .
R R' = H, TMS
(CH2)z-0 \ CONHOH
\ /
CO2Me (ex¨ /
If? (CH2)x ¨0 NH --"IN (CH2)y_0ONHOH
R
(CH2)z-0 \
\
CONHOH

[00165] The overall process of adding a chelating compound of the present invention to a resin support by means of an amine linkage is shown in Scheme 21.

[00166] Scheme 21 N-methylpyrrolidone Nal1 Insoluble Matrix! 1 -Insoluble Matrix, II
CI RHN-ligand precursor N¨Ligand precursor modified resin NH2OR, Me0H, Me0Na Insoluble Matrix 441 N¨Ligand R' = H, TMS

CO2Me (CH2)x-0 CO2Me RHN (CH2)y-0 (CH2)z-0 CO2Me h(CH2)x-0CI N-methylpyrrolidonCe 2Me RN c (CH2)y-0 CO2Me Nal Insoluble (cH2)z-0 Matrix CO2Me CONHOH

NH2OR', Me0H, Me0Na (;)_(_) (cHox-0 R ' = H, TMS RN (CH2)y¨OCONHOH

(CH2)z-0 CONHOH

1001671 The overall process of adding a chelating compound of the present invention to a resin support by means of an amide linkage is shown in Scheme 22.

[00168] Scheme 22 Insoluble Matrix' 0 Et3N, THE
Insoluble Matrixl Cl RHN-ligand precursor N¨Ligand precursor modified resin NH2OR', Me0H, Me0Na ' Insoluble Matrix,' 111 R' = H, TMS

N¨Ligand CO2Me Insoluble rA
Matrix w (CH2)x--0 _(:)CO2Me RHN (CH2)y (CH2)z-0 CO2Me CO2Me =
0 (CH2)x-0 " THF, Et3N

RN -c (CH2)y-07 CO2Me Cl (CH2)z-0 CO2Me /CONHOH
NH2OR', Me0H, Me0Na /
R' = H, TMS =
RN (cH2)x-0(CH2)y-0 CONHOH

(CH2)z-0 CONHOH

[00169] For certain applications it may be desirable to elongate the linker by adding polyethylene glycol units between the resin support and the ligand in order to increase the rate of metal binding to the resin-bound ligand. These elongated linkers are added using commercially available amine capped polyethylene glycols of variable length, with the use of a urea functional group to covalently bind the ligand and linker moieties.

NH2 N=C=0 CO2Me (CH2)x-0 CO2Me ,N
(CH2)y-0 /c 0/ ' (CH2)z-0 NH
CO2Me Insoluble CO2Me Matrix w 4. 0 (CH2)x-0 (CH2)y_oCO2Me H H
(CH2)z-0 CO2Me a II
(cH2)x¨o/

(CH2)y-0 H H
(CH2)z-0 Other commercially available amine-capped polyethyleneglycols include the compound H2 )0 \

[00171]
which gives chelating resins with the structures shown below Insoluble Matrix l= 1¨NH
O

NR2¨Ligand or Insoluble Matrix, 111 [00172] The immobilized, tethered chelators of the present invention comprise the chelating compounds identified above bound to a resin support through an appropriate linkage. The immobilized, tethered chelators of the present invention may be generally described as having the following formula:
R6¨N¨R3 [00173] wherein R6 =

Insoluble g_ Insoluble Matrix Matrix !I
Insoluble(¨)_ Insoluble Matrix / CH2 NH Matrix )¨CH2¨NH-_ Insoluble Matrix Insoluble MatrixNH--0,,8 H

Insoluble c¨)_c?H JCH3 9 Matrix S¨N 0 n = 10-12 [00174] R2= hydrogen, methyl, ethyl; n-propyl or isopropyl and [00175] R3 =

a) N-OH

(C1-12)X-0 R4 (C1-12))/-0 'OH
(CH2)z-0 0 () Ho'N-R4 [00176] wherein x, y, and z vary independently from 1 to 4, X = CH2 and 0, and R4 =

hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent.

b) 0 ,OH

(CH2)x-0 0 (CH2))/-01--- N-R4 (CH2)z¨O\ OH

)7-N)R4 [00177] wherein x, y, and z vary independently from 1 to 4, and R4 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent c) HO, 0 (CH2)z-0 HdpH
NX

[00178] wherein x and y vary independently from 1 to 4, X = CH2 and 0, and R4 =

hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent.

d) 0 pH

(CH2)x-0 0 (CI-12)y -0/N-N
(CH2)z-0 HO /OH 1 [00179] wherein x and y vary independently from 1 to 4, X = CH2 and 0, and R4 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent.

e) (CH2)x-0--(CH2)y-CON(OH)-CH2(CH2),CONR4OH

(CH2)x-0-(CH2)y-CON(OH)-CH2(CH2),CONR4OH

[00180] wherein x varies from 1-4, y varies from 1-2, and z varies independently from 2 to 8, R4 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent, and R5 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent.

(CH2)x--0¨(cH2)y¨CoN(0H)¨CH2(CF12),CH2N(COR6)0H
c R5 (CH2)x--0¨(CH2)y¨CoN(oH)¨cH2(CH2),CH2N(COR6)0H

[00181] wherein x varies from 1-4, y varies from 1-2, and z varies independently from 2 to 8, R5 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent, and R6 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent,or Ph or similar aryl substituent [00182] Example 16 , . ., [00183] General Synthesis of Tripodal ligands.

[00184] A general approach to the amino triol scaffolds (24a-d to 27a-d) employs the chemistry of nitroalkanes. Starting with 2-nitroethanol (23a), 3-nitropropanol (23b), 4-nitrobutanol (23c) or 5-nitropentanol (23d), two identical alkenol chains can be added with 1 carbon atom (24a-d), 2 carbon atoms (25a-d), 3 carbon atoms (26a-d), or 4 carbon atoms (27a-d). In all cases, reduction of the nitro group with hydrogen over Ti Raney nickel will yield the amino compounds. Protection of the nitroalkenols (23a-d) with a TBS
group (series B) will yield amino triols with one chain differentiated for further reaction.

[00185] Scheme 24 0 (cH2)x¨OR
(cHox¨OR ,,,,,,, 1. Pd(0), --',--K,J
1. Na2CO3, H20, CH20 02N(CH2)x¨OR
, H2N c (cHoy¨OH
H2N (CH2)y OH , A R = H 2.T1 Raney Nickel 2. T1 Raney Nickel, (CH2)z¨OH
(CH2)z¨OH B R = SiMe2t-Bu H2, Me0H
H2, Me0H
23a x = 1 24ax= y=z=1 1. t-BuOK
THF 23b x = 2 's= 1. '-'- CHO 27ax=1,y=z=4 24bx= 2,y=z= 1 23c x = 3 s=,, 2. NaBH4 27bx=2,y=z=4 24c x= 3,y=z= 1 o 23d x= 4 24dx= 4,y =z= 1 µ-3. T1 Raney Nickel 27c x = 3, y = z = 4 2. Ti Raney Nickel '1-42, Me0H 27dx=y=z=4 H2, Me0H
(CH2)x¨OR
(CH2)x¨OR

H2N c (CH2)y¨OH

(CH2)y¨OH 26ax=1,y=z=3 25a x = 1, y = z = 2 (0H2)z¨OH (CH2)z¨OH 26b x = 2, y = z = 3 25bx=y=z= 2 26cx=y=z= 3 25cx=3,y=z=2 AR=H
26d x= 4, y =z= 3 25d x= 4, y = z = 2 B R = SiMe2t-Bu [00186] The tripodal ligands with different length side arms can be prepared by the reactions of aminomethyl triol scaffolds (24-27)] with acrylonitrile then HC1/Me0H (28-31), or ethyl diazoacetate (36-39). The resulting esters (28-31 and 36-39) are converted to the hydroxamates (32-35 and 40-43) with hydroxylamine or TMSONE12.

[00187] Scheme 25 , COX
/
/
(CH2)x-OH^-,- (CH2)x-0 X = OMe TMSONH2 or 1. KOH, dioxane, ON ,--,,,COX
NH2OH.HCI
H2N (CH2)y¨OH 2. HCl, Me0H, reflux ' H2N
(CH2)y-0 Me0H
(CH2)z¨OH (CH2)z-0 X = NHOH ' NaOH
\
Series A X= OMe28a-d to 31a-d \
23a-d to 27a-d X= NHOH,32a-d to35a-d COX

[00188] Scheme 26 /----cox (cH2)x---OH (cH2)x--0 x = OEt TMSONH2 or 1. Rh2(0Ac)4, N2CHCO2Et NH2OH.HCI
H2N c (CH2)y¨OH ' H2N c (CH2)y¨OCOX
Me0H

(CH2)z¨OH(CH2)z-0 X = NHOH * NaOH

Series A
23a-d to 27a-d X= OEt, 36a-d to 39a-d X= NHOH, 40a-d to 43a-d [00189] Example 17 [00190] Synthesis of ligand 8 [00191] Scheme 27 1.(Boc)20, Me0H' BocHN
1.CF3002H, 0H2012, H2N OH it, 20 h, 68%
C)."--0O2Et it, 73%
.
, 0-"---CO2Et OH 2. Rh2(0Ac)4, 2. TsCI, Et3N, ethyl diazoacetate, 0 26c OH CH2Cl2, it, 6 h, 73% \--0O2Et CH2Cl2, 40 C, 15 h, 91%
Intermediate 23 TsHN Cr¨''CO2Et NH2OTMS, TsHN
C)---CONHOH

`-' CO2Et Me0H, it, 30 h 'µC)----CONHOH
0 80% 0 \--0O2Et \---CONHOH

' Intermediate 24 Ligand 8 [00192] Boc- protection of the amino group of amino triol 26c followed by rhodium acetate catalyzed alkylation of hydroxyl groups with ethyl diazoacetate gave intermediate 23. Boc- deprotection and reprotection of the amine by a tosyl group proceeded smoothly to give intermediate 24. Reaction of tri-ester (Intermediate 24) with 0-trimethylsily1 hydroxylamine produced the ligand 8. The ligand 8 is a colorless solid and was purified by recrystalization from ethanol/isopropanol mixture (1:1) and its structure was determined by x-ray crystallography.
100193] To a stirred solution of aminotriol 26c (1.65g, 8.06 mmol) in dry Me0H
(33 mL) was added (Boc)20 (1.86g, 8.5 mmol) and the mixture was stirred at room temperature for 20 h. The solvent was evaporated under reduced pressure and the residue was purified by column chromatography (Si02, CH2C12/Me0H gradient) to give the product (1.681 g, 68%) which was crystallized from C112C12 as white crystals: IR (neat) 3453, 3400, 3307, 3228, 2947, 2872, 1689 cm-1; 'H NMR (Me0D) 8 (ppm) 3.43 (t, J= 6.6 Hz, 6H), 1.57-1.51 (m, 6H), 1.42-1.35 (m, 6H), 1.32 (s, 911); 13C (Me0D) 8 (ppm) 156.5, 79.3, 63.4, 57.9, 32.4, 28.8, 27.5.
[00194] To a stirred solution of Boc protected amino triol (0.6 g, 1.96 mmol) and Rh2(0Ac)4 (0.043 g, 0.098 mmol) in CH2C12 (6 mL) was added a solution of ethyl diazoacetate (1.05 ml, 9.98 mmol) in CH2C12 (50 ml) over a period of 1 h (syringe pump).
After the addition was complete, the mixture was stirred for 5 h at room temperature. The solvent was removed under reduced pressure and the crude product was purified by column chromatography (Si02, hexane/Et0Ac gradient) to give intermediate 23 (0.8 g, 73%): IR
(neat) 3356, 2952, 2875, 1749, 1729, 1640 cm-1, 1FINMR (CDC13) 8 (ppm) 4.20 (q, J¨ 7.1 Hz, 6H), 4.05 (s, 611), 3.49 (t, J= 6.3 Hz, 6H), 1.63-1.57 (m, 12H), 1.39 (s, 911), 1.27 (t, J-7.1 Hz, 911); 13C (CDC13) 5 (ppm) 170.6, 154.3 78.7 72.1, 68.5, 60.9, 56.9, 31.8, 28.6, 23.7, 14.4; HRMS (FAB) calcd for C271-149N011Na [M+Nar: 586.32037. Found: 586.31950.
[00195] Intermediate 23 (1.147 g, 2.03 mmol) was in a 1:1 mixture of CF3CO2H
(2.3 mL, 31 mmol) and CH2C12 (2.3 mL) and the resulting solution was stirred at room temperature. The reaction was monitored by TLC (50% Et0Ac in hexane). When the reaction was complete, the solvent was evaporated under reduced pressure. The residue was dissolved in CH2C12 (10 mL) and a saturated solution of Na2CO3 was added dropwise while cautiously shaking the flask until CO2 evolution ceased. The layers were separated and the aqueous layer was extracted with twice more with CH2C12 (2x20 mL). The combined organic layers were dried over Na2SO4 and evaporated under reduced pressure to give the crude product, which was purified by column chromatography (Si02, CH2C1,/Me0H
gradient) to give the free amine as a colorless oil (0.683 g, 73%): IR (neat) 3436, 2945, 2864, 1744, 1634, cm-1; 1H NMR (CDC13) 8 (ppm) 4.19 (q, J= 7.1 Hz, 6H), 4.05 (s, 6H), 3.52 (t, J= 5.7 Hz, 6H), 1.65-1.54 (m, 12H), 1.24 (t, J= 7.1 Hz, 9H); 13C (CDC13) 8 (ppm) 170.6, 72.1, 68.5, 61.0, 54.7, 35.2, 23.7, 14.3.
[00196] To the stirred solution of the free amine (0.68 g, 1.46 mmol) and tosyl chloride (0.66 g, 3.5 mmol) in CH2C12 (13 ml) was added Et3N (0.40 mL, 2.87 mmol) and the mixture was heated at 40 C for 17 h. The solvent was evaporated under reduced pressure and the residue was dissolved in CH2C12 (25 ml) and washed with water (2x25 mL). The aqueous layer was extracted with CH2Cl2(2x25 mL) and the combined organic layers was dried over Na2SO4 and concentrated under reduced pressure. The resiude was purified by column chromatography (Si02, hexane/Et0Ac gradient) to give intermediate 24 as a gummy solid (0.826 g, 91%): IR (neat) 3278, 2948, 2877, 1747 cm-1, 1H NMR (CDC13) 8 (ppm) 7.68 (d, J
= 8.2 Hz, 2H), 7.17 (d, J= 8.2 Hz, 2H), 4.10 (q, J= 7.1 Hz, 6H), 3.88 (s, 6H), 3.24 (t, J= 5.9 Hz, 6H), 2.31 (s, 3H), 1.48-1.38 (m, 12H), 1.18 (t, J= 7.1 Hz, 9H); 13C
(CDC13) 8 (ppm) 170.3, 142.6, 140.7, 129.3, 126.7, 72.4, 68.1, 61.9, 61.0, 32.7, 23.2, 21.3, 14.1; HRMS
(FAB) calcd for C29H47N011SNa [M+Nar: 640.27673. Found: 640.27700 [00197] To a solution of the triester (intermediate 24) (0.54 g, 1.46 mmol) in Me0H (6 mL) was added NH2OTMS (1.00 mL, 8.17 mmol) and the mixture was stirred at room temperature for 30 h. The solvent was evaporated under reduced pressure to give a foamy solid. Recrystallization from a 1:1 mixture of Et0H and iPrOH yielded ligand 8 as a white crystalline solid (0.55 g, 80%): IR (neat) 3256, 2953, 2872, 1642 cm "1; 1H
NMR (D20) 8 (ppm) 7.81 (d, J= 8.3 Hz, 2H), 7.44 (d, J= 8.3 Hz, 2H), 3.99 (s, 6 H), 3.35 (t, J= 5.9 Hz, 611), 2.43 (s, 3H), 1.52 (m, 12H); 13C (Me0D) 8 (ppm) 169.2, 144.4, 142.7, 130.7, 128.0, 72.9, 70.1, 63.0, 33.7, 24.2, 21.5; HRMS (FAB) calcd for C23H39N4011S [M+H]:
579.23358.
Found: 579.23320. The product structure was confirmed X-ray crystallography.
[00198] Example 18 [00199] Synthesis of Ligand 9 [00200] Scheme 28 K2CO3, H20, 02N OH T-1 Raney Nickel,, H2NOH

