Browsing by Subject "Hyperpolarization"

Recent work has shown that hyperpolarized magnetic resonance spectroscopy (HP-MRS) can trace in vivo metabolism of biomolecules and is therefore extremely promising for diagnostic imaging. The most severe challenge this technique faces is the short signal lifetime for hyperpolarization, which is dictated by the spin-lattice (T1) relaxation. In this thesis we show with theory, simulation and experiment that the long-lived nuclear spin states in chemically equivalent or near equivalent spin systems offer a solution to this problem. Spin polarization that has lifetime much longer than T1 (up to 70-fold) has been demonstrated with pulse sequence techniques that are compatible with clinical imaging settings. Multiple classes of molecules have been demonstrated to sustain such long-lived hyperpolarization.

The purpose of this thesis is to report technological developments in contrast mechanisms for MRI. The search for new forms of contrast is on-going, with the hope that new contrast mechanisms and new contrast agents will provide unique insights into various molecular processes and disease states. In this thesis, we will describe new contrast mechanisms developed by manipulating the inherent physics of the system, as well as the development of exogenous contrast agents. More specifically, we will describe the application of iMQCs (intermolecular multiple quantum coherences) to thermometry and structural imaging, and the unique information provided from these studies. We will also describe methods for migrating iMQC-based pulse sequences from a Bruker research console onto a clinical GE console, thus enabling the application of iMQCs to humans. We will describe the development of hyperpolarized contrast agents which have the potential to provide an unprecedented level of molecular contrast to MRI and the development of techniques to enhance the lifetime of these hyperpolarized contrast agents. Finally, we will discuss a new type of T2 -weighted imaging which significantly improves the refocusing of CPMG-type sequences.

Hyperpolarized magnetic resonance spectroscopic imaging (HP-MRSI) enables non-invasive visualization of metabolism and physiological activities in real-time. Hyperpolarized agents developed to date are primarily 13C-labeled metabolites with short polarization lifetimes of less than a minute, limiting the imaging assay to fast metabolic pathways. To expand on the applications of HP-MRSI, we proposed the use of a versatile 15N-molecular tagging strategy.

This dissertation reports our exploration and application of highly adaptable 15N-molecular handles for development of hyperpolarized molecular imaging probes. Towards this goal, we have investigated 15N2-diazirines and 15N3-azides as biocompatible HP tags with long polarization lifetimes. Several 15N-tagged biological molecules were prepared, including amino acid, glucose, and drug molecules. Hyperpolarization with a d-DNP method demonstrated high signal enhancements (over 400,000-fold) and long 15N relaxation lifetimes (T1) of average 3–4 minutes in aqueous solutions, which warrants a long MR imaging window.

Moreover, we have rationally designed and synthesized novel 15N-labeled reaction-based probes for sensing hydrogen peroxide (H2O2) and nitric oxide (NO) as biomarkers for oxidative stress. We were able to observe the 15N-signal from our 15N-labeled H2O2 sensing probe in vivo using an animal model, which presents exciting progress in the field. Additionally, we explored various molecular probe designs to pursue the most practical 15N-labeled gamma-glutamyl transferase (GGT) sensor. These reaction-based redox and enzyme sensing probes showed favorable hyperpolarization and bioimaging properties. Our work on innovative de novo chemical probes highlights the unprecedented HP-MRSI applications for imaging disease biomarkers.

There are two fundamental limitations in magnetic resonance: the poor signal amplitude and the short duration before the system return to equilibrium. Hyperpolarization methods solve the problem of signal amplitude, however, the duration of the hyperpolarized signal is still limited by the spin-lattice relaxation time, T1. Disconnected eigenstates provide a mechanism by which hyperpolarization can be stored for several times T1. This thesis contributes to the knowledge of these states in four important ways. First, the decay of hyperpolarized magnetization of gas is simulated in lung tissue with a contrast agent, yielding insights about the optimal field strength for imaging. Second, I show that it is possible to rapidly discover and characterize disconnected eigenstates by showing that they can be measured without synthesizing the isotopically labeled compound. Third, I extend the spin systems that can support disconnected eigenstates by expanding the theory to include spin-1 nuclei. Finally, I show that disconnected states with long lifetimes can be populated in conjunction with hyperpolarization techniques to simultaneously yield large signal amplitudes for long durations.

