Systems and methods for hyperpolarized nuclear magnetic resonance spectroscopy and magnetic resonance imaging.

Abstract

This disclosure describes systems and methods for enhancing the spin lattice (T1) relaxation time in a hyperpolarized molecule of interest during the period between a hyperpolarization procedure and the performance of a nuclear magnetic resonance (NMR) or magnetic resonance imaging (MRI) experiment. The systems and methods generally involve exposing the hyperpolarized molecule to a relatively low magnetic field during the hyperpolarization procedure and / or any purification procedure following the hyperpolarization procedure. At such relatively low magnetic fields, hyperpolarizable nuclei may exhibit relatively long T1 relaxation time constants at near physiological pH. The T1 relaxation time can be further enhanced by performing the hyperpolarization procedure and / or purification procedure in a deuterated water (D2O) solvent and / or in a solution containing a scavenger such as ethylenediaminetetraacetic acid (EDTA). Such systems and methods can generate long T1 relaxation times such that the hyperpolarized molecule retains substantial polarization during the NMR / MRI experiment.

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims priority to U.S. Provisional Patent Application No. 63 / 524,242, filed Jun. 30, 2023, entitled "SYSTEMS AND METHODS FOR HYPERPOLARIZED NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY AND MAGNETIC RESONANCE IMAGING", which is hereby incorporated by reference in its entirety for all purposes.

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

Background Art

[0003] Nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI) are techniques with important applications in chemistry, biology, and medical imaging. Despite their success, it is recognized that magnetic resonance applications often have limitations due to the small nuclear polarization of analytes (typically on the order of 10 -5 . This small nuclear polarization can lead to limited sensitivity compared to other analytical techniques such as mass spectrometry.

Summary of the Invention

[0004] By increasing nuclear spin polarization beyond the thermal equilibrium value, magnetic resonance sensitivity can be improved. Nuclear spin polarization can be increased using known techniques such as para-hydrogen-induced polarization (PHIP), PHIP-sidearm hydrogenation (PHIP-SAH), PHIP nuclear Overhauser effect system (PHIPNOESYS) reversible exchange signal amplification (SABRE), and dynamic nuclear polarization (DNP). Using these techniques, the nuclear spin polarization of a material can be increased significantly. For example, the nuclear spin polarization of a material can be increased by more than 10,000 times in some cases. Enhanced nuclear spin polarization can result in a proportional increase in the NMR / MRI signal. This enhanced polarization decays over time due to the relaxation time of the nuclear spin in the polarized molecule, but for many molecules, the relaxation time can be in the order of seconds, during which the increase in polarization can result in a dramatic increase in NMR / MRI signal sensitivity. By enabling such a dramatic increase in NMR / MRI signal sensitivity, increased nuclear spin polarization can lead to new applications such as in vivo imaging of metabolism using metabolites with increased nuclear spin polarization in MRI scanners, accelerating NMR spectroscopic investigations and enabling the visualization of molecular dynamics and structures that were previously invisible.

[0005] The accompanying drawings, including portions of this specification, illustrate several embodiments and, together with this specification, serve to illustrate specific principles and features of the disclosed embodiments. These are shown in the drawings as follows: [Brief explanation of the drawing]

[0006] [Figure 1] Figure 1 shows a first exemplary process for generating polarized bio-related contrast agents according to various embodiments. [Figure 2] Figure 2 shows a second exemplary process for generating polarized bio-related contrast agents according to various embodiments. [Figure 3] Figure 3 shows a third exemplary process for generating polarized bio-related contrast agents according to various embodiments. [Figure 4]Figure 4 shows exemplary compositions containing a target hyperpolarized molecule having hypernuclear spin polarization in a solution, according to various embodiments. [Figure 5] Figure 5 shows exemplary carbon-13 (13C) spin lattice (T1) relaxation times associated with hyperpolarized pyruvate in water (H2O) and deuterated water (D2O) solvents at pH 7 according to various embodiments. [Figure 6] Figure 6 shows exemplary 13C T1 relaxation times associated with hyperpolarized pyruvate in D2O solvent at two different pH values ​​(5.5 and 7) under various magnetic fields, according to various embodiments. [Figure 7] Figure 7 shows exemplary 13C T1 relaxation times related to hyperpolarized pyruvate in D2O solvent, in the absence and presence of ethylenediaminetetraacetic acid (EDTA), under various magnetic fields according to various embodiments. [Modes for carrying out the invention]

[0007] Herein, exemplary embodiments are described in detail and discussed with respect to the accompanying drawings. Unless otherwise defined, technical and / or scientific terms have the meanings generally understood by those skilled in the art. The disclosed embodiments are described in sufficient detail to enable those skilled in the art to carry out the disclosed embodiments. Naturally, other embodiments may be used and modifications may be made without departing from the scope of the disclosed embodiments. Thus, the materials, methods and examples are illustrative and not necessarily intended to be limiting.

[0008] Recent research in the fields of NMR and MRI has demonstrated that NMR and MRI signals associated with various target molecules (e.g., bio-related contrast agents) can be dramatically enhanced using various so-called hyperpolarization techniques. This signal enhancement allows for improved spectroscopic analysis of the target molecules. For example, this signal enhancement enables improved spectroscopic analysis of bio-related contrast agents as they are metabolized by various tissues at different locations in the body. Analysis of metabolic information determined by such spectroscopic contrast can enable non-invasive determination of the health status of tissues in the body. For example, abnormal metabolism of bio-related contrast agents may indicate diseases such as cancer at several locations in the body.

[0009] Existing techniques for hyperpolarizing target molecules include dissolved DNP, PHIP, PHIP-SAH, PHIPNOESYS, and SABRE. In PHIP and PHIP-SAH, a derivative of the target molecule containing double or triple bonds (e.g., a precursor) is reacted with parahydrogen to form a parahydrogenated derivative of the precursor. The spin order is then transferred from the added proton via the parahydrogenation reaction to the target nucleus (e.g., a carbon-13 or nitrogen-15 nucleus) contained within the target molecule. In PHIP-SAH, the parahydrogenated derivative of the precursor is cleaved (e.g., hydrolyzed) to obtain the target polarized molecule. The target molecule is then purified and used in NMR or MRI procedures.

[0010] PHIPNOESYS utilizes PHIP or PHIP-SAH to generate a hyperpolarized material (e.g., a starting compound) and transfer the polarization from the starting compound to the material used in NMR spectroscopy (e.g., a target compound or molecule of interest). This polarization transfer from the starting compound to the target compound proceeds via the intermolecular nuclear Overhauser effect (NOE). PHIPNOESYS has been shown to increase the signal of NMR spectroscopy by up to approximately 2,000 times, allowing NMR spectroscopy to be applied at significantly lower concentrations than otherwise achievable.

[0011] In SABRE, the target molecule itself forms a coordination complex with a polarized migration catalyst or catalyst precursor and a parahydrogen. The spin order is then transferred from the parahydrogen to the target nucleus within the target molecule via the coordination complex. The target molecule is then selectively purified and used in NMR or MRI procedures.

[0012] Hyperpolarization techniques can dramatically increase NMR / MRI signals, but this increase is typically relatively short-lived and often decreases with a T1 relaxation time constant of less than tens of seconds. Each unit time T1 passage leads to a decrease in the NMR / MRI signal by a multiple of 1 / e, where e is Euler's number and has a value of approximately 2.718. Therefore, after passing through T1, the hyperpolarized NMR / MRI signal decreases to 1 / e (or about 36.79%) of its initial value. After passing through 2T1, the hyperpolarized NMR / MRI signal decreases to 1 / e of its initial value. 2 (Or it decreases to about 13.53%). After passing through 3T1, the hyperpolarization NMR / MRI signal is 1 / e of its initial value. 3 (Or it decreases to about 4.98%). Many hyperpolarization techniques require a time-consuming purification step before performing the NMR / MRI experiment, so the hyperpolarized NMR / MRI signal can decay significantly between the time the hyperpolarization procedure and the actual time the NMR / MRI experiment is performed, resulting in a much weaker NMR / MRI signal than desired. Therefore, there is a need for systems and methods to enhance the T1 relaxation time in the hyperpolarized molecule during the period between the hyperpolarization procedure and the performance of the NMR / MRI experiment.