02N OH CH20 (2 eq.) H2, Et0H, 45 psi 23c 30 C, 3 h, 84% OH OH rt, 20 h, 95 %
OH OH
intermediate 25 interemediate 26 1. Me0H, dry HCI
acrylonitrile, dioxane H2N reflux, 15 h, 74 %TsHN CO2Me KOH (cat.), rt, 24 h, 49 % 0 2. TsCI, Et3N, CH2Cl2, 40 C, 15 h 75%CN \--0O2Me intermediate 27 intermediate 28 Me0H, KOH (cat.), 0 0¨\--CONHOH
amberlyst-15, rt, 7 h, 88 A) CONHOH ligand 9 [00201] The base catalyzed addition of two equivalents of formaldehyde to 3-nitropropanol (Griesser, H.; Ohrlein, R.; Ehrler, R.; Jager, V. Synthesis 1999, 77, 236) 23c gave intermediate 25. The nitro group was reduced to an amine by hydrogenation using T-1 Raney nickel as a catalyst and hydrogen at 45 psi. The amino triol intermediate 26 was alkylated with acrylonitrile to give trinitrile. Treatment with HCI in methanol at reflux gave the aminotriester, which protected by tosylation to give intermediate 28.
Reaction of intermediate 28 with 0-trimethylsily1 hydroxylamine produced the ligand 9.

[00202] To a solution of 3-nitropropanol (6.70 g, 63.7 mmol) (Griesser, H.; Ohrlein, R.; Ehrler, R.; Jager, V. Synthesis 1999, 77, 236 ) in H20 (7 mL) were added 37 weight %

formaldehyde solution (10 mL, 128 mmol) and solid K2CO3.3/2H20 (21.0 g, 127 mmol) and the mixture was stirred at room temperature for 1 h (reaction was exothermic) and then at 30 C for lh. The reaction was monitored by TLC (10% Me0H in CHC13). 20% aqueous solution was added drop wise with stirring until the effervescence of CO2 ceased. The resulting mixture was washed with CH2C12 (2x30 mL) to remove some impurities and the aqueous layer was evaporated under reduced pressure. The residue was triturated with hot Et0H (3 x 50 mL) and the Et0H was filtered and evaporated under reduced pressure to yield intermediate 25 as a thick oil (8.85 g, 84%). IR (neat) 3378, 2949, 2888 cm-1;

(D20) 8 (ppm) 4.05 (ABq, A6 = 22.2 Hz, J = 12.4 Hz, 4H) 3.73 (t, J = 6.6 Hz, 2H), 2.26 (t, J
= 6.6 Hz, 2H); 13C NMR (D20) 6 (ppm) 93.9, 62.1, 57.0, 33.3.
[00203] Freshly prepared T-1 Raney nickel (3.6 g) was transferred to a Parr hydrogenation flask as a slurry in absolute Et0H (45 mL). Intermediate 25 (4.80 g, 29.1 mmol) was dissolved in absolute Et0H (45 mL) and transferred to the hydrogenation flask.
The resulting mixture was shaken on a Parr hydrogenator under 45 psi of H2 at room temperature for 20 h. The flask was removed from the hydrogenator and flushed with argon for 20 min. The catalyst (pyrophoric when dry) was then filtered through a short pad of celite, which was never allowed to dry. The celite was washed with Et0H (3 x 30 mL). The Et0H was evaporated to give intermediate 26 a viscous brown gel (traces of Et0H was always present) (3.7 g, 95%) which was used in the next step without further purification. IR
(neat) 2931, 2875 cm-1; 111 NMR (D20) 5 (ppm) 3.72 (t, J= 7.2 Hz, 2H), 3.47 (s, 4H), 1.65 (t, J = 7.2 Hz, 2H); 13C NMR (D20) 8 (ppm) 64.7, 56.9, 54.8, 35.2 [00204] To a stirred solution of intermediate 26 (3.65 g, 27.0 mmol) and KOH
pellets (0.4 g) in 1,4-dioxane (13 mL) was added acrylonitrile (6.3 mL, 94 mmol) drop wise over a period of 1 h. Once the addition was complete, the mixture was stirred at room temperature for 24 h. The solvent was evaporated under reduced pressure to yield thick liquid residue which was dissolved in CH2C12 (50 mL) and washed with H20 (50 mL). The aqueous layer was re-extracted with additional portions of CH2C12 (2x30 mL). The combined fractions were dried over Na2SO4 and evaporated under reduced pressure and the residue was purified by column chromatography (SiO2, 5% Et0H in Et0Ac) to give intermediate 27 as a thick brown oil (3.89 g, 49%): IR (neat) 3371, 3307, 2873, 2245 cm-1; 11-1 NMR
(CDC13) 5 (ppm) 3.68 (t, J = 6.0 Hz, 4H), 3.65 (t, J = 6.1 Hz, 2H), 3.62 (t, J = 6.2 Hz, 2H), 3.41 (ABq, A8 = 19.5 Hz, J- 8.7 Hz, 4H), 2.60 (t, J = 6.0 Hz, 4H), 2.59 (t, J = 6.0 Hz, 2H), 1.72 (t, J=
6.1 Hz, 2H); '3C NMR (CDC13) 8 (ppm) 118.2, 75.1, 67.5, 66.0, 65.8, 54.7, 34.6, 19.1, 19.0;
HRMS (FAB) C,41-123N403 [M+Hr calcd 295.17703, found 295.17720 [00205] Dry HC1 gas was bubbled into a solution of intermediate 27 (1.43 g, 4.86 mmol) in Me0H (12 ml) until it was saturated. The resulting mixture was heated at reflux for h. Saturated Na2CO3 was added drop wise while stirring the mixture until CO2 effervescence ceased. The solution was extracted with Et0Ac (3x30 mL). The combined organic layers were washed with brine, dried over Na2SO4 and evaporated to obtain the the amino triester as a liquid (1.41 g, 74%), which was used in the next step without further purification. IR (neat) 2951, 2875, 1732 cm-I; NMR (CDC13) 8 (ppm) 3.74- 3.63 (m, 15H), 3.53 (t, J= 6.5 Hz, 2H), 3.26 (ABq, AS = 13.8 Hz, J= 8.9 Hz, 4H), 2.55 (t, J= 6.3 Hz, 6H), 2.03 (br s, 2H), 1.62 (t, J= 6.4 Hz, 2H); 13C NMR (CDC13) 6 (ppm) 172.3, 172.2, 75.1, 67.4, 66.8, 66.3, 54.7, 51.8, 51.7, 35.1, 35.0, 34.5; HRMS (FAB) Ci7H32N09 [M+1-11+ calcd 394.20770, found 394.20860 [00206] To a stirred solution of the amino triester (1.40 g, 3.56 mmol) and TsC1 (1.60 g, 8.40 mmol) in CH2C12 (32 mL) was slowly added Et3N (1.0 mL, 7.2 mmol) and the resulting mixture was heated at reflux for 20 h. The solvent was evaporated under reduced pressure and the residue was dissolved in CH2C12 (50 mL), washed with water (2x25 mL), dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by column chromatography (Si02, hexanes/Et0Ac gradient) to give intermediate 28 as a thick oil (1.42 g, 74%): IR (neat) 3287, 2953, 2876, 1732 cm-1; 111 NMR
(CDC13) 6 (ppm) 7.73 (d, J= 8.3 Hz, 2H), 7.24 (d, J= 8.3 Hz, 2H), 5.58 (s, 1H), 3.67 (s, 3H), 3.64 (s, 6H), 3.58 (t, J= 6.3 Hz, 2H), 3.50-3.36 (m, 10H), 2.52 (t, J= 6.3 Hz, 2H), 2.39 (t, J= 6.3 Hz, 4H), 2.38 (s, 3H), 1.88 (t, J= 6.0 Hz, 2H); 1-3C NMR (CDC13) 8 (ppm) 172.1, 172.0, 142.8, 140.6, 129.3, 126.9, 72.1, 66.9, 66.5, 66.2, 61.6, 51.8, 51.7, 34.8, 34.6, 32.6, 21.5; HRMS
(FAB) C24H38N011S [M+Hr calcd 548.21655, found 548.21570 [00207] To a solution of intermediate 28 (0.5 g, 0.91 mmol) in dry Me0H
(5.5 mL) was added NH2OTMS (0.67 mL, 5.48 mmol) and the resulting solution was stirred at room temperature. The reaction was monitored by TLC (50% Et0Ac in hexanes). No reaction was observed after 3 h. KOH (0.3 g) was added and stirring was continued for an additional 45 min. Amberlyst-15 (2.6 g, washed with dry Me0H) was added to the reaction mixture and stirring was continued for 1 h. The mixture was filtered and the filtrate was evaporated under reduced pressure to obtain ligand 9 as a solid (0.44 g, 88%): IR (neat) 3215, 2876, 1638 cm-1;
11-1 NMR (D20) 8 (ppm) 7.80 (d, J= 8.2 Hz, 2H), 7.47 (d, J= 8.2 Hz, 2H), 3.64 (t, J= 6.0 Hz, 2H), 3.48-3.52 (m, 6H), 3.39 (s, 4H), 2.44 (s, 3H), 2.40 (t, J= 6.0 Hz, 2H), 2.32 (t, J=

5.8 Hz, 4H), 1.85 (t, J = 6.7 Hz, 2H); 13C NMR (Me0D) 8 (ppm) 171.0, 170.9, 144.4, 141.9, 123.5, 127.9, 72.5, 67.6, 67.3, 62.5, 54.9, 34.4, 33.3, 21.5; HRMS (FAB) C211-[M+Hf calcd 551.20227, found 551.20260 [00208] Example 19 [00209] General Synthesis of Macrocyclic Ligands.

[00210] A general approach to macrocyclic ligands begins with mono protected aminotriols (series B, 24ad-27ad). The free hydroxyls are alkylated with t-butyl acrylate under basic conditions. The hydroxyl protecting is removed and the hydroxyl is alkylated methyl acrylate. Finally, the amine is protected with a Cbz group to give compounds 44a-d to 47a-d. The t-butyl groups are removed selectively with TFA and the resulting diacids are converted into the acid chlorides using oxalyl chloride. Macrocycles are prepared by reaction of diacid chlorides with benzyl protected hydroxylamines under high dilution conditions. The remaining methyl is reacted with 0-benzyl hydroxylamine and then benzyl groups are removed via hydrogenolysis to yield the hydroxamic acids 50a-d to 53a-d.

[00211] Scheme 29.

