Systems and methods for producing hyperpolarized materials

Abstract

A method is disclosed for increasing the concentration of a hyperpolarized molecule of interest in a solution. The hyperpolarized molecule of interest is prepared in an organic solution. A portion of the organic solution is rapidly evaporated to increase its concentration in the solution while maintaining sufficient nuclear spin polarization. The hyperpolarized molecule of interest is then used in an MRI or NMR experiment. A method is disclosed for increasing nuclear spin polarization in a molecule of interest polarized by the SABRE method. An at least partially deuterated molecule of interest is placed in a magnetic field having an average field strength of at most about 2 T. The molecule of interest forms a coordination complex with a SABRE catalyst or catalyst precursor and parahydrogen in the solution. An oscillating magnetic field is applied to transfer spin order from the parahydrogen to the molecule of interest, thereby increasing the nuclear spin polarization in the molecule of interest.

Description

[Technical Field]

[0001] cross reference This application claims priority to U.S. Provisional Patent Application No. 63 / 387,416, filed December 14, 2022, and U.S. Provisional Patent Application No. 63 / 478,437, filed January 4, 2023, each of which is incorporated by reference herein in its entirety for all purposes.

[0002] Technical Field The disclosed embodiments relate generally to the production of hyperpolarized materials for use in nuclear magnetic resonance, magnetic resonance imaging, or similar applications. [Background technology]

[0003] Nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI) are techniques with important applications in chemistry, biology, and medical imaging. Despite their success, nuclear magnetic resonance applications require minute nuclear polarization of the analyte (typically 10 -5 It is recognized that there is a limit due to the small nuclear polarization (on the order of 1000 Å). This small nuclear polarization can result in limited sensitivity compared to other analytical techniques such as mass spectrometry.

[0004] Increasing nuclear spin polarization beyond the thermal equilibrium value can improve magnetic resonance sensitivity. Nuclear spin polarization can be increased using known techniques such as dynamic nuclear polarization (DNP), parahydrogen-induced polarization (PHIP), PHIP-sidearm hydrogenation (PHIP-SAH), and signal amplification by reversible exchange (SABRE). Using these techniques, the nuclear spin polarization of a material can be increased by more than 10,000 times. Enhanced nuclear spin polarization can result in a proportional increase in NMR / MRI signal. This enhanced polarization decays over time due to the relaxation time of nuclear spins in polarized molecules. However, for many molecules, relaxation times can be on the order of seconds to minutes, during which increased polarization can result in a dramatic increase in NMR / MRI signal sensitivity. By enabling such dramatic increases in NMR / MRI signal sensitivity, increased nuclear spin polarization can enable new applications, such as in vivo metabolic imaging using metabolites with increased nuclear spin polarization in MRI scanners, accelerating NMR spectroscopy investigations and enabling the visualization of previously invisible molecular dynamics and structures. Summary of the Invention

[0005] Disclosed embodiments include methods for performing a magnetic resonance imaging (MRI) or nuclear magnetic resonance (NMR) procedure using hyperpolarized molecules of interest. The method may include (a) obtaining an organic solution having hyperpolarized molecules of interest dissolved therein at a first concentration, (b) evaporating at least a portion of the organic solution to thereby produce an organic solution having hyperpolarized molecules of interest dissolved therein at a second concentration greater than the first concentration, and (c) performing the MRI or NMR procedure using the hyperpolarized molecules of interest. The first concentration may be less than 100 millimolar (mM), 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, or less. The second concentration can be at least 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, 200 mM, or more.

[0006] Disclosed embodiments include compositions for use in magnetic resonance imaging (MRI) or nuclear magnetic resonance (NMR) procedures. The compositions can include a solution and hyperpolarized molecules of interest dissolved therein. The compositions can be produced by (a) obtaining an organic solution having hyperpolarized molecules of interest dissolved therein at a first concentration, and (b) evaporating at least a portion of the organic solution, thereby producing an organic solution having hyperpolarized molecules of interest dissolved therein at a second concentration greater than the first concentration. The first concentration can be less than 100 millimolar (mM), 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, or less. The second concentration can be at least 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, 200 mM, or more.

[0007] Disclosed embodiments include a method for increasing nuclear spin polarization in a molecule of interest. The method may include: (a) obtaining a molecule of interest containing at least one deuterium atom; (b) placing the molecule of interest in a magnetic field having an average field strength (B) of at most about 2 Tesla (T); (c) forming a coordination complex between the molecule of interest, a signal amplification by reversible exchange (SABRE) catalyst or catalyst precursor, and para-hydrogen; and (d) applying an oscillating magnetic field to the coordination complex, thereby transferring spin order from the para-hydrogen to the molecule of interest, thereby increasing the nuclear spin polarization of at least one atom in the molecule of interest. The oscillating magnetic field may have a maximum field strength of about 0.1 μT to about 10 millitesla (mT).

[0008] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of embodiments of the disclosure as claimed.

[0009] The accompanying drawings, which form a part of this specification, illustrate some embodiments and, together with the description, serve to explain the principles and features of the disclosed embodiments. [Brief explanation of the drawings]

[0010] [Figure 1] FIG. 1 illustrates an exemplary method for increasing the concentration of hyperpolarized molecules of interest in a solution, according to disclosed embodiments. [Figure 2] FIG. 2 illustrates a composition featuring an increased concentration of hyperpolarized molecules of a molecule of interest in solution, according to disclosed embodiments. [Figure 3] FIG. 3 illustrates an exemplary method for increasing nuclear spin polarization in a molecule of interest polarized by the SABRE method, according to a disclosed embodiment. [Figure 4] FIG. 4 illustrates a composition characterized by increased nuclear spin polarization in a molecule of interest polarized by the SABRE method, according to a disclosed embodiment. [Figure 5]FIG. 5 illustrates an exemplary increase in solution concentration using manual extraction of the solution, according to disclosed embodiments. [Figure 6] FIG. 6 illustrates an exemplary increase in solution concentration using automated extraction of the solution, according to disclosed embodiments. [Figure 7] FIG. 7 shows an exemplary solution concentration increase using automated extraction of a solution with deuterium and carbon labeled DMAD according to disclosed embodiments. [Figure 8] FIG. 8 shows exemplary C NMR spectra of [1-C]pyruvate-d3 and [2-C]pyruvate-d3 polarized using the SLIC-SABRE method described herein and the conventional gold standard SABRE-SHEATH method, according to disclosed embodiments. [Figure 9] FIG. 9 shows exemplary C polarization levels for protonated and deuterated [1-C]pyruvate and [2-C]pyruvate polarized using the SLIC-SABRE method and SABRE-SHEATH described herein, according to disclosed embodiments. [Figure 10] FIG. 10 shows exemplary polarization accumulations of protonated and deuterated [1-C]pyruvate and [2-C]pyruvate polarized using the SLIC-SABRE method and SABRE-SHEATH described herein, according to disclosed embodiments. [Figure 11] FIG. 11 shows exemplary T and Tρ values ​​for protonated and deuterated [1-C]pyruvate and [2-C]pyruvate, according to disclosed embodiments. DETAILED DESCRIPTION OF THE INVENTION

[0011] Exemplary embodiments will now be described in detail and discussed with reference to the accompanying drawings. In some instances, the same reference numbers are used throughout the drawings, and the following description refers to the same or similar parts. Unless otherwise defined, technical and / or scientific terms have the meanings commonly understood by those skilled in the art. The disclosed embodiments are described in sufficient detail to enable those skilled in the art to practice the disclosed embodiments. It will be understood that other embodiments may be utilized, and changes may be made without departing from the scope of the disclosed embodiments. Accordingly, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

[0012] Nuclear magnetic resonance (NMR) spectroscopy can be used for applications ranging from determining chemical structures in synthetic intermediates to determining atomic-level structure and dynamics in proteins and nucleic acids. Magnetic resonance imaging (MRI) can be used for applications such as noninvasive imaging of the internal structure of biological samples (such as tissues or organs) with submillimeter spatial resolution. However, NMR spectroscopy / MRI can be limited in sensitivity due to the small size of nuclear magnetic moments, combined with the corresponding small polarization at thermal equilibrium. This limited sensitivity can prevent the use of NMR spectroscopy / MRI in some applications and can make other applications of NMR spectroscopy / MRI impractically time- or material-intensive.

[0013] NMR / MRI sensitivity can be increased by using higher magnetic fields and optimized detection systems. However, an alternative approach is to increase NMR / MRI sensitivity by increasing nuclear spin polarization to levels significantly higher than thermal equilibrium. Such hyperpolarization techniques can often increase NMR / MRI sensitivity by a factor significantly greater than increasing the magnetic field or using optimized detection systems. Furthermore, such hyperpolarization techniques may enable new NMR spectroscopy / MRI applications, such as the observation of low-gamma nuclei or low-concentration analytes or molecular imaging.

[0014] Nuclear spin polarization can be increased using a variety of techniques, including dynamic nuclear polarization (DNP), parahydrogen-induced polarization (PHIP), PHIP-sidearm hydrolysis (PHIP-SAH), PHIP nuclear Overhauser effect system (PHIPNOESYS), signal amplification by reversible exchange (SABRE), spin-exchange optical pumping (SEOP), optically initialized electronic triplet states (also called photoexcited triplet states, PETS), and other suitable methods. Among these techniques, parahydrogen-based methods such as PHIP, PHIP-SAH, PHIPNOESYS, and SABRE are particularly promising because they can be implemented with high throughput using relatively low-cost equipment.

[0015] For example, recent research in NMR and MRI has demonstrated that NMR and MRI signals associated with various biocontrast agents can be enhanced by several orders of magnitude using PHIP or PHIP-SAH. Such dramatic signal enhancement enables spectroscopic analysis of biocontrast agents as they are metabolized by various tissues at different locations within the body. Analysis of metabolic information determined by such spectroscopic imaging can enable noninvasive determination of the health of tissues within the body. For example, abnormal metabolism of biocontrast agents can indicate diseases such as cancer at several locations within the body.

[0016] In PHIP and PHIP-SAH, a derivative (e.g., precursor) of a molecule of interest is reacted with para-hydrogen to form the para-hydrogenated form of the derivative. Spin order is then transferred from the proton added via the para-hydrogenation reaction to a nucleus of interest (e.g., carbon-13 nucleus) contained within the molecule of interest. In PHIP, the para-hydrogenated form of the derivative is chemically identical to the molecule of interest and is distinguished from it only by the spin order resulting from the para-hydrogenation reaction. In PHIP-SAH, the para-hydrogenated form of the derivative is cleaved (e.g., hydrolyzed) to yield the hyperpolarized molecule of interest.

[0017] In SABRE, the molecule of interest itself forms a coordination complex with a polarization transfer catalyst or catalyst precursor and parahydrogen. Spin order is then transferred from the parahydrogen to a nucleus of interest within the molecule of interest via the coordination complex. The molecule of interest is then optionally purified and used in an NMR or MRI procedure.

[0018] PHIPNOESYS utilizes PHIP or PHIP-SAH to generate hyperpolarized material (e.g., a source compound) and transfer the polarization from the source compound to a material (e.g., a target compound) used in NMR spectroscopy. The transfer of polarization from the source compound to the target compound proceeds via the intermolecular Nuclear Overhauser Effect (NOE). PHIPNOESYS has been shown to increase the NMR spectroscopy signal by up to approximately 2,000-fold, allowing NMR spectroscopy to be applied at significantly reduced concentrations than would otherwise be achievable.

[0019] However, parahydrogen-based methods such as PHIP, PHIP-SAH, PHIPNOESYS, and SABRE may be limited in the concentration of molecules that can be hyperpolarized. In some cases, higher concentrations of hyperpolarized molecules are required, such as when injecting hyperpolarized molecules into human subjects during hyperpolarized MRI experiments. For example, a hyperpolarized MRI molecular imaging experiment may require the injection of a hyperpolarized biorelevant contrast agent having a concentration of about 100 millimolar (mM) to about 200 millimolar (mM). However, PHIP, PHIP-SAH, or SABRE may only be able to produce a hyperpolarized biorelevant contrast agent at a concentration of, for example, about 30 mM to about 50 mM. Therefore, there is a need for methods and systems that increase the concentration of hyperpolarized molecules in solution.

[0020] Furthermore, current SABRE techniques can be limited by the relatively short longitudinal (T1) relaxation times of protons in molecules of interest. These relatively short relaxation times can limit the polarization that can be transferred to molecules of interest. Therefore, a need exists for methods and systems that increase the nuclear spin polarization in molecules of interest polarized by the SABRE technique.

[0021] Disclosed embodiments increase the concentration of hyperpolarized molecules of interest in a solution. The hyperpolarized molecules of interest are generally prepared in an organic solution at a specific concentration. At least a portion of the organic solution is rapidly evaporated to increase the concentration of the hyperpolarized molecules of interest in the solution while maintaining sufficient nuclear spin polarization to perform MRI or NMR experiments. The hyperpolarized molecules of interest are then dissolved in an aqueous solution, enabling the use of the hyperpolarized molecules of interest in MRI or NMR experiments.

[0022] Disclosed embodiments further increase the nuclear spin polarization in a molecule of interest polarized by the SABRE method. The molecule of interest is at least partially deuterated by replacing at least one proton in the molecule of interest with a deuterium atom. The molecule of interest is placed in a magnetic field having an average field strength of at most about 2 Tesla (T). The molecule of interest forms a coordination complex with a SABRE catalyst or catalyst precursor and parahydrogen in solution. An oscillating magnetic field is then applied to the coordination complex to transfer spin order from the parahydrogen to the molecule of interest, thereby increasing the nuclear spin polarization in the molecule of interest.