[0013] The systems and methods presented herein relate to the T1 relaxation time (e.g.,) in the hyperpolarization molecule of interest during the period between the hyperpolarization procedure and the performance of the NMR / MRI experiment. 13This allows for enhancement of the C T1 relaxation time. Such enhancement may enable the use of hyperpolarized molecules in NMR / MRI experiments, but the hyperpolarized molecules retain the high degree of nuclear spin polarization that is conferred upon them during the hyperpolarization procedure. The system and method generally involves subjecting the hyperpolarized molecules to a relatively low magnetic field (e.g., less than 1 Tesla (T)) during the hyperpolarization procedure and / or any purification procedure that follows the hyperpolarization procedure. In such a relatively low magnetic field, the hyperpolarizable nuclei of the hyperpolarized molecules (e.g., 13 C) may exhibit a relatively long T1 relaxation time constant (e.g., 60 seconds or more) at near physiological pH values ​​(e.g., pH values ​​of 5-9). Since such a T1 relaxation time is relatively long compared to the time required to carry out the purification procedure, the hyperpolarized molecule retains a relatively large proportion of its initial polarization during the purification procedure and therefore during the NMR / MRI experiment. The T1 relaxation time can be further enhanced by carrying out the hyperpolarization procedure and / or purification procedure in solution. For example, the T1 relaxation time can be enhanced by performing hyperpolarization and / or purification procedures in a D2O solvent and / or in a solution containing a scavenger such as ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2',2'',2'''-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid (DOTA), 1,7,10,16-tetraoxa-4,13-diazacyclooctadecane (Kryptofix® 22), 1,4,7,10,13,16-hexaazacyclooctadecane (Hexacyclone), crown ether, etc. The system and method can be applied to any solution-based hyperpolarization procedure, including but not limited to parahydrogen-based hyperpolarization procedures such as PHIP, PHIP-SAH, PHIPNOESYS, and / or SABRE.

[0014] Where used herein, unless otherwise specifically stated, the term “or” encompasses all possible combinations, except where impractical. For example, the phrase “A or B” means, unless otherwise stated or unless such meaning is impractical, element A alone, element B alone, and combinations of elements A and B. In another embodiment, the phrase “A, B, or C” means, unless otherwise specified or unless such meaning is impractical, element A alone, element B alone, element C alone, combinations of elements A and B without element C, combinations of elements A and C without element B, combinations of elements B and C without element A, and combinations of elements A, B, and C.

[0015] Figure 1 shows a first exemplary process 100 for generating a hyperpolarized molecule of a target having hypernuclear spin polarization according to various embodiments. In 110, the target molecule or a derivative of the target molecule is obtained. In some embodiments, the target molecule or a derivative of the target molecule includes hyperpolarizable nuclei (i.e., hyperpolarization-sensitive nuclei using hyperpolarization procedures such as DNP, PHIP, PHIP-SAH, PHIPNOESYS, or SABRE). In some embodiments, the hyperpolarizable nuclei are 13 C or nitrogen-15 15 Includes spin-1 / 2 nuclei such as N).

[0016] In these embodiments, derivatives of the molecule in question include precursors of the molecule in question. For example, in some embodiments, derivatives of the molecule in question are chemically identical to the molecule in question, except that the derivative contains at least one double bond and the molecule in question contains a single bond in another way, or the derivative contains at least one triple bond and the molecule in question contains a single or double bond in another way. As another example, in some embodiments, derivatives of the molecule in question contain cleavable side arms bonded to the molecule in question. In some embodiments, the molecule in question includes any bio-relevant contrast agent described herein. In some embodiments, the molecule in question includes any derivative of any bio-relevant contrast agent described herein or a precursor of any bio-relevant contrast agent. In some embodiments, the molecule in question includes carboxylates, carbon-13 labeled carboxylates, partially or fully deuterated carboxylates, or carbon-13 labeled and partially or fully deuterated carboxylates. In some embodiments, the molecule in question or a derivative of the molecule in question is contained in a solution. Therefore, in some embodiments, derivatives of the molecule in question include derivatives of carboxylate salts, carbon-13 labeled derivatives of carboxylates, partially or completely deuterated derivatives of carboxylate salts, or carbon-13 labeled and partially or completely deuterated derivatives of carboxylates. In some embodiments, the molecule in question or a derivative of the molecule in question is contained in a solution.

[0017] In 120, a first magnetic field is applied to the target molecule or a derivative of the target molecule. In some embodiments, the first magnetic field can be up to approximately 1 Tesla (T), 900 millitesla (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 m T, 2mT, 1mT, 900 microtesla (μT), 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. In some embodiments, the first magnetic field 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 These values ​​are 900μT, 1mT, 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, 1T or higher. In some embodiments, the first magnetic field is within the range defined by any two of the aforementioned values. For example, in some embodiments, the first magnetic field is in the range of approximately 1 μT to approximately 1 T, approximately 1 μT to approximately 100 mT, approximately 1 μT to approximately 10 mT, approximately 1 μT to approximately 1 mT, approximately 1 μT to approximately 100 μT, approximately 1 μT to approximately 10 μT, approximately 10 μT to approximately 1 T, approximately 10 μT to approximately 100 mT, approximately 10 μT to approximately 10 mT, approximately 100 μT to approximately 1 T, approximately 100 μT to approximately 100 mT, approximately 100 μT to approximately 10 mT, approximately 1 mT to approximately 1 T, approximately 1 mT to approximately 100 mT, approximately 1 mT to approximately 10 mT, approximately 10 mT to approximately 1 T, approximately 10 mT to approximately 100 mT, or approximately 100 mT to approximately 1 T.

[0018] In 130, the target molecule or a derivative of the target molecule is subjected to a nuclear spin hyperpolarization procedure. In some embodiments, the nuclear spin hyperpolarization procedure includes the PHIP procedure, the PHIP-SAH procedure, the PHIPNOESYS procedure, or the SABRE procedure. In some embodiments, the nuclear spin hyperpolarization procedure generates the target hyperpolarized molecule. In some embodiments, the nuclear spin hyperpolarization procedure imparts a first nuclear spin polarization to the hyperpolarizable nucleus. In some embodiments, the first nuclear spin polarization exhibits polarization of at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, or greater, at most about 50%, 40%, 30%, 20%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, or within the range defined by any two of the aforementioned values.

[0019] At 140, a second magnetic field is applied to the target hyperpolarized molecule. In some embodiments, the second magnetic field can be up to approximately 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, 900μT, 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. In some embodiments, the second magnetic field 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, 900μT, 1mT, 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, 1T, or greater. In some embodiments, the second magnetic field is within the range defined by any two of the aforementioned values. For example, in some embodiments, the second magnetic field is approximately 1 μT to approximately 1 T, approximately 1 μT to approximately 100 mT, approximately 1 μT to approximately 10 mT, approximately 1 μT to approximately 1 mT, approximately 1 μT to approximately 100 μT, approximately 1 μT to approximately 10 μT, approximately 10 μT to approximately 1 T, approximately 10 μT to approximately 100 mT, approximately 10 μT to approximately 10 mT, approximately 10 μT to approximately 1 mT, approximately 100 μT to approximately 1 T, approximately 100 μT to approximately 10 mT, approximately 100 μT to approximately 10 mT, approximately 10 mT to approximately 1 T, approximately 1 mT to approximately 100 mT, approximately 1 mT to approximately 100 mT, approximately 10 mT to approximately 1 T, or approximately 100 mT to approximately 1 T.

[0020] In some embodiments, the solution containing the hyperpolarizing molecule is characterized by a pH of at least about 5, 6, 7, 8, 9 or greater, and at most about 9, 8, 7, 6, 5 or less, or within a range defined by any two of the aforementioned values. For example, in some embodiments, the solution is characterized by a pH of about 5 to about 9, about 5 to about 8, about 5 to about 7, about 5 to about 6, about 6 to about 9, about 6 to about 8, about 6 to about 7, about 7 to about 9, about 7 to about 8, or about 8 to about 9. In some embodiments, the solution contains a D2O solvent.

[0021] In some embodiments, the hyperpolarizable nucleus has a T1 relaxation time of at least about 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, 190 seconds, 200 seconds, or longer in solution with a second magnetic field and pH. In some embodiments, the hyperpolarizable nucleus has a T1 relaxation time of up to about 200 seconds, 190 seconds, 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, or shorter in solution with a second magnetic field and pH. In some embodiments, the hyperpolarizable nucleus has a T1 relaxation time within a range defined by any two of the aforementioned values ​​in solution with a second magnetic field and pH. In some embodiments, the long T1 relaxation time described herein is obtained using the desired fully or partially deuterated molecule. In some embodiments, the long T1 relaxation time described herein is obtained using the desired non-deuterated molecule.