CO2Me (CH2)x¨OR 1. KOH, dioxane, CO2t-Bu (CH2)x-0 H2N ( (CH2)y¨OH 2. HF.Py, CH2C12 CbzNH (CH2)y¨OCO2t-Bu (CH2)z¨OH 3. KOH, dioxane, (CH2)z-0 Series B 4. CbzCI CO2Me 24a-d to 27a-d 44a-d to 47a-d CO2t-Bu CONHOH

1. TFA0 (CH2)x 2. DMF, (C0C 3. BnONH2 ¨012)2, CH3CN
NH2 c (CH2)y high dilution 4. H2, Pd.0 OBn OBn 2OH, Hd ) H¨N
50a-d to 53a-d AX=CH2,BX=0 0 X-- 48, X = CH2 49, X = 0 [00212] Example 20 [00213] General Synthesis of Tetrahydroxamic Acids Type A

[00214] Tetrahydroxamates, designed to mimic the structure of DF0-13, can be prepared from protected amino diacids. The amino diol scaffold for the diacids are prepared using chemistry similar to that for the aminotriols (para 00199). Starting with nitroalkanes (54), two identical alkenol chains can be added with 1 carbon atom (55), 2 carbon atoms (56), 3 carbon atoms (57), or 4 carbon atoms (58). In all cases, reduction of the nitro group with hydrogen over Ti Raney nickel will yield the amino compounds.

[00215] Scheme 30 0 (CH2)4-0H
(CH2)¨OH 1. Pd(0), 1. Na2003, H20, CH20 02N R H2N ( R
H2N R 2. Ti Raney nickel 2. Ti Raney Nickel, H2(CH2)4-0H
(CH2)-0H H2, Me0H 54 R = Me, Et,etc 55 1. t-BuOK 58 THF 1. CHO

2, NaBH4 2. T1 Raney Nickel 3. Ti Raney Nickel, H2, Me0H H2, Me0H

(CH2)2-0H (CH2)3-0H

(CH2)2-0H
56 57 (CH2)3-0H

The diesters with different length side arms can be prepared by the reactions of aminodiol scaffolds (55-58)] with acrylonitrile then HCl/Me0H (59-62) or diazoacetate (67-70). The resulting esters (59-62 and 67-70) are converted to the acids (63-66 and 71-74) by hydrolysis with lithium hydroxide.

Scheme 31 COR' (CH2)x-OH(CH2)x-0 59-62 R' = OMe H2N R H2N 1. KOH, dioxane, CN R
LiOH
(CH2)x¨OH 2. HCI, Me0H, reflux (CH2)x-0 63-66 R = OH

55-58 COR' (CH2)x-OH (CH2)x-0 67-70 R' = OEt H2N R 1. Rh2(0Ac)4, N2CHCO2Et"*" H2N R
LiOH
(CH2)x¨OH (CH2)x-0 71-74 R' = OH -= ---55-58 \¨COR' For series A, the acids are coupled with a protected monoacyl dihydroxylamine (78). The oximes are made by oxidizing diols to dialdehydes and condensing with 0 benzyl (or other protected) hydroxylamine to give an oxime. Reduction of the oxime with sodium cyanoborohydride under acidic conditions give the protected dihydroxylamine which is monoacylated with one equivalent acetic anhydride.

Scheme 32 1. PCC, Celite, CH2Cl2, rt, 2h, HCI.Me0H (2N), HOCH2(CH2),CH2OH BnON=CH2(CH2),CH2N-0Bn 75 2. BnONH2.HCI, 76 Me0H, rt, 3 h, Py, Et0H, NaBH3CN, ref lux, 4 h Ac20 (1 eq.), DMAP, BnONHCH2(CH2)zCH2NHOBn BnONHCH2(CH2)zCH2N(Ac)0Bn 77 Py, CH2Cl2, rt, 2 h 78 The acids are coupled with two equivalents of protected monoacyl dihydroxylamine (78) using dicyclohexyl carbodiimide (DCC) and then the benzyl groups are removed via hydrogenolysis to give the tetrahydroxamic acids 79 Scheme 33 (CH2)x-0¨(CH2)y¨COOH

1. DCC, HOBt, R
Py, DMAP, (CH2)z-0¨(CH2)y¨COOH CH2Cl2, 0 C to it, 63-66 or 71-74 15h 2. H2, Pd/C (10%), BnONHCH2(CH2),CH2N(Ao)0Bn Me0H, rt, 1 h 78 (CH2)x-0¨(CH2)y¨CONH(OH)CH2(CH2),C1-12NH(Ac)OH

R

(CH2)z-0¨(CH2)y¨CONH(OH)CH2(CH2),CH2NH(Ac)OH

[00216] Example 21 [00217] (Type A) Tetrahydroxamate Ligand 10 [00218] PCC oxidation of 1,9-nonanediol gave nonanedial, which was reacted with 0-benzylhydroxylamine hydrochloride and pyridine in refluxing ethanol to produce the dioxime intermediate 29 Sodium cyanoborohydride reduction of dioxime gave benzyl protected bis-hydroxylamine intermediate 30. Acetylation of one of the hydroxylamine nitrogen using one equivalent of acetic anhydride led to the formation of intermediate 31.

[00219] Scheme 34 1. PCC, Celite, CH2Cl2, rt, 2h, NaBH3CN, 66%
HCI.Me0H (2N), HO OH
r N
Bn0 µ0Bn Me0H, rt, 3 h, 1,9-nonanediol L. B nONH2.HCI, 76%
Py, Et0H, intermediate 29 reflux at 79 C, 4 h 67%

Ov Ac20 (1 eq.), DMAP, N, ,N
BnO,N OBn Py, CH2Cl2, it, 2 h, 30 % BnO µ0Bn intermediate 30 intermediate 31 =

[00220] The dicarboxylic acid (intermediate 32) was prepared (Scheme 35) by the base catalyzed hydrolysis of intermediate 17 (R. A. Yokel, W. R. Harris, C. D.
Spilling and C.-G.

Zhan (2011) US Patent 7,932,326).

[00221] Scheme 35 i) Li0H.H20, Aq. THF (50%), 000 to rt TsHNi ii) 2N HC1 (to pH 1) TsHN7 intermediate 17 intermediate 32 [00222] Finally, the two partners 32 and 31 were coupled using DCC (Scheme 36) and deprotected by hydrogenation with H2 over 10% Pd on C to obtain ligand 10.

[00223] Scheme 36 1, DCC, HOBt, Py, DMAP, 0 TsNN/ CH2C12, 000 to it' OH
intermediate 32 15 h 7 OH
CD Tsl-INKcy¨-CONN,OH
2. H2, Pd/C (10 /o), OH 7 BnO ,N \OBn Me0H, it, 1 h Ligand 10 intermediate 31 [00224] To a solution of PCC (32.33 g, 150.0 mmol) and with a suspension of celite (11.0 g) in CH2C12 (300 mL) was added 1,9-nonanediol (10.0 g, 62.4 mmol). This solution was stirred at room temperature and the reaction was monitored by TLC (1:1 hexanes/Et0Ac). After 2 h the reaction was complete. The reaction mixture was passed through a short column of silica and celite, which was washed with CH2C12 (5x50 mL). The solvent evaporated under reduced pressure and the residue was purified by column chromatography (Si02 hexanes/Et0Ac gradient) to give the dialdehyde (A. Ozane, L

Pouysegu, D. Depernet, B. Francois, S. Quideau Organic Letter 2003, 5, 2903-2906) as a colorless liquid (6.4 g, 66%): 1H NMR (CDC13) 8 (ppm) 9.73 (t, J= 1.8 Hz, 2H), 2.40 (td, J

= 7.3, 1.8 Hz, 4H), 1.58 (m, 4H), 1.32 (br s, 6H); 13C NMR (CDC13) 5 (ppm) 202.8, 43.9, 29.2, 29.0, 22Ø
[00225] To a solution of nonanedial (5.60 g, 35.8 mmol) and 0-benzylhydroxylamine hydrochloride (15.0 g, 94.0 mmol) in Et0H (120 mL) was added pyridine (7.6 mL, 94 mmol) drop wise. The resulting solution was heated at reflux for 4 h. The solvent was evaporated under reduced pressure and the residue was dissolved in CH2C12 (200 mL) and washed with water (3x100 mL). The aqueous layer was re-extracted with CH2C12 (2x100 mL).
The combined organic layers were dried over Na2SO4, evaporated under reduced pressure, and the residue was purified by column chromatography (Si02 hexanes) to give intermediate 29 as a colorless liquid (8.01 g, 61%) as a 1.5:1 mixture of geometric isomers.
IR (neat) 3030, 2925, 2856 cm-1; 1H NMR (CDC13) 5 (ppm) 7.46 (t, J 6.2 Hz, 0.6H), 7.39-7.31 (m, 5H), 6.69 (t, J= 5.4 Hz, 0.4H), 5.13 (s, 0.8 H), 5.08 (s, 1.2H), 2.39 (app q, 0.8H), 2.20 (app q, 1.2H), 1.48 (m 2H), 1.32 ( m, 3H); 13C NMR (CDC13) 8 (ppm) 152.6, 151.7, 138.3, 137.8, 128.5 (x2), 128.3, 128.0, 127.9, 127.8, 75.8, 75.6, 29.6, 29.3, 29.1, 29.0, 26.7, 26.2, 25.9;
HRMS (FAB) C23H31N202 [M+H] calcd 367.23856, found 367.23830. One of the isomers was isolated and characterized completely. 11-1 NMR (CDC13) 5 (ppm) 7.39 (t, J= 6.2 Hz, 2H), 7.31-7.24 (m, 10H), 5.00 (s, 4H), 2.13 (app q, 4H), 1.41 (quin, J= 6.8 Hz, 4H), 1.25 (m, 6H); 13C NMR (CDC13) 8 (ppm) 151.7, 137.9, 128.5, 128.4, 128.0, 127.9, 75.6, 29.6, 29.1, 29.0, 26.7.
[00226] To a solution of intermediate 29 (0.11 g, 0.3 mmol) and NaCNBH3 (0.042 g, 0.66 mmol) in Me0H (2 mL) was added 2N HC1 in Me0H drop wise until the solution pH
was between 3 and 4. The resulting mixture was stirred for 3 h at room temperature. The solvent was evaporated under reduced pressure and the solid residue was dissolved in water (2 mL) and 6 N KOH solution was added drop wise to adjust the solution pH to >9. The aqueous solution was extracted with CH2C12 (3 x10 ml), and combined organic extracts were washed with brine (20 mL), dried over Na2SO4 and evaporated under reduced pressure. The residue was purified by column chromatography (Si02hexanes/Et0Ac gradient) to give the intermediate 30 as a colorless liquid (0.083 g, 76%): IR (neat) 3028, 2924, 2852, 1453, 1363 cm-1; 111 NMR (CDC13) 5 (ppm) 7.30-7.38 (m, 10H), 4.73 (s, 4H), 2.94 (t, J=
7.0 Hz, 4H), 1.52 (app quin, J=6.7 Hz, 4H), 1.30 (br s, 10H); 13C NMR (CDC13) 8 (ppm) 138.2, 128.5 (x2), 127.9, 76.3, 52.3, 29.6, 27.4, 27.3.
[00227] To a solution of intermediate 30 (0.40 g, 1.08 mmol) in CH2C12 (15 mL) was added DMAP (0.13 g, 1.06 mmol), pyridine (0.18 mL, 2.22 mmol), and Ac20 (0.102 mL, 1.08 mmol) sequentially and the resulting mixture was stirred at room temperature for 2 h.
The mixture was diluted with CH2C12 (20 mL) was added and washed with saturated NaHCO3 (20 mL). Organic layer was dried over Na2SO4 and evaporated under reduced pressure and the solid residue was purified by column chromatography (Si02, hexanes/Et0Ac gradient) to give intermediate 31 as a colorless liquid (0.120 g, 30%): IR
(neat) 3031, 2927, 2854, 1660 cm-1; 1H NMR (CDC13) 8 (ppm) 7.31-7.39 (m, 10H), 5.56 (br s, 1H), 4.82 (s, 2H), 4.71 (s, 2H), 3.63 (t, J= 6.9 Hz, 2H), 2.93 (t, J= 7.1 Hz, 2H), 2.10 (s, 3H), 1.64 (m, 2H), 1.50 (m, 211), 1.29 (br s, 10H); 13C NMR (CDC13) 5 (ppm) 154.2, 138.2, 134.7, 129.3, 129.0, 128.9, 128.5 (x2), 127.9, 76.4, 76.3, 52.3, 29.6, 29.4, 27.5, 27.3, 27.0, 26.9, 20.7; HRMS (FAB) C25H37N203 [M+H] calcd 413.28040, found 413.2776 [00228] To a solution of intermediate 17 (3.00 g, 6.96 mmol) in aqueous THF
(1:1, 30 ml) at 0 C, was added Li0H.H20 (1.17 g, 27.9 mmol) and the resulting mixture was stirred for 1 h. After 1 h, the flask was allowed to warm to room temperature while stirring was continued. When all starting material was consumed (TLC 1:1 Et0Ac/hexanes), the reaction was quenched with 2N HC1 and solution pH was adjusted to 1. The mixture was filtered (to remove LiC1) and extracted with CH2C12 (3x50 mL). The combined organic extracts were washed with brine, dried over Na2SO4 and evaporated under reduced pressure to give the di-acid as a white powder (2.67 g, 95%) which was used in the next step without further purification. IR (neat) 3300, 3051, 2925, 2875, 1697 cm -1; 111 NMR (D20) 6 (ppm) 7.84 (d, J= 8.3 Hz, 2H), 7.45 (d, J= 8.3 Hz, 2H), 3.60-3.53 (m, 411), 3.37 (ABq, A5 =
22.8 Hz, J=
Hz, 4H), 2.53 (t, J= 6.0 Hz, 4H), 2.44 (s, 3H), 1.14 (s, 311); 13C NMR (Me0D) 8 (ppm) 175.6, 144.3, 142.5, 130.5, 128.0, 74.8, 68.0, 60.1, 35.7, 21.5, 19.4; HRMS
(FAB) C17H26N08S [M+Hr calcd 404.13790, found 404.13880.