[0023] Hyperpolarized and Parahydrogen As used in this disclosure, "polarization" refers to an imbalance in electron or nuclear spin orientation. In some embodiments, polarization can be the normalized, approximate difference between the number of spins in a first direction minus the number of spins in the opposite direction. As a non-limiting example, 200,000 1 Assuming a nuclear spin of H, a polarization of 2% can correspond to 102,000 spins in one direction and 98,000 spins in the opposite direction. In some embodiments, "hyperpolarization" can include polarization of a species (e.g., nuclear, selective, or the like) that exceeds the typical polarization level of that species observed at thermal equilibrium upon exposure to a particular magnetic field. As a non-limiting example, 1 A sample in a 1 T magnetic field at thermal equilibrium with a H nuclear spin polarization greater than 0.000341% is substantially higher (e.g., at least one order of magnitude higher) than the 0.000341% thermal equilibrium polarization.1 As a further non-limiting example, the hologram may be hyperpolarized to have a H nuclear spin polarization of greater than 0.000257%. 13 A sample in a magnetic field of 3 T at thermal equilibrium with a spin polarization of C can be hyperpolarized. 15 A sample in a magnetic field of 3 T at thermal equilibrium with a spin polarization of N can be hyperpolarized.

[0024] Parahydrogen is a form of molecular hydrogen. In this form of molecular hydrogen, the two proton spins are in a singlet state. In some embodiments, parahydrogen can be formed in gaseous or liquid form. For example, in some embodiments, parahydrogen can be generated in gaseous form by flowing hydrogen gas through a chamber using a catalyst. In some embodiments, the hydrogen gas can be subjected to low temperatures, such as temperatures of at most about 100K, 90K, 80K, 70K, 60K, 50K, 40K, 30K, 20K, 10K, 9K, 8K, 7K, 6K, 5K, 4K, 3K, or lower. In some embodiments, the catalyst can be iron oxide. In some embodiments, the hydrogen gas can contain both parahydrogen and orthohydrogen, and the low temperature can bring the hydrogen gas to thermodynamic equilibrium within the chamber, during which a population of parahydrogen grows.

[0025] In some embodiments, the gas may be generated at a first location and then transported to a second location for use. In some embodiments, the first location may be a chamber, which may be part of a container, bottle, holder, or other area capable of holding a gas or liquid. Such a chamber may be maintained at an appropriate pressure, temperature, or combination thereof. In some embodiments, the first location may refer to a physical location, such as a room, a laboratory, a specific warehouse, a hospital, or other location where parahydrogen may be generated.

[0026] In some embodiments, the generated para-hydrogen may be transported in a chamber different from the chamber in which the para-hydrogen was generated. The chamber transporting the para-hydrogen gas may be maintained at a suitable pressure or temperature and may be transported by vehicle or person. In some embodiments, transporting the para-hydrogen may involve moving the para-hydrogen from one container to a different container. In some embodiments, transporting the para-hydrogen may involve moving the para-hydrogen within the same location, such as from one part of a room to another part of a room. In some embodiments, transporting the para-hydrogen may involve moving the para-hydrogen from one room in a building to another room in the same building or to a nearby building. In some embodiments, transporting the para-hydrogen may involve moving the para-hydrogen to another part of the same city or to another location in a different city. For example, transporting the para-hydrogen may involve bringing the para-hydrogen near a polarizer or an NMR / MRI machine. In another example, in some embodiments, transporting the para-hydrogen includes packaging or shipping the para-hydrogen in an appropriate container.

[0027] Methods for increasing the concentration of hyperpolarized molecules of interest in solution 1 illustrates an exemplary method 100 for increasing the concentration of hyperpolarized molecules of interest in a solution, according to disclosed embodiments. Method 100 utilizes an evaporation procedure to increase the concentration of hyperpolarized molecules of interest in an organic solution.

[0028] In step 110, an organic solution is obtained. In some embodiments, the organic solution contains hyperpolarized molecules of interest dissolved therein. In some embodiments, the organic solution includes at least one organic molecule selected from the group consisting of methanol, ethanol, n-propanol, isopropanol, and acetone. In some embodiments, the hyperpolarized molecules of interest are hyperpolarized via a hyperpolarization procedure such as PHIP, PHIP-SAH, SABRE, PHIPNOESYS, DNP, or SEOP. In some embodiments, the hyperpolarized molecules of interest contain at least one nucleus having a nuclear spin polarization of at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more. In some embodiments, at least one nucleus has a nuclear spin polarization of at most about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, hi some embodiments, at least one nucleus has a nuclear spin polarization that is within a range defined by any two of the foregoing values.

[0029] In some embodiments, the hyperpolarized molecules of interest are dissolved in an organic solution at a first concentration. In some embodiments, the first concentration is at most about 100 millimolar (mM), 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, or less. In some embodiments, the first concentration is at least about 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, or more. In some embodiments, the first concentration is within a range defined by any two of the foregoing values. For example, in some embodiments, the first concentration is about 10 mM to about 100 mM, about 10 mM to about 90 mM, about 10 mM to about 80 mM, about 10 mM to about 70 mM, about 10 mM to about 60 mM, about 10 mM to about 50 mM, about 10 mM to about 40 mM, about 10 mM to about 30 mM, about 10 mM to about 20 mM, about 20 mM to about 10 0mM, approximately 20mM to approximately 90mM, approximately 20mM to approximately 80mM, approximately 20mM to approximately 70mM, approximately 20mM to approximately 60mM, approximately 20mM to approximately 50mM, approximately 20mM to Approximately 30mM, approximately 30mM to approximately 100mM, approximately 30mM to approximately 90mM, approximately 30mM to approximately 80mM, approximately 30mM to approximately 70mM, approximately 30mM to approximately 60mM, approximately 30m M ~ about 50mM, about 30mM - about 40mM, about 40mM - about 100mM, about 40mM - about 90mM, about 40mM - about 80mM, about 40mM - about 70mM, about 40mM to about 60mM, about 40mM to about 50mM, about 50mM to about 100mM, about 50mM to about 90mM, about 50mM to about 80mM, about 50mM to about 70mM , about 50 mM to about 60 mM, about 60 mM to about 100 mM, about 60 mM to about 90 mM, about 60 mM to about 80 mM, about 60 mM to about 70 mM, about 70 mM to about 100 mM, about 70 mM to about 90 mM, about 70 mM to about 80 mM, about 80 mM to about 100 mM, about 80 mM to about 90 mM, or about 90 mM to about 100 mM.

[0030] In step 120, at least a portion of the organic solution is evaporated. In some embodiments, evaporating the portion of the organic solution increases the concentration of the hyperpolarized compound of interest in the organic solution. That is, in some embodiments, evaporating the portion of the organic solution produces an organic solution having the hyperpolarized compound of interest dissolved therein at a second concentration. In some embodiments, the second concentration exceeds the first concentration.

[0031] In some embodiments, the second concentration exceeds the first concentration by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, or more. In some embodiments, the second concentration exceeds the first concentration by at most about 400%, 375%, 350%, 325%, 300%, 275%, 250%, 225%, 200%, 175%, 150%, 125%, 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, or less. In some embodiments, the second concentration exceeds the first concentration by an amount that is within a range defined by any two of the foregoing values. For example, in some embodiments, the second concentration is about 10% to about 400%, about 10% to about 350%, about 10% to about 300%, about 10% to about 250%, about 10% to about 200%, about 10% to about 150%, about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 10% to about 30%, about 10% to about 20%, about 20% to about 400%, about 20% to about 350%, about 20% to about 300%, about 20% to about 250%, about 20% to about 200%, about 20% to about 150%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 2 0% to about 30%, about 30% to about 400%, about 30% to about 350%, about 30% to about 300%, about 30% to about 250%, about 30% to about 200%, about 30% to about 150%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, about 30% to about 50%, about 30% to about 40%, about 40% to about 400%, about 40% to about 3 50%, approximately 40% to approximately 300%, approximately 40% to approximately 250%, approximately 40% to approximately 200%, approximately 40% to approximately 150%, approximately 40% to approximately 100%, approximately 40% to approximately 90%, approximately 40% to approximately 80%, approximately 40% to approximately 70%, approximately 40% to approximately 60%, approximately 40% to approximately 50%, approximately 50% to approximately 400%, approximately 50% to approximately 350%, approximately 50% to approximately 300%, approximately 50% to approximately 250%, approximately 50% to approximately 200%,Approximately 50% to approximately 150%, approximately 50% to approximately 100%, approximately 50% to approximately 90%, approximately 50% to approximately 80%, approximately 50% to approximately 70%, approximately 50% to approximately 60%, approximately 60% to approximately 400%, approximately 60% to approximately 350%, approximately 60% to approximately 300%, approximately 60% to approximately 250%, approximately 60% to approximately 200%, approximately 60% to approximately 150%, approximately 60% to approximately 100%, approximately 60% to approximately 90%, approximately 60% to approximately 80%, approximately 60% to approximately 70%, approximately 70 % to approximately 400%, approximately 70% to approximately 350%, approximately 70% to approximately 300%, approximately 70% to approximately 250%, approximately 70% to approximately 200%, approximately 70% to approximately 150%, approximately 70% to approximately 100%, approximately 70% to approximately 90%, approximately 70% to approximately 80%, approximately 80% to approximately 400%, approximately 80% to approximately 350%, approximately 80% to approximately 300%, approximately 80% to approximately 250%, approximately 80% to approximately 200%, approximately 80% to approximately 150%, approximately 80% to approximately 100%, approximately 80% to about 90%, about 90% to about 400%, about 90% to about 350%, about 90% to about 300%, about 90% to about 250%, about 90% to about 200%, about 90% to about 150%, about 90% to about 100%, about 100% to about 400%, about 100% to about 350%, about 100% to about 300%, about 100% to about 250%, about 100% to about 200%, about 100% to about 150%, about 150% to about 400%, about 1 50% to about 350%, about 150% to about 300%, about 150% to about 250%, about 150% to about 200%, about 200% to about 400%, about 200% to about 350%, about 200% to about 300%, about 200% to about 250%, about 250% to about 400%, about 250% to about 350%, about 250% to about 300%, about 300% to about 400%, about 300% to about 350%, or about 350% to about 400%.

[0032] In some embodiments, the second concentration is at least about 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, 200 mM, 225 mM, 250 mM, 275 mM, 300 mM, 325 mM, 350 mM, 375 mM, 400 mM, 425 mM, 450 mM, 475 mM, 500 mM. In some embodiments, the second concentration is at most about 500 mM, 475 mM, 450 mM, 425 mM, 400 mM, 375 mM, 350 mM, 325 mM, 300 mM, 275 mM, 250 mM, 225 mM, 200 mM, 190 mM, 180 mM, 170 mM, 160 mM, 150 mM, 140 mM, 130 mM, 120 mM, 110 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, or less. In some embodiments, the second concentration is within a range defined by any two of the foregoing values. For example, in some embodiments, the second concentration is about 50 mM to about 500 mM, about 50 mM to about 450 mM, about 50 mM to about 400 mM, about 50 mM to about 350 mM, about 50 mM to about 300 mM, about 50 mM to about 250 mM, about 50 mM to about 200 mM, about 50 mM to about 150 mM, about 50 mM to about 100 mM, Approximately 50mM to approximately 90mM, approximately 50mM to approximately 80mM, approximately 50mM to approximately 70mM, approximately 50mM to approximately 60mM, approximately 60mM to approximately 500mM, approximately 60mM to approximately 45 0mM, approximately 60mM to approximately 400mM, approximately 60mM to approximately 350mM, approximately 60mM to approximately 300mM, approximately 60mM to approximately 250mM, approximately 60mM to approximately 200mM, approximately 60mM to about 150mM, about 60mM to about 100mM, about 60mM to about 90mM, about 60mM to about 80mM, about 60mM to about 70mM, about 70mM to about 50 0mM, about 70mM to about 450mM, about 70mM to about 400mM, about 70mM to about 350mM, about 70mM to about 300mM, about 70mM to about 250mM, about 70mM to about 200mM, about 70mM to about 150mM, about 70mM to about 100mM, about 70mM to about 90mM, about 70mM to about 80mM, about 80mM to about 5 00mM, about 80mM to about 450mM, about 80mM to about 400mM, about 80mM to about 350mM, about 80mM to about 300mM, about 80mM to about 250mM,Approximately 80mM to approximately 200mM, approximately 80mM to approximately 150mM, approximately 80mM to approximately 100mM, approximately 80mM to approximately 90mM, approximately 90mM to approximately 500mM, approximately 90mM to approximately 450mM, approximately 90mM to approximately 400mM, approximately 90mM to approximately 350mM, approximately 90mM to approximately 300mM, approximately 90mM to approximately 250mM, approximately 90mM to approximately 200mM, approximately 90mM to approximately 150mM, approximately 90mM to approximately 100mM, approximately 100 mM to about 500 mM, about 100 mM to about 450 mM, about 100 mM to about 400 mM, about 100 mM to about 350 mM, about 100 mM to about 300 mM, about 100 mM to about 250 mM, about 100 mM to about 200 mM, about 100 mM to about 150 mM, about 150 mM to about 500 mM, about 150 mM to about 450 mM, about 150 mM to about 400 mM, about 150 mM to about 350 mM, about 50mM to about 300mM, about 150mM to about 250mM, about 150mM to about 200mM, about 200mM to about 500mM, about 200mM to about 450mM, about 200mM to about 400mM, about 2 00mM to about 350mM, about 200mM to about 300mM, about 200mM to about 250mM, about 250mM to about 500mM, about 250mM to about 450mM, about 250mM to about 400mM, about 25 The concentration is 0 mM to about 350 mM, about 250 mM to about 300 mM, about 300 mM to about 500 mM, about 300 mM to about 450 mM, about 300 mM to about 400 mM, about 300 mM to about 350 mM, about 350 mM to about 500 mM, about 350 mM to about 450 mM, about 350 mM to about 400 mM, about 400 mM to about 500 mM, about 400 mM to about 450 mM, or about 450 mM to about 500 mM.