[0022] In some embodiments, the solution further comprises a scavenger. In some embodiments, the scavenger further increases the T1 relaxation time. Although not bound by theory, it is assumed that the scavenger captures, chelates, binds to, or otherwise removes contaminants from the solution, which would result in a reduction in the T1 relaxation time. Such contaminants may include, but are not limited to, molecular oxygen (O2), paramagnetic molecules, ions, and metals. In some embodiments, the scavenger is selected from the group consisting of EDTA, DTPA, DOTA, Kryptofix® 22, Hexacyclen, crown ethers, and the like. In some embodiments, the solution contains the scavenger in a concentration of at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or greater, and at most about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, or within the range defined by any two of the aforementioned values. For example, in some embodiments, the scavenger is present at a concentration of about 1% to about 10%, about 1% to about 9%, about 1% to about 8%, about 1% to about 7%, about 1% to about 6%, about 1% to about 5%, about 1% to about 4%, about 1% to about 3%, about 1% to about 2%, about 2% to about 10%, about 2% to about 9%, about 2% to about 8%, about 2% to about 7%, about 2% to about 6%, about 2% to about 5%, about 2% to about 4%, about 2% to about 3%, about 3% to about 10%, about 3% to about 9%, about 3% to about 8%, about 3% to about 7%, The percentages are approximately 3% to 6%, 3% to 5%, 3% to 4%, 4% to 10%, 4% to 9%, 4% to 8%, 4% to 7%, 4% to 6%, 4% to 5%, 5% to 10%, 5% to 9%, 5% to 8%, 5% to 7%, 5% to 6%, 6% to 10%, 6% to 9%, 6% to 8%, 6% to 7%, 7% to 10%, 7% to 9%, 7% to 8%, 8% to 7%, 7% to 10%, 7% to 9%, 7% to 8%, 8% to 10%, 8% to 9%, or 9% to 10%.

[0023] In step 150, the target hyperpolarized molecules are subjected to a purification procedure. In some embodiments, the purification procedure produces the desired purified hyperpolarized molecules by reducing the concentration of contaminants in the solution. In some embodiments, the purification procedure includes one or more elements selected from the group consisting of catalytic filtration, catalyst capture, solvent washing, solvent exchange, liquid-liquid exchange, multi-step liquid-liquid exchange, distillation, evaporation, inert gas foaming, crystallization, and redissolution.

[0024] In some embodiments, hyperpolarizable nuclei relax at a T1 relaxation rate during the purification procedure, resulting in a decrease in the nuclear spin polarization of the hyperpolarizable nuclei during the purification procedure. Thus, in some embodiments, the purification procedure imparts a second nuclear spin polarization to the hyperpolarizable nuclei. Generally, the second nuclear spin polarization is smaller than the first nuclear spin polarization. However, the use of one, two, three, or four of the relatively low magnetic field, pH, scavenger, and D2O solvent described herein may provide a longer T1 relaxation time than otherwise achievable. Thus, hyperpolarizable nuclei may undergo significantly less T1 relaxation using Method 100 than otherwise achievable. In this way, hyperpolarizable nuclei may have a significantly higher second nuclear spin polarization than otherwise achievable.

[0025] In some embodiments, the second nuclear spin polarization is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, or greater, and at most about 50%, 40%, 30%, 20%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, or within the range defined by any two of the aforementioned values. In some embodiments, the second nuclear spin polarization is a percentage of the first nuclear spin polarization that is about 90%, 80%, 70%, 60%, 50% or less of the first nuclear spin polarization, about 50%, 60%, 70%, 80%, 90% or less of the nuclear spin polarization, or within the range defined by any two of the aforementioned values. In other words, in some embodiments, the purification procedure reduces the nuclear spin polarization of hyperpolarizable nuclei by an amount defined by a maximum of about 50%, 40%, 30%, 20%, 10%, or less, or at least about 10%, 20%, 30%, 40%, 50%, or more, or any two of the aforementioned values.

[0026] In step 160, the target hyperpolarizing molecule is administered to the target. In some embodiments, the target includes humans. In some embodiments, the target includes animals such as non-human primates, monkeys, horses, dogs, cats, rats, and mice.

[0027] In step 170, a magnetic resonance spectroscopy (MRS) procedure is performed on the subject. For example, in some embodiments, a metabolic imaging MRS procedure is performed on the subject.

[0028] In some embodiments, method 100 (i.e., all of steps 110, 120, 130, 140, 150, 160, and 170) is performed for a period of time that is at most about 60 seconds, 50 seconds, 40 seconds, 30 seconds, 20 seconds, 10 seconds or less, at least about 10 seconds, 20 seconds, 30 seconds, 40 seconds, 50 seconds, 60 seconds or more, or within a range defined by any two of the preceding values.

[0029] Figure 2 shows a second exemplary process 200 for generating a hyperpolarized molecule of a target having hypernuclear spin polarization, according to various embodiments. In 210, the target molecule or a derivative of the target molecule is obtained. In some embodiments, the target molecule includes any target molecule described in this disclosure with respect to method 100 of Figure 1. In some embodiments, the derivative of the target molecule includes any derivative of any target molecule described in this disclosure with respect to method 100 of Figure 1. In some embodiments, the target molecule or a derivative of the target molecule includes any hyperpolarizable nucleus described in this disclosure with respect to method 100 of Figure 1. In some embodiments, the target molecule or a derivative of the target molecule is dissolved in any target solution described in this disclosure with respect to method 100 of Figure 1. In some embodiments, the solution is characterized by any pH described in this disclosure with respect to method 100 of Figure 1. In some embodiments, the hyperpolarizable nucleus is characterized by any T1 relaxation time described in this disclosure with respect to method 100 of Figure 1 in any second magnetic field described in this disclosure with respect to method 100 of Figure 1.

[0030] In step 220, the molecule of interest or a derivative thereof is subjected to any nuclear spin hyperpolarization procedure described herein with respect to method 100 of Figure 1. In some embodiments, the nuclear spin hyperpolarization procedure generates the hyperpolarized molecule of interest, as described herein with respect to method 100 of Figure 1. In some embodiments, the nuclear spin hyperpolarization procedure imparts any first nuclear spin polarization described herein with respect to method 100 of Figure 1 to the hyperpolarizable nucleus.

[0031] In some embodiments, a first magnetic field is applied before step 220. In some embodiments, the first magnetic field includes any first magnetic field described herein with respect to method 100 of Figure 1.

[0032] In step 230, the hyperpolarized molecule of interest is subjected to any purification procedure described herein with respect to method 100 in Figure 1. In some embodiments, the purification procedure produces the purified hyperpolarized molecule as described herein with respect to method 100 in Figure 1. In some embodiments, the purification procedure imparts any second nuclear spin polarization as described herein with respect to method 100 in Figure 1 to the hyperpolarizable nucleus.

[0033] In some embodiments, a second magnetic field is applied before step 230. In some embodiments, the second magnetic field includes any second magnetic field described herein with respect to method 100 in Figure 1.

[0034] At step 240, the target hyperpolarizing molecule is administered to the target. In some embodiments, the target includes any target described herein with respect to method 100 in Figure 1.

[0035] At 250, the MRS procedure is performed on the subject. In some embodiments, the MRS procedure includes any MRS procedure described herein with respect to method 100 in Figure 1.

[0036] In some embodiments, Method 200 (i.e., all of steps 210, 220, 230, 240, and 250) may be performed at any time described in this disclosure with respect to Method 100 in Figure 1.

[0037] Figure 3 shows a third exemplary process 300 for generating a hyperpolarized molecule of a target having hypernuclear spin polarization, according to various embodiments. In 310, the target molecule or a derivative of the target molecule is obtained. In some embodiments, the target molecule includes any target molecule described in this disclosure with respect to method 100 of Figure 1. In some embodiments, the derivative of the target molecule includes any derivative of any target molecule described in this disclosure with respect to method 100 of Figure 1. In some embodiments, the target molecule or a derivative of the target molecule includes any hyperpolarizable nucleus described in this disclosure with respect to method 100 of Figure 1.

[0038] In 320, the molecule of interest or a derivative of the molecule of interest is subjected to any nuclear spin hyperpolarization procedure described herein with respect to method 100 of Figure 1. In some embodiments, the nuclear spin hyperpolarization procedure generates the hyperpolarized molecule of interest, as described herein with respect to method 100 of Figure 1. In some embodiments, the nuclear spin hyperpolarization procedure imparts any first nuclear spin polarization described herein with respect to method 100 of Figure 1 to the hyperpolarizable nucleus.

[0039] In some embodiments, a first magnetic field is applied before step 320. In some embodiments, the first magnetic field includes any first magnetic field described herein with respect to method 100 of Figure 1.

[0040] In step 330, the hyperpolarized molecule of interest is subjected to any purification procedure described herein with respect to method 100 of Figure 1. In some embodiments, the purification procedure produces the purified hyperpolarized molecule as described herein with respect to method 100 of Figure 1. In some embodiments, the purification procedure imparts any second nuclear spin polarization as described herein with respect to method 100 of Figure 1 to the hyperpolarizable nucleus.

[0041] In some embodiments, a second magnetic field is applied before step 330. In some embodiments, the second magnetic field includes any second magnetic field described herein with respect to method 100 of Figure 1.