[00229] Ligand 10. To a solution of intermediate 31(0.40 g, 0.97 mmol) and DMAP
(0.16 g, 1.31 mmol) in CH2C12 (10 mL) and pyridine (0.11 mL, 1.4 mmol) was added a solution of intermediate 32 (0.18 g, 0.45 mmol) and HOBt (0.14 g, 1.03 mmol) in CH2C12 (10 mL) and the mixture was cooled to 0 C. DCC (0.2 g, 0.97 mmol) was added and the mixture was stirred for 1 h at 0 C, then it was allowed to warm to room temperature and was stirred for an additional 15 h. The solvent was evaporated under reduced pressure and the residue was purified by column chromatography (Si02, hexanes/Et0Ac gradient) to give the tetrabenzyl tetrahydroxamate as a thick colorless oil (0.342 g, 65%): IR
(neat) 3033, 2930, 2856, 1650, 1601 cm-1; 1H NMR (CDC13) 5 (ppm) 7.75 (d, J= 8.2 Hz, 2H), 7.45-7.33 (m, 20H), 7.17 (d, J= 8.2 Hz, 2H), 5.74 (s, 1H), 4.78 (s, 4H), 4.76 (s, 4H), 3.63-3.59 (m, 12H), 3.30 (ABq, M = 52.5 Hz, J= 9.2 Hz, 4H), 2.66-2.55 (m, 4H), 2.25 (s, 3H), 2.04 (s, 6H), 1.58 (m, 8H), 1.23 (br s, 20H), 1.08 (s, 3H); 13C NMR (CDC13) 8 (ppm) 172.2, 172.0, 142.4, 140.8, 134.4, 134.3, 129.9, 129.3, 129.2, 129.0, 128.7, 128.6, 128.5, 127.0, 126.7, 126.2, 77.6, 76.2, 76.0, 73.7, 66.7, 58.6, 45.2, 32.5, 29.2, 29.0, 26.7, 26.5, 21.3, 20.4, 18.1; HRMS
(FAB) C67H94N5012S [M+H] calcd 1192.66199, found 1192.66110 [00230] To a solution of tetrabenzyl tetrahydroxamate (0.34 g, 0.28 mmol) in Me0H
(11 mL) was added 10% Pd/C (0.074 g). Flask was then evacuated and flushed with H2 from two balloons and the mixture was stirred under H2 at room temperature until the starting material was consumed ('TLC, 70% Et0Ac in hexanes). The reaction flask was purged with Ar and the mixture was filtered. The filtrate was evaporated under reduced pressure to give ligand 10 as a foamy solid (0.22 g, 91%): IR (neat) 3360, 3060, 2928, 2857, 1611 cm-1; 11-1 NMR (Me0D) 5 (ppm) 7.68 (d, J= 8.2 Hz, 2H), 7.23 (d, J= 8.2 Hz, 2H), 3.52-3.46 (m, 12H), 3.27-3.17 (m, 4H overlapping with Me0H peak), 2.58 (t, J= 6.0 Hz, 4H), 2.31 (s, 3H), 1.99 (s, 6H), 1.51 (br s, 8H), 1.21 (br s, 20H), 1.00 (s, 3H); 13C NMR (Me0D) 8 (ppm) 173.6, 173.5, 144.2, 142.6, 130.5, 128.0, 75.0, 68.0, 60.1, 48.9, 33.9, 30.6, 30.4, 27.8, 21.6, 20.3, 19.5; HRMS (FAB) C39H70N50/2S [M+H] calcd 832.4742, found 832.4725 [00231] Example 22 [00232] Tetrahydroxamate ligand 11 , =

[00233] Ligand 11 is prepared from amino diester intermediate 17.
The amine is protected with Cbz to give intermediate 33. Hydrolysis with lithium hydroxide yields diacid intermediate 34 which is coupled with intermediate 31 using DCC. Global deprotection of the Cbz and benzyl protecting groups gives ligand 11.

[00234] Scheme 37 0 ----0O2Me H2N/ CbzHNK0 intermediate 17 intermediate 33 i) Li0H.H20, DCC, HOBt, Py, DMAP, Aq. THF (50%), 0 C to it CH2Cl2, 0 C to it, 15 h ii) 2N HCI (to pH 1) CbzHNK0-------0O2H
(2.2 eq.) 0y intermediate 34 H
BnO" N \OBn intermediate 31 OH

H2, Pd/C (10%), 0 -OH
Me0H, rt, 1 h X 7 N .0H

Ligand 11 [00235] To a stirred solution of intermediate 17 (R. A. Yokel, W. R.
Harris, C. D.

Spilling and C.-G. Zhan (2011) US Patent 7,932,326) (2.0 g, 7.2 mmol) in THF
(57 mL) was added 10% aqueous Na2CO3 (57.0 mL, 53.7 mmol). After 15 minutes at room temperature CbzCl (1.13 mL, 7.93 mmol) was added and stirring was continued for an additional 5 h. The layers were separated and the aq. layer was extracted with Et0Ac (2x30 mL).
The combined organic extracts were washed with water and brine, dried over Na2SO4 and evaporated under reduced pressure. The residue was purified by column chromatography (SiO2, hexane/Et0Ac gradient) to give intermediate 33 as a colorless oil (1.9 g, 65%): IR (neat) 3366, 2951, 2875, 1731 cm-1; 11-1 NIVIR (300 MHz, CDC13) 8 (ppm) 7.33-7.31 (m, 5H), 5.28 (s, 1H), 5.01 (s, 2H), 3.67 (t, J= 6.2 Hz, 4H), 3.64 (s, 6H), 3.47 (ABq, A8 =
42.9 Hz, J= 9.0 Hz, 4H), 2.53 (t, J= 6.2 Hz, 411), 1.29 (s, 311); 13C NMR (300 MHz, CDC13) 8 (ppm) 172.0, 155.1, 137.4, 128.5, 128.0, 72.9, 66.8, 66.1, 55.7, 51.7, 34.8, 19.8; FIRMS
(FAB) C20H30N08 [M+H] calcd 412.19714, found 412.19750 [00236] To a solution of intermediate 33 (7.40 g, 18.0 mmol) in 50%
aqueous THF (75 mL) was added LiOH (1.68 g, 70.1 mmol) at 0 C and the resulting mixture was stirred for 1 h. After 1 h, the flask was allowed to warm to room temperature with continued stirring.
Once the starting material was consumed (TLC 1:1 Et0Ac/hexanes), the reaction was quenched with 2N HC1 to adjust the solution pH to 2. The mixture was extracted with CH2C12 (3x100 mL) and the combined extracts were washed with brine, dried over Na2SO4, and evaporated under reduced pressure to give intermediate 34 as a colorless gel (6.8 g, quant) which was used in the next step without further purification. IR (neat) 3033, 2939, 2878, 1709 cm -1; 11-1 NMR (300 MHz, CDC13) 8 (ppm) 10.28 (br s, 211), 7.35-7.29 (m, 511), 5.34 (br s, 1H), 5.04 (s, 211), 3.68 (t, J 6.2 Hz, 4H), 3.49 (ABq, A8 = 39.6 Hz, J= 8.9 Hz, 4H), 2.57 (t, J= 6.2 Hz, 4H), 1.31 (s, 3H); 13C NMR (300 MHz, CDC13) 8 (ppm) 177.3, 155.3, 136.5, 128.5 (x2), 128.3, 128.1, 128.0, 77.3, 72.8, 66.4, 55.7, 34.6, 19.0; FIRMS
(FAB) Ci8H26N08 [M+Hr calcd 384.1658, found 384.1648 [00237] To a solution of intermediate 31(1.1 g, 2.66 mmol) and DMAP (0.44 g, 3.60 mmol) in CH2C12 (50 mL) and pyridine (0.29 mL 3.58 mmol), was added intermediate 34 (0.47 g, 1.22 mmol) and HOBt (0.37 g, 2.71 mmol) in CH2C12 (50 mL). The mixture was cooled to 0 C and DCC (0.58 g, 2.81 mmol) was added. The mixture was stirred for 1 h at 0 C, then allowed to warm to room temperature and stirred for additional 15 h.
The solvent was concentrated under reduced pressure and the solids were removed by filtration (DCC
urea) and washed with CH2C12. The filtrate was evaporated under reduced pressure and the residue was purified by column chromatography (SiO2' Et0Ac/hexanes gradient) to give the Cbz tetrabenzyl tetrahydroxylamine as a white solid product (0.80 g, 56%): IR
(neat) 3029, 2930, 2856, 1719, 1649 cm-1; NMR (300 MHz, CDC13) 8 (ppm) 7.36-7.25 (m, 5H), 5.49 (s, 1H), 5.00 (s, 211), 4.79 (s, 414), 4.78 (s, 4H), 3.71 (t, J= 6.2 Hz, 4H), 3.60-3.55 (m, 8H), 3.49 (ABq, A5 = 45.8 Hz, J= 9.1 Hz, 4H), 2.64 (t, J= 5.9 Hz, 4H), 2.07 (s, 6H), 1.60-1.58 , .
(m, 811), 1.32 (s, 3H), 1.24 (s, 20H); 13C NMR (300 MHz, CDC13) 6 (ppm) 172.4, 172.2, 155.1, 136.8, 134.5, 129.1 (x2), 128.9, 128.8, 128.7, 128.4, 127.9, 127.8, 77.4, 76.3, 76.2, 73.2, 67.1, 65.9, 55.7, 45.3, 33.9, 32.7, 29.4, 29.3, 29.2, 29.1, 26.8, 26.7, 26.6, 20.5, 19.0;
HRMS (FAB) C68H94N5012 [M+H] calcd 1172.68994, found 1172.68620.

[00238] To a solution of Cbz tetrabenzyl tetrahydroxylamine (0.50 g, 0.43 mmol) in Me0H (16 mL) was added 10% Pd/C (0.11 g) under argon. The flask was evacuated and flushed with 112 (balloons) and then mixture was stirred under H2 at room temperature until the starting material was consumed (TLC analysis 70% Et0Ac in hexanes). The reaction mixture was then flushed with Ar and filtered. The filtrate was evaporated under reduced pressure to obtain ligand 11 as a sticky solid (0.29 g, quant): IR (neat) 3133 (broad), 2927, 2854, 1607 cm-1; 111 NMR (300 MHz, Me0D) 8 (ppm) 3.52 (t, J= 6.1 Hz, 4H), 3.37 (t, J =-7.1 Hz, 4H), 3.35 (t, J= 7.1 Hz, 4H), 3.18 (ABq, M = 26.4 Hz, J= 9.6 Hz, 4H), 2.53 (t, J-6.1 Hz, 4H), 1.85 (s, 6H), 1.38 (m, 8H), 1.09 (m, 20H), 0.92 (s, 3H); 13C NMR
(300 MHz, Me0D) 8 (ppm) 173.5, 173.4, 74.9, 68.3, 56.5, 48.9, 34.8, 33.8, 30.7, 30.4, 27.8, 26.8, 26.2, 20.4 (x2); HRMS (FAB) C32H64N5010 [M+11]+ calcd 678.46533, found 678.46620 [00239] Example 23 [00240] General Synthesis of Tetrahydroxamic Acids Type B
[00241] The synthesis of type B tetrahydroxamate ligands begins with selective reduction of oc,co diacid mono esters 80 (Z > 4) using borane in THF to give the hydroxy esters, which are reoxidized to give the ester aldehydes 81. The aldehydes are condensed with 0-benzyl (or other protected) hydroxylamine to give oximes 82. Reduction with sodium cyanoborohydride under acidic conditions will yield the hydroxylamine esters 83.
[00242] Scheme 38 , , 1. BH3.THF

BnONH2.HCI,t, THF, -18 C to r Py, Et0H, 4 h, 75 %
, HO2C(CH2)zCO2Me - 0=CH(CH2)zCO2Me 80 2. PCC, silica, CH2C12, at 4 h,, rt, 2 h 86 %
81 89%

NaBH3CN, BnON=CH(CH2),CO2Me BnONHCH2(CH2),CO2Me 2 N HCI. Me0H, , 82 Me0H, rt, 97%

The hydroxylamine esters 83 are condensed with diacids (63-66 or 71-74) to benzyl protected dihydroxamic acids 84. Reaction of the ester with 0-trimethylsily1 hydroxylamine in Me0H and removal of the benzyl groups via hydrogenolysis will yield the type B

tetrahydroxamates 85.

Scheme 39 (CH2)x¨o¨(cH2)y¨CooH

R DCC, HOBt, (CH2)x-0¨(CH2)y¨CONH(OBn)-CH2(CH2)zCO2Me (CH2)z-0¨(CH2)y¨COOH
c - c R
Py, DMAP, CH2-I2' (63-66 or 71-74) 0 C to rt, 82 % (CH2)z-0¨(CH2)y¨CONH(OBn) CH2(CH2)zCO2Me BnONHCH2(CH2),CO2Me 1. TMSONH2, Me0H, (CH2)x-0¨(CH2)y¨CONH(OH)CH2(CH2),CONHOH
rt, 20 h R

2. H2, PcI/C (10%), ''. C
(CH2)z-0¨(CH2)y¨CONH(OH)CH2(CH2)zCONHOH
Me0H, rt [00243] Example 24 [00244] (Series B) Tetrahydroxamte Ligand 12 [00245] The acid group of monomethyl azelate was selectively reduced with BH3 to give an alcohol (Scheme 40), which was reoxidized with PCC to form the aldehyde intermediate 35. The aldehyde was reacted with 0-benzylhydroxylamine hydrochloride in the refluxing ethanol to obtain the oxime ester intermediate 36, which was reduced to the benzyl protected hydroxylamine ester intermediate 37.