[0033] In some embodiments, evaporating a portion of the organic solution comprises performing at least one evaporation step. In some embodiments, the at least one evaporation step is selected from the group consisting of flowing an inert gas (e.g., nitrogen or argon gas) over the organic solution, subjecting the organic solution to a vacuum, and heating the organic solution. In some embodiments, the organic solution is heated to a temperature of at least about 30 degrees Celsius (°C), 40°C, 50°C, 60°C, 70°C, 80°C, 90°C, 100°C, or higher. In some embodiments, the organic solution is heated to a temperature of at most about 100°C, 90°C, 80°C, 70°C, 60°C, 50°C, 40°C, 30°C, or lower. In some embodiments, the organic solution is heated to a temperature that is within a range defined by any two of the preceding values. In some embodiments, the at least one evaporation step comprises at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more evaporation steps. In some embodiments, the at least one evaporation step includes at most about 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 evaporation steps. In some embodiments, the at least one evaporation step includes several evaporation steps within a range defined by any two of the foregoing values. For example, in some embodiments, the evaporation steps may be from about 1 to about 10, from about 1 to about 9, from about 1 to about 8, from about 1 to about 7, from about 1 to about 6, from about 1 to about 5, from about 1 to about 4, from about 1 to about 3, from about 1 to about 2, from about 2 to about 10, from about 2 to about 9, from about 2 to about 8, from about 2 to about 7, from about 2 to about 6, from about 2 to about 5, from about 2 to about 4, from about 2 to about 3, from about 3 to about 10, from about 3 to about 9, from about 3 to about 8, from about 3 to about 7, from about 3 to about 6, from about 3 to about 6, or from about 3 to about 8. About 5, about 3 to about 4, about 4 to about 10, about 4 to about 9, about 4 to about 8, about 4 to about 7, about 4 to about 6, about 4 to about 5, about 5 to about 10, about 5 to about 9, about 5 to about 8, about 5 to about 7, about 5 to about 6, about 6 to about 10, about 6 to about 9, about 6 to about 8, about 6 to about 7, about 7 to about 10, about 7 to about 9, about 7 to about 8, about 8 to about 10, about 8 to about 9, or about 9 to about 10 evaporation steps are performed.

[0034] In step 130, an NMR or MRI procedure is performed using the hyperpolarized molecule of interest. In some embodiments, the NMR or MRI procedure is performed using an organic solution. In some embodiments, the NMR or MRI procedure is performed using a different organic solution in which the hyperpolarized molecule of interest has been dissolved (e.g., via a precipitation and redissolution procedure or a solvent exchange procedure). In some embodiments, the NMR or MRI procedure is performed using an aqueous solution in which the hyperpolarized molecule of interest has been dissolved (e.g., via a precipitation and redissolution procedure or a solvent exchange procedure). Examples of precipitation and redissolution procedures are provided, for example, in WO2022 / 018514 and WO2022 / 269350, each of which is incorporated by reference herein in its entirety for all purposes. An example of a solvent exchange procedure is provided in WO2022 / 269350, which is incorporated by reference herein in its entirety for all purposes.

[0035] For example, in some embodiments, prior to step 130, the organic solution is mixed with water or a different organic solvent, thereby producing an aqueous or organic solution having the hyperpolarized compound dissolved therein at a third concentration. In some embodiments, the third concentration is at least about 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, 200 mM, 225 mM, 250 mM, 275 mM, 300 mM, 325 mM, 350 mM, 375 mM, 400 mM, 425 mM, 450 mM, 475 mM, or 500 mM. In some embodiments, the third concentration is at most about 500 mM, 475 mM, 450 mM, 425 mM, 400 mM, 375 mM, 350 mM, 325 mM, 300 mM, 275 mM, 250 mM, 225 mM, 200 mM, 190 mM, 180 mM, 170 mM, 160 mM, 150 mM, 140 mM, 130 mM, 120 mM, 110 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, or less. In some embodiments, the third concentration is within a range defined by any two of the foregoing values. For example, in some embodiments, the third concentration is about 50 mM to about 500 mM, about 50 mM to about 450 mM, about 50 mM to about 400 mM, about 50 mM to about 350 mM, about 50 mM to about 300 mM, about 50 mM to about 250 mM, about 50 mM to about 200 mM, about 50 mM to about 150 mM, about 50 mM to about 100 mM, about 50 mM to about 90 mM, about 50 mM to about 80 mM, about 50 mM to about 70 mM, about 50 mM to about 60 mM, about 60 mM to about 500 mM, about 60 mM to about 450 mM, about 60 mM to about 400 mM, about 60 mM to about 65 ... 0mM to about 350mM, about 60mM to about 300mM, about 60mM to about 250mM, about 60mM to about 200mM, about 60mM to about 150mM, about 60mM to about 100mM, about 60mM to about 90mM, about 60mM to about 80mM, about 60mM to about 70mM, about 7 0mM to about 500mM, about 70mM to about 450mM, about 70mM to about 400mM, about 70mM to about 350mM, about 70mM to about 300mM, about 70mM to about 250mM, about 70mM to about 200mM, about 70mM to about 150mM, about 70mM to about 100mM,Approximately 70mM to approximately 90mM, approximately 70mM to approximately 80mM, approximately 80mM to approximately 500mM, approximately 80mM to approximately 450mM, approximately 80mM to approximately 400mM, approximately 80mM to approximately 350mM, approximately 80mM to approximately 300mM, approximately 80mM to approximately 2 50mM, about 80mM to about 200mM, about 80mM to about 150mM, about 80mM to about 100mM, about 80mM to about 90mM, about 90mM to about 500mM, about 90mM to about 450mM, about 90mM to about 400mM, about 9 0 mM to about 350 mM, about 90 mM to about 300 mM, about 90 mM to about 250 mM, about 90 mM to about 200 mM, about 90 mM to about 150 mM, about 90 mM to about 100 mM, about 100 mM to about 500 mM, about 100 mM to about 450 mM, about 100 mM to about 400 mM, about 100 mM to about 350 mM, about 100 mM to about 300 mM, about 100 mM to about 250 mM, about 100 mM to about 200 mM, about 100 mM to about 150 mM, about 150 mM to About 500mM, about 150mM to about 450mM, about 150mM to about 400mM, about 150mM to about 350mM, about 150mM to about 300mM, about 150mM to about 250mM, about 150mM to about 200mM, about 200mM ~about 500mM, about 200mM to about 450mM, about 200mM to about 400mM, about 200mM to about 350mM, about 200mM to about 300mM, about 200mM to about 250mM, about 250mM to about 500mM, about 250mM to about 450 mM, about 250 mM to about 400 mM, about 250 mM to about 350 mM, about 250 mM to about 300 mM, about 300 mM to about 500 mM, about 300 mM to about 450 mM, about 300 mM to about 400 mM, about 300 mM to about 350 mM, about 350 mM to about 500 mM, about 350 mM to about 450 mM, about 350 mM to about 400 mM, about 400 mM to about 500 mM, about 400 mM to about 450 mM, or about 450 mM to about 500 mM.

[0036] In some embodiments, method 100 is performed rapidly. In some embodiments, the rapid performance of method 100 allows the concentration of the molecule of interest to increase while preventing significant loss of nuclear spin polarization in the molecule of interest. In some embodiments, method 100 (or one, two, or three of steps 110, 120, and 130) is performed within a total period of at most about 180 seconds, 170 seconds, 160 seconds, 150 seconds, 140 seconds, 130 seconds, 120 seconds, 110 seconds, 100 seconds, 90 seconds, 80 seconds, 70 seconds, 60 seconds, 50 seconds, 40 seconds, 30 seconds, 20 seconds, 19 seconds, 18 seconds, 17 seconds, 16 seconds, 15 seconds, 14 seconds, 13 seconds, 12 seconds, 11 seconds, 10 seconds, 9 seconds, 8 seconds, 7 seconds, 6 seconds, 5 seconds, 4 seconds, 3 seconds, 2 seconds, 1 second, or less. In some embodiments, method 100 (or one, two, or three of steps 110, 120, and 130) is performed within a period of at least about 1 second, 2 seconds, 3 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 11 seconds, 12 seconds, 13 seconds, 14 seconds, 15 seconds, 16 seconds, 17 seconds, 18 seconds, 19 seconds, 20 seconds, 30 seconds, 40 seconds, 50 seconds, 60 seconds, 70 seconds, 80 seconds, 90 seconds, 100 seconds, 110 seconds, 120 seconds, 130 seconds, 140 seconds, 150 seconds, 160 seconds, 170 seconds, 180 seconds, or more. In some embodiments, method 100 (or one, two, or three of steps 110, 120, and 130) is performed within a period of time that is within a range defined by any two of the preceding values.

[0037] In some embodiments, the rapid performance of method 100 results in relatively low loss of nuclear spin polarization in the molecules of interest. For example, in some embodiments, immediately after step 110, at least one nucleus of the hyperpolarized molecules of interest has a first nuclear spin polarization. In some embodiments, before step 130, at least one nucleus of the hyperpolarized molecules of interest has a second nuclear spin polarization. In some embodiments, the second nuclear spin polarization is reduced by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or less from the first nuclear spin polarization. That is, in some embodiments, the second nuclear spin polarization is 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or 50% or more of the first nuclear spin polarization. In some embodiments, the second nuclear spin polarization is reduced from the first nuclear spin polarization by 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, or 5% or more. That is, in some embodiments, the second nuclear spin polarization is 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% or less of the first nuclear spin polarization. In some embodiments, the second nuclear spin polarization is reduced from the first nuclear spin polarization by an amount that is within a range defined by any two of the aforementioned values.For example, in some embodiments, the second nuclear spin polarization is about 50% to about 95%, about 50% to about 90%, about 50% to about 85%, about 50% to about 80%, about 50% to about 75%, about 50% to about 70%, about 50% to about 65%, about 50% to about 60%, about 50% to about 55%, about 55% to about 95%, about 55% to about 90%, about 55% to about 85%, about 55% to about 80%, about 55% to about 75%, about 55% to about 70%, about 55% to about 65%, about 55% to about 60%, about 60% to about 95%, about 60% to about 90%, about 60% to about 85%, or about 60% to about 80% of the first nuclear spin polarization. , about 60% to about 75%, about 60% to about 70%, about 60% to about 65%, about 65% to about 95%, about 65% to about 90%, about 65% to about 85%, about 65% to about 80%, about 65% to about 75%, about 65% to about 70%, about 70% to about 95%, about 70% to about 90%, about 70% to about 85%, about 70% to about 80%, about 70% to about 75%, about 75% to about 95%, about 75% to about 90%, about 75% to about 85%, about 75% to about 80%, about 80% to about 95%, about 80% to about 90%, about 80% to about 85%, about 85% to about 95%, about 85% to about 90%, or about 90% to about 95%.

[0038] In some embodiments, method 100 includes generating a hyperpolarized molecule of interest. In some embodiments, the hyperpolarized molecule of interest is generated before step 110. In some embodiments, the hyperpolarized molecule of interest is generated using at least one hyperpolarization procedure selected from the group consisting of PHIP, PHIP-SAH, PHIPNOESYS, and SABRE. For example, in some embodiments, the hyperpolarized molecule of interest is generated using a method described in any of WO2022 / 157534, WO2022 / 018514, WO2021 / 198776, WO2022 / 269350, US8,154,284, WO2022 / 162466, and PCT / IB2023 / 059050, each of which is incorporated by reference in its entirety for all purposes.

[0039] In some embodiments, hyperpolarized molecules of interest include any hyperpolarized biologically relevant imaging agent described herein, hi some embodiments, hyperpolarized molecules of interest are selected from the group consisting of dimethyl maleate, pyruvate, glutamate, glutamine, lactate, acetate, acetoacetate, zymonate, alanine, fructose, fumarate, bicarbonate, urea, dehydroascorbate, alpha-ketoglutarate, dihydroxyacetone, glucose, ascorbate, and their conjugate acids.

[0040] Compositions characterized by increased concentrations of hyperpolarized molecules of interest in solution 2 illustrates a composition 200 featuring an increased concentration of hyperpolarized molecules of a molecule of interest in a solution, according to disclosed embodiments. In the example shown, composition 200 includes solution 210 and hyperpolarized molecules of interest 220. In some embodiments, solution 210 includes any solution described herein with respect to FIG. 1. In some embodiments, hyperpolarized molecules of interest 220 include any hyperpolarized molecules of interest described herein with respect to FIG. 1. In some embodiments, the solution is for use in any NMR or MRI procedure described herein with respect to FIG. 1, such as any PHIP, PHIP-SAH, PHIPNOESYS, or SABRE procedure described herein with respect to FIG. 1.

[0041] In some embodiments, composition 200 is produced using method 100 described herein with respect to FIG. 1 . That is, in some embodiments, composition 200 is produced by (a) obtaining an organic solution having hyperpolarized molecules of interest dissolved therein at a first concentration, and (b) evaporating at least a portion of the organic solution, thereby producing an organic solution having hyperpolarized molecules of interest dissolved therein at a second concentration that is higher than the first concentration. In some embodiments, the organic solution comprises any organic solution described herein with respect to FIG. 1 . In some embodiments, the first concentration comprises any first concentration described herein with respect to FIG. 1 . In some embodiments, the second concentration comprises any second concentration described herein with respect to FIG. 1 . In some embodiments, (b) comprises evaporating at least the portion by performing any evaporation procedure described herein with respect to FIG. 1 . In some embodiments, (a) and (b) are performed for any total period of time described herein with respect to FIG. 1 . In some embodiments, immediately after (a), at least one nucleus of the hyperpolarized molecules of interest has any first respective spin polarization described herein with respect to Figure 1. In some embodiments, immediately before use of the composition in an MRI or NMR procedure, at least one nucleus of the hyperpolarized molecules of interest has any second nuclear spin described herein with respect to Figure 1. In some embodiments, the composition is further produced, prior to (a), by producing the hyperpolarized molecules of interest using at least one hyperpolarization procedure described with respect to Figure 1. In some embodiments, the composition is further produced, following (b), by mixing the organic solution with water, thereby producing an aqueous solution having the hyperpolarized compound dissolved therein at any third concentration described herein with respect to Figure 1. In some embodiments, the aqueous solution is used in an NMR or MRI procedure.