[0042] At 340, the target hyperpolarizing molecule is administered to the target. In some embodiments, the target includes any target described herein with respect to method 100 in Figure 1.

[0043] At 350, the MRS procedure is performed on the subject. In some embodiments, the MRS procedure includes any MRS procedure described herein with respect to method 100 in Figure 1.

[0044] In some embodiments, the target molecule, a derivative of the target molecule, or the target hyperpolarizable molecule is dissolved in a solution for any of steps 310, 320, and 330. In some embodiments, the solution is characterized by any pH as described herein with respect to method 100 of Figure 1. In some embodiments, the hyperpolarizable nucleus is characterized by any T1 relaxation time as described herein with respect to method 100 of Figure 1 in a solution at any second magnetic field as described herein with respect to method 100 of Figure 1.

[0045] In some embodiments, Method 300 (i.e., all of steps 310, 320, 330, 340, and 350) is carried out for any time period described herein with respect to Method 100 in Figure 1.

[0046] Figure 4 shows exemplary compositions 400 comprising hyperpolarized molecules of a target having hypernuclear spin polarization in a solution, according to various embodiments. As shown in Figure 4, composition 400 comprises solvent 410 and hyperpolarized molecules of a target 420 dissolved in the solvent. In some embodiments, the hyperpolarized molecules of a target 420 comprises any hyperpolarizable nuclei as described herein with respect to Method 100 of Figure 1. In some embodiments, the hyperpolarizable nuclei are characterized by any second magnetic field as described herein with respect to Method 100 of Figure 1, any T1 relaxation time as described herein with respect to Method 100 of Figure 1, and any pH as described herein with respect to Method 100 of Figure 1.

[0047] In some embodiments, the target hyperpolarizable molecule 420 includes any target hyperpolarizable molecule described herein with respect to method 100 of Figure 1. In some embodiments, the hyperpolarizable nuclei of the target hyperpolarizable molecule 420 have nuclear spin polarization. In some embodiments, the nuclear spin polarization in the target molecule includes any second nuclear spin polarization described herein with respect to method 100 of Figure 1.

[0048] In some embodiments, solvent 410 includes D2O solvent.

[0049] In some embodiments, composition 400 further comprises a scavenger. In some embodiments, the scavenger comprises any scavenger described herein in relation to method 100 of Figure 1. In some embodiments, composition 400 comprises a scavenger at any concentration described herein in relation to method 100 of Figure 1.

[0050] Hyperpolarization and parahydrogen As used in this disclosure, hyperpolarization describes a state in which the absolute difference between a spin state in one state (e.g., spin-up, a nuclear spin state, a proton spin state, a carbon-13 spin state, etc.) and a group of spin states in another state (e.g., spin-down) exceeds the absolute difference of the corresponding difference in thermal equilibrium.

[0051] Parahydrogen can be used as a polarization source, consistent with the disclosed embodiments. Parahydrogen is a form of molecular hydrogen in which two proton spins are in a singlet state, as described herein. The disclosed embodiments are not limited to a specific method for producing parahydrogen. Parahydrogen can be formed in gaseous or liquid form. In some embodiments, parahydrogen is produced in gaseous form by flowing hydrogen gas through a chamber at a low temperature using a catalyst (e.g., iron oxide or another suitable catalyst). The hydrogen gas may contain both parahydrogen and orthohydrogen. The low temperature can cause the hydrogen gas to reach thermodynamic equilibrium in the chamber, increasing the population of parahydrogen.

[0052] The disclosed embodiments are not limited to a specific parahydrogen production or use location. Parahydrogen may be produced 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 a suitable pressure or temperature. In some embodiments, the first location may be a physical location such as a room, laboratory, a specific warehouse, a hospital, or other location where parahydrogen is produced.

[0053] The disclosed embodiments are not limited to any particular parahydrogen transport method. The generated parahydrogen may be transported in a chamber that may be different from the chamber in which the parahydrogen was generated. The chamber in which the parahydrogen gas is transported may be maintained at a suitable pressure or temperature that can be transported by vehicle or by person. Transporting parahydrogen may include moving parahydrogen from one container to a different container. Transporting parahydrogen may include moving parahydrogen within the same location, such as from one part of a room to another part of a room. Transporting parahydrogen may include moving parahydrogen from one room in a building to a different room in the same building, or to a nearby building. Transporting parahydrogen may include moving parahydrogen to a different location in a different part of the same city, or to a different city. Transporting parahydrogen may include transporting parahydrogen near a polarizer, NMR device, or MRI device. Transporting parahydrogen may include packaging or transporting parahydrogen in a suitable container.

[0054] In some embodiments, the collective difference between two spin states is the difference between the two spin states divided by the total collective of the two spin states. The collective difference can be expressed as a fractional collective difference or a percentage collective difference. In some embodiments, the fractional collective difference is at least about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or greater, and at most about 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1 or less, or within the range defined by any two of the aforementioned values.

[0055] Hydrogen gas can exhibit a collective difference between proton spin states that significantly exceeds the collective difference between proton spin states in thermal equilibrium. Parahydrogen can have a large collective difference between either the singlet spin state or the triplet spin state. For example, in the case of Iz1Iz2, there is a large collective difference between the spin states |↑>|↓> and |↑>|↑>. The collective difference between proton spin states can be at least about 0.1 (e.g., a 10% difference in spin states - 55% of parahydrogen molecules in a sample are in the singlet state and 45% are in the triplet state), 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or greater, and at most about 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or less, or within the range defined by any two of the aforementioned values.

[0056] Target molecules and bio-related contrast agents The disclosed embodiments include systems and methods for generating and utilizing target molecules having clinically significant polarization, concentration, volume, or purity. In some embodiments, the method is for preparing a target molecule. In some embodiments, the target molecule is suitable for use in NMR or MRI procedures. In some embodiments, the target molecule increases NMR or MRI signals and signal-to-noise ratio (SNR). In some embodiments, the target molecule is suitable for use in solution NMR spectroscopy. In some embodiments, the target molecule is a chemical compound. In some embodiments, the target molecule is a metabolite (e.g., a molecule having biological relevance such as an amino acid, sugar, its derivatives, etc.) suitable for use in NMR metabolomics applications. In some embodiments, the target molecule is suitable for in vitro probing of the metabolism of cell cultures or other biological tissues. In some embodiments, the target molecule is used in an NMR probe to investigate transient effects that require high signal enhancement due to hyperpolarization, such as proton exchange between water and biomolecules. In some embodiments, the target molecule is a small molecule or metabolite suitable for injection into cells, tissues, or organisms for detection in an MRI scan. In some embodiments, the target molecule is introduced into a chamber for further analysis by NMR or MRI procedures. In some embodiments, the target molecule is enriched with one or more deuterium ([ 2 2 H) or carbon-13 ([ 13 13 C) atoms.

[0057] In accordance with the disclosed embodiments, the molecule of interest may be configured to include a bio-related contrast agent. In some embodiments, the bio-related contrast agent may be suitable for use in NMR or MRI procedures. In some embodiments, the bio-related contrast agent may increase the NMR or MRI signal or signal-to-noise ratio (SNR). In some embodiments, the bio-related contrast agent may be suitable for use in solution NMR spectroscopy. In some embodiments, the bio-related contrast agent may be a metabolite (e.g., a bio-related molecule such as an amino acid, sugar, or its derivative) suitable for use in NMR metabolomics applications. In some embodiments, the bio-related contrast agent is used for perfusion or contrast-enhanced contrast in MRI scans. In some embodiments, the bio-related contrast agent may be suitable for in vitro probing of the metabolism of cell cultures or other biological tissues. In some embodiments, the bio-related contrast agent is used for in vitro probing of the metabolism of cell cultures or other biological tissues. In some embodiments, the bio-related contrast agent may be used with NMR probes to investigate transient effects requiring high signal enhancement due to hyperpolarization, such as proton exchange between water and biomolecules. In some embodiments, the bio-relevant contrast agent may be a small molecule or metabolite suitable for injection into cells, tissues, or organisms for detection in MRI scans. In some embodiments, the bio-relevant contrast agent is introduced into a chamber for further analysis by NMR or MRI procedures. In some embodiments, the bio-relevant contrast agent may be one or more 2 H or 13 It is concentrated by carbon atoms.

[0058] In some embodiments, the bio-relevant contrast agent is pirubate, lactate, alpha-ketoglutarate, bicarbonate, fumarate, urea, dehydroascorbate, glutamate, glutamine, acetate, dihydroxyacetone, acetacetate, glucose, ascorbate, zymonate, alanine, fructose, imidazole, nicotinamide, nitroimidazole, pyrazinamide, isoniazid, any of the above conjugate acids, natural and unnatural amino acids, their esters, or any of the above.2 H, 13 C, or 15 This includes versions rich in N. In some embodiments, the bio-required contrast agent comprises pirubate, lactate, or alpha-ketoglutarate. In some embodiments, the bio-required contrast agent comprises pirubate. In some embodiments, the bio-required contrast agent comprises lactate. In some embodiments, the bio-required contrast agent comprises alpha-ketoglutarate (e.g., ethylalpha-ketoglutarate).

