This is the first time that P-gp activity at the human BBB has been measured. The modest inhibition of human BBB P-gp by cyclosporine has implications for P-gp-based drug interactions at the human BBB. Our method for imaging P-gp activity can be used to identify multidrug-resistant tumors or to determine the contribution of P-gp polymorphism, inhibition, or induction to interindividual variability in drug response.

To examine inhibitor binding to an iron site resembling that of the metalloenzyme nitrile hydratase (NHase), a coordinatively unsaturated, five-coordinate FeIII thiolate complex was synthesized, and its reactivity examined. Ferricinium hexafluorophosphate induced oxidation of gem-dimethyl-protected [FeIIS2 Me2N3(Pr,Pr)] affords the chiral, five-coordinate complex [FeIIIS2 Me2N3(Pr,Pr)]+ (2) in reasonable yields. The magnetic properties and EPR of 2 are consistent with an S = 1/2 ground state. This unusual spin state, in conjunction with the low coordination number, of 2 result in unusually short Fe−S bonds (2.15(2) Å). Ligand constraints distort the S−Fe−N angles in 2 and create an open (132.3(1)°) reactive site. Azide binds to this site to afford a model for the azide-inhibited form of NHase [FeIIIS2 Me2N3(Pr,Pr)(N3)] (3). In MeOH azide binds reversibly, whereas in MeCN it binds irreversibly. This demonstrates that the secondary coordination sphere (i.e., the solvent, or possibly a protein binding pocket) can have a dramatic influence on the substrate binding properties of a metal complex. A variable-temperature equilibrium study in MeOH afforded thermodynamic parameters (ΔH = −5.2 ± 0.9 kcal/mol and ΔS = −12.4 ± 0.4 eu) for the binding of this inhibitor. The electronic spectrum of 3 displays an intense band at 708 (1600) nm similar to that (710 (∼1200) nm) of the pH = 7.3 form of NHase, and other six-coordinate cis-dithiolate ligated FeIII complexes synthesized by our group. EPR parameters for 3 (g = 2.23, 2.16, 1.99) are nearly identical to those of the azide-inhibited form of NHase (g = 2.23, 2.14, 1.99), suggesting that (1) the iron site of our model closely resembles that of the enzyme, and (2) azide binds directly to the metal ion in NHase. Reactivity is oxidation-state dependent, and the reduced analogue of 2, [FeIIS2N3(Pr,Pr)] (4), reversibly binds CO, but not azide, whereas oxidized 2 binds azide, but not CO.

spectrometer. The observing frequencies were 250.161 (lH), 47.704 V7Se), and 78.917 MHz (125Te) on the WM-250 spectrometer and 500.138 ( ), 95.383 (77Se), 130.885 (123Te), and 157.792 MHz (125Te) on the AM-500 spectrometer. Free induction decays were typically accumulated in 16 K or 32K memories. Spectral width settings of 25-100 kHz were employed, yielding data point resolutions of 3.0-6.1 Hz and acquisition times of 0.328-0.655 s, respectively. No relaxation delays were applied. The number of free induction decays accumulated depended upon the concentration and sensitivity of the nucleus under consideration with 10000-300000 scans being typical for these samples. Pulse widths for bulk magnetization tip angles corresponding to 90® were 2 ( ), 20 (77Se), and 25 µ$ (l25Te) on the WM-250 spectrometer and 2 ( ), 12 (77Se), 26 (123Te), and 10-18 µ$ (l25Te) on the AM-500 spectrometer. Line-broadening parameters used in the exponential multiplication of the free-induction decays were 5-20 Hz for the heavier isotopes; however, values on the order of 100-350 Hz were used for the broad lines. In order to enhance the resolution of some satellite peaks, the free induction decay was transformed with the use of a Gaussian fit rather than the conventional Lorentzian fit. In these instances, Gaussian broadenings of between 0.1 and 0.5 Hz and a line broadening greater than or equal to the negative of the data point resolution were applied.The respective nuclei were referenced to neat samples of (CH3)2Se(77Se), (CH3)2Te (,23Te and 125Te), and (CH3)4Si ( ) at 24 ®C. The chemical shift convention used is that a positive (negative) sign signifies a chemical shift to high (low) frequency of the reference compound.Acknowledgment. We wish to thank the Natural Sciences and Engineering Research Council of Canada for support of this work and McMaster University and the Ontario Ministry of Colleges and Universities for the award of scholarships to M.B.