[0042] Method for increasing nuclear spin polarization in molecules of interest polarized by the SABRE method - Patent Application 20070122997 3 illustrates an exemplary method 300 for increasing nuclear spin polarization in a molecule of interest polarized by the SABRE method, according to disclosed embodiments. Method 300 utilizes partial or full deuteration of the molecule of interest and / or places the molecule of interest in a relatively low magnetic field during the SABRE polarization procedure.

[0043] In step 310, a molecule of interest is obtained. In some embodiments, the molecule of interest is to be polarized via a SABRE polarization procedure. In some embodiments, the molecule of interest is dissolved in a solution. In some embodiments, the solution comprises an organic solution. In some embodiments, the organic solution comprises methanol, ethanol, a methanol-water mixture, an ethanol-water mixture, pyridine, chloroform, dichloromethane, acetone, dimethyl sulfoxide (DMSO), a sulfur-containing molecule, or any mixture thereof. In some embodiments, the molecule of interest contains at least one carbon-13 ( 13 C) or nitrogen-15( 15 N) atoms.

[0044] In some embodiments, the molecule of interest is a deuterium ( 2 H) enriched molecules. That is, in some embodiments, the molecules of interest are proton ( 1The molecule of interest comprises at least one deuterium atom at a chemical site on the molecule of interest where a proton (H) would typically be expected. In some embodiments, the molecule of interest comprises at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more deuterium atoms at chemical sites on the molecule of interest where a proton would typically be expected. In some embodiments, the molecule of interest comprises at most about 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 deuterium atom at chemical sites on the molecule of interest where a proton would typically be expected. In some embodiments, the molecule of interest is partially deuterated (i.e., contains fewer deuterium atoms than all chemical sites on the molecule of interest where a proton would typically be expected). In some embodiments, the molecule of interest is fully deuterated (i.e., contains deuterium atoms at all chemical sites on the molecule of interest where a proton would typically be expected).

[0045] In some embodiments, the molecule of interest comprises a partially or fully deuterated chemical analog of any biologically relevant imaging agent, hi some embodiments, the molecule of interest comprises a partially or fully deuterated chemical analog of pyruvate, alpha-ketoglutarate, Z-4-methyl-2-oxopent-3-enedioc acid (Z-OMPD), zymonate, urea, azidothymidine (AZT), metronidazole, trimethylphenylammonium (TMPA), pyridine, nicotinamide, diazirine tag, or their conjugate acids.

[0046] In step 320, the molecule of interest is positioned (i.e., placed) within a magnetic field. In some embodiments, the magnetic field has an average field strength B. In some embodiments, B is at least about 1 microtesla (μT), 2 μT, 3 μT, 4 μT, 5 μT, 6 μT, 7 μT, 8 μT, 9 μT, 10 μT, 20 μT, 30 μT, 40 μT, 50 μT, 60 μT, 70 μT, 80 μT, 90 μT, 100 μT, 200 μT, 300 μT, 400 μT, 500 μT, 600 μT, 700 μT, 800 μT, 900 μT, 1 millitesla (mT), 2 μT, 3 μT, 4 μT, 5 μT, 6 μT, 7 μT, 8 μT, 9 μT, 1 ... Tesla (mT), 2mT, 3mT, 4mT, 5mT, 6mT, 7mT, 8mT, 9mT, 10mT, 20mT, 30mT, 40mT, 50mT, 60mT, 70mT, 80mT, 90mT, 100mT, 200mT, 300mT, 400mT, 500mT, 600mT, 700mT, 800mT, 900mT, 1 Tesla (T), 2T, 3T, 4T, 5T, or more. In some embodiments, B0 is at most about 5T, 4T, 3T, 2T, 1T, 900mT, 800mT, 700mT, 600mT, 500mT, 400mT, 300mT, 200mT, 100mT, 90mT, 80mT, 70mT, 60mT, 50mT, 40mT, 30mT, 20mT, 10mT, 9mT, 8mT, 7mT, 6mT, 5mT, 4mT , 3 mT, 2 mT, 1 mT, 900 μT, 800 μT, 700 μT, 600 μT, 500 μT, 400 μT, 300 μT, 200 μT, 100 μT, 90 μT, 80 μT, 70 μT, 60 μT, 50 μT, 40 μT, 30 μT, 20 μT, 10 μT, 9 μT, 8 μT, 7 μT, 6 μT, 5 μT, 4 μT, 3 μT, 2 μT, 1 μT, or less. In some embodiments, B is within a range defined by any two of the foregoing values.For example, in some embodiments, B0 is from about 1 μT to about 1 T, from about 1 μT to about 500 mT, from about 1 μT to about 100 mT, from about 1 μT to about 50 mT, from about 10 μT to about 1 T, from about 10 μT to about 500 mT, from about 10 μT to about 100 mT, from about 10 μT to about 50 mT, from about 50 μT to about 1 T, from about 50 μT to about 500 mT, from about 50 μT to about 1 T B0 is 0 mT, about 50 μT to about 50 mT, about 100 μT to about 1 T, about 100 μT to about 500 mT, about 100 μT to about 100 mT, about 100 μT to about 50 mT, about 500 μT to about 1 T, about 500 μT to about 500 mT, about 500 μT to about 100 mT, about 1 mT to about 1 T, about 1 mT to about 500 mT, or about 1 mT to about 100 mT. In some embodiments, B0 is generated using a magnetic shield (such as a mu-metal magnetic shield) and a magnetic field source such as a solenoid, other electromagnetic coil, or permanent magnet.

[0047] In step 330, a coordination complex is formed between the molecule of interest, the SABRE catalyst or catalyst precursor, and parahydrogen. In some embodiments, the coordination complex is formed in a solution as described herein with respect to step 320. In some embodiments, the SABRE catalyst or catalyst precursor comprises an iridium (Ir) catalyst or catalyst precursor, such as [IrCl(COD)(IMes)], where COD is cis,cis-1,5-cyclododecadiene and IMes is a cobalt (Co) catalyst or catalyst precursor, such as 1,3-bis(2,4,6-trimethylphenyl)imidazol-2-ylidine or (MesCCC)Co-py. In some embodiments, step 330 is performed in a solution comprising the molecule of interest, the SABRE catalyst or catalyst precursor, and parahydrogen. In some embodiments, the parahydrogen is mixed into the solution using a mixing mechanism. In some embodiments, the mixing mechanism comprises a gas-liquid exchange mechanism. For example, the gas-liquid exchange mechanism can be a bubbler or a diffusion system. In some embodiments, the mixing mechanism includes a membrane adapted to allow diffusion of molecular hydrogen. In some embodiments, para-hydrogen is mixed into the solution at a pressure of at least about 1 bar, 2 bar, 3 bar, 4 bar, 5 bar, 6 bar, 7 bar, 8 bar, 9 bar, 10 bar, 20 bar, 30 bar, 40 bar, 50 bar, 60 bar, 70 bar, 80 bar, 90 bar, 100 bar, or more. In some embodiments, para-hydrogen is mixed into the solution at a pressure of at most about 100 bar, 90 bar, 80 bar, 70 bar, 60 bar, 50 bar, 40 bar, 30 bar, 20 bar, 10 bar, 9 bar, 8 bar, 7 bar, 6 bar, 5 bar, 4 bar, 3 bar, 2 bar, 1 bar, or less. In some embodiments, the para-hydrogen is mixed into the solution at a pressure that is within the range defined by any two of the aforementioned values. For example, in some embodiments, the para-hydrogen is mixed into the solution at a pressure of 1 bar to 50 bar.

[0048] In step 340, an oscillating magnetic field is applied to the coordination complex. In some embodiments, the oscillating magnetic field has a maximum field strength B1 and is applied to the coordination complex. In some embodiments, applying the oscillating magnetic field transfers spin order from parahydrogen to the molecule of interest. In some embodiments, transferring spin order from parahydrogen to the molecule of interest increases the nuclear spin polarization of at least one nucleus in the molecule of interest. In some embodiments, at least one nucleus is 13 C nucleus or 15 Contains N nuclei.

[0049] In some embodiments, B1 is at least about 0.1 μT, 0.2 μT, 0.3 μT, 0.4 μT, 0.5 μT, 0.6 μT, 0.7 μT, 0.8 μT, 0.9 μT, 1 μT, 2 μT, 3 μT, 4 μT, 5 μT, 6 μT, 7 μT, 8 μT, 9 μT, 10 μT, 20 μT, 30 μT, 40 μT, 50 μT, 60 μT, 70 μT, 80 μT, 90 μT, 100 μT, 200 μT, 300 μT, 400 μT, 500 μT, 600 μT, 700 μT, 800 μT, 900 μT, 1 mT, 2 mT, 3 mT, 4 mT, 5 mT, 6 mT, 7 mT, 8 mT, 9 mT, 10 mT, or more. In some embodiments, B1 is at most about 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 μT, 800 μT, 700 μT, 600 μT, 500 μT, 400 μT, 300 μT, 200 μT, 100 μT, 90 μT, 80 μT, 70 μT, 60 μT, 50 μT, 40 μT, 30 μT, 20 μT, 10 μT, 9 μT, 8 μT, 7 μT, 6 μT, 5 μT, 4 μT, 3 μT, 2 μT, 1 μT, 0.9 μT, 0.8 μT, 0.7 μT, 0.6 μT, 0.5 μT, 0.4 μT, 0.3 μT, 0.2 μT, 0.1 μT, or less. In some embodiments, B1 is within a range defined by any two of the aforementioned values. For example, in some embodiments, B1 is between about 0.1 μT and about 10 μT, about 0.1 μT and about 9 μT, about 0.1 μT and about 8 μT, about 0.1 μT and about 7 μT, about 0.1 μT and about 6 μT, about 0.1 μT and about 5 μT, about 0.1 μT and about 4 μT, about 0.1 μT and about 3 μT, about 0.1 μT and about 2 μT, about 0.1 μT and about 1 μT, about 0.1 μT and about 0.9 μT, about 0.1 μT and about 0.8 μT, about 0.1 μT and about 0.7 μT, about 0.1 μT and about 0.6 μT, about 0.1 μT and about 0.5 μT, or about 0. .1μT to approx. 0.4μT, approx. 0.1μT to approx. 0.3μT, approx. 0.1μT to approx. 0.2μT, approx. 0.2μT to approx. 10μT, approx. 0.2μT to approx. 9μT, approx. 0.2μT to approx. 8μT, approx. 0.2μT to approx. 7μT, approx. 0.2μT to approx. 6μT, approx. 0.2μT T ~ about 5μT, about 0.2μT - about 4μT, about 0.2μT - about 3μT, about 0.2μT - about 2μT, about 0.2μT - about 1μT, about 0.2μT - about 0.9μT, about 0.2μT - about 0.8μT, about 0.2μT - about 0.7μT, about 0.2μT - about 0.6 μT, approximately 0.2 μT to approximately 0.5 μT, approximately 0.2 μT to approximately 0.4 μT, approximately 0.2 μT to approximately 0.3 μT, approximately 0.3 μT to approximately 10 μT, approximately 0.3 μT to approximately 9 μT, approximately 0.3 μT to approximately 8 μT, approximately 0.3 μT to approximately 7 μT, approximately 0.3 μT to approximately 6 μT, approximately 0.3 μT to approximately 5 μT, approximately 0.3 μT T ~ approximately 4 μT, approximately 0.3 μT ~ approximately 3 μT, approximately 0.3 μT ~ approximately 2 μT, approximately 0.3 μT ~ approximately 1 μT, approximately 0.3 μT ~ approximately 0.9 μT, approximately 0.3 μT ~ approximately 0.8 μT, approximately 0.3 μT ~ approximately 0.7 μT, approximately 0.3 μT ~ approximately 0.6 μT, approximately 0.3 μT ~ approximately 0.5 μT, approximately 0.3 μT ~ approximately 0.4 μT μT, approximately 0.4μT to approximately 10μT, approximately 0.4μT to approximately 9μT, approximately 0.4μT to approximately 8μT, approximately 0.4μT to approximately 7μT, approximately 0.4μT to approximately 6μT, approximately 0.4μT to approximately 5μT, approximately 0.4μT to approximately 4μT, approximately 0.4μT to approximately 3μT, approximately 0.4μT to approximately 2μT, approximately 0.4μT to approximately 1μT, Approximately 0.4 μT to approximately 0.9 μT, approximately 0.4 μT to approximately 0.8 μT, approximately 0.4 μT to approximately 0.7 μT, approximately 0.4 μT to approximately 0.6 μT, approximately 0.4 μT to approximately 0.5 μT, approximately 0.5 μT to approximately 10 μT, approximately 0.5 μT to approximately 9 μT, approximately 0.5 μT to approximately 8 μT, approximately 0.5 μT to approximately 7 μT, approximately 0.5 μT ~approx. 6μT, approx. 0.5μT~approx. 5μT, approx. 0.5μT~approx. 4μT, approx. 0.5μT~approx. 3μT, approx. 0.5μT~approx. 2μT, approx. 0.5μT~approx. 1μT, approx. 0.5μT~approx. 0.9μT, approx. 0.5μT~approx. 0.8μT, approx. 0.5μT~approx. 0.7μT, approx. 0.5μT~approx. 0.6μT, approx. 0 0.6μT to approximately 10μT, approximately 0.6μT to approximately 9μT, approximately 0.6μT to approximately 8μT, approximately 0.6μT to approximately 7μT, approximately 0.6μT to approximately 6μT, approximately 0.6μT to approximately 5μT, approximately 0.6μT to approximately 4μT, approximately 0.6μT to approximately 3μT, approximately 0.6μT to approximately 2μT, approximately 0.6μT to approximately 1μT, approximately 0.6μT ~approx. 0.9μT, approx. 0.6μT~approx. 0.8μT, approx. 0.6μT~approx. 0.7μT, approx. 0.7μT~approx. 10μT, approx. 0.7μT~approx. 9μT, approx. 0.7μT~approx. 8μT, approx. 0.7μT~approx. 7μT, approx. 0.7μT~approx. 6μT, approx. 0.7μT~approx. 5μT, approx. 0.7μT~approx. 4μT, approx. 0. 7μT to approximately 3μT, approximately 0.7μT to approximately 2μT, approximately 0.7μT to approximately 1μT, approximately 0.7μT to approximately 0.9μT, approximately 0.7μT to approximately 0.8μT, approximately 0.8μT to approximately 10μT, approximately 0.8μT to approximately 9μT, approximately 0.8μT to approximately 8μT, approximately 0.8μT to approximately 7μT, approximately 0.8μT to approximately 6μT, approximately 0.8μT to about 5μT, about 0.8μT to about 4μT, about 0.8μT to about 3μT, about 0.8μT to about 2μT, about 0.8μT to about 1μT, about 0.8μT to about 0.9μT, about 0.9μT to about 10μ T, about 0.9μT to about 9μT, about 0.9μT to about 8μT, about 0.9μT to about 7μT, about 0.9μT to about 6μT, about 0.9μT to about 5μT, about 0.9μT to about 4μT, about 0.9μT to about 3μT, about 0.9μT to about 2μT, about 0.9μT to about 1μT, about 1μT to about 10μT, about 1μT to about 9μT, about 1μT to about 8μT, about 1μT to about 7μT, about 1μT to about 6μT, about 1μT T ~ approx. 5 μT, approx. 1 μT ~ approx. 4 μT, approx. 1 μT ~ approx. 3 μT, approx. 1 μT ~ approx. 2 μT, approx. 2 μT ~ approx. 10 μT, approx. 2 μT ~ approx. 9 μT, approx. 2 μT ~ approx. 8 μT, approx. ~Approx. 6μT, approx. 2μT ~ approx. 5μT, approx. 2μT ~ approx. 4μT, approx. 2μT ~ approx. 3μT, approx. 3μT ~ approx. 10μT, approx. 3μT ~ approx. 9μT, approx. 3μT ~ approx. 8μT, approx. 3μT ~ approx. 7μT, approx. 3μT ~ Approximately 6μT, approximately 3μT to approximately 5μT, approximately 3μT to approximately 4μT, approximately 4μT to approximately 10μT, approximately 4μT to approximately 9μT, approximately 4μT to approximately 8μT, approximately 4μT to approximately 7μT, approximately 4μT to approximately 6μT, approximately 4μT to approximately 5 μT, about 5 μT to about 10 μT, about 5 μT to about 9 μT, about 5 μT to about 8 μT, about 5 μT to about 7 μT, about 5 μT to about 6 μT, about 6 μT to about 10 μT, about 6 μT to about 9 μT, about 6 μT to about 8 μT, about 6 μT to about 7 μT, about 7 μT ​​to about 10 μT, about 7 μT ​​to about 9 μT, about 7 μT ​​to about 8 μT, about 8 μT to about 10 μT, about 8 μT to about 9 μT, or about 9 μT to about 10 μT.