[0059] In some embodiments, the bio-relevant contrast agent contains at least one non-hydrogen polarized nucleus (i.e., a non-hydrogen nuclear spin). In some embodiments, the non-hydrogen nucleus contains at least one spin 1 / 2 atom. In some embodiments, the non-hydrogen nuclear spin is 13 C or 15 Contains N. In some embodiments, the bio-related contrast agent is at least partially isotope-labeled with non-hydrogen nuclear spin. That is, in some embodiments, the bio-related contrast agent is at least partially enriched with non-hydrogen nuclear spin compared to analogs of the bio-related contrast agent characterized by its natural abundance of non-hydrogen nuclear spin. In some embodiments, the bio-related contrast 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 greater, at most about 99%, 98%, 97%, 96%. The non-hydrogen nucleus spins are enriched to characterize them in abundances of %, 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, or within the range defined by any two of the aforementioned values.

[0060] In some embodiments, non-hydrogen nuclear spins are NMR-inactive (i.e., spin-0) nuclei of bio-related imaging agent analogs characterized by their natural abundance (e.g., 12 C) or quadrupolecular (i.e., spin > 1 / 2) nuclei (e.g., nitrogen-14, 14 Substitute N). For example, with its natural abundance 13 Pyruvate analogs containing C have the structure H3C-C * (=O)-C * One of the C's in OOH * So, approximately 98.9% 12 C and approximately 1.1% 13 It may contain C. As a bio-reactive contrast agent, pyruvate contains one or both C. * in any abundance as described herein 13 To include C, instead 13 It can be isotope enriched in 1C. When used herein, * C and C * teeth, 12 C or 13 This lists the carbon that may be any of the 1C carbon isotopes. As another example, its natural abundance is 15 Urea analogs containing N have the structure H2N * -C(=O)- * Any of the N in NH2 * Approximately 99.6% 14 N and approximately 0.4% 15 It may contain N. As a bio-reactive contrast agent, urea contains one or both N. * in any abundance as described herein 15 Instead, include N 15 It can be isotope enriched with N. When used herein, * N and N * teeth, 14 N or 15 Describes nitrogen, which can be any of the nitrogen isotopes of N.

[0061] In some embodiments, the molecule in question is partially deuterated (i.e., it contains fewer deuterium atoms than all the chemical sites on the molecule in question where protons are typically expected). In some embodiments, the molecule in question is fully deuterated (i.e., it contains deuterium atoms in all chemical sites on the molecule in question where protons are typically expected).

[0062] Precursor of the target molecule In some embodiments, the target hyperpolarizing molecule (such as a hyperpolarizing bio-related imaging agent as described herein) is generated via a PHIP process between a parahydrogen, a precursor of the target molecule, and a PHIP catalyst as described herein. In some embodiments, the precursor of the target molecule contains a carbon-carbon triple bond at a position where the target molecule contains a carbon-carbon double bond or a carbon-carbon double bond.

[0063] In some embodiments, the hyperpolarized molecule of interest (such as a highly polarized bio-imaging agent as described herein) is generated via a PHIP-SAH process between parahydrogen, a precursor of the molecule of interest, and a PHIP-SAH catalyst as described herein. In some embodiments, such precursors include the molecule of interest (e.g., a bio-imaging agent) and a side arm. In some embodiments, the molecule of interest is covalently bonded to the side arm. In some embodiments, the molecule of interest is attached to the side arm via a moving part, such as a PHIP moving part which is part of the side arm. In some embodiments, the side arm is cleaved from the precursor (e.g., via hydrolysis) after parahydrogenation and spin order transfer to the molecule of interest. Examples of PHIP-SAH precursors are described, for example, in CT Publication WO2021198776, entitled “SYSTEMS AND METHODS FOR GENERATION OF HYPERPOLARIZED MATERIAL,” filed on March 31, 2021; PCT Publication WO2022200859, entitled “SYSTEMS AND METHODS FOR GENERATION OF HYPERPOLARIZATED MATERIALS,” filed on March 23, 2022; and PCT Publication WO2023026252, entitled “SYSTEMS AND METHODS FOR GENERATION OF HYPERPOLARIZED MATERIALS,” filed on August 26, 2022. Each of these documents is incorporated herein by reference in its entirety for all purposes.

[0064] The sidearms can be para-hydrogenated using para-hydrogen (for example, by mixing the precursor and para-hydrogen). In some embodiments, the hydrogenation produces an Iz1Iz2 order, which is a lower energy state between |↑>|↓> and |↓>|↑>, or a singlet spin order of two hydrogen spins, depending on whether the hydrogenation is performed in a low or high magnetic field.

[0065] In some embodiments, the precursor is selected after hydrogenation and other optional chemical reactions to be suitable for use in NMR or MRI applications where the molecule of interest is hyperpolarized. In some embodiments, additional chemical reactions after hydrogenation can be used to separate the molecule of interest from the precursor. Such additional chemical reactions may include, for example, cleavage of the side arms of the precursor by hydrolysis. For example, the molecule of interest may be a metabolite molecule, so that the precursor may be a derivative of a metabolite molecule. The molecule of interest may be polarized using the PHIP-SAH method (i.e., para-hydrogenation of the side arms and subsequent polarization transfer to the molecule of interest). After hydrogenation and polarization transfer, the bonds in the precursor (e.g., ester bonds) may be hydrolyzed to produce the polarized molecule of interest and separate side arm elements.

[0066] As used herein, hydrolysis is defined as the cleavage of a molecule via a nucleophilic substitution reaction involving the addition of the element water. Hydrolysis may also be carried out under anhydrous conditions in the presence of hydroxide ions.

[0067] Parahydrogenation In accordance with the disclosed embodiments, a precursor of the molecule of interest (such as any precursor described herein) may be parahydrogenated by combining the precursor, parahydrogen, and a hydrogenation catalyst (such as any fluorinated PHIP or PHIP-SAH catalyst described herein). The disclosed embodiments are not limited to any particular method for producing a parahydrogenated precursor. In some embodiments, the precursor is added to a mixture containing parahydrogen. In some embodiments, the parahydrogen gas is added to a solution containing the precursor (for example, the parahydrogen gas may be bubbled in such a solution). In the hydrogenation of the precursor, the parahydrogen may create an Iz1Iz2 order, a preferred group of lower energy states between |↑>|↓>, |↓>|↑>, or a singlet spin order on two hydrogen spins in the precursor.

[0068] The precursor may have unsaturated bonds (such as unsaturated carbon-carbon double bonds or unsaturated carbon-carbon triple bonds) that can be hydrogenated by parahydrogen gas. After combining the precursor with parahydrogen, a proportion of the precursor can be hydrogenated, which is at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or greater, and at most about 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, or less, or within the range defined by any two of the aforementioned values.

[0069] In some embodiments, the parahydrogenated precursor has a gregacy difference of at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 40%, 45%, 50%, or greater in the parahydrogenated proton spin state, and at most about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, or within the range defined by any two of the aforementioned values. For example, in some embodiments, the group differences are 10%~15%, 10%~20%, 10%~25%, 10%~30%, 10%~35%, 10%~40%, 10%~45%, 10%~50%, 15%~20%, 15%~25%, 15%~30%, 15%~35%, 15%~40%, 15%~45%, 15%~50%, 20%~25%, 20%~30%, 2 The percentages are 0%~35%, 20%~40%, 20%~45%, 20%~50%, 25%~30%, 25%~35%, 25%~40%, 25%~45%, 25%~50%, 30%~35%, 30%~40%, 30%~45%, 30%~50%, 35%~40%, 35%~45%, 35%~50%, 40%~45%, 40%~50%, or 45%~50%. In some embodiments, the collective difference is between spin states containing parahydrogenated protons and spin states containing other nuclear spins, e.g., additional protons on the compound. In some embodiments, the parahydrogenated precursor includes a side arm, and the parahydrogenated spins may be located on the side arm.

[0070] In some embodiments, the concentration of the hydrogenation catalyst during hydrogenation is at least about 0.1 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, or greater than that, maximum The values ​​are approximately 100mM, 90mM, 80mM, 70mM, 60mM, 50mM, 40mM, 30mM, 20mM, 10mM, 9mM, 8mM, 7mM, 6mM, 5mM, 4mM, 3mM, 2mM, 1mM, 0.9mM, 0.8mM, 0.7mM, 0.6mM, 0.5mM, 0.4mM, 0.3mM, 0.2mM, 0.1mM, or smaller, or within the range defined by any two of the aforementioned values.