Applications of hyperpolarized spin order are likely to be in complex geophysical or biological structures. Understanding the effect of the inhomogeneous fields created when such structures are placed in a magnetic field on hyperpolarized spin order is a necessity to characterize the experimental signal. An example case of hyperpolarized 3He and 129Xe diffusing through lung tissue is examined. In particular a Monte Carlo simulation tool, combined with a magnetic field map of the inhomogeneous field created by mouse lung tissue, is used to determine the dephasing rate of hyperpolarized 3He and 129Xe in the presence of SuperParamagnetic Iron Oxide Nanoparticles (SPION). Contributions to the dephasing rate include the inhomogeneous field, the SPION magnetic field, and dephasing caused by collisions with the confining geometry. The sensitivity of either gas to SPION increases with increasing SPION concentration and decreasing field strength.

There are some general rules about what makes for a disconnected eigenstate (or singlet state) with a long lifetime. However, no systematic experimental study has been undertaken due to the cost and time-constraints of synthesizing the labeled species for study. I show that synthesis is not a barrier for characterizing the long-lived states. Instead the lifetimes may be determined by using the naturally occurring doubly-labeled isotopomer. I verified this method with two compounds, diphenyl acetylene (DPA) and diethyl oxylate (DEO). The former was determined to have a singlet lifetime TS = 251.40 ±3.16 s from the synthesized species, while the naturally occurring isotopomer yielded a lifetime TS = 202 ±55.30 s, both substantially longer than the spin-lattice relaxation time, T1 = 1.63 ±0.01s. In DEO, the lifetime from the disconnected eigenstate was determined to be TS = 14.62 ±0.76 s (synthesized), TS = 19.32 ±3.16 s (naturally occurring). This method is applied to a range of compounds ranging from simple four-spin systems, such as diacetylene (TS = 48.80 ±22.74 s, T1 = 18.66 ±1.16 s) to eight spin systems in dimethylmaleic anhydride (TS = 27.25 ±3.39 s, T1 = 9.38 ±0.43 s). Additionally, a family of compounds including naphthalene (TS = 4.37 ±0.34 s, T1 = 11.33 ±4.89 s), biphenyl (TS = 3.09 ±0.66 s, T1 = 4.69 ±0.10 s), and DPA show that the rotation of the phenyl rings and intermolecular dipole-dipole relaxation can be critical to the relaxation dynamics.

One particular method of accessing the disconnected eigenstate involves coupling a chemically equivalent spin-1/2 pair asymmetrically to an auxiliary spin-1/2 pair. I demonstrate that the disconnected state may still be accessed when the auxiliary nuclei are spin-1. This has two distinct advantages. When the auxiliary nuclei change from proton to deuterium, the couplings are reduced by a factor of ~6.5 which prevents the disconnected state from relaxing as rapidly back to equilibrium. This is demonstrated in diacetylene-d2 and DPA-d10, where the singlet lifetime was extended by a factor of ~1.7 via deuteration (TS,1H = 49 ±23 s, TS,2H = 83 ±30 s for diacetylene and TS,1H = 274 ±6.1 s, TS,2H = 479 ±83 s for DPA). Additionally, by reducing the coupling strength, deuteration allows additional structural moieties to be explored, such as RDC=CDR. One such structure is explored in trans-ethylene-d2, where the singlet character of the protons can be accessed by the reduced coupling to the deuterium. Additionally, this allows for a relatively strong deuterium-deuterium scalar coupling, requiring modification to the theory. This is carried out analytically, and implications for the relaxation properties are performed using a spin-dynamics numerical simulation. The lifetime of the disconnected state was determined to be TS = 30.2 ±12.3 s, compared to the T1 = 1.1 ±0.2 s at high concentration (270 mM), and increasing to TS = 117. ±9.80 s at low concentration (52 mM). The variation in long lifetime is attributed to intermolecular dipole-dipole relaxation.

Ultimately, the gains in lifetime from using disconnected eigenstates provide a means to the practical implementation of hyperpolarization in a wider range of experiments. A recent hyperpolarization method, Signal Amplification By Reversible Exchange in Shield Enables Alignment Transfer to Heteronuclei (SABRE-SHEATH) is shown to directly hyperpolarize long lived spin order in a diazirine containing molecule. Diazirine rings are three member N=N-C groups that can replace a methylene group and serve as a versatile MR and optical molecular tag. Hyperpolarization is accomplished by bubbling parahydrogen through a solution containing the diazirine and an iridium catalyst. Due to the chemical inequivalence of the 15N of the diazirine, hyperpolarization of longitudinal magnetization and singlet character could be observed by transfer to the high field spectrometer. Signal enhancements of over 14,000 were observed. The magnetic field strength required for buildup of magnetization and singlet character was derived and is in agreement with the experiment. The magnetization lifetime was observed to be T1 = 5.75 ±0.18 minutes and independent of field strength, while the lifetime of the singlet character was observed to be as long as TS = 30.1 ±13.4 minutes at low field (3 Gauss).