[0050] In some embodiments, the oscillating magnetic field is generated by a waveform generator, which may further include one or more computing units, processors, controllers, associated memory, PCs, computer servers, or any device capable of conducting computations using inputs and generating outputs.

[0051] The oscillating magnetic field can be applied to the coordination complex using an RF coil. The RF coil can be disposed around the chamber in which the SABRE polarization transfer was performed. The RF coil can have one or more channels. Such channels can be paths for applying RF signals to the coordination complex. At least one channel can be provided for each different type of nuclear spin species. In some embodiments, at least one proton ( 1 H) channel, and / or another nuclear spin species (e.g., 2 H, 3 H, 13 C. 15 N, 19 F, 31 In some embodiments, there may be at least one channel for a specific species (e.g., P, or other suitable species). 1 The RF waveforms applied to the H channel and each of the channels for the other nuclear spin species may be different. In some embodiments, at least one proton channel is 1 In some embodiments, the H spin is selectively addressed by the 1 RF waveforms on the H channel, and / or other nuclear spin species (e.g., 2 H, 13 C, and / or 15 Each of the channels for (N channels) is configured to apply a polarization transfer sequence, such as PulsePol, ADAPT, PH-INEPT, Goldman's sequence, S2M, S2hM, or ESOTERIC.

[0052] In some embodiments, any one, two, three, or four of steps 310, 320, 330, and 340 are performed with a magnetic shield, such as a mu-metal magnetic shield.

[0053] In some embodiments, method 300 imparts nuclear spin polarization to at least one nucleus of a molecule of interest, hi some embodiments, the nuclear spin polarization is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more. In some embodiments, the nuclear spin polarization is at most about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, hi some embodiments, the nuclear spin polarization is within a range defined by any two of the foregoing values.

[0054] In some embodiments, method 300 is performed at a temperature of at least about -30 degrees Celsius (°C), -25°C, -20°C, -15°C, -10°C, -5°C, 0°C, 5°C, 10°C, 15°C, 20°C, or higher, at most about 20°C, 15°C, 10°C, 5°C, 0°C, -5°C, -10°C, -15°C, -20°C, -25°C, -30°C, or lower, or within a range defined by any two of the foregoing values.

[0055] In some embodiments, the SABRE catalyst or catalyst precursor can be extracted from the solution, or undesired solutes can be removed from the solution. The disclosed embodiments are not limited to any particular method of separating the SABRE catalyst or catalyst precursor. In some embodiments, separation is performed by liquid-liquid separation. In some embodiments, the SABRE catalyst or catalyst precursor (or undesired solutes) can be extracted from the solution by precipitation or solidification. Such precipitation or solidification can be induced to reduce the solubility of the SABRE catalyst or catalyst precursor in the solution. In some embodiments, the solution can be modified (e.g., by changing the solution's pH, temperature, adding another solvent or solvents, or the like). In some embodiments, electromagnetic stimulation (e.g., optical radiation such as ultraviolet light or optical radiation at another suitable wavelength or wavelengths) or mechanical stimulation (e.g., ultrasound, stirring, or another suitable mechanical stimulation), adding another solute or solvent to the solution, or applying another suitable method. In some embodiments, after precipitation, the SABRE catalyst or catalyst precursor can be separated from the solution (e.g., using a filter, centrifuge, or another suitable method).

[0056] In some embodiments, the molecule of interest may be used in NMR or MRI applications. In some embodiments, at least a portion of the molecule of interest may be injected into a subject or patient for use in MRI imaging of the subject or patient. In various embodiments, at least a portion of the molecule of interest may be used in NMR spectroscopy.

[0057] Compositions characterized by increased nuclear spin polarization in molecules of interest polarized by the SABRE method 4 illustrates a composition 400 featuring increased nuclear spin polarization in a molecule of interest polarized by the SABRE method, according to disclosed embodiments. In the example shown, composition 400 includes a solution 410 and a hyperpolarized molecule of interest 420. In some embodiments, solution 410 includes any solution described herein with respect to FIG. 3. In some embodiments, hyperpolarized molecule of interest 420 includes any hyperpolarized molecule of interest described herein with respect to FIG. 3. In some embodiments, the solution is for use in any NMR or MRI procedure described herein with respect to FIG. 3, such as any SABRE procedure described herein with respect to FIG. 3.

[0058] In some embodiments, composition 400 is produced using method 300 described herein with respect to Figure 3. That is, in some embodiments, composition 400 is produced by (a) obtaining a molecule of interest that includes at least one deuterium atom, (b) placing the molecule of interest in a magnetic field having an average field strength (B) of at most about 2 Tesla (T), (c) forming a coordination complex between the molecule of interest, a signal amplification by reversible exchange (SABRE) catalyst or catalyst precursor, and para-hydrogen, and (d) applying an oscillating magnetic field to the coordination complex, thereby transferring spin order from the para-hydrogen to the molecule of interest, thereby increasing the nuclear spin polarization of at least one atom in the molecule of interest.

[0059] In some embodiments, the molecule of interest includes any molecule of interest described herein with respect to FIG. 3. In some embodiments, the average magnetic field strength is any average magnetic field strength described herein with respect to FIG. 3. In some embodiments, the SABRE catalyst or catalyst precursor includes any SABRE catalyst or catalyst precursor described herein with respect to FIG. 3. In some embodiments, the oscillating magnetic field includes any oscillating magnetic field described herein with respect to FIG. 3. In some embodiments, (a)-(d) are performed with a magnetic shield. In some embodiments, (c) is performed in a solution including the molecule of interest, the SABRE catalyst or catalyst precursor, and parahydrogen. In some embodiments, the composition is generated by bubbling parahydrogen into the solution. In some embodiments, the parahydrogen is bubbled into the solution at any of the pressures described herein with respect to FIG. 3. In some embodiments, following (d), the nuclear spin polarization in the molecule of interest is any nuclear spin polarization described herein with respect to FIG. 3.

[0060] Bio-related contrast agents Disclosed embodiments include methods for producing and utilizing biorelevant imaging agents with clinically relevant polarization, concentration, volume, or purity. In some embodiments, the methods are for preparing NMR materials (also referred to herein as "molecules of interest"). In some embodiments, the NMR materials are suitable for use in NMR or MRI operations. In some embodiments, the NMR materials increase NMR or MRI signal and signal-to-noise ratio (SNR). In some embodiments, the NMR materials are suitable for use in solution NMR spectroscopy. In some embodiments, the NMR materials are chemical compounds. In some embodiments, the NMR materials are metabolites (e.g., molecules with biorelevance, such as amino acids, sugars, and derivatives thereof), such as metabolites suitable for use in NMR metabolomics applications. In some embodiments, the NMR materials are suitable for in vitro probing of metabolism in cell cultures or other biological tissues. In some embodiments, the NMR materials are used in NMR probes to investigate transient effects where high signal enhancement due to hyperpolarization is required, such as proton exchange between water and biomolecules. In some embodiments, the NMR material is a small molecule or metabolite suitable for injection into a cell, tissue, or organism for detection in an MRI scan. In some embodiments, the NMR material is introduced into a chamber for further analysis by NMR or MRI operation. In some embodiments, the NMR material is one or more deuterium ( 2 H), carbon-13( 13 C), or nitrogen-15( 15 N) atoms.

[0061] Consistent with disclosed embodiments, the NMR material may include a biorelevant contrast agent. In some embodiments, the biorelevant contrast agent may be suitable for use in NMR or MRI operations. In some embodiments, the biorelevant contrast agent may increase the NMR or MRI signal or signal-to-noise ratio (SNR). In some embodiments, the biorelevant contrast agent may be suitable for use in solution NMR spectroscopy. In some embodiments, the biorelevant contrast agent may be a metabolite (e.g., a molecule with biorelevance, such as an amino acid, sugar, or derivative thereof), such as a metabolite suitable for use in NMR metabolomics applications. In some embodiments, the biorelevant contrast agent is used for perfusion or contrast-enhanced imaging in MRI scans. In some embodiments, the biorelevant contrast agent may be suitable for in vitro probing of the metabolism of cell cultures or other biological tissues. In some embodiments, the biorelevant contrast agent is used for in vitro probing of the metabolism of cell cultures or other biological tissues. In some embodiments, the biorelevant contrast agent may be used in NMR probes to investigate transient effects where high signal enhancement due to hyperpolarization is required, such as proton exchange between water and biomolecules. In some embodiments, the biorelevant imaging agent may be a small molecule or metabolite suitable for injection into a cell, tissue, or organism for detection in an MRI scan. In some embodiments, the biorelevant imaging agent may be introduced into a chamber for further analysis by NMR or MRI operation. In some embodiments, the biorelevant imaging agent may be one or more 2 H, 13 C, or 15 It is enriched in N atoms.

[0062] In some embodiments, the biologically relevant imaging agent is pyruvate, lactate, alpha-ketoglutarate, bicarbonate, fumarate, urea, dehydroascorbate, glutamate, glutamine, acetate, dihydroxyacetone, acetoacetate, glucose, ascorbate, zymonate, alanine, fructose, imidazole, nicotinamide, nitroimidazole, pyrazinamide, isoniazid, conjugate acids of any of the above, natural and unnatural amino acids, esters thereof, or any of the foregoing. 2 H, 13 C, or nitrogen-15( 15 In some embodiments, the biologically relevant imaging agent comprises pyruvate, lactate, alpha-ketoglutarate. In some embodiments, the biologically relevant imaging agent comprises pyruvate. In some embodiments, the biologically relevant imaging agent comprises lactate. In some embodiments, the biologically relevant imaging agent comprises alpha-ketoglutarate (e.g., ethyl alpha-ketoglutarate).

[0063] In some embodiments, the biologically relevant imaging agent comprises at least one non-hydrogen nuclear spin. In some embodiments, the non-hydrogen nucleus comprises at least one spin 1 / 2 atom. In some embodiments, the non-hydrogen nuclear spin is 13 C or 15N. In some embodiments, the biologically relevant imaging agent is at least partially isotopically labeled with non-hydrogen nuclear spins. In some embodiments, the biologically relevant imaging agent is at least partially enriched in non-hydrogen nuclear spins compared to an analog of the biologically relevant imaging agent characterized by non-hydrogen nuclear spins at their natural abundance. In some embodiments, the biologically relevant imaging agent is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or at most about 99%, 98%, 97%, 96%, 98%, 99%, or more. %, 95%, 94%, 93%, 92%, 91%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less abundance, or an abundance within a range defined by any two of the foregoing values.