[0071] In some embodiments, the parahydrogenation process is carried out in an organic solvent. In some embodiments, the organic solvent includes acetone. In some embodiments, the parahydrogenation process is carried out in a fluorinated solvent.

[0072] The disclosed embodiments may include methods implemented by the disclosed system for generating a hyperpolarized molecule of interest. The methods of this disclosure may include mixing a solution (e.g., by a mixing mechanism) comprising a precursor and a hydrogenation catalyst for the molecule of interest. The mixing mechanism may be a device for introducing, holding, and facilitating a blend, mixture, or solution of two or more materials. In some embodiments, the mixing mechanism is located within a chamber, and the mixing occurs within the chamber. In some embodiments, the solutions are mixed at a location away from the chamber. The solution may be in an amount of at least about 1 milliliter (mL), 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 20 mL, 30 mL, 40 mL, 50 mL, 60 mL, 70 mL, 80 mL, 90 mL, 100 mL, or more, and may be in an amount of at most about 100 mL, 90 mL, 80 mL, 70 mL, 60 mL, 50 mL, 40 mL, 30 mL, 20 mL, 10 mL, 9 mL, 8 mL, 7 mL, 6 mL, 5 mL, 4 mL, 3 mL, 2 mL, 1 mL, or less, or within the range defined by any two of the aforementioned values.

[0073] In some embodiments, the mixing mechanism is a gas-liquid exchange mechanism. For example, the gas-liquid exchange mechanism may be a bubbler or a diffusion system. In some embodiments, the mixing mechanism includes a membrane adapted to allow the diffusion of molecular hydrogen. In some embodiments, mixing may be carried out using a spray chamber, in which the solution is sprayed into a chamber filled with pressurized parahydrogen.

[0074] In some embodiments, the catalyst is any molecule, complex, or particle system described in this disclosure that catalyzes hydrogenation. In some embodiments, the catalyst includes a homogeneous metal catalyst such as a rhodium complex or a ruthenium complex. Rhodium complexes may be used for the preparation and activation of precursor molecules and parahydrogen. In some embodiments, heterogeneous metal catalysts are attached to nanoparticles.

[0075] Various embodiments of this disclosure describe introducing a solution comprising a precursor of the molecule of interest and a hydrogenation catalyst into a chamber configured to hold the solution during polarization transfer. In some embodiments, the solution is mixed within the chamber. In some embodiments, the solution is hydrogenated within the chamber. In some embodiments, the chamber is located within a magnetic shield (e.g., a mu-metal shield). The magnetic shield can reduce the influence of the Earth's magnetic field (or other external magnetic fields) and allows modulation of the amplitude of the low-level magnetic field applied to the solution. Therefore, placing the solution within a chamber may include placing the solution within a magnetic shield.

[0076] As described herein, in some embodiments, parahydrogenation occurs before polarization transfer (e.g., before amplitude modulation, such as a magnetic field applied to a solution). In some embodiments, parahydrogenation occurs during polarization transfer. For example, parahydrogen can be combined with the solution during modulation of the magnetic field amplitude (e.g., flowed through the solution or bubbled).

[0077] In some embodiments, parahydrogen gas is combined with a solution in a hydrogenation chamber under pressure. The pressure may be at least about 10 bar, 15 bar, 20 bar, 30 bar, 50 bar, or greater, and at most about 50 bar, 30 bar, 20 bar, 15 bar, 10 bar, or less, or within the range defined by any two of the aforementioned values. In some embodiments, parahydrogen is combined with a solution in a metal chamber capable of withstanding the pressure. Parahydrogen may be combined with the solution over a time interval (or the dissolution of parahydrogen may occur in less than a time interval). The time interval may be at most about 90 seconds, 60 seconds, 30 seconds, 20 seconds, 10 seconds, 9 seconds, 8 seconds, 7 seconds, 6 seconds, 5 seconds, 4 seconds, 3 seconds, 2 seconds, 1 second, or less, and at least about 1 second, 2 seconds, 3 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 20 seconds, 30 seconds, 60 seconds, 90 seconds, or longer, or within the range defined by any two of the aforementioned values. In some embodiments, hydrogenation is carried out or occurs within the time interval.

[0078] Polarization shift using high-frequency waveforms

[0079] In some embodiments, the concentration of the precursor or target molecule in the solution before polarization transfer is at least about 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 200 mM, 300 mM, 400 mM, 500 mM, 600 mM, 700 mM, 800 mM, 900 mM, 1,000 mM, or greater. The maximum value is approximately 1,000 mM, 900 mM, 800 mM, 700 mM, 600 mM, 500 mM, 400 mM, 300 mM, 200 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, or smaller, or within the range defined by any two of the aforementioned values. The amount of solution should be at least approximately 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 20 mL, 30 mL, 40 mL, 50 mL, 60 mL, 70 mL, 80 mL, 90 mL, 100 mL, 200 mL, 300 mL, 400 mL, 500 mL, 600 mL, 700 mL, 800 mL, 900 mL, 1,000 mL, 2,000 mL, or larger, up to approximately 2,000 mL. 000mL, 900mL, 800mL, 700mL, 600mL, 500mL, 400mL, 300mL, 200mL, 100mL, 90mL, 80mL, 70mL, 60mL, 50mL, 40mL, 30mL, 20mL, 10mL, 9mL, 8mL, 7mL, 6mL, 5mL, 4mL, 3mL, 2mL, 1mL, or smaller, or within the range defined by any two of the aforementioned values.

[0080] Various embodiments of this disclosure describe applying a polarized transitional magnetic perturbation aimed at generating a magnetic field around a solution. In some embodiments, the magnetic field is at least about 0.1 Gauss (G), 0.2G, 0.3G, 0.4G, 0.5G, 0.6G, 0.7G, 0.8G, 0.9G, 1G, 2G, 3G, 4G, 5G, 6G, 7G, 8G, 9G, 10G, 20G, 30G, 40G, 50G, 60G, 70G, 80G, 90G, 100G, 200G, 300G, 400G, 500G, 600G, 700G, 800G, 90 0G, 1,000G, 2,000G, 3,000G, 4,000G, 5,000G, 6,000G, 7,000G, 8,000G, 9,000G, 10,000G, 20,000G, 30,000G, 40,000G, 50,000G, 60,000G, 70,000G, 80,000G, 90,000G, 100,000G, 200,000G, or larger, up to approximately 200 ,000G, 100,000G, 90,000G, 80,000G, 70,000G, 60,000G, 50,000G, 40,000G, 30,000G, 20,000G, 10,000G, 9,000G, 8,000G, 7,000G, 6,000G, 5,000G, 4,000G, 3,000G, 2,000G, 1,000G, 900G, 800G, 700G, 600G, 500G, The magnetic field has an intensity of 400G, 300G, 200G, 100G, 90G, 80G, 70G, 60G, 50G, 40G, 30G, 20G, 10G, 9G, 8G, 7G, 6G, 5G, 4G, 3G, 2G, 1G, 0.9G, 0.8G, 0.7G, 0.6G, 0.5G, 0.4G, 0.3G, 0.2G, 0.1G, or less, or within the range defined by any two of the aforementioned values. In some embodiments, the magnetic field has an intensity of 0.1G to 200,000G around the solution. The magnetic perturbation can be generated by an electromagnet or a permanent magnet. The magnetic field can be applied to the sample in pulse or continuous wave (CW) form. The magnetic perturbation may be static or vary over time.

[0081] A signal generator may be configured to generate one or more radio frequency (RF) waveforms that can be applied to a sample to shift its polarization. The signal generator may include another computing unit, processor, controller, connected memory, PC, computer service, or any device capable of performing computational operations and generating outputs using inputs. In some embodiments, the RF coil may emit or "apply" a pulse sequence containing a first RF waveform. In some embodiments, the RF coil may have one or more channels. A channel may be a path for the RF signal. At least one channel may be provided for each different type of NMR spectroscopy. In some embodiments, 1 H has at least one channel, 2 H, 13 C, 15 N, 19 F, and 31 One, two, three, four, or five of P have at least one channel. For example, the first RF waveform is transmitted through one or more high-frequency coils (RF coils) arranged around the sample. 1 It can be applied to the H channel. In some embodiments, the second RF waveform is applied to the RF coil. 13 It is applied to the C channel. In some embodiments, 1 H channel and 13 The RF waveform on the C channel is configured to have a polarization shift sequence applied, such as PH-INEPT, Goldman's sequence, S2M, S2hM, SLIC, ADAPT, or ESOTERIC.

[0082] In some embodiments, the RF waveform is configured to support polarization movement even in the presence of a large proton full width half-maximum (FWHM). Such RF waveforms may include a pulse sequence that may contain tens to hundreds of RF pulses. The sequence may be configured so that the pulses protect against the detrimental effects of magnetic field inhomogeneity on polarization movement.