[00246] Scheme 40 1. BH3.THF

0 0 THF, -18 C to rt, BnONH2.HCI, 4 h, 75 % 0 0 Py, Et0H, HO OMe 2. PCC, silica, CH2Cl2, OMe ref lux at , 4 h, Monomethyl Azelate rt, 2 h 86 % intermediate 35 89%

BnO,N
0 NaBH3CN, 0 OMe 2 N HC1. r BnO, N Me0H, OMe intermediate 36 Me0H, it, 97% intermediate 37 [00247] DCC mediated coupling of dicarboxylic acid intermediate 17 with intermediate 37 produced the benzyl protected hydroxamic acid intermediate 38 (Scheme 41), which was purified by chromatography. The ester groups were then converted to the hydroxamic acids by reaction with 0-trimethylsily1 hydroxylamine. The benzyl protected hydroxamic acid groups were deprotected using hydrogenolysis to give ligand 12.

[00248] Scheme 41 OBn 0 TsHNK0--------0O2H I DCC, HOBt, 7 OMe intermediate 32 0 Py, DMAP, CH2Ci2 TsHN/ Me, BnO,N 0 C to it, 82 % OBn 70 OMe intermediate 38 intermediate 37 OCON OH
1. TMSONH2, Me0H, 7H
it, 20 h TsH N-OH
2. H2, Pd/C (10%), OH 0 Me0H, it ligand12 [00249] To a solution of intermediate 35 (Kai, K.; Takeuchi, J.;
Kataoka, T.;

Yokoyama, M.; Watanabe, N. Tetrahedron 2008, 64, 6760 ) (3.40 g, 18.2 mmol) and 0-benzylhydroxylamine hydrochloride (3.79 g, 23.7 mmol) in Et0H (73 mL) was added pyridine (3.84 mL, 47.4 mmol) drop wise. The resulting solution was heated at reflux for 3 h.
The solvent was evaporated under reduced pressure and the residue was triturated with Et0Ac (5x20 mL). The Et0Ac fractions were combined and filtered, then evaporated under reduced pressure. The residue was purified by column chromatography (Si02 hexanes/Et0Ac gradient) to give intermediate 36 as a thick colorless liquid (4.7 g, 89%) as the mixture of two geometric isomers: IR (neat) 3027, 2929, 2856, 1736, cm-1;

(CDC13) 8 (ppm) 7.44 (t, J = 6.2 Hz, 0.6H), 7.37-7.27 (m, 5H), 6.67 (t, J =
5.5 Hz, 0.4H), 5.11 (s, 0.8H), 5.06 (s, 1.2H), 3.67 (s, 3H), 2.37 (m, 0.6H), 2.31 (m, 2H), 2.17 (m, 1.4H), 1.62 (m, 2H), 1.47 (m, 2H), 1.31 (m, 6H); '3C NMR (CDC13) 8 (ppm) 174.4, 152.6, 151.7, 138.3, 137.8, 128.5 (x 2), 128.4, 128.0, 127.9 (x 2), 75.8, 75.6, 51.6, 34.2, 29.6, 29.3, 29.1, 29.0, 26.7, 26.3, 25.9, 25Ø
1002501 One of the isomers was isolated and characterized, but isomerized soon on standing: 'H NMR (CDC13) 8 (ppm) 7.44 (t, J= 6.2 Hz, 1H), 7.37-7.27 (m, 5H), 5.06 (s, 2H), 3.67 (s, 3H), 2.30 (t, J= 7.4Hz, 2H), 2.17 (app q, J= 6.4 Hz, 2H), 1.62 (m, 2H), 1.47 (m, 2H), 1.31 (br s, 6H).
[00251] To a solution of intermediate 36 (4.70 g, 16.1 mmol) and NaCNBH3 (1.12 g, 17.8 mmol) in Me0H (100 mL) was added 2N HC1 in Me0H drop wise at room temperature until the solution pH (checked by universal indicator) was adjusted to 3-4, then the solution was stirred for 3 h at room temperature. The solvent was evaporated under reduced pressure to give solid residue, which was dissolved in water (100 mL). 6 N KOH solution was added drop wise to adjust the solution the pH to >9. The aqueous solution was extracted with CH2C12 (3x100 mL). The combined organic layers were washed with brine, dried over Na2SO4 and evaporated under reduced pressure. The residue was purified by column chromatography (Si02 2% Et0Ac in hexanes) to give the hydroxylamine intermediate 37 as a colorless liquid (4.6 g, quant): IR (neat) 3022, 2928, 2854, 1736 cm-1; NMR
(CDC13) 8 (ppm) 7.36-7.27 (m, 5H), 4.71 (s, 2H), 3.67 (s, 3H), 2.92 (t, J = 7.0 Hz, 2H), 2.30 (t, J = 7.4 Hz, 2H), 1.62 (m, 2H), 1.50 (m, 2H), 1.30 (br s, 8H); 13C NMR (CDC13) 8 (ppm) 174.5, 138.1, 128.5, 127.9, 76.3, 52.3, 51.6, 34.2, 29.4, 29.3, 29.2, 27.4, 27.2, 25.1; HRMS (FAB) C17H28NO3 [M+H]+ calcd 294.20691, found 294.20750 [00252] To a solution of intermediate 37 (0.10 g, 0.34 mmol) and DMAP (0.06 g, 0.49 mmol) in CH2C12 (4 mL) and pyridine (0.04 mL, 0.49 mmol) was added intermediate 32 (0.06 g, 0.15 mmol) and HOBt (0.05 g, 0.37 mmol) in CH2C12 (4 mL). The mixture was cooled to 0 C and DCC (0.07 g, 0.34 mmol) was added. The mixture was stirred for 1 h at 0 C, then it allowed to warm to room temperature and stirred for additional 20 h. The solvent was evaporated under reduced pressure and the liquid residue was purified by column chromatography (Si02 hexanes/EtoAc gradient) to give intermediate 38 as a thick colorless oil (0.142 g, 60%): IR (neat) 3269, 3028, 2930, 2857, 1734, 1653 crn-1; 111 NMR (CDC13) 8 (ppm) 7.77 (d, J= 8.2 Hz, 2H), 7.41-7.36 (m, 10H), 7.22 (d, J= 8.2 Hz, 2H), 5.65 (s, 1H), 4.82 (s, 4H), 3.66-3.63 (m, 14H), 3.34 (ABq, A8 = 87 Hz, J= 9.2 Hz, 4H), 2.67-2.57 (m, 4H), 2.38 (s, 3H), 2.28 (t, J= 7.5 Hz, 4H), 1.62-1.57 (m, 8H), 1.28 (m, 16H), 1.09 (s, 3H);
13C NMR (CDC13) 8 (ppm) 174.4, 172.5, 142.8, 141.0, 134.6, 129.5, 129.3, 129.1, 128.9, 127.0, 76.5, 74.0, 76.1, 58.9, 51.6, 45.6, 34.2, 34.8, 29.3, 29.2, 26.8, 25.0, 21.6, 18.3; HRMS
(FAB) C511176N3012S [M+H] calcd 954.51495, found 954.51690.
[00253] To a solution of intermediate 38 (2.03 g, 2.13 mmol) in dry Me0H (45 mL) was added KOH (0.80 g, 14.3 mmol), NH2OTMS (1.22 mL, 9.38 mmol) and the resulting solution was stirred at room temperature for 15 h. Amberlyst-15 (6 g, washed with dry Me0H) was added to the reaction mixture and stirred for additional 1 h. The mixture was filtered and the filtrate was evaporated under reduced pressure to obtain the dibenzyl tetrhydroxamate as a white foamy solid (2.01 g, quant.) which was used without further purification in the next step: IR (neat): 3258, 2927, 2857, 1632, 1454, 1110 cm-1; 1H NMR
(Me0D) 8 (ppm) 7.63 (d, J= 8.2 Hz, 2H), 7.15 (d, J= 8.2 Hz, 2H), 4.76 (s, 4H), 3.55 (app t, J= 5.9 Hz, 4H), 3.42 (app t, J= 5.8 Hz, 4H), 3.23-3.12 (ABq, 4H over laps with Me0D), 2.47 (app t, J= 5.8 Hz, 4H), 2.24 (s, 3H), 2.13 (t, J= 7.4 Hz, 1.4H), 1.94 (t, J= 7.3 Hz, 2.3H), 1.48 (m, 811), 1.17 (m, 16H), 0.95 (s, 3H); HRMS (FAB) C49H74N5012S
[M+H] calcd 956.50543, found 956.50500 , [00254] To a solution of dibenzyl tetrahydroxamate (2.0 g, 2.1 mmol) in Me0H
(100 mL) under argon was added 10% Pd/C (0.22 g). The flask was then evacuated, flushed with H2 (balloons) and the mixture was stirred under H2 at room temperature for 3h.
The reaction mixture was filtered and the filtrate was evaporated under reduced pressure to give ligand 12 as a foamy solid (1.3 g, 81%): IR (neat) 3500-2600 (broad), 2927, 2856, 1613 cm-1; 1H NMR
(Me0D) 8 7.58 (d, J= 8.0 Hz, 2H), 7.14 (d, J= 8.0 Hz, 2H), 3.44-3.36 (m, 8 H), 3.16-3.09 (m, 4H, overlaps with CD30D), 2.47 (t, J = 5.7 Hz, 4H), 2.21 (s, 3H), 1.89 (t, J= 7.2 Hz, 4H), 1.41 (br. S, 8H), 1.12 (br. S, 16H), 0.89 (s, 3H); 13C NMR (Me0D) 8 173.6, 173.1, 144.3, 142.5, 130.5, 128.0, 74.9, 68.0, 60.1, 52.1, 34.9, 33.8, 30.3, 30.1, 27.8, 27.7, 26.8, 26.1, 21.6, 19.4; HRMS (FAB) C35H62N5012S [M+Hr calcd 776.41150, found 776.41130.
[00255] Example 25 [00256] Preparation of Urea-linked Tris Hydroxamic Acid Resin 6 [00257] Scheme 42 .
, --CD
Method A
¨0 /
/
j H2N¨fo Triphosgene ).
N
=
tO
, CH2Cl2, aq NaHCO3 intermediate 2 intermediate 39 NCO Triphosgene Method B
j 4--/
intermediate 41 \--NH
\--0 HNt 0 0 HN
, tO 0 4-7 1) NH2OTMS
HN¨i 0 10H-OH
N
HN¨
31r C)v H
2) aq. AcOH
Resin 6 ,I,N,OH

(Method A:1.04 meq/g) intermediate 40 (Method B: 0.54 meq/g) (Method A:1.23 meq/g) (Method B: 1.23 meq/g) Method A ¨ Amine resin and ruus Isocyanate Route [00259]
To 12.26 g (32.3 mmol) of the free amine intermediate 2 in 130 mL of CH2C12 was added 130 rnL of saturated' NaHCO3. The stirred mixture was treated PORTION WISE
with 3.2 g (10.8 mmol) of triphosgene. This reaction is fast and results in a lot of gas formation. After stirring for 15 min, the organic phase was separated. The aqueous layer was extracted twice with CH2C12, combined organic extracts dried and concentrated to afford intermediate 39 (12.2 g, 93%) as a clear, dark oil: IR(ATR) 2955, 2878, 2245 (NCO), 1733 (CO), 1437; 111 NMR (CDC13, 300 MHz) 8 2.52 (t, 3H, 6.3 Hz), 3.39 (s, 6H), 3.62 (s, 9H), 3.68 (t, 3H, 6.3 Hz); 13C NMR (CDC13, 75 MHz) 8 34.7, 51.6, 63.7, 67.0, 71.1, 127.2 (CNO), 171.8.