[0064] In some embodiments, the non-hydrogen nuclear spins are NMR inactive (i.e., spin 0) nuclei of an analog of a biologically relevant imaging agent that characterizes the non-hydrogen nuclear spins at their natural abundance (e.g., 12 C or quadrupolar (i.e., spin >1 / 2) nuclei (e.g., nitrogen-14, 14 N) at their natural abundance. 13 The analogue of pyruvate that characterizes C has the structure H3C-C * (=O)-C * OOH either C * So, about 98.9% 12 C and about 1.1% 13 As a biorelevant imaging agent, pyruvate may contain one or both of the C * at any abundance described herein 13 To include C, instead 13 As used herein, * C and C * teeth,12 C or 13 C describes carbon, which can be any of the carbon isotopes. 15 The analogue of urea that characterizes N is the structure H2N * -C(=O)- * Any N in NH2 * Approximately 99.6% 14 N and about 0.4% 15 As a biorelevant imaging agent, urea may contain one or both N * at any abundance described herein 15 To include N, instead 15 As used herein, * N and N * teeth, 14 N or 15 Describes nitrogen, which can be any of the nitrogen isotopes N. [Example]

[0065] Example 1: Increased concentration of a molecule of interest in solution Experiments were performed using dimethylacetylenedicarboxylic acid (DMAD, also known as dimethyl 2-butynedioate). DMAD is a precursor that, when hydrogenated, yields dimethyl maleate (DMM). The goal of the initial experiments was to demonstrate that it is possible to reduce the solution concentration by at least two-fold (e.g., 2-2.5-fold) while retaining sufficient nuclear spin polarization and having enough solution for automated injection into an NMR spectrometer.

[0066] In the first experiment, evaporation was performed using a manual procedure. 0.5 M DMAD precursor was dissolved in acetone-d6. 1 milliliter (mL) of precursor was injected into the reactor. The solution was heated to 50 degrees Celsius (°C). The solution was purged with nitrogen (N2) gas at 4 bar for 3 seconds. The precursor was then hydrogenated in the solution at 10 bar for 5 seconds. The solution was purged again with N2 gas for 3 seconds. The reactor was manually opened to vacuum just before the heater on the reactor increased the temperature for boil-off. The reactor was then closed to vacuum, vented, and the remaining solution was manually extracted. The composition was characterized by NMR.

[0067] Figure 5 shows the results of a manual test with 1 mL of precursor solution, manual extraction of the remaining solution from the reactor, and characterization of the evaporation by NMR. As shown in Figure 5, evaporation allowed the concentration of DMAD to be increased by 2.4 to 7.8 times, depending on the temperature, vacuum, and time conditions.

[0068] In the second experiment, evaporation was performed using an automated system. 0.2 M DMAD precursor was dissolved in acetone-d6. 2 mL of precursor was injected into the reactor. The solution was heated to 50 °C. The solution was purged with N2 gas at 4 bar for 3 seconds. The precursor was then parahydrogenated in the solution using parahydrogen gas at 10 bar for 5 seconds. The solution was purged again with N2 gas for 3 seconds. A heater increased the temperature for boil-off. An audio frequency sweep was applied to convert the singlet state polarization to magnetization. The system opened the reactor to vacuum for boil-off. The reactor was closed to vacuum, vented, and the solution was injected into the NMR spectrometer for measurement.

[0069] Figure 6 shows the results of an automated test using 2 mL of precursor solution. As shown in Figure 6, evaporation allowed the concentration of DMAD to be increased by 1.7 to 2.415 times, depending on the temperature, vacuum, and time conditions.

[0070] Figure 7 shows deuterium and carbon labeled DMAD (d6, 13C) Results of an automated test using 2 mL of precursor solution. As shown in Figure 7, evaporation allowed for a 2.405- to 2.59-fold increase in the concentration of DMAD (indicated by the white bars) while maintaining a nuclear spin polarization of 1.3% to 2.6% (indicated by the solid bars), depending on the temperature, vacuum, and time conditions.

[0071] Example 2: Increased nuclear spin polarization in molecules of interest polarized by the SABRE method We investigated methods to increase the hyperpolarization of pyruvate produced by the SABRE process. 13 The polarization transfer to C is mediated by indirect dipolar coupling (J-coupling) between nuclei in the transient complex. For SABRE, the ultra-low magnetic field regime (e.g., average magnetic field magnitude of several hundred nanoteslas relative to the sample) plays a crucial role. In these magnetic fields, the nuclear spin energy states of the molecules are coincident, and the J-coupling interactions effectively perturb the eigenstates such that energy level crossings are avoided (i.e., the system energy state displays level anticrossings (LACs)). When the SABRE reaction is carried out at LACs, the target 13 The C spins are spontaneously hyperpolarized. This LAC-dependent method is called SABRE-SHEATH (SABRE in the shield allows alignment transfer to heteronuclei), and [1- 13 More recently, pulsed static magnetic fields have been used to induce this effect, resulting in the polarization of [1-C]pyruvate. 13 C]pyruvate as well as SABRE-SHEATH 13 C polarization (P 13C ) delivered to the target. Pulsed radio frequency (rf) magnetic fields can be employed for polarization transfer. Spin-lock-induced crossover by continuous rf irradiation in Tesla magnetic fields (SLIC) has also been applied with SABRE and was called low-irradiation high-Tesla generation (LIGHT-) or SLIC-SABRE. However, the P achieved by high-field SLIC-SABRE 13C was found to be lower than that obtained via SABRE-SHEATH.13C This may arise from the fact that SLIC at high (e.g., hundreds of millitesla to several tesla) magnetic fields faces significant challenges, such as often unfavorable relaxation dynamics due to chemical shift anisotropy (CSA), singlet-to-triplet mixing of parahydrogen, and the need for selective excitation of bound and free SABRE species due to larger chemical shift dispersions.

[0072] In stark contrast, at much lower magnetic fields (e.g., a few microteslas), the difference between bound and free SABRE species is 13 The difference in the chemical shift of C is negligible, favoring relaxation times. Therefore, we used SLIC-SABRE in a microtesla magnetic field to measure [1- 13 C]pyruvate-d3 and [2- 13 [C]pyruvate-d3 was rapidly hyperpolarized and P 13C =22% and P 13C =6% 13 C polarization was obtained. 13C To achieve this level, we performed the SABRE reaction by bubbling parahydrogen gas through a solution of approximately 30 mM pyruvate-d3 in CD3OD at 50 μT and approximately 3-7°C in the presence of a spin-lock-induced crossover (SLIC) rf magnetic field. 13 used with C abundance (1.1%) to minimize sample-to-sample variability, 13 P between the C-labeled sample and the natural abundance sample 13C No difference in values ​​was observed. After the SLIC process, an adiabatic 90° pulse was used to measure the densities along a 50 μT magnetic field. 13 The C polarization is aligned vertically and the sample is taken from the SABRE setup. 、13 The system was then transferred to an 80 MHz benchtop NMR system for C signal acquisition. A detailed description of the setup and method is presented below.

[0073] FIG. 8 shows the polarized [1- 13C]pyruvate-d3 and [2- 13 Illustrative of [C]pyruvate-d3 13 As shown in Figure 8, the SLIC-SABRE method dramatically outperforms the SABRE-SHEATH method, yielding pyruvate polarization levels that are 2-6 times higher than any other previous SABRE process.

[0074] To better understand why SLIC-SABRE exhibits such high efficacy in microtesla fields, we performed experiments using both protonated and deuterated pyruvate, and using both SLIC-SABRE and SABRE-SHEATH. Figure 9 shows the results of protonated and deuterated [1- 13 C]pyruvate and [2- 13 Illustrative examples for [C]pyruvate 13 C polarization level. 13 C]pyruvate and [2- 13 [C]pyruvate had similar P for each polarization protocol 13C The value is [1- 13 C]pyruvate, 12% vs. 9%, [2- 13 For [C]pyruvate, the results were 2% in both cases. However, we found that SLIC-SABRE performed much better for pyruvate-d3. P using SABRE-SHEATH (0.4 μT) 13C is [1- 13 C]pyruvate-d3 and [2- 13 C]pyruvate-d3 was 4% and 0.2%, but P using SLIC-SABRE 13C increased to 22% and 6%, respectively.

[0075] Without limiting the scope of this disclosure, the inventors believe that these results occur because deuterium nuclei have two distinct effects on relaxation. First, at very low magnetic fields, deuterium 13The C2 carbon is strongly coupled to the C spin and acts as a quadrupolar relaxation sink. This is the case in SABRE-SHEATH, and is particularly detrimental to the C2 carbon, which has a stronger J-coupling to deuterium than the C1 carbon. Second, deuterium is more sensitive to the C2 carbon because the magnetic moment of deuterium is smaller than the magnetic moment of the proton. 13 This effect is seen in the higher 13 C polarization and more favorable polarization accumulation kinetics. To support this hypothesis, we investigated the measured T and T 1ρ The values ​​are also presented, and T at 50 μT 1ρ We find that for all investigated pyruvate isotopes (shown in Figure 11), T is longer than T at 0.4 μT, ultimately allowing for effective polarization accumulation. Therefore, T under SABRE-SHEATH and T under SLIC-SABRE conditions are 1ρ For pyruvate-d3 isotopes, which are almost identical to each other, much higher polarizations are achieved using SLIC-SABRE. We envision that further analysis of polarization transfer in the SABRE exchange complex during SLIC and experimental parameter optimization (such as reaction temperature, static magnetic field, and SLIC rf field) will enable even greater signal enhancement.

[0076] In particular, deuterium isotope labeling can also prolong T relaxation at clinical fields. The longer polarization lifetime obtained by deuteration can facilitate the purification, quality assurance, and post-injection high P of hyperpolarized pyruvate or other biorelevant contrast agents. 13C Importantly, no deuterium isotope effect on metabolic conversion kinetics was observed for pyruvate, implying that metabolic MRI pyruvate-d3 can be successfully used in place of non-deuterated pyruvate. In particular, isotopically enriched [1- 13 C]pyruvate-d3 and [2- 13Both [C]pyruvate-d3 and [C]pyruvate-d3 are commercially available, and no further chemical modification is required for SABRE. This is in stark contrast to parahydrogen-based hyperpolarization approaches, which require suitable unsaturated molecular precursors. Additionally, due to the low cost and simplicity of the SABRE hardware, this approach is widely accessible to researchers. Given recent advances in rapidly extracting SABRE-polarized pyruvate from methanol into catalyst-free aqueous solutions, the first preclinical metabolic MRI studies using this technique are expected to be performed soon.

[0077] For all experiments, samples containing 30 mM sodium pyruvate or sodium pyruvate-d3 in methanol-d4, 6 mM IrIMes(cod)Cl, and 40 mM dimethyl sulfoxide (DMSO) were prepared. After mixing the components, the samples were sonicated for 5 minutes (min). If residual particles were visible in the solution, they were filtered through a 0.45 μm polytetrafluoroethylene (PTFE) syringe filter. The solution was then degassed by bubbling N2 gas through it. IrIMes(cod)Cl SABRE catalysts were synthesized according to known procedures. Each sample was activated by bubbling parahydrogen gas through it at 10 bar for 10 minutes.

[0078] The sample was loaded into an NMR pressure tube and connected to a gas system that allowed automatic injection of gas. For the generation of hyperpolarization, the sample was placed in an apparatus consisting of a mu-metal shield (to shield from external magnetic fields), a through-hole solenoid (to provide a static magnetic field along the z-axis B0), and a coil aligned along the x-axis (B1) (to generate a linearly polarized oscillating magnetic field along the x-axis).

[0079] For the SABRE-SHEATH experiment, a static magnetic field of 400 nanotesla (nT) was applied for a bubbling time t bThe magnetic field was then rapidly increased to 30 μT to stop the SABRE hyperpolarization process and allow sample extraction through the mu-metal shield. For SLIC-SABRE, a static magnetic field of 50 μT was applied to the sample. To facilitate polarization transfer, an oscillating B1 field of amplitude 1.8 μT was applied during the bubbling of parahydrogen. 13 A magnetic field was applied to the sample at the Larmor frequency of the C spins (535.25 Hz). During this time, a rotating magnetic field in the xy plane was generated. The magnetization was then rotated parallel to the z-axis using a half-pass adiabatic 90-degree pulse with a starting amplitude of 4 μT. The pulse amplitude was linearly decayed from 4 μT to 0 within 2 seconds. Simultaneously, the frequency of the B1 magnetic field was linearly increased to 50 Hz.

[0080] All experiments were automated by the Qudi software package for experimental control. After the accumulation of hyperpolarization, 13 For measurement of C NMR spectra, samples were manually transferred to a Bruker F80 benchtop NMR spectrometer (Bruker).

[0081] After completion of the SABRE experiment, the concentrations of free and bound pyruvate in solution were determined as follows: pyruvate and pyruvate-d3, respectively. 1 H NMR or 2 The nuclear spin polarization of hyperpolarized samples was determined by H NMR. For pyruvate, an external reference standard consisting of 40 mM DMSO in methanol was used. For pyruvate-d3, the integral of the CD3 line of the solvent was used as the reference. 13 C was determined by calibration against methanol (99%). Polarization P hyp , P hyp =S hyp / S ref ·[C ref ] / [C hyp ]·P ref The calculation was performed according to the following formula: where S hyp , S ref , [C ref ], and [C hyp ] are the signal and concentration of the hyperpolarized and reference samples, respectively. Pref is the thermal equilibrium Boltzmann polarization of the reference sample in the 1.9 T measurement field of the F80 spectrometer at room temperature.

[0082] For the measurement of T1 at 1.88 T, the hyperpolarized sample was inserted into the F80 NMR spectrometer. 13 C NMR spectra were recorded every 10 seconds using a 10 degree flip angle for excitation. 13 The dissipation of the C signal was subsequently corrected before fitting the data. For T measurements at 0.4 μT, the sample was hyperpolarized using SABRE-SHEATH or SLIC-SABRE for the generation of the hyperpolarized signal. Note that SLIC-SABRE was used for the hyperpolarization of deuterated pyruvate isotopes due to its higher polarization yield. The parahydrogen supply to the sample was then stopped and the sample was left for an additional 20 seconds to minimize the effect of any residual accumulation of SABRE magnetization during the relaxation delay. The magnetic field was then reduced to 0.4 μT and the sample was placed in a 0.4 μT magnetic field. 13 The samples were allowed to relax for different times before being transferred to an NMR spectrometer for measurement of the C NMR spectra. This procedure was repeated at each reported time point. 1ρ The data were measured in the same way, but now the sample was held in an external magnetic field of 50 μT, the magnetization was reversed along the z-axis of the static magnetic field, and the sample was 13 Before transfer to NMR for acquisition of C spectra, the magnetization was spin-locked with an amplitude of B1 = 1.86 μT for different times as reported.