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Nitrile hydratase is the first and only current example of a metalloenzyme containing a single non-heme Fe in a mixed N/Sligated coordination site.'-3 Its function is to hydrolyze nitriles to the corresponding amides in organisms which can live on R-CN as their sole C and N source. On the basis of EPR? MCD,5 resonance Raman, EXAFS,* and ENDOR' studies, the active site of nitrile hydratase is proposed to contain a six-coordinate, low-spin (S = ' / 2 ) Fe3+ ion ligated by two ciscysteinates, three N's, and a water: N The spectroscopic properties of nitrile hydratase are pHdependent, and two distinct forms (pH -7 and pH = 9) have been ~haracterized.~.' The high-pH form6 appears to be identical to the "substrate-bound'' form on the basis of its spectroscopic properties.4 Two impediments to the synthetic modeling of this enzyme have been the ease with which ferric ions oxidize thiolates to disulfides and the propensity of metal thiolates to o l i g~m e r i z e .~.~ The small number of reported monomeric Fe-(111) thiolate complexes attests to this.I0-l3 Mixed N-/SR--(1) Jin, H.; Tumer, I. M., Jr.; Nelson, M. J.; Gurbiel, R. J.; Doan, P. E.; Hoffman, B. M. J . Am. Chem. Soc. 1993, 115, 5290-1. (2) Nelson, M. J.; Jin, H.; Turner. I. M., Jr.; Grove, G.; Scarrow, R. C.; Brennan. B. A.: Que, L., Jr. J. Am. Chem. Soc. 1991, 113, 7072-3. (3) Nagasawa. T.; Ryuno, K.; Yamada, H. Biochem. Biophys. Res. Commun. 1986, 139, 1305-12. (4) Sugiura, Y.; Kuwahara, J.; Nagasawa, T.; Yamada, H. J. Am. Chem. (5) Johnson, M. K.: et al. Manuscript in preparation. (6) Jin. H.; Brennan, B. A,; Nelson, M. J.; et. al. Manuscript in preparation. (7) Honda, J.; Kandori, H.; Okada, T.; Nagamune, T.; Shichida. Y.; Sasabe, H.; Endo, I. ligated Fe(II1) systems are even rarer.I4-l6 Low-spin complexes of this type are extremely rare; most Fe(II1) compounds are S = 5 / 2 h i g h -~p i n '~. '~ or, at best, exist in a S = 5 / 2 -' / 2 spin equi1ibri~m.I~ Previously reported model compounds approximate the coordination geometry and metric parameters of the nitrile hydratase active site but do not reproduce the spin state and electronic spectral proper tie^.'^ Herein we report the synthesis, structure, and properties of a stable low-spin Fe(II1) thiolate complex which is remarkably similar to nitrile hydratase in terms of its electronic and geometric structure.Thiolate-ligated [Fe*11(AMIT)2]Cl (1) was synthesizedz0 via the Schiff-base condensation of ethylenediamine with a-mercaptoacetone at an Fe3+ template. This approach has provided access to a variety of mixed aminekhiolate S,Nyligated transition-metal complexes by our g r o~p .~' -*~ Although we have not yet obtained X-ray-quality crystals of l, (13) Sellmann, D.; Geck, M.; Knoch, F.; Ritter, G.; Dengler, J. J. Am. Chem. (14) Beissel, T.; Buerger, K. S.; Voigt, G.; Wieghardt, K.; Butzlaff, C.: (15) Marini, P. J.; Murray, K. S.; West, B. 0. J. Chem. Soc., Dalron Trans. (16) Fallon. G. D.; Gatehouse, B. M. J. Chem. Soc., Dalton Trans. 1975, (17) Greenwood, N. N.; Eamshaw, A. In Chemisrv of the Elemenrs; ...

ChemInform Abstract Amide ligands (I) (preparation described) and (III), modified by the presence of the diarylboryl substituents, are used to synthesize low-coordinate metal complexes of unusual structure. X-ray analyses are carried out for (IIIa) (P1, Z=2), (VIa) (Pbca, Z = 8), (VIIIb) (P21/c, Z=4), (Xa), (XI) and (XIIa) (all P1, Z=2). The transition-metal complexes are all high-spin with nominally two-coordinate geometries. However, they all display some deviation from linearity, the strongest N-M-N bending is observed in (VIII). In all the complexes the B-N bond lengths are consistent with π-bonds between boron and nitrogen. The neutral ligands are thus B-N analogues of the alkenyls and behave, for the most part, as one-electron donors. Instead of association through bridging at the nitrogen centers, moderate interactions between the metal and the ipso carbons of the B(Mst)2 groups are found for (VIIIb), (Xb) and (XI), leading to pseudo-square-planar coordination in the first and pseudotetrahedral environment in the latter two cases.

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A series of four structurally related cis-dithiolate-ligated Fe(III) complexes, [Fe III (DITpy) (3 and 4). The crystallographically determined mean Fe-S bond distances in 1-4 range from 2.196 to 2.232 Å and are characteristic of low-spin Fe(III)-thiolate complexes. The low-spin S = ½ ground state was confirmed by both EPR and magnetic susceptibility measurements. The electronic spectra of these complexes are characterized by broad absorption bands centered near ~700 nm that are consistent with ligand-to-metal chargetransfer (CT) bands. The complexes were further characterized by cyclic voltammetry measurements, and all possess highly negative Fe(III)/Fe(II) redox couples (~ −1 V vs SCE, saturated calomel electrode) indicating that alkyl thiolate donors are effective at stabilizing Fe(III) centers. Both the redox couple and the 700 nm band in the visible spectra show solvent-dependent shifts that are dependent upon the H-bonding ability of the solvent. The implications of these results with respect to the active site of the iron-containing nitrile hydratases are also discussed.