[00260] To 2 g (4.4 mmol) of aminomethyl resin (Aldrich 564095; Macroporous 30-mesh, 2.2 meq/g) was added 10 mL of CH2C12 followed by 2.3 mL of diisopropylethyl amine (DIEA). The mixture was treated with 5.3 g (13.2 mmol) of intermediate 39 and allowed to agitate at rt overnight. The resin was subsequently filtered and washed three times each with CH2C12, Me0H, H20, saturated NaHCO3, H20, Me0H and Et20. After filtration the resin was dried under reduced pressure overnight at rt to afford 3.11 g of a light tan resin intermediate 40 IR(ATR) 3382 (br), 3023, 2924, 1737 (C=0), 1680; Elemental Analysis: C, 74.31; H, 7.53; N, 3.44; EA shows loading of 1.23 meq/g based on N analysis.
[00261] A suspension of 2.74 g (3.37 mmol) of urea-triester resin intermediate 40 in 17 mL of Me0H was prepared in the 60 mL peptide reactor. The mixture was treated with 3.02 mL of NH2OTMS followed by 1.34 g of KOH in 10 mL of Me0H. The mixture was allowed to agitate by rocking overnight. The mixture was filtered and washed three times each with Me0H, H20, Me0H and H20. The resin was treated with 10% aq. acetic acid and allowed to agitate for one hour. The resin was filtered, washed three times each with Me0H, H20, Et20 and dried under reduced pressure to give 2.85 g of the trihydroxamic acid resin 6 IR(ATR) 3205, 2919, 1636, 1550. Elemental Analysis: C, 66.02; H, 7.08; N, 7.28; EA shows loading of 1.04 meq/g based on N analysis.
[00262] Method B ¨ Isocyanate resin and TRIS Amine Route [00263] To 2 g (4.4 mmol) of aminomethyl resin (Aldrich 564095; macroporous 30-mesh, 2.2 meq/g) was added 10 mL of CH2C12 followed by 770 uL of DIEA and 1.3 g (4.4 mmol) of triphosgene. The mixture was allowed to rock in a peptide reactor for 15 min and subsequently filtered and washed with CH2C12. The resin was suspended in 10 mL
CH2C12, treated with 2.31 mL of DIEA and 5.0 g (13.2 mmol) of aminetriester intermediate 2 and allowed to rock overnight. Filtration followed by washing three times each with CH2C12, Me0H, H20, satr NaHCO3, H20, Me0H and Et20 afforded intermediate 40. The product was dried in vacuo at rt overnight to give 2.35 g of an off-white resin:
IR(ATR) 3300 (br, weak), 3025, 2923, 2260 (very weak, residual isocyanate), 1738, 1679, 1601.
Elemental Analysis: C, 81.06; H, 7.62; N, 3.41; EA shows loading of 1.23 meq/g based on N analysis.
[00264] A suspension of 2.2 g (2.7 mmol) of urea-triester resin intermediate 40 in 10 mL of Me0H was prepared in the 60 mL peptide reactor. The mixture was treated with 1.2 mL of NH2OTMS followed by 0.56 g of KOH in 8 mL of Me0H. The mixture was allowed to rock overnight. The mixture was filtered and washed three times each with Me0H and H20. The resin was treated with 10% aq. acetic acid and allowed to rock for one hour. The resin was filtered, washed three times each with H20, Me0H, Et20 and dried under reduced pressure to give 2.2 g of the trihydroxamic acid resin 6: IR(ATR) 3311, 3023, 2920, 1651, 1600. Elemental Analysis: C, 80.46; H, 7.51; N, 3.75; Elemental analysis shows loading of 0.54 meq/g based on N analysis.
1002651 Example 26 1002661 Preparation of Polyalkoxy Tether-linked Tris Hydroxamic Acid Resin 7 , 0-1-0\---\

1) CH2Cl2, TEA, triphosgene NH
p. (a___ j HN- NH2 4-1NH2 2) CH2Cl2, TEA, 0 0(0}-120H200H20H20H2NH2)2 intermediate 42(0.95 meq/g) \

070 o / 0-1¨
ro o d---\__o 0 /
o/
\ Y-0 HN NH HN HN---\0____\
...z.- 4 ¨/ o -i o o \
intermediate 39 0 , intermediate 43 (0.71 meq/g) o 0 0 o \--NHpH
1) NH2OTMS 0i 2) aq. AcOH
HN---"No\
HN-i NH , 0 0 e 0 HN-OH

Resin 7 (0.70 meq/g) rENII,OH

[00267] To 5 g (11 mmol) of aminomethyl resin (Sigma-Aldrich cat#
564095; 2.2 meq/g) in 25 mL CH2C12 in the 60 mL peptide reactor was added 1.53 mL of triethylamine.

The mixture was treated portionwise with 3.26 g (11 mmol) of triphosgene.
After 15 min the mixture was filtered and washed three times with CH2C12. The resin was suspended in 25 mL CH2C12 and treated with 4.6 mL FLA and 7.23 nth of 4,7,10-trioxa-1,13-tridecanediamine. After rocking for two days, the mixture was filtered and washed three times each with CH2C12, Me0H, H20, Me0H, Et20 and allowed to dry under a stream of N2 for 1h. The resin was dried overnight in vacuo at rt to afford 6.177 g of an off-white resin intermediate 42. IR (ATR) 3560 (br, weak), 2920, 2880, 1655. Elemental Analysis: C, 80.14; H, 8.18; N, 4.01; EA shows loading of 0.95 meq/g based on N analysis.
[00268] To 6.14 g of resin intermediate 42 (5.8 mmol) was added 30 mL of followed by 2.3 mL of diisopropylethylamine (DIEA). The mixture was treated with 5.3 g of isocyanate intermediate 39 and allowed to agitate by rocking for 2 days at rt.
The resin was filtered and washed three times each with CH2C12, Me0H, H20, satr NaHCO3, H20, Me0H
and Et20. After filtration the resin was dried in vacuo overnight at rt to afford 5.5 g of a light tan resin intermediate 43: IR(ATR) 3322 (br), 3023, 2920, 2865, 1736 (C=0), 1685, 1655.
Elemental Analysis: C, 78.52; H, 7.84; N, 3.99; EA shows loading of 0.71 meq/g based on N
analysis.
[00269] A suspension of 5.5 g of triester resin intermediate 43 (3.8 mmol) in 15 mL
of Me0H was prepared in a 60 mL peptide reactor. The mixture was treated with 4.2 mL of NH2OTMS followed by a solution of 1.92 g of KOH in 20 mL of Me0H. The mixture was allowed to rock overnight at rt. The mixture was subsequently filtered and washed three times each with Me0H and H20. The resin was treated with 10% aq. acetic acid and allowed to rock for one hour. The resin was filtered, washed three times each with H20, Me0H, Et20 and dried in vacuo at rt to give 5.96 g of the trihydroxamic acid resin 7:
IR(ATR) 3200, 2915, 1670, 1650, 1552. Elemental Analysis: C, 76.24; H, 7.77;
N, 4.68; EA
shows loading of 0.67 meq/g based on N analysis.
[00270] Example 27 [00271] A urea linked unsymmetrical resin 8 [00272] Scheme 44 H2NO CO2Me L.A.)C12, CH2C12 OCN-0 CO2Me Q/NH2 \O--\
N---0O2Me aq NaHCO3 iPr2NEt, CH2C12 --0O2Me CO2Me intermediate 44 intermediate 45 CO2Me H H H H

?v CO2Me 11TMS-ONH2 r.1 CONHOH

CO2Me Me0H, KOH CONHOH
intermediate 46 OC 2Me resin 8 \--CONHOH

[00273] To a vigorously stirred solution of amine triester intermediate 44 (12.0 g, 30.5 mmol) in CH2C12 (100 mL) and saturated NaHCO3 (100 mL) was added triphosgene (3.08 g, 36.0 mmol) in small portions. Once the addition was complete, the mixture was stirred for an additional 15 min., then the layers were separated and the aqueous layer was extracted with CH2C12 (2x100 mL). The combined organic extracts were dried over Na2SO4, evaporated under reduced pressure, and the residue was purified by column chromatography (Si02, hexanes/Et0Ac gradient) to give the isocyanate intermediate 45 as a colorless oil (5.14 g, 41%): IR (neat) 2957, 2874, 2244, 1733 cm-1; 1H N1VIR (300 MHz, CDC13) 8 3.70 (t, J= 6.3 Hz, 4H), 3.64 (s, 9H), 3.63 (t, J= 6.3 Hz, 2H), 3.50 (t, J= 6.3 Hz, 2H), 3.39(s, 4H), 2.54 (t, J= 6.2 Hz, 4H), 2.52 (t, J = 6.2 Hz, 2H), 1.72 (t, J- 6.4 Hz, 2H); 13C NMR
(300 MHz, CDC13) 6 172.1, 171.9, 126.6, 73.4, 66.8, 66.7, 66.2, 62.7, 51.8, 51.7, 34.9, 34.8, 33.8;

HRMS (FAB) calcd for CI8H301\1010 [M + Hr: 420.1870. Found 420.1885.

[00274] To a suspension of aminomethyl resin (Aldrich 564095, macroporous 30-60 mesh) (1.84 g, 2.2 meq/g, 4.05 mmol of -NH2), in CH2C12 (15 mL) was added diisopropylethyl amine (2.12 ml, 12.2 mmol) followed by the isocyanate intermediate 45 (5.10 g, 12.2 mmol). The suspension was shaken using orbital shaker overnight at room temperature. The resin was filtered and washed 3 times each with CH2C12, Me0H, H20, saturated NaHCO3, H20, Me0H and Et20. It was dried under reduced pressure to obtain pale yellow resin intermediate 46 (2.86 g). Wt. added to the resin = 1.02 g; IR
(AIR): 3393, 3026, 2912, 2869, 1736 (CO), 1673 cm-1; Elemental analysis C = 75.50%, H =
7.59%, N =

3.35%, loading = 0.87 meq/g (based on %N); %C indicates 76% conversion of available NH2 groups of the resin.

[00275] A suspension of the urea triester resin intermediate 46 (2.68 g, 1.14 meq/g -maximum loading, 3.06 mmol) in Me0H (20 mL) was shaken for 15 minutes. A
solution of KOH (1.54 g, 27.59 mmol) in Me0H (5 mL) was added followed by NH2OTMS (3.37 mL, 27.6 mmol) and the mixture was shaken for 20 h using orbital shaker. The resin was filtered and washed 3 times each with Me0H, H20. The resin was then suspended in 10%
aqueous AcOH (20 mL) and for shaken for 30 minutes. The resin was filtered and washed 3 times with 10 % aqueous CH3CO2H, H20, Me0H and Et20 and dried under reduced pressure to give a light yellow colored resin 8 (2.7 g). Wt. gained by the resin = 0.02 g;
IR (A'TR): 3221, 3025, 2920, 1641 (C=0 hydroxamate, sharp), 1551 cm-i; Elemental analysis C =
71.48%, H

= 7.36%, N = 4.65%, loading = 0.58 meq/g (based on %N). , [00276] Example 28 [00277] Scheme 45 ON

HO

orj OH acrylonitrile Me0H, HCI 0 H N-0 i KOH, H tO 0 46% 2 acetonitrile OH 1.,CN

C) 0 TRIS

CN .r0,, intermediate 1 intermediate 2 o OH
,-0 ),--Nil-1 Method A
/

0 1) NH2OTMS

o/
clCa HNtO 0 2) aq. AcOH

t Nal, NMP0 0 Method B LõJLN-OH

1) NH2OH, Me0H, Me0Na C) H

=,..,1y0.., 2) aq. AcOH H

intermediate 47 (0.51 meq/g) 8 OH

Resin 9 0 Method A (0.46 meq/g) Method B (0.49 meq/g) [00278] Tris[2-(cyanoethoxy)methyllmethylamine (intermediate 1) by an improved method. A stirred suspension of tris-(hydroxymethyl)aminomethane (127.0 g) in acetonitrile (500 mL) in a 2L round bottom flask equipped with an overhead stirrer was treated with KOH (5.0 g). Acrylonitrile (207 mL) was added to the stirred suspension over a few minutes. After lh, an exotherm was observed to 36 C. After 3h, the mixture was a homogeneous, slightly orange solution and showed complete conversion based on analysis. After a total of 4 h, the mixture was concentrated under reduced pressure to afford intermediate 1 (237.8 g, 89%) as a light tan oil. IR(neat) 3504, 3288, 2857, 2250(CN); r1D23 = 1.4687 1H NMR (CDC13, 300 MHz) M.45 (brs, 2H), 2.53 (t, 6.0 Hz, 6H), 3.34 (s, 6H), 3.59 (t, 6.0 Hz, 6H); 13C NMR (CDC13, 75 MHz) 618.8, 56.0, 65.7, 72.5, 188.1.
This compound has been previously prepared, however the yield appears to be improved with acetonitrile solvent.
[00279] A 10 mL oven dried peptide reactor was charged with Merrifield resin (2.0 g) (Marcroporous, 100-200 mesh, 150-75 urn, 1.2 mmol/g, SigmaAldrich 564087, Lot #05629MC, washed and dried) and anhydrous NaI (400 mg). The mixture was suspended in a solution of intermediate 2 (2.93 g) in of anhydrous NMP (12 mL) and agitated for 7 days at rt. The mixture was sampled at regular intervals and the reaction progress monitored by IR
(ATR). After a total of 7 days the reaction was complete. The product was collected by filtration and washed sequentially three times each with DMA, H20, Me0H and Et20 to give a light, tan resin intermediate 47: IR(ATR) 3026, 2921, 1739 (C=0), 1602;
Elemental Analysis: C, 83.03; H, 7.59; N, 0.72; EA shows loading of 0.51 meq/g (53%
conversion of benzylchloride groups). The resin was kept in the peptide reactor and used directly in the next step.
[00280] Preparation of Trishydroxamic Acid Resin 9 using Method A. A
suspension of the ester resin intermediate 47 in anhydrous Me0H (5 mL) was treated with NH2OTMS (2.42 mL), followed by a solution of KOH (1.11 g) in anhydrous Me0H (5 mL).
The mixture was agitated overnight, then the resin was washed three times each with Me0H
and H20, treated with 10% aq. AcOH and agitated for lh. The mixture was filtered and treated a 2nd time with 10% AcOH for 30 min. The resin was filtered, washed three times each with Me0H, H20, Et20 and dried under reduced pressure to give of the trihydroxamic acid resin 9 (1.84 g): IR(ATR) 3210, 3026, 2921, 1652, 1602, 1493; Elemental Analysis: C, 80.33; H, 7.46; N, 2.55; EA shows loading of 0.46 meq/g.