[0083] To quantify the error in the SABRE experiment using the setup and sample preparation, we typically repeated the SLIC-SABRE hyperpolarization of pyruvate-d3 N = 5 times. Each sample was prepared independently. The pyruvate concentration was determined and the polarization quantified as described in the Experimental section above. We found an average [1- 13 C]pyruvate-d3 polarization was found.

[0084] The foregoing description has been presented for purposes of illustration. It is not exhaustive and is not limited to the precise forms or embodiments disclosed. Modifications and adaptations of the embodiments will be apparent from consideration of the specification and practice of the disclosed embodiments. For example, while the described implementations include hardware, systems and methods consistent with the present disclosure can be implemented using both hardware and software. Further, while certain components are described as being coupled to each other, such components may be integrated with each other or distributed in any suitable manner.

[0085] Moreover, while exemplary embodiments are described herein, the scope includes any and all embodiments having equivalent elements, modifications, omissions, combinations (e.g., of aspects across various embodiments), adaptations, or alterations based on this disclosure. Claim elements should be construed broadly based on the language used in the claims and not limited to the examples described herein or during prosecution of the application, which examples should be construed as non-exclusive. Furthermore, the steps of the methods of the present disclosure can be modified in any manner, including rearranging steps or inserting or deleting steps.

[0086] The features and advantages of the present disclosure will be apparent from the detailed specification, and accordingly, the appended claims are intended to cover all systems and methods that fall within the true spirit and scope of the present disclosure. As used herein, the indefinite articles "a" and "an" mean "one or more." Similarly, the use of a plural term does not necessarily imply a plurality, unless the use is ambiguous in a given context. Furthermore, since numerous modifications and variations will readily occur from a study of this disclosure, it is not desired to limit the disclosure to the exact construction and operation illustrated and described. Accordingly, all suitable modifications and equivalents may be resorted to, falling within the scope of the present disclosure.

[0087] As used herein, unless specifically stated otherwise, the term "or" includes all possible combinations unless impracticable. For example, if a component is stated to include A or B, the component may include A, or B, or A and B, unless specifically stated otherwise or impracticable. As a second example, if a component is stated to include A, B, or C, the component may include A, or B, or C, or A and B, or A and C, or B and C, or A and B and C, and C.

[0088] Enumeration of Embodiments Embodiment 1. A method for performing a magnetic resonance imaging (MRI) or nuclear magnetic resonance (NMR) procedure using a hyperpolarized molecule of interest, the method comprising: (a) obtaining an organic solution having hyperpolarized molecules of interest dissolved therein at a first concentration; (b) evaporating at least a portion of the organic solution, thereby producing an organic solution having the hyperpolarized molecules of interest dissolved therein at a second concentration greater than the first concentration; (c) performing an MRI or NMR procedure using the hyperpolarized molecule of interest.

[0089] Embodiment 2. The method of embodiment 1, wherein the first concentration is less than 100 millimolar (mM), 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, or less.

[0090] Embodiment 3. The method of embodiment 1 or 2, wherein the second concentration is at least 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, 200 mM, or more.

[0091] Embodiment 4. The method of any one of embodiments 1-3, wherein the organic solution comprises at least one organic molecule selected from the group consisting of methanol, ethanol, n-propanol, isopropanol, and acetone.

[0092] Embodiment 5. The method of any one of embodiments 1-4, wherein (b) comprises evaporating at least a portion by performing at least one evaporation procedure selected from the group consisting of flowing an inert gas over the organic solution, subjecting the organic solution to a vacuum, and heating the organic solution.

[0093] Embodiment 6. The method of any one of embodiments 1-5, wherein (a)-(b) are carried out for a total period of at most about 150 seconds, 140 seconds, 130 seconds, 120 seconds, 110 seconds, 100 seconds, 90 seconds, 80 seconds, 70 seconds, 60 seconds, 50 seconds, 40 seconds, 30 seconds, 20 seconds, 19 seconds, 18 seconds, 17 seconds, 16 seconds, 15 seconds, 14 seconds, 13 seconds, 12 seconds, 11 seconds, 10 seconds, 9 seconds, 8 seconds, 7 seconds, 6 seconds, 5 seconds, 4 seconds, 3 seconds, 2 seconds, 1 second, or less.

[0094] Embodiment 7. The method of any one of embodiments 1-6, wherein immediately after (a), at least one nucleus of the hyperpolarized molecule of interest has a first respective spin polarization, and immediately before (c), at least one nucleus of the hyperpolarized molecule of interest has a second nuclear spin polarization, and the second nuclear spin polarization is reduced from the first nuclear spin polarization by no more than 10%, 20%, 30%, 40%, or 50%.

[0095] Embodiment 8. The method of any one of embodiments 1-7, further comprising, prior to (a), generating the hyperpolarized molecule of interest using at least one hyperpolarization procedure selected from the group consisting of parahydrogen-induced polarization (PHIP), PHIP side-arm hydrogenation (PHIP-SAH), PHIP nuclear Overhauser effect system (PHIPNOESYS), and signal amplification by reversible exchange (SABRE).

[0096] Embodiment 9. The method of any one of embodiments 1-8, wherein the hyperpolarized molecule of interest is selected from the group consisting of dimethyl maleate, pyruvate, glutamate, glutamine, lactate, acetate, acetoacetate, zymonate, alanine, fructose, fumarate, bicarbonate, urea, dehydroascorbate, alpha-ketoglutarate, dihydroxyacetone, glucose, ascorbate, and their conjugate acids.

[0097] Embodiment 10. The method of any one of embodiments 1 to 9, wherein (c) comprises performing an MRI or NMR procedure using an organic solution.

[0098] The method of any one of embodiments 1-9, further comprising, following embodiment 11.(b), mixing the organic solution with water, thereby producing an aqueous solution having the hyperpolarized molecules of interest dissolved therein at a third concentration.

[0099] Embodiment 12. The method of embodiment 11, wherein (c) comprises performing an MRI or NMR procedure using an aqueous solution.

[0100] Embodiment 13. The method of embodiment 11 or 12, wherein the third concentration is at least 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, 200 mM, or more.

[0101] Embodiment 14. A composition for use in a magnetic resonance imaging (MRI) or nuclear magnetic resonance (NMR) procedure, comprising: a composition comprising a solution and hyperpolarized molecules of interest dissolved therein, (a) obtaining an organic solution having hyperpolarized molecules of interest dissolved therein at a first concentration; (b) evaporating at least a portion of the organic solution, thereby producing an organic solution having the hyperpolarized molecules of interest dissolved therein at a second concentration that is greater than the first concentration.

[0102] Embodiment 15. The composition of embodiment 14, wherein the first concentration is less than 100 millimolar (mM), 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, or less.

[0103] Embodiment 16. The composition of embodiment 14 or 15, wherein the second concentration is at least 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, 200 mM, or more.

[0104] Embodiment 17. The composition of any one of embodiments 14-16, wherein the organic solution comprises at least one organic molecule selected from the group consisting of methanol, ethanol, n-propanol, isopropanol, and acetone.

[0105] Embodiment 18. The method of any one of embodiments 14-17, wherein (b) comprises evaporating at least a portion by performing at least one evaporation procedure selected from the group consisting of flowing an inert gas over the organic solution, subjecting the organic solution to a vacuum, and heating the organic solution.

[0106] Embodiment 19. The composition of any one of embodiments 14-18, wherein (a)-(b) are carried out for a total period of at most about 150 seconds, 140 seconds, 130 seconds, 120 seconds, 110 seconds, 100 seconds, 90 seconds, 80 seconds, 70 seconds, 60 seconds, 50 seconds, 40 seconds, 30 seconds, 20 seconds, 19 seconds, 18 seconds, 17 seconds, 16 seconds, 15 seconds, 14 seconds, 13 seconds, 12 seconds, 11 seconds, 10 seconds, 9 seconds, 8 seconds, 7 seconds, 6 seconds, 5 seconds, 4 seconds, 3 seconds, 2 seconds, 1 second, or less.

[0107] Embodiment 20. The composition of any one of embodiments 14-19, wherein immediately after (a), the hyperpolarized molecules of interest have a first respective spin polarization, and immediately prior to use of the composition in an MRI or NMR procedure, at least one nucleus of the hyperpolarized molecules of interest has a second nuclear spin polarization, and the second nuclear spin polarization is reduced from the first nuclear spin polarization by no more than 10%, 20%, 30%, 40%, or 50%.

[0108] Embodiment 21. The composition of any one of embodiments 14-20, wherein prior to (a), the composition is further produced by producing the hyperpolarized molecule of interest using at least one hyperpolarization procedure selected from the group consisting of parahydrogen-induced polarization (PHIP), PHIP side-arm hydrogenation (PHIP-SAH), PHIP nuclear Overhauser effect system (PHIPNOESYS), and signal amplification by reversible exchange (SABRE).

[0109] Embodiment 22. The composition of any one of embodiments 14-21, wherein the hyperpolarized molecule of interest is selected from the group consisting of dimethyl maleate, pyruvate, glutamate, glutamine, lactate, acetate, acetoacetate, zymonate, alanine, fructose, fumarate, bicarbonate, urea, dehydroascorbate, alpha-ketoglutarate, dihydroxyacetone, glucose, ascorbate, and their conjugate acids.

[0110] Embodiment 23. The composition of any one of embodiments 14 to 22, wherein the MRI or NMR procedure is performed using an organic solution.

[0111] Embodiment 24. The composition of any one of embodiments 14-22, wherein the composition is further formed by, subsequent to (b), mixing the organic solution with water, thereby forming an aqueous solution having the hyperpolarized molecules of interest dissolved therein at a third concentration.

[0112] Embodiment 25. The composition of embodiment 24, wherein the MRI or NMR procedure is performed using an aqueous solution.

[0113] Embodiment 26. The composition of embodiment 24 or 25, wherein the third concentration is at least 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, 200 mM, or more.

[0114] Embodiment 27. A method for increasing nuclear spin polarization in a molecule of interest, comprising: (a) obtaining a molecule of interest that contains at least one deuterium atom; (b) placing the molecule of interest in a magnetic field having an average field strength (B) of at most about 2 Tesla (T); (c) forming a coordination complex between the molecule of interest, a signal amplification by reversible exchange (SABRE) catalyst or catalyst precursor, and parahydrogen; (d) applying an oscillating magnetic field to the coordination complex, thereby transferring spin order from the para-hydrogen to the molecule of interest, thereby increasing the nuclear spin polarization of at least one atom in the molecule of interest.

[0115] Embodiment 28 The method of embodiment 27, wherein the molecule of interest comprises a deuterated chemical analog of a biologically relevant imaging agent.

[0116] Embodiment 29. The method of embodiment 28, wherein the biologically relevant imaging agent is selected from pyruvate, alpha-ketoglutaric acid, Z-4-methyl-2-oxopent-3-enedioic acid (Z-OMPD), zymonate, urea, azidothymidine (AZT), metronidazole, trimethylphenylammonium (TMPA), pyridine, nicotinamide, diazirine tag, and their conjugate acids.

[0117] Embodiment 30. The molecule of interest comprises at least one carbon-13( 13 C) atom or nitrogen-15( 15 30. The method of any one of embodiments 27-29, comprising N) atoms.

[0118] Embodiment 31. The method of any one of embodiments 27-30, wherein the molecule of interest comprises at least two deuterium atoms or at least three deuterium atoms.

[0119] Embodiment 32. The average magnetic field strength is at least about 1 μT, 2 μT, 3 μT, 4 μT, 5 μT, 6 μT, 7 μT, 8 μT, 9 μT, 10 μT, 20 μT, 30 μT, 40 μT, 50 μT, 60 μT, 70 μT, 80 μT, 90 μT, 100 μT, 200 μT, 300 μT, 400 μT, 500 μT, 600 μT, 700 μT, 800 μT, 900 μT, 1 mT (mT), 2mT, 3mT, 4mT, 5mT, 6mT, 7mT, 8mT, 9mT, 10mT, 20mT, 30mT, 40mT, 50mT, 60mT, 70mT, 80mT, 90mT, 100mT, 300mT, 400mT, 500mT, 600mT, 700mT, 800mT, 900mT, 1T, or more.

[0120] Embodiment 33. The average magnetic field strength is at most about 1T, 900mT, 800mT, 700mT, 600mT, 500mT, 400mT, 300mT, 200mT, 100mT, 90mT, 80mT, 70mT, 60mT, 50mT, 40mT, 30mT, 20mT, 10mT, 9mT, 8mT, 7mT, 6mT, 5mT, 4mT, 3mT, 2mT, 1mT, 900 33. The method of any one of embodiments 27-32, wherein the concentration is 1 μT, 800 μT, 700 μT, 600 μT, 500 μT, 400 μT, 300 μT, 200 μT, 100 μT, 90 μT, 80 μT, 70 μT, 60 μT, 50 μT, 40 μT, 30 μT, 20 μT, 10 μT, 9 μT, 8 μT, 7 μT, 6 μT, 5 μT, 4 μT, 3 μT, 2 μT, 1 μT, or less.

[0121] Embodiment 34 The method of any one of embodiments 27-33, wherein the SABRE catalyst or catalyst precursor comprises an iridium (Ir) complex or a cobalt (Co) complex.

[0122] Embodiment 35. The method of embodiment 34, wherein the SABRE catalyst or catalyst precursor comprises [IrCl(COD)(IMes)].

[0123] Embodiment 36. The method of any one of embodiments 27 to 35, wherein the oscillating magnetic field has a maximum field strength of at most about 10 mT.