The combination of these experiments – understanding lifetimes in inhomogeneous magnetic fields that will be encountered in experiment, identification of disconnected eigenstates with long lifetimes via the naturally occurring isotopomer and extending these lifetimes even further with deuteration, and finally, the direct generation of long-lived hyperpolarized spin order – allows a measurement that required hyperpolarized spin order for the enhanced signal amplitude, to be carried out.

N-Heterocycles are ubiquitous in biologically active natural products and pharmaceuticals. Yet, new syntheses and modifications of N-heterocycles are continually of interest for the purposes of expanding chemical space, finding quicker synthetic routes, better pharmaceuticals, and even new handles for molecular labeling. There are several iterations of molecular labeling; the decision of where to place the label is as important as of which visualization technique to emphasize.

Piperidine and indole are two of the most widely distributed N-heterocycles and thus were targeted for synthesis, functionalization, and labeling. The major functionalization of these scaffolds should include a nitrogen atom, while the inclusion of other groups will expand the utility of the method. Towards this goal, ease of synthesis and elimination of step-wise transformations are of the utmost concern. Here, the concept of electrophilic amination can be utilized as a way of introducing complex secondary and tertiary amines with minimal operations.

Molecular tags should be on or adjacent to an N-heterocycle as they are normally the motifs implicated at the binding site of enzymes and receptors. The labeling techniques should be useful to a chemical biologist, but should also in theory be useful to the medical community. The two types of labeling that are of interest to a chemist and a physician would be positron emission tomography (PET) and magnetic resonance imaging (MRI).

Coincidentally, the 3-positions of both piperidine and indole are historically difficult to access and modify. However, using electrophilic amination techniques, 3-functionalized piperidines can be synthesized in good yields from unsaturated amines. In the same manner, 3-labeled piperidines can be obtained; the piperidines can either be labeled with an azide for biochemical research or an 18F for PET imaging research. The novel electrophiles, N-benzenesulfonyloxyamides, can be reacted with indole in one of two ways: 3-amidation or 1-amidomethylation, depending on the exact reaction conditions. Lastly, a novel, hyperpolarizable 15N2-labeled diazirine has been developed as an exogenous and versatile tag for use in magnetic resonance imaging.

Magnetic resonance techniques are among the most powerful methods for characterization. However, they inherently suffer from an intrinsically low signal-to-noise due to the weak interaction of the nuclear spin with external magnetic fields. Hyperpolarization methods circumvent this limitation by deriving non-equilibrium spin polarization from an external source of spin order, dramatically increasing the magnetic resonance signals. Signal Amplification By Reversible Exchange, or SABRE, is a relatively new and promising method that derives spin hyperpolarization from parahydrogen, the singlet spin isomer of dihydrogen, allowing it to operate at a fraction of the cost of other hyperpolarization methods. A target molecule and parahydrogen transiently bind an organometallic complex, during which time polarization is transferred from the parahydrogen to target nuclei. The reversible nature of this interaction makes the hyperpolarization method readily scalable, giving SABRE the potential to supplant older, more expensive techniques and bring hyperpolarization technology to a broader audience. However, the current demonstrations of SABRE generate polarizations that are about an order of magnitude away from the upper theoretical limits of the technique, and variants of this experiment have been limited in target scope by the underlying physics. To address these limitations, this dissertation returns to examine the theoretical underpinnings of SABRE, and in doing so re-interrogates the unification of chemical exchange and quantum dynamics. We derive exact formulations of the chemical exchange interaction in both the magnetic resonance limits, which is used to construct a physically exhaustive computational model for SABRE. This model then facilitates in silico exploration of the system and permits us to address experimental limitations of the method. In particular, this dissertation utilizes simulations to develop and expand the capabilities of SABRE performed at arbitrarily high magnetic fields. We show that the limitations in the scope of SABRE imposed by the spin physics under these conditions may be removed, culminating in the first demonstration of simultaneous hyperpolarization of multiple components. This expansion is a key step towards translating SABRE into areas of conventional magnetic resonance such as biomolecular NMR and metabonomics. The theoretical framework presented here provides access to new routes for optimizing SABRE hyperpolarization, and we demonstrate that sequences may be developed to generate up to a five-fold increase in performance. Finally, we extend the theoretical treatment of chemical exchange within the Lindblad formalism to obtain exact master equations that are valid in any physical limit.