[0083] In some embodiments, the polarization pulse sequence is, for example, when the chemical shift difference is greater than the J-coupling between them, two non-equivalents 1 It is configured to transfer spin order from H hydrogenation spins. ESOTHERIC may be, for example, a pulse sequence suitable for polarization transfer in this regime.

[0084] In some embodiments, the pulse sequence is equivalent when, for example, the chemical shift difference is smaller than the J-coupling between them. 1These pulse sequences are configured to transfer spin order from the H hydrogen spin. , 9mT, 10mT, 20mT, 30mT, 40mT, 50mT, 60mT, 70mT, 80mT, 90mT, 100mT, 200mT, 300mT, 400mT, 500mT, 600mT, 700mT, 800mT, 900mT, 1,000mT, 2,000mT, 3,000mT, 4,000mT, 5,000mT, 6,000mT, or greater than that, up to approximately 6,000mT, 5,000mT, 4,000mT, 3,000mT, 2,000mT, 1,000mT, 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, These sequences may be used in magnetic fields having strengths of 0.9mT, 0.8mT, 0.7mT, 0.6mT, 0.5mT, 0.4mT, 0.3mT, 0.2mT, 0.1mT, 0.09mT, 0.08mT, 0.07mT, 0.06mT, 0.05mT, 0.04mT, 0.03mT, 0.02mT, 0.01mT, or less, or within the range defined by any two of the aforementioned values. Examples of such sequences may be Goldman sequences (M. Goldman, H. Johannesson, CRPhys. 2005, 6, 575-581; this reference is incorporated herein by reference in relation to pulse sequence configurations for transferring spin order), singlet-to-heteronuclear magnetization (S2hM) sequences, or other sequences used in singlet NMR (e.g., ADAPT, SLIC, etc.).

[0085] In some embodiments, the magnetic shield is applied to a magnetic field applied to a solution of at least about 0mG, 0.1mG, 0.2mG, 0.3mG, 0.4mG, 0.5mG, 0.6mG, 0.7mG, 0.8mG, 0.9mG, 1mG, 2mG, 3mG, 4mG, 5mG, 6mG, 7mG, 8mG, 9mG, 10mG, 20mG, 30mG, 40mG, 50mG, 60mG, 70mG, 80mG, 90mG, 100mG, or greater, up to about 100 The magnetic shield is configured to maintain a magnetic field that is within the range defined by any two of the above values ​​when applied to a solution with a magnetic field of mG, 90mG, 80mG, 70mG, 60mG, 50mG, 40mG, 30mG, 20mG, 10mG, 9mG, 8mG, 7mG, 6mG, 5mG, 4mG, 3mG, 2mG, 1mG, 0.9mG, 0.8mG, 0.7mG, 0.6mG, 0.5mG, 0.4mG, 0.3mG, 0.2mG, or 0.1mG or less, or when applied to a solution with a magnetic field of any two of the above values. The magnetic shield can maintain the magnetic field strength in the polarization chamber even when a polarization waveform is applied to one or more high-frequency coils, even at such amplitudes.

[0086] In accordance with the disclosed embodiments, the RF waveform may be applied to a solution containing a parahydrogenated precursor.

[0087] Polarization shift using magnetic field modulation In some embodiments, polarization-transferring magnetic perturbations are carried out within a magnetic shield (e.g., a mu-shield) to achieve a homogeneous low magnetic field. The magnetic shield operates in a microtesla (μT) magnetic field, which is below the Earth's magnetic field. 13This enables the performance of polarization transfer to the C nuclear spin. Low magnetic fields include at least about 0mG, 0.1mG, 0.2mG, 0.3mG, 0.4mG, 0.5mG, 0.6mG, 0.7mG, 0.8mG, 0.9mG, 1mG, 2mG, 3mG, 4mG, 5mG, 6mG, 7mG, 8mG, 9mG, 10mG, 20mG, 30mG, 40mG, 50mG, 60mG, 70mG, 80mG, 90mG, 100mG, or greater, up to about 100mG, 9 It may be 0mG, 80mG, 70mG, 60mG, 50mG, 40mG, 30mG, 20mG, 10mG, 9mG, 8mG, 7mG, 6mG, 5mG, 4mG, 3mG, 2mG, 1mG, 0.9mG, 0.8mG, 0.7mG, 0.6mG, 0.5mG, 0.4mG, 0.3mG, 0.2mG, 0.1mG, or smaller, or within the range defined by any two of the aforementioned values.

[0088] In such a magnetic field, polarization is related to proton spin, 2 H, 13 C, 15 N, 19 F, and 31 The polarization is moved by utilizing level pseudocrossing (LAC) between P and other spin species of interest. In some embodiments, the magnetic field may be adjusted to a specific magnetic field strength relative to the LAC, as performed, for example, in the SABRE-SHEATH experiment. In various embodiments, the magnetic field strength may be adjusted over time to enable robust polarization movement in larger volume samples. For example, the magnetic field strength may be swept through the LAC conditions. Alternatively or additionally, the sample may be physically moved within the magnetic field. Such modulation can relax constraints on magnetic field homogeneity and magnetic field offset. Thus, robust polarization movement can be achieved in larger volumes with greater efficiency. Furthermore, relaxing constraints on magnetic field homogeneity and magnetic field offset may allow the use of less complex, more precise, or more expensive polarization systems.

[0089] The lower limit of magnetic field modulation is at least approximately -10μT, -9μT, -8μT, -7μT, -6μT, -5μT, -4μT, -3μT, -2μT, -1.9μT, -1.8μT, -1.7μT, -1.6μT, -1.5μT, -1.4μT, -1.3μT, -1.2μT, -1.1μT, -1μT, -0.9μT, -0.8μT, -0.7μT, -0.6μT, -0.5μT, -0.4μT, -0.3μT, -0.2μT, -0.1μT, or greater, up to approximately -0.1μT. The values ​​may be -0.2μT, -0.3μT, -0.4μT, -0.5μT, -0.6μT, -0.7μT, -0.8μT, -0.9μT, -1μT, -1.1μT, -1.2μT, -1.3μT, -1.4μT, -1.5μT, -1.6μT, -1.7μT, -1.8μT, -1.9μT, -2μT, -3μT, -4μT, -5μT, -6μT, -7μT, -8μT, -9μT, -10μT, or less, or within the range defined by any two of the aforementioned values. The upper limits of modulation are at least approximately 0.1μT, 0.2μT, 0.3μT, 0.4μT, 0.5μT, 0.6μT, 0.7μT, 0.8μT, 0.9μT, 1μT, 1.1μT, 1.2μT, 1.3μT, 1.4μT, 1.5μT, 1.6μT, 1.7μT, 1.8μT, 1.9μT, 2μT, 3μT, 4μT, 5μT, 6μT, 7μT, 8μT, 9μT, 10μT, or greater, with a maximum of approximately 10μT and 9μT. , 8μT, 7μT, 6μT, 5μT, 4μT, 3μT, 2μT, 1.9μT, 1.8μT, 1.7μT, 1.6μT, 1.5μT, 1.4μT, 1.3μT, 1.2μT, 1.1μT, 1μT, 0.9μT, 0.8μT, 0.7μT, 0.6μT, 0.5μT, 0.4μT, 0.3μT, 0.2μT, 0.1μT, or smaller, or within the range defined by any two of the aforementioned values.

[0090] The magnetic field is applied to volumes of at least approximately 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 20 mL, 30 mL, 40 mL, 50 mL, 60 mL, 70 mL, 80 mL, 90 mL, 100 mL, 200 mL, 300 mL, 400 mL, 500 mL, 600 mL, 700 mL, 800 mL, 900 mL, 1,000 mL, 2,000 mL, or larger, with a maximum of approximately 2,000 mL, 1,000 mL, 90 mL. Such amplitudes may have over volumes of 0 mL, 800 mL, 700 mL, 600 mL, 500 mL, 400 mL, 300 mL, 200 mL, 100 mL, 90 mL, 80 mL, 70 mL, 60 mL, 50 mL, 40 mL, 30 mL, 20 mL, 10 mL, 9 mL, 8 mL, 7 mL, 6 mL, 5 mL, 4 mL, 3 mL, 2 mL, 1 mL, or smaller, or within the range defined by any two of the aforementioned values. Modulation may be performed over a certain duration. The duration may be at least approximately 100 milliseconds (ms), 200 milliseconds, 300 milliseconds, 400 milliseconds, 500 milliseconds, 600 milliseconds, 700 milliseconds, 800 milliseconds, 900 milliseconds, 1 second (s), 2 seconds, 3 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 20 seconds, 30 seconds, or longer, and at most approximately 30 seconds, 20 seconds, 10 seconds, 9 seconds, 8 seconds, 7 seconds, 6 seconds, 5 seconds, 4 seconds, 3 seconds, 2 seconds, 1 second, 900 milliseconds, 800 milliseconds, 700 milliseconds, 600 milliseconds, 500 milliseconds, 400 milliseconds, 300 milliseconds, 200 milliseconds, 100 milliseconds, or shorter, or within the range defined by any two of the aforementioned values.