, , , ., [00281] Preparation of Trishydroxamic Acid Resin 9 using Method B. A
mixture of 900 mg of hydroxylamine hydrochloride in 30 mL of 0.5_M sodium methoxide in methanol was stirred at rt for 15 mm. The mixture was filtered to remove NaC1 salts and added directly to 2.0 g of triester resin intermediate 47 (1.24 mmol) in a flask equipped with an overhead stirrer, heating mantle and kept under nitrogen. The stirred mixture was heated to 45 C and allowed to stir for 3 days. After filtration the resin was washed three times each with methanol, 10% aq. acetic acid, H20, methanol, and ethyl ether. The resin was dried in vacuo overnight to afford an amber resin 9: IR(ATR) 3210, 2921, 1652, 1602.
Elemental Analysis: C, 78.95; H, 7.43; N, 2.74; EA shows loading of 0.49 meq/g based on N
analysis.
[00282] Example 29 [00283] Resin linked tetra hydroxamic acid [00284] Scheme 46 0 0 THP-ONH2 THPO.,N

OMe Et0H, Py I OMe intermediate 35 intermediate 48 NaBH3CN, Me0H THPO,N 0 OMe DCC, DMAP, CH2Cl2 HCI intermediate 49 CbzHN/\--0-----CO2H
intermediate 34 OTHP
0 --CON CO2Me 1) Me0H, Pd.C, H2 CbZHN000NCOMe 2) COCl2, CH2Cl2, aq NaHCO3 OTHP
intermediate 50 OTHP CO2Me (ar---NH2 OC~OCOCO2Me CH2Cl2, 1Pr2NEt OTHP
intermediate 51 OTHP
0 0 CO2Me 1) TMS-ONH2, Me0H, KOH
NNOCONH OTHP CO2Me 2) Me0H, Ts0H
intermediate 52 OH

ar N OH CONHOH siH
resin 10 1002851 To a solution of aldehyde intermediate 35 (Kai, K.;
Takeuchi, J.; Kataoka, T.;

Yokoyama, M.; Watanabe, N. Tetrahedron 2008, 64, 6760 ) (2.89 g, 15.5 mmol) and 0-(tetrahydro-2H- pyran-2-yl)hydroxylamine (2.0 g, 17.1 mmol) in Et0H (62 mL) at room temperature was added pyridine (1.88 mL, 23.3 mmol) drop wise. The resulting solution was heated at reflux for 4 h. Solvent was evaporated under reduced pressure and the residue was dissolved in CH2C12 (100 mL) and washed with water (2x100 mL). The aqueous layer was re-extracted with CH2C12 (2x100 mL). The combined organic layers were dried over Na2SO4 and evaporated reduced pressure. The crude product was purified by column chromatography (SiO2hexanes) to give the oxime intermediate 48 as a colorless liquid (3.70 g, 84%) and as a mixture of geometric isomers: IR (neat) 2933, 2856, 1736 cm-1; 'H NMR (CDC13) 8 (ppm) 7.42 (t, J= 6.3 Hz, 0.4H), 7.71 (t, J= 5.5 Hz, 0.611), 5.19 (m, 1.2H), 5.15 (m, 0.8H), 3.89-3.84 (m, 1H), 3.62 (s, 3H), 3.59-3.53 (m, 1H), 2.35 (m, 1.5H), 2.26 (m, 2H), 2.18 (m, 0.5H), 1.90-1.40 (m, 10H), 1.29-1.26 (m, 6H); 13C NMR (CDC13) 8 (ppm) 174.2, 153.5, 152.9, 100.6, 100.5, 63.2, 63.1, 51.4, 34.0, 29.5, 29.1, 29.0, 28.9 (x 3), 26.6, 26.1, 25.8, 25.2(x 2), 24.9, 20.1, 20.0; HRMS (FAB) C15H28N04 [M+Hr calcd 286.20184, found 286.20110 [00286] To a solution of intermediate 48 (3.60 g, 12.31 mmol) and NaCNBH3 (0.95 g, 15.1 mmol) in Me0H (100 mL) was added 2N HC1 in Me0H drop wise until the solution pH
was adjusted to 4 (pH was never allowed to go down from 4). The mixture was stirred for 3 h at rt. The solvent was evaporated under reduced pressure to give solid residue which was dissolved in water (100 mL) and then 6 N KOH solution was added drop wise to adjust the solution pH to >9. The aqueous mixture was extracted with CH2C12 (3x100 mL).
The organic layers were combined and washed with brine, dried over Na2SO4 and evaporated under reduced pressure. The crude product was purified by column chromatography (SiO2 hexanes/Et0Ac gradient) to give intermediate 49 as a colorless liquid (2.80 g, 78%): IR
(ATR, neat) 2931, 2854, 1736 cm-1; 111 NMR (CDC13) 8 (ppm) 5.53 (s, 1H), 4.73-4.71 (m, 1H), 3.89-3.82 (m, 1H), 3.59 (s, 3H), 3.53-3.46 (m, 111), 2.96-2.82 (m, 2H), 2.23 (t, J= 7.4 Hz, 211), 1.75-1.39 (m, 10H), 1.25 (m, 8H); 13C NMR (CDC13) 5 (ppm) 174.2, 101.4, 63.1, 52.2, 51.4, 34.0, 29.3, 29.2, 29.1, 29.0, 27.2, 27.1, 25.3, 24.9, 20.2; HRMS
(FAB) C15H30N04 [M+Hr calcd 288.21747, found 288.21780 [00287] To a stirred solution of intermediate 49 (2.30 g, 8.00 mmol) and DMAP
(1.33 g, 10.9 mmol) in CH2C12 (70 mL) and pyridine (0.88 mL, 10.9 mmol) was added intermediate 34 (1.39 g, 3.63 mmol) in CH2C12 and the mixture was cooled to 0 C. DCC
(1.65 g, 8.00 mmol) was added and the mixture was stirred for 1 h at 0 C. The mixture was then allowed to warm to room temperature and was stirred for additional 15 h.
The solvent =

was concentrated under reduced pressure, filtered and washed with CH2C12. The filtrate was evaporated under reduced pressure and the residue was purified by column chromatography (Si02hexanes/Et0Ac gradient) to give intermediate 50 as a colorless oil (2.50 g, 76%): IR
(neat) 3329, 2930, 2856, 1732, 1655 cm-1; 1H NMR (CDC13) 8 (ppm) 7.33-7.24 (m, 511), 5.51 (s, 1H), 4.99 (s, 2H), 4.87 (s, 2H), 3.92-3.87 (m, 2H), 3.78-3.68 (m, 6H), 3.61 (s, 6H), 3.58-3.37 (m, 8H), 2.65-2.60 (m, 4H), 2.25 (t, J= 7.5 Hz, 4H), 1.76-1.54 (m, 20H), 1.29 (s, 3H), 1.24 (br s, 1611); 13C NMR (CDC13) 8 (ppm) 174.1, 172.7, 155.1, 136.7, 136.6, 128.3 (x 2), 127.8, 127.7, 104.3, 77.4, 73.2, 73.1, 67.9, 67.0, 65.8, 63.4, 55.7, 55.6, 51.3, 50.1, 48.6, 48.1, 35.5, 33.9, 32.9, 32.5, 30.6, 29.0 (x 2), 28.9, 28.8 (x 2), 26.6, 26.5, 26.0, 25.6, 25.1, 19.8, 19.1; HRMS (FAB) C48H79N30i4Na [M+Na] calcd 944.5460, found 944.5488.
[00288] To a solution of intermediate 50 (2.50 g, 2.70 mmol) in Me0H (200 mL) under argon was added 10% Pd/C (0.275 g). The flask was evacuated then flushed with 112 (balloons) and resulting the mixture was stirred under 112 at room temperature until TLC
analysis (75% Et0Ac in hexanes) indicated that the starting material was consumed. The reaction flask was then purged with argon and filtered. The filtrate was evaporated under reduced pressure to give the amine as thick oil (2.1 g, 100%) which was used directly in the next step. To a stirred biphasic mixture of the amine (2.1 g, 2.7 mmol) in CH2C12 (30 mL) and saturated NaHCO3 (30 mL) was added triphosgene (0.26 g, 0.89 mmol) in small portions. After 20 min after the addition of triphosgene the layers were separated and the aqueous layer was washed with CH2C12 (2x30 mL). The combined organic layers were dried over Na2SO4 and evaporated under reduced pressure to give the isocyanate intermediate 51 as a thick colorless oil product (1.7 g, 78 %) which was found to be pure enough to use in the next step without further purification. IR (neat) 2932, 2856, 2241, 1736, 1655 crn-1; 1H NMR
(CDC13) 8 (ppm) 4.89 (br s, 211), 3.96-3.89 (m, 2H), 3.81-3.72 (m, 6H), 3.62 (s, 6H), 3.60-3.53 (m, 4H), 3.41-3.32 (m, 4H), 2.68- 2.61 (m, 4H), 2.26 (t, J= 7.4 Hz, 4H), 1.78-1.57 (m, 20H), 1.26 ( br s, 16H), 1.18 (s, 3H); 13C NMR (CDC13) 8 (ppm) 174.3, 172.7, 126.8, 104.5, 77.4, 75.3, 75.2, 75.1, 68.2, 67.3, 63.6, 61.2, 61.1, 61.0, 55.0, 51.5, 50.2, 48.3, 35.7, 34.1, 33.1, 32.6, 30.8, 29.5, 29.2, 29.1, 26.9, 26.8, 26.2, 25.5, 25.4, 22.5, 19.8;
HRMS (FAB) C411-171N3013Na [M+Nar calcd 836.48846, found 836.48570.

[00289] To a suspension of the aminomethyl resin (1.1 g, 2.2 meq/g, 2.42 mmol of ¨
N112, Aldrich 564095, macroporous 30-60 mesh) in CH2C12 (17 mL) and diisopropyl ethyl amine (2.66 mL, 15.3 mmol) was added the isocyanate intermediate 51(5.0 g, 6.10 mmol), and the resulting mixture was shaken on orbital shaker for 24 h at room temperature. The resin was filtered and washed 3 times each with CH2CH2, Me0H, H20, saturated NaHCO3, H20, Me0H and Et20. It was then dried under reduced pressure to obtain light colored resin intermediate 52 (1.51 g). Wt. added to the resin = 0.41 g. IR (ATR): 3377, 3022, 2924, 2848, 1734 (C=0 ester), 1655 (C=0 hydroxamate cm-1. Elemental analysis C
81.23%, H
7.96%, N 3.67%. loading = 0.28 meq/g (based on %N). Change in %C indicates 43%

conversion of available NH2 groups of the resin.
[00290] To a suspension of the resin intermediate 52 (1.51 g, 0.30 meq/g, 0.45 mmol) in Me0H (10 mL) was added KOH (0.15 g, 2.7 mmol) and NH2OTMS (0.33 mL, 2.7 mmol) and the mixture was shaken for 20 h on an orbital shaker. The resin was isolated by filtration and washed 3 times with Me0H. The resin was re-suspended in a solution of Ts0H.H20 (0.52 g, 2.7 mmol) in Me0H (10 mL) and shaken for for 3 h. The resin was isolated by filtration and washed 3 times each with Me0H, H20, Me0H and Et20 and dried under reduced pressure to give the product as pale yellow colored resin 10 (1.73 g).
Wt. gained by the resin = 0.22 g. IR (ATR): 3382 (br), 3500-2500 (br), 3024, 2922, 2854, 1646 (C=0 hydroxamate) cm'. Elemental analysis C 75.74%, H 7.51%, N 3.68%. Loading =
0.19 meq/g (based on %N).
[00291] Example 30 [00292] Binding of Al to resin 9 [00293] The compounds and compositions of the present invention are useful in a method of removing a trivalent metal ion such as Al3+ from an aqueous solution. This is accomplished by treating the aqueous solution with the invention, which consists of a resin to which the chelating agent is attached by a covalent bond to form a chelating resin.

[00294] To demonstrate the ability of resin 9 to bind A13 , a 240 mg portion of the resin was suspended in three different aqueous solutions: (1) an aqueous solution buffered at pH
6.06 by 0.1 M MES (4-morpholineethanesulfonic acid); (2) a solution of gluconate that had been adjusted to pH 6.42 by the addition of tetramethylammonium hydroxide; and (3) a commercial sample of calcium(gluconate)2, which had a pH of 6.07. Each solution was stirred by a magnetic overhead stirrer to keep the resin suspended in solution with significant mechanical damage as might result from use of a stir bar.
, [00295] At periodic times, the stirring was stopped for 1-2 minutes to let the resin settle, and a 100 uL aliquot was removed from the sample solution. Five I, of concentrated, metal-free nitric acid was added immediately to each aliquot removed to stabilize the Al3+ in solution. Samples were collected and subsequently analyzed by inductively coupled plasma-mass spectrometry to determine the Al concentration. Figure 6 shows plots of the fraction of original Al concentration remaining in the extracted solutions as a function of the extraction time.
[00296] The pH of the sample solutions was measured before and immediately after the completion of the extraction experiment. The addition of the resin and the extraction produce essentially no change in the pH of any of the solutions tested.
[00297] In the extraction of Al3+ from the MES buffer, the only competition to binding to the chelating resin is the hydrolysis of Al3+ to a mixture of Al-hydroxide complexes.
Figure 6 shows that the concentration of Al3+ decreases to ¨0. In the case of both gluconate and calcium gluconate solutions, chelation of Al3+ by gluconate is competitive with the binding of Al3+ to the resin. Nevertheless, resin 9 removes approximately 90%
of the Al from both of these solutions.
[00298] Figure 7 shows the extraction of three solutions of commercial calcium(gluconate)2. The figure includes data from two duplicate extractions of the calcium(gluconate)2 by 240 mg portions of resin 9. The resin removes ¨90% of the total Al.
The rate of Al removal from the solution can be fit to a single-exponential function to give an apparent first-order rate constant for Al removal of 4.2 hr-1, which corresponds to a half-life for Al removal of only 10 minutes. The solid line in Figure 7 is the least squares fit of one of the two data sets to the single-exponential function.