[0124] Embodiment 37. The oscillating magnetic field is at least about 0.1 μT, 0.2 μT, 0.3 μT, 0.4 μT, 0.5 μT, 0.6 μT, 0.7 μT, 0.8 μT, 0.9 μT, 1 μT, 2 μT, 3 μT, 4 μT, 5 μT, 6 μT, 7 μT, 8 μT, 9 μT, 10 μT, 20 μT, 30 μT, 40 μT, 50 μT, 60 μT 37. The method of embodiment 36, wherein the maximum magnetic field strength is 70 μT, 80 μT, 90 μT, 100 μT, 200 μT, 300 μT, 400 μT, 500 μT, 600 μT, 700 μT, 800 μT, 900 μT, 1 mT, 2 mT, 3 mT, 4 mT, 5 mT, 6 mT, 7 mT, 8 mT, 9 mT, 10 mT, or more.

[0125] Embodiment 38. The oscillating magnetic field is at most about 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 μT, 800 μT, 700 μT, 600 μT, 500 μT, 400 μT, 300 μT, 200 μT, 100 μT, 90 μT, 80 μT, 70 μT, 60 μT, 50 μT, 40 38. The method of embodiment 36 or 37, wherein the maximum magnetic field strength is 0.9 μT, 0.8 μT, 0.7 μT, 0.6 μT, 0.5 μT, 0.4 μT, 0.3 μT, 0.2 μT, 0.1 μT, or less.

[0126] Embodiment 39. The method of any one of embodiments 27 to 38, further comprising performing (a) to (d) within a magnetic shield.

[0127] Embodiment 40. The method of embodiment 39, wherein the magnetic shield comprises a mu-metal magnetic shield.

[0128] Embodiment 41. The method of any one of embodiments 27-40, wherein (c) is carried out in a solution comprising the molecule of interest, a SABRE catalyst or catalyst precursor, and parahydrogen.

[0129] Embodiment 42 The method of embodiment 41, further comprising bubbling parahydrogen into the solution.

[0130] Embodiment 43. The method of embodiment 42, wherein parahydrogen is bubbled into the solution at a pressure of from 1 bar to 50 bar.

[0131] Embodiment 44. The method of any one of embodiments 27-43, wherein following (d), the nuclear spin polarization in the molecule of interest is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or more.

[0132] Embodiment 45. The method of any one of embodiments 27 to 44, wherein the average magnetic field strength is from about 1 μT to about 50 mT.

[0133] Embodiment 46. A composition for use in a magnetic resonance imaging (MRI) or nuclear magnetic resonance (NMR) procedure, comprising: a composition comprising a solution and hyperpolarized molecules of interest dissolved therein, (a) obtaining a molecule of interest that contains at least one deuterium atom; (b) placing the molecule of interest in a magnetic field having an average field strength (B) of at most about 2 Tesla (T); (c) forming a coordination complex between the molecule of interest, a signal amplification by reversible exchange (SABRE) catalyst or catalyst precursor, and parahydrogen; (d) applying an oscillating magnetic field to the coordination complex, thereby transferring spin order from parahydrogen to the molecule of interest, thereby increasing the nuclear spin polarization of at least one atom in the molecule of interest.

[0134] Embodiment 47. The composition of embodiment 46, wherein the molecule of interest comprises a deuterated chemical analog of a biologically relevant imaging agent.

[0135] Embodiment 48. The composition of embodiment 47, wherein the biologically relevant imaging agent is selected from pyruvate, alpha-ketoglutaric acid, Z-4-methyl-2-oxopent-3-enedioic acid (Z-OMPD), zymonate, urea, azidothymidine (AZT), metronidazole, trimethylphenylammonium (TMPA), pyridine, nicotinamide, diazirine tag, and their conjugate acids.

[0136] Embodiment 49. The molecule of interest has at least one carbon-13( 13 C) atom or nitrogen-15( 15 49. The composition of any one of embodiments 46-48, comprising a N) atom.

[0137] Embodiment 50. The composition of any one of embodiments 46-49, wherein the molecule of interest comprises at least two deuterium atoms or at least three deuterium atoms.

[0138] Embodiment 51. The average magnetic field strength is at least about 1 μT, 2 μT, 3 μT, 4 μT, 5 μT, 6 μT, 7 μT, 8 μT, 9 μT, 10 μT, 20 μT, 30 μT, 40 μT, 50 μT, 60 μT, 70 μT, 80 μT, 90 μT, 100 μT, 200 μT, 300 μT, 400 μT, 500 μT, 600 μT, 700 μT, 800 μT, 900 μT, 1 millitesla ( 51. The composition of any one of embodiments 46-50, wherein the hydroxybenzoate is 2mT, 3mT, 4mT, 5mT, 6mT, 7mT, 8mT, 9mT, 10mT, 20mT, 30mT, 40mT, 50mT, 60mT, 70mT, 80mT, 90mT, 100mT, 300mT, 400mT, 500mT, 600mT, 700mT, 800mT, 900mT, 1T, or more.

[0139] Embodiment 52. The average magnetic field strength is at most about 1 T, 900 mT, 800 mT, 700 mT, 600 mT, 500 mT, 400 mT, 300 mT, 200 mT, 100 mT, 90 mT, 80 mT, 70 mT, 60 mT, 50 mT, 40 mT, 30 mT, 20 mT, 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 μ 52. The composition of any one of embodiments 46-51, wherein the solubility of ...

[0140] Embodiment 53. The composition of any one of embodiments 46-52, wherein the SABRE catalyst or catalyst precursor comprises an iridium (Ir) complex or a cobalt (Co) complex.

[0141] Embodiment 54. The composition of embodiment 53, wherein the SABRE catalyst or catalyst precursor comprises [IrCl(COD)(IMes)].

[0142] Embodiment 55. The composition of any one of embodiments 46-54, wherein the oscillating magnetic field has a maximum field strength of at most about 10 mT.

[0143] Embodiment 56. The oscillating magnetic field is at least about 0.1 μT, 0.2 μT, 0.3 μT, 0.4 μT, 0.5 μT, 0.6 μT, 0.7 μT, 0.8 μT, 0.9 μT, 1 μT, 2 μT, 3 μT, 4 μT, 5 μT, 6 μT, 7 μT, 8 μT, 9 μT, 10 μT, 20 μT, 30 μT, 40 μT, 50 μT, 60 μT, 56. The composition of embodiment 55, having a maximum magnetic field strength of 70 μT, 80 μT, 90 μT, 100 μT, 200 μT, 300 μT, 400 μT, 500 μT, 600 μT, 700 μT, 800 μT, 900 μT, 1 mT, 2 mT, 3 mT, 4 mT, 5 mT, 6 mT, 7 mT, 8 mT, 9 mT, 10 mT, or more.

[0144] Embodiment 57. The oscillating magnetic field is at most about 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 μT, 800 μT, 700 μT, 600 μT, 500 μT, 400 μT, 300 μT, 200 μT, 100 μT, 90 μT, 80 μT, 70 μT, 60 μT, 50 μT, 40 57. The composition of embodiment 55 or 56, having a maximum magnetic field strength of 0.9μT, 0.8μT, 0.7μT, 0.6μT, 0.5μT, 0.4μT, 0.3μT, 0.2μT, 0.1μT, or less.

[0145] Embodiment 58. The composition of any one of embodiments 46-57, wherein the composition is produced by carrying out (a)-(d) within a magnetic shield.

[0146] Embodiment 59. The composition of embodiment 58, wherein the magnetic shield comprises a mu-metal magnetic shield.

[0147] Embodiment 60. The composition of any one of embodiments 46 to 59, wherein (c) is carried out in a solution comprising the molecule of interest, a SABRE catalyst or catalyst precursor, and parahydrogen.

[0148] Embodiment 61. The composition of embodiment 60, wherein the composition is produced by bubbling parahydrogen into a solution.

[0149] Embodiment 62. The composition of embodiment 61, wherein parahydrogen is bubbled into the solution at a pressure of from 1 bar to 50 bar.

[0150] Following embodiment 63.(d), the composition of any one of embodiments 46-62, wherein the nuclear spin polarization in the molecule of interest is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or more.

[0151] Embodiment 64. The composition of any one of embodiments 46 to 63, wherein the average magnetic field strength is from about 1 μT to about 50 mT.

Claims

1. 1. A method for increasing nuclear spin polarization in a molecule of interest, comprising: (a) obtaining a molecule of interest that includes at least one deuterium atom; (b) subjecting the molecule of interest to an average magnetic field strength (B) of at most about 2 Tesla (T); ０ ) in a magnetic field having a (c) forming a coordination complex between the molecule of interest, a signal amplification by reversible exchange (SABRE) catalyst or catalyst precursor, and parahydrogen; (d) applying an oscillating magnetic field to the coordination complex, thereby transferring spin order from the para-hydrogen to the molecule of interest, thereby increasing the nuclear spin polarization of at least one atom in the molecule of interest.

2. The method of claim 1 , wherein the molecule of interest comprises a deuterated chemical analog of a biologically relevant imaging agent.

3. 3. The method of claim 2, wherein the biorelevant imaging agent is selected from pyruvate, alpha-ketoglutaric acid, Z-4-methyl-2-oxopenta-3-enedioic acid (Z-OMPD), zymonate, urea, azidothymidine (AZT), metronidazole, trimethylphenylammonium (TMPA), pyridine, nicotinamide, diazirine tag, and their conjugate acids.

4. The molecule of interest has at least one carbon-13 ( １３ C) atom or nitrogen-15 ( １５ The method of any one of claims 1 to 3, wherein the aryl group comprises a substituted or unsubstituted N atom.

5. The method of any one of claims 1 to 4, wherein the molecule of interest comprises at least two deuterium atoms or at least three deuterium atoms.

6. The average magnetic field strength is at least about 1 μT, 2 μT, 3 μT, 4 μT, 5 μT, 6 μT, 7 μT, 8 μT, 9 μT, 10 μT, 20 μT, 30 μT, 40 μT, 50 μT, 60 μT, 70 μT, 80 μT, 90 μT, 100 μT, 200 μT, 300 μT, 400 μT, 500 μT, 600 μT, 700 μT, 800 μT, 900 μT, 1 millitesla (mT ), 2mT, 3mT, 4mT, 5mT, 6mT, 7mT, 8mT, 9mT, 10mT, 20mT, 30mT, 40mT, 50mT, 60mT, 70mT, 80mT, 90mT, 100mT, 300mT, 400mT, 500mT, 600mT, 700mT, 800mT, 900mT, 1T, or more.

7. the average magnetic field strength is at most about 1 T, 900 mT, 800 mT, 700 mT, 600 mT, 500 mT, 400 mT, 300 mT, 200 mT, 100 mT, 90 mT, 80 mT, 70 mT, 60 mT, 50 mT, 40 mT, 30 mT, 20 mT, 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 μT, 7. The method of any one of claims 1 to 6, wherein the concentration is 800μT, 700μT, 600μT, 500μT, 400μT, 300μT, 200μT, 100μT, 90μT, 80μT, 70μT, 60μT, 50μT, 40μT, 30μT, 20μT, 10μT, 9μT, 8μT, 7μT, 6μT, 5μT, 4μT, 3μT, 2μT, 1μT, or less.

8. The method of any one of claims 1 to 7, wherein the SABRE catalyst or catalyst precursor comprises an iridium (Ir) or cobalt (Co) complex.

9. 9. The method of claim 8, wherein the SABRE catalyst or catalyst precursor comprises [IrCl(COD)(IMes)].

10. The method according to any one of claims 1 to 9, wherein the oscillating magnetic field has a maximum field strength of at most about 10 mT.

11. the oscillating magnetic field is at least about 0.1 μT, 0.2 μT, 0.3 μT, 0.4 μT, 0.5 μT, 0.6 μT, 0.7 μT, 0.8 μT, 0.9 μT, 1 μT, 2 μT, 3 μT, 4 μT, 5 μT, 6 μT, 7 μT, 8 μT, 9 μT, 10 μT, 20 μT, 30 μT, 40 μT, 50 μT, 60 μT, 70 μT 11. The method of claim 10, wherein the magnetic field strength is 1 mT, 80 μT, 90 μT, 100 μT, 200 μT, 300 μT, 400 μT, 500 μT, 600 μT, 700 μT, 800 μT, 900 μT, 1 mT, 2 mT, 3 mT, 4 mT, 5 mT, 6 mT, 7 mT, 8 mT, 9 mT, 10 mT, or more.

12. The oscillating magnetic field is at most about 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 μT, 800 μT, 700 μT, 600 μT, 500 μT, 400 μT, 300 μT, 200 μT, 100 μT, 90 μT, 80 μT, 70 μT, 60 μT, 50 μT, 40 μT, 12. The method of claim 10 or 11, wherein the magnetic field has a maximum magnetic field strength of 30 μT, 20 μT, 10 μT, 9 μT, 8 μT, 7 μT, 6 μT, 5 μT, 4 μT, 3 μT, 2 μT, 1 μT, 0.9 μT, 0.8 μT, 0.7 μT, 0.6 μT, 0.5 μT, 0.4 μT, 0.3 μT, 0.2 μT, 0.1 μT, or less.

13. The method of any one of claims 1 to 12, further comprising performing (a)-(d) within a magnetic shield.

14. The method of claim 13 , wherein the magnetic shield comprises a mu-metal magnetic shield.

15. 15. The method of any one of claims 1 to 14, wherein (c) is carried out in a solution comprising the molecule of interest, the SABRE catalyst or catalyst precursor, and the parahydrogen.

16. 16. The method of claim 15, further comprising bubbling the parahydrogen into the solution.

17. 17. The method of claim 16, wherein the parahydrogen is bubbled into the solution at a pressure of from 1 bar to 50 bar.

18. 18. The method of any one of claims 1-17, wherein following (d), the nuclear spin polarization in the molecule of interest is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or more.

19. The method of any one of claims 1 to 18, wherein the average magnetic field strength is from about 1 µT to about 50 mT.

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