Inorg. Chem. 1997, 36, 3493-3498
Ion-Pairing Interactions between Co(en)33+ and the TmDOTP5- †
23Na
3493
NMR Frequency Shift Reagent
Jimin Ren,‡ Charles S. Springer, Jr.,§ and A. Dean Sherry*,‡,| Department of Chemistry, The University of Texas at Dallas, P.O. Box 830688, Richardson, Texas 75083-0688, The Mary Nell and Ralph B. Rogers Magnetic Resonance Center, Department of Radiology, University of Texas Southwestern Medical Center, 5801 Forest Park Road, Dallas, Texas 75235-9085, Chemistry Department, Brookhaven National Laboratory, Upton, New York 11973-5000, and Department of Chemistry, State Universities of New York, Stony Brook, New York 11794-3400 ReceiVed February 6, 1997X
Three new formulations of TmDOTP5- (DOTP8- ) 1,4,7,10-tetraazacyclododecane-1,4,7,11-tetrakis(methylenephosphonate)) have been prepared in an effort to develop a low-osmolality form of the 23Na frequency shift reagent (SR). Equally concentrated (0.32 M) solutions of (MegH)4HTmDOTP (Meg ) N-methylglucamine or meglumine), Na4HTmDOTP, and [Co(en)3]4/3HTmDOTP have solution osmolalities of 1245, 1040, and 707 mOsm/ kg, respectively, comparable to the ionic and non-ionic gadolinium-based MRI contrast agent preparations in clinical use. An analysis of 23Na and 59Co frequency shifts induced by TmDOTP5- indicated that Co(en)33+ can form both 1:1 and 2:1 adducts with TmDOTP5- with (log) binding constants of 3.1 ( 0.4 and 2.5 ( 0.4, respectively. These values were comparable with those obtained by analysis of the 1H frequency shifts observed for Co(en)33+ upon binding to HoDOTP5-. The 1H shifts of Co(en)33+ signals induced by YbDOTP5- at pH 7.4 were fitted best by a 1:1 binding model with a conditional binding constant of 3.1 ( 0.2. The 59Co and 1H limiting frequency shifts of Co(en)33+ could be fitted with a dipolar shift model in which the Co atom of the Co(en)33+ cation is located 5.0 ( 0.3 Å from the Ln atom of the LnDOTP5- chelate, and with an angle of 40 ( 0.2° between the Co-Ln vector and the 4-fold symmetry axis of the LnDOTP5- complex. Ion pairing of Co(en)33+ and TmDOTP5- was significant enough in both saline and human blood plasma to reduce the effectiveness of the 23Na frequency SR. Comparisons between all formulations suggested that Na HTmDOTP represents the best 4 compromise of lower osmolality with minimal reduction of SR shift potency.
Introduction Na+
The maintenance of a cytolemmal gradient is a primary function of the ubiquitous enzyme Na+-K+-ATPase. As pump dysfunction has been implicated in numerous diseases including hypertension and sepsis,1 there is considerable interest in developing methods to monitor intra- and extracellular Na+ concentrations in intact tissue.2-6 In comparison to other techniques for quantitating [Na+] in intact tissues, 23Na NMR in combination with a paramagnetic shift reagent (SR) is one of the more promising methods because it can be minimally invasive and it offers measurement of intracellular sodium with optimal temporal resolution. Paramagnetic SRs used for this purpose are typically highly charged, anionic species that ion* Author to whom correspondence should be sent at either address. Telephone: 972-883-2907 or 214-648-5877. Fax: 972-883-2925 or 214648-5881. E-mail:
[email protected]. † A portion of this work was presented at the 221st National Meeting of the American Chemical Society, New Orleans, March 1996. ‡ The University of Texas at Dallas. § Brookhaven National Laboratory and State Universities of New York. || University of Texas Southwestern Medical Center. X Abstract published in AdVance ACS Abstracts, July 1, 1997. (1) Garcia, N. M.; Horton, J. W. Shock 1994, 1, 354-358. (2) Springer, C. S., Jr. Annu. ReV. Biophys. Biophys. Chem. 1987, 16, 375-399. (3) Sherry, A. D.; Geraldes, C. F. G. C. In Lanthanide Probes in Life, Chemical and Earth Sciences: Theory and Practice; Bu¨nzli, J.-C. G., Choppin, G. R., Eds.; Elsevier: Amsterdam, 1989; pp 93-126. (4) Balschi, J. A.; Bittl, J. A.; Springer, C. S., Jr.; Ingwall, J. S. NMR Biomed. 1990, 3, 47-58. (5) Hutchison, R. B.; Shapiro, J. I. Concepts Magn. Reson. 1991, 3, 215236. (6) Bansal, N.; Germann, M. J.; Seshan, V.; Shires, G. T., III; Malloy, C. R.; Sherry, A. D. Biochemistry 1993, 32, 5638-5643.