1002991 For the purpose of comparison, another aliquot of the same commercial calcium(gluconate)2 solution was extracted with the commercial chelating resin Chelex .
This resin consists of polystyrene beads to which the chelating agent iminodiacetic acid has been linked by covalent bonds. This resin is Widely used in a variety of applications to remove metal ions from solution. Figure 7 shows that the Chelex resin is not able to remove any significant fraction of Al3+ from the calcium(gluconate)2 solution. This poor extraction reflects the avidity with which gluconate binds Al3+, and demonstrates a strong chelating agent is required to compete with gluconate to remove Al3+ from the solution.
[00300] The Al-binding constants for gluconate have been reported (R.J.
Motekaitis and A.E. Martell, Inorg. Chem. 1984, 23: 18-23). Given these binding constants for Al-gluconate, the percentage of Al remaining in a gluconate solution in equilibrium with resin-9 can be used to estimate the effective Al binding constant of the trihydroxamate chelating agent covalently bound to the resin. The removal of 90% of the Al corresponds to an Al3+-resin binding constant of log K = 20.6.
[00301] The immobilized chelating agents described in this document may be used in a number of difference devices. Figures 8a-8d disclose three different embodiments of cartridge 10 filled with any of the immobilized chelating agents described in this document.
Each cartridge 10 contains a sealed body 12 having a first Luer Lock fitting 14 at the inlet end and a second Luer Lock fitting 16 at the outlet end. The resin 18 including the immobilized chelating agent is held in the body 12. A membrane 20 covers the outlet 22 thereby preventing any particles that might be released from the resin 18 from exiting the cartridge 10. A mesh, sieve, frit or membrane 24 covers the inlet 26 and functions to maintain the resin 18 in position in the body 12 of the cartridge 10.
1003021 The body 12 and resin 18 contained therein may have a cylindrical shape. The Figure 8a and 8d embodiment has a relatively intermediate length and diameter.
The Figure 8b embodiment has a relatively long length and relatively narrow diameter. The Figure 8c embodiment has a relatively short length and a relatively large diameter.
[00303] Each of the embodiments has a different diameter and length which affects the flow characteristic of any fluid passing through the cartridge 10 and the resin 18 holding the immobilized chelating agent. The embodiment chosen for use will depend upon the application.
[00304] All of the embodiments 8a-8c provide a straight flow path from the inlet 26 to the outlet 22. While not illustrated, it should be appreciated that the cartridge 10 may be substantially any shape including non-linear, arcuate, even coiled. Further, while the illustrated cartridges 10 are all symmetrical it should be appreciated that nonsymmetrical shapes could be provided. In addition, while all the illustrated cartridges 10 are cylindrical in cross section, substantially any other shape of cross section may be provided including but not limited to frustoconical, helical, square, hexagon and T-shaped.
[00305] The cartridge 10 may be used with a continuous flow of solution passing through the cartridge or in a stop-flow mode where solution is introduced into the cartridge, stopped for a time to allow the chelating agent to act, and then expelled from the cartridge.
[00306] Figures 9a and 9b both show vessels 50 that hold a packet 52 containing resin beads 54 holding the immobilized chelating agent. The packet 52 is made from a semi-permeable membrane similar, for example, to the paper used in the production of tea bags.
[00307] In the figure 9a embodiment, the vessel 50 is opened and a solution to be treated and a packet 52 containing resin beads 54 are both introduced into the vessel. The vessel 50 is closed and the solution is agitated by magnetic stirrer (not shown) or other means to facilitate interaction and contact between the solution which freely passes through the packet 52 and the chelating agent that is immobilized on the resin beads 54 sealed in the packet.
[00308] In the embodiment illustrated in Figure 9b, the vessel 50 includes an inlet 56 and an outlet 58. The packet 52 is placed in the open vessel 50 and the lid 60 of the vessel 50 is then closed. A pump (not shown) is then actuated to pump solution through the vessel by means of the inlet 56 and outlet 58. Solution flow may be continuous or stop-flow mode in the manner described above. Agitation of solution within the vessel 50 may also be provided. Depending on the application, the vessel 50 may be a sterile vessel.
[00309] In an alternative approach, the resin beads 54 holding the chelating agent may be placed directly in the vessel 50 without a packet (see Figure 9c).
Membranes may be provided over the inlet and outlet to insure the resin beads and any particles they might release are maintained in the vessel 50. Lure Lock fittings may also be provided at the inlet and outlet if desired.
[00310] Figure 10 is a plot showing the mean percentage of aluminum removed from a 10% calcium gluconate injection USP as a function of flow rate using the device illustrated in Figure 8a and the resin and immobilized chelating agent of resin 9. The device had an internal cavity diameter of 6 mm and a length of 20 mm. It held approximately 245 mg of resin and immobilized chelating agent at a loading rate of 0.55 mmoles/gram resin.
[00311] As noted above, the chelating agents described in this document are particularly useful in removing aluminum from solutions including calcium gluconate. It should be appreciated, however, that they are also very useful in a multitude of other applications involving the separation of trivalent metal ions from a solution.
Such applications include but are not limited to: (1) removal of Al from dialysis fluids and biological products such as albumin and other pharmaceutical solutions in which it is a contaminant, such as phosphates; (2) treatment of metal overload of any trivalent hard or intermediate acid, according to the HSAB theory (for example iron from blood transfusions in beta-thalassemia); (3) as a complexing agent for MR1 contrast enhancement (for example with gadolinium); (4) as a treatment for poisoning with tri- and tetravalent metal ions, including radioactive elements that workers may be inadvertently exposed to and are potential chemical warfare agents (dirty bomb components) (for example americium, cerium, and plutonium); (5) extraction of metal ions from solution either to isolate the pure metal or as a pre-concentration step prior to some sort of elemental analysis ; (6) environmental remediation by complexing and removing toxic metals from contaminated water and/or soils;
and (7) use as a complexing agent for radionuclides of metals such as Ga or In for use as diagnostic imaging agents or as therapeutic radiopharmaceuticals.
[00312] The invention has been described herein with reference to certain preferred embodiments. However, as obvious variations will become apparent to those skilled in the art, the invention is not to be considered as limited thereto.

Claims (14)

1. A compound of the formula:
wherein R1 = hydrogen, sulfonamide, urea, carboxamide or benzyl, R2 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl and R3 =

wherein x, y, and z vary independently from 1 to 4, X = CH2 and O, and R4 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

wherein x, y, and z vary independently from 1 to 4, and R4 = hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

wherein x and y vary independently from 1 to 4, X = CH2 and O, and R4 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

wherein x and y vary independently from 1 to 4, X = CH2 and O, and R4 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

wherein x varies from 1-4, y varies from 1-2, and z varies independently from

2 to 8, hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

R4 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent, and R5 =

wherein x varies from 1-4, y varies from 1-2, and z varies independently from 2 to 8, R5 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent, and R6 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl, or Ph or aryl.

2.
The compound of claim 1 wherein R3 =
wherein x=1, y=1, z=1 and R4=H.

3. A compound of the formula:

wherein R1 = hydrogen, sulfonamide, urea, carboxamide or benzyl, R2 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl and R7 =

hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl and Ph or aryl.
wherein x, y, and z vary independently from 1 to 4, X = CH2 and O, and R8 =

hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl and Ph or aryl.

wherein x, y, and z vary independently from 1 to 4, X = CH2 and O, and R8 =

4. A compound of the formula:

where R1 =

R2 = hydrogen, methyl, ethyl; n-propyl or isopropyl and R3 =

hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.
wherein x, y, and z vary independently from 1 to 4, X = CH2 and O, and R4 =

wherein x, y, and z vary independently from 1 to 4, and R4 = hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

wherein x and y vary independently from 1 to 4, X = CH2 and O, and R4 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

wherein x and y vary independently from 1 to 4, X = CH2 and O, and R4 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.
e) 101 wherein x varies from 1-4, y varies from 1-2, and z varies independently from 2 to 8, R4 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent, and R5 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

wherein x varies from 1-4, y varies from 1-2, and z varies independently from 2 to 8, R5 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent, and R6 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl, or Ph or aryl.

5. A compound of the formula:

wherein R6 =

<MG>

R2 = hydrogen, methyl, ethyl; n-propyl or isopropyl and R3 =

wherein x, y, and z vary independently from 2 to 4 and R4 = hydrogen or C1 -straight or branched alkyl;

wherein x, y, and z vary independently from 2 to 4, and R4 = hydrogen or C1 -straight or branched alkyl;

wherein R4 = hydrogen or C1 - C10 straight or branched alkyl;

wherein R4 = hydrogen or C1 - C10 straight or branched alkyl;

wherein n = 2 or 3, R5 = hydrogen or methyl, and R4 = hydrogen or C1 - C10 straight or branched alkyl;

wherein R4 = hydrogen or C1 - C10 straight or branched alkyl;

wherein R4 = hydrogen or C1 - C10 straight or branched alkyl; and alkyl.
wherein le = hydrogen or methyl and R4 = hydrogen or C1 - C10 straight or branched

6. A method of removing trivalent and tetravalent metal ions from a solution, comprising contacting said solution containing said trivalent metal ions with a chelating agent selected from a group consisting of :

a compound of the formula:

wherein R1 = hydrogen, sulfonamide, urea, carboxamide or benzyl, R2 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl and R3 =

wherein x, y, and z vary independently from 1 to 4, X = CH2 and O, and R4 =

wherein x, y, and z vary independently from 1 to 4, and R4 = hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

wherein x and y vary independently from 1 to 4, X = CH2 and O, and R4 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

wherein x and y vary independently from 1 to 4, X = CH2 and O, and R4 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

wherein x varies from 1-4, y varies from 1-2, and z varies independently from 2 to 8, hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.
R4 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent, and R5 =

wherein x varies from 1-4, y varies from 1-2, and z varies independently from 2 to 8, R5 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent, and R6 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl, or Ph or aryl.

7. A method of removing trivalent and tetravalent metal ions from a solution, comprising contacting said solution containing said trivalent and tetravalent metal ions with a chelating agent having the formula:
where R1 =hydrogen, sulfonamide, urea, carboxamide or benzyl, R2=hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl and R3=

wherein x=1, y=1, z=1 and R4=H.

8. A method of removing trivalent and tetravalent metal ions from a solution, comprising contacting said solution containing said trivalent and tetravalent metal ions with a chelating agent selected from a group consisting of:

R3 =

wherein R1 =

R2 = hydrogen, methyl, ethyl; n-propyl or isopropyl and a.) wherein x, y, and z vary independently from 2 to 4 and R4 = hydrogen or C1 -straight or branched alkyl;

wherein x, y, and z vary independently from 2 to 4, and R4 = hydrogen or C1 -straight or branched alkyl;

wherein R4 = hydrogen or C1 - C10 straight or branched alkyl;

wherein R4 = hydrogen or C1 - C10 straight or branched alkyl;

wherein n = 2 or 3, R5 = hydrogen or methyl, and R4 = hydrogen or C1 - C10 straight or branched alkyl;

wherein R4 = hydrogen or C1 - C10 straight or branched alkyl;

wherein R4 = hydrogen or C1 - C10 straight or branched alkyl; and branched alkyl.wherein R5 = hydrogen or methyl and R4 = hydrogen or C1 - C10 straight or

9. A device for separating trivalent and tetravalent metal ions from a solution, comprising a body carrying a resin holding an immobilized chelating agent selected from a group of chelating agents having the formula:

wherein R1 = hydrogen, sulfonamide, urea, carboxamide or benzyl, R2 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl and R3 =

wherein x, y, and z vary independently from 1 to 4, X = CH2 and O, and R4 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

wherein x, y, and z vary independently from 1 to 4, and R4 = hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

wherein x and y vary independently from 1 to 4, X = CH2 and O, and R4 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

methyl, ethyl, n-propyl, isopropyl or alkyl.
wherein x and y vary independently from 1 to 4, X = CH2 and O, and R4 =
hydrogen, wherein x varies from 1-4, y varies from 1-2, and z varies independently from 2 to 8, hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl.

R4 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent, and R5 =

wherein x varies from 1-4, y varies from 1-2, and z varies independently from 2 to 8, R5 = hydrogen, methyl, ethyl, n-propyl, isopropyl or similar alkyl substituent, and R6 =
hydrogen, methyl, ethyl, n-propyl, isopropyl or alkyl, or Ph or aryl.

10. The device of claim 9, wherein said body comprises a cartridge.

11. The device of claim 10, wherein said cartridge includes an inlet and an outlet.

12. The device of claim 9, wherein said body comprises a vessel.

13. The device of claim 12, wherein said vessel includes an inlet and an outlet.

14. A device for separating trivalent and tetravalent metal ions from a solution, comprising a body carrying a resin holding an immobilized chelating agent selected from a group of chelating agents having the formula:

R3 =

wherein R1 = hydrogen or tosylate, R2 = hydrogen, methyl, ethyl, n-propyl, or isopropyl and wherein x, y, and z vary independently from 2 to 4, and R4= hydrogen or C1-C10 straight or branched alkyl;

wherein x, y, and z vary independently from 2 to 4 and R4 = hydrogen or C1-Cl0 straight or branched alkyl;

wherein R4 = hydrogen or C1-C10 straight or branched alkyl;

wherein R4 = hydrogen or C1-C10 straight or branched alkyl;

wherein n = 2 or 3 and R5 = hydrogen or methyl, and R4 = hydrogen or C1-C10 straight or branched alkyl;

wherein R4 = hydrogen or C1-C10 straight or branched alkyl;

wherein R4 = hydrogen or C1-C10 straight or branched alkyl; and wherein R5 = hydrogen or methyl and R4 = hydrogen or C1-C10 straight or branched alkyl.