S0020-1669(97)00128-6 CCC: $14.00
pair with extracellular sodium, [Na+e], in a labile equilibrium, and thereby induce a frequency shift that separates its signal from the intracellular Na+ resonance.7-9 Any candidate SR for in ViVo use must produce a large hyperfine frequency shift at relatively low dose (preferably 1 (where Co(en)33+ is greater than 95% bound) using the reduced Solomon-Bloembergen equation:26 (27) Bleaney, B. J. Magn. Reson. 1972, 8, 91-100. (28) Ren, J.; Sherry A. D. J. Magn. Reson. Ser. B 1996, 111, 178-182.
3498 Inorganic Chemistry, Vol. 36, No. 16, 1997
T1-1 ) (4/3)(µ0/4π)2µeff2γ2β2τs/〈r6〉
Ren et al.
(9)
where µ0/4π is the magnetic susceptibility parameter for a vacuum, µeff is the effective magnetic moment of the Ln3+ ion, γ is the magnetogyric ratio of 1H, β is the Bohr magneton, and τs is the electron-spin correlation time of LnDOTP5-. Using τs values of 0.54 and 0.28 ps determined previously28 for HoDOTP5- and YbDOTP5- and measured 1H T1’s of 3.0 ( 0.3 and 12.2 ( 0.7 ms for Co(en)33+, respectively ([HoDOTP5-]: [Co(en)33+] ) 1.6 and [YbDOTP5-]:[Co(en)33+] ) 1.4), an average Ln-H distance of 5.0 ( 0.3 Å can be estimated for these two ion-paired complexes. Substituting this 〈r〉 value into eq 8 gives an average angle (〈θ〉) of 40 ( 2° for the Co(en)33+ protons in the ion-paired complex. The validity of these 〈r〉 and 〈θ〉 values can be further checked by the 59Co frequency shift induced by TmDOTP5-. Using a DLnA20〈r2〉Tm value of 18 970 Å3 ppm for TmDOTP5-,28 together with a Tm-Co distance of 5.0 Å and a 〈θ〉 value of 40°, the calculated 59Co limiting shift was 131 ppm, in good agreement with the experimental result (135 ppm). The 〈r〉 and 〈θ〉 values found for Co(en)33+ are similar to those previously estimated for Cs+ (ionic radius: 1.67 Å) bound to TmDOTP5- where the binding occurs at the outer oxygen axial oxygens of the phosphonate groups.21 The success of the assumption that the cobalt and the hydrogen nuclei of the cation are at the same averaged location may reflect rapid inter-ion pair exchange and/or rapid intra-ion pair rotation of the countercation. Summary Three new, excess-salt-free formulations of the 23Na shift reagent TmDOTP5- have been prepared and characterized. The
osmolalities of these depended upon the identity and charge of the countercation, with the osmolalities decreasing in the order MegH+ > Na+ > Co(en)33+. The tripositive cation Co(en)33+ forms an ion-pair complex(log K ) 2.5-3.1) with TmDOTP5at pH 7.4, which is somewhat stronger than that with the smaller monovalent ion, Na+ (log K ) 1.6-2.5).13 Although the sugar cation meglumineH+ reportedly binds to GdDOTP5- with a higher affinity (log K ) 2.96)29 than Na+, our osmolality measurements indicate that MegH+ more fully dissociates from TmDOTP5- than does Na+ at equivalent concentrations. Measured 59Co and 1H frequency shifts indicate that Co(en)33+ forms an ion pair with TmDOTP5- in a geometrical position similar to that of Na+ on the charged surface of the SR, but at a considerably longer distance from the paramagnetic center. Finally, the SR formulation Na4HTmDOTP appears to represent the best compromise of lower osmolality with minimal reduction of SR potency. Acknowledgment. Support of this research by grants from the Robert A. Welch Foundation (AT-584) and the NIH Biotechnology Research Program (RR02584) is gratefully acknowledged. We also thank Dr. Z. Kovacs for synthesis of H8DOTP. C.S.S. acknowledges an NIH grant (R01-GM32125) and Contract DE-AC02-76CH00016 with the DOE and its office of Health and Environmental Research to BNL, as well as early discussions with Mr. Christian Labadie and Professor Albert Haim. IC970128Q (29) Aime, S.; Botta, M.; Terreno, E.; Anelli, P. L.; Uggeri, F. Magn. Reson. Med. 1993, 30, 583-591.