[0091] Therefore, the rate of change of the magnetic field amplitude is at least approximately 0.01 μT / sec, 0.015 μT / sec, 0.02 μT / sec, 0.025 μT / sec, 0.03 μT / sec, 0.035 μT / sec, 0.04 μT / sec, 0.045 μT / sec, 0.055 μT / sec, 0.06 μT / sec, 0.065 μT / sec, 0.07 μT / sec, 0.075 μT / sec, 0.08 μT / sec, 0.085 μT / sec, 0.09 μT / sec 1 μT / sec, 0.095 μT / sec, 0.1 μT / sec, 0.15 μT / sec, 0.2 μT / sec, 0.25 μT / sec, 0.3 μT / sec, 0.35 μT / sec, 0.4 μT / sec, 0.45 μT / sec, 0.5 μT / sec, 0.55 μT / sec, 0.6 μT / sec, 0.65 μT / sec, 0.7 μT / sec, 0.75 μT / sec, 0.8 μT / sec, 0.85 μT / sec, 0.9 μT / sec, 0.95 μT / sec, 1 μT / sec, or greater than that. Large: approximately 1 μT / sec, 0.95 μT / sec, 0.9 μT / sec, 0.85 μT / sec, 0.8 μT / sec, 0.75 μT / sec, 0.7 μT / sec, 0.65 μT / sec, 0.6 μT / sec, 0.55 μT / sec, 0.5 μT / sec, 0.45 μT / sec, 0.4 μT / sec, 0.35 μT / sec, 0.3 μT / sec, 0.25 μT / sec, 0.2 μT / sec, 0.15 μT / sec, 0.1 μT / sec, 0.095 μT / sec, 0.09 μT / sec, 0.0 The rate of change of the magnetic field amplitude may be 8 μT / sec, 0.075 μT / sec, 0.07 μT / sec, 0.065 μT / sec, 0.06 μT / sec, 0.055 μT / sec, 0.05 μT / sec, 0.045 μT / sec, 0.04 μT / sec, 0.035 μT / sec, 0.03 μT / sec, 0.025 μT / sec, 0.02 μT / sec, 0.015 μT / sec, 0.01 μT / sec, or less, or within the range defined by any two of the aforementioned values. The upper limit of the rate of change of the magnetic field amplitude may be determined by the capabilities of the equipment used to perform the sweep.

[0092] In some embodiments, when the magnetic field is within the upper and lower limits disclosed above, the spatial deviation of the magnetic field across the modulated volume is less than about half (or a quarter, or an eighth, or a tenth) of the amplitude of the magnetic field. For example, when the magnetic field strength is less than 2 μT (or greater than -2 μT), the spatial deviation of the magnetic field across the modulated volume may be less than 1 μT. As an additional example, when the magnetic field strength is less than 10 μT (or greater than -10 μT), the spatial deviation of the magnetic field across the modulated volume may be less than 5 μT. The spatial deviation can be measured, for example, by taking at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500 or more spatially randomly sampled or spatially equally distributed measurements of the magnetic field in the volume and calculating the standard deviation of the sampled magnetic field measurements. Such homogeneity can be achieved within a large homogeneous magnetic shield, for example, by having a large puncture solenoid valve through the magnetic shield, or by using a large Helmholtz coil with a large homogeneous region for generating magnetic field amplitude modulation. In some embodiments, the modulation is a sweep of the magnetic field. In some embodiments, the magnetic field amplitude modulation includes non-adiabatic jumps, monotonic amplitude changes, or a combination thereof.

[0093] In some embodiments, following the polarization transfer step, the target molecule (for example, the target molecule) 13 C or 15The non-hydrogen nuclear spins of N) have 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 greater, and a maximum of about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or within the range defined by any two of the aforementioned values. For example, in some embodiments, following the polarization transfer step, the non-hydrogen nucleus spins of the target molecule are 10%~15%, 10%~20%, 10%~25%, 10%~30%, 10%~35%, 10%~40%, 10%~45%, 10%~50%, 15%~20%, 15%~25%, 15%~30%, 15%~35%, 15%~40%, 15%~45%, 15%~50%, 20%~25%, 20% It has nuclear spin polarization of %~30%, 20%~35%, 20%~40%, 20%~45%, 20%~50%, 25%~30%, 25%~35%, 25%~40%, 25%~45%, 25%~50%, 30%~35%, 30%~40%, 30%~45%, 30%~50%, 35%~40%, 35%~45%, 35%~50%, 40%~45%, 40%~50%, or 45%~50%.

[0094] In some embodiments, this polarization is achieved for solution volumes of at least about 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 20 mL, 30 mL, 40 mL, 50 mL, 60 mL, 70 mL, 80 mL, 90 mL, 100 mL, 200 mL, 300 mL, 400 mL, 500 mL, or larger, up to about 500 mL, 400 mL, 300 mL, 200 mL, 100 mL, 90 mL, 80 mL, 70 mL, 60 mL, 50 mL, 40 mL, 30 mL, 20 mL, 10 mL, 9 mL, 8 mL, 7 mL, 6 mL, 5 mL, 4 mL, 3 mL, 2 mL, 1 mL, or smaller, or within the range defined by any two of the aforementioned values.

[0095] In some embodiments, after polarization transfer, a portion of the collective difference in the parahydrogenated proton spin state is the target of the molecule in question (e.g., 13 C or 15 N) Transferred to the polarization of nuclear spin. This portion may be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or greater, at most about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or within the range defined by any two of the aforementioned values. For example, in some embodiments, this portion is 10%~15%, 10%~20%, 10%~25%, 10%~30%, 10%~35%, 10%~40%, 10%~45%, 10%~50%, 15%~20%, 15%~25%, 15%~30%, 15%~35%, 15%~40%, 15%~45%, 15%~50%, 20%~25%, 20%~30%, The percentages are 20%~35%, 20%~40%, 20%~45%, 20%~50%, 25%~30%, 25%~35%, 25%~40%, 25%~45%, 25%~50%, 30%~35%, 30%~40%, 30%~45%, 30%~50%, 35%~40%, 35%~45%, 35%~50%, 40%~45%, 40%~50%, or 45%~50%.

[0096] In some embodiments, magnetic field modulation includes non-adiabatic jumps in the magnetic field. Non-adiabatic jumps may be performed for magnetic fields where level pseudocrossing occurs, involving proton spins and aproton spins. Considering J-coupling between nuclear spins in the system, this value can be analytically calculated or identified by plotting the energy levels of the Hamiltonian for different magnetic fields and identifying the LAC. In some embodiments, the duration for which the magnetic field amplitude is in the LAC condition is at most about 5 seconds, 4 seconds, 3 seconds, 2 seconds, 1 second, 0.9 seconds, 0.8 seconds, 0.7 seconds, 0.6 seconds, 0.5 seconds, 0.4 seconds, 0.3 seconds, 0.2 seconds, 0.1 seconds, or less, at least about 0.1 seconds, 0.2 seconds, 0.3 seconds, 0.4 seconds, 0.5 seconds, 0.6 seconds, 0.7 seconds, 0.8 seconds, 0.9 seconds, 1 second, 2 seconds, 3 seconds, 4 seconds, 5 seconds, or longer, or within the range defined by any two of the aforementioned values.

[0097] In some embodiments, modulating the magnetic field amplitude involves monotonically changing the magnetic field amplitude (or monotonically over each of a limited number of intervals, such as increasing intervals of 1 to 10 and / or decreasing intervals of 1 to 10). In some embodiments, modulating the magnetic field amplitude involves linearly changing the magnetic field amplitude. The initial and final magnetic field amplitudes of the sweep, and the total duration of the sweep, may be optimized for the molecule of interest. In some embodiments, the magnetic field amplitude during the sweep is within lower and upper limits. The lower limit is at least approximately -2μT, -1.9μT, -1.8μT, -1.7μT, -1.6μT, -1.5μT, -1.4μT, -1.3μT, -1.2μT, -1.1μT, -1μT, -0.9μT, -0.8μT, -0.7μT, -0.6μT, -0.5μT, -0.4μT, -0.3μT, -0.2μT, -0.1μT, or greater, up to approximately -0.1μT, -0. The values ​​may be 2μT, -0.3μT, -0.4μT, -0.5μT, -0.6μT, -0.7μT, -0.8μT, -0.9μT, -1μT, -1.1μT, -1.2μT, -1.3μT, -1.4μT, -1.5μT, -1.6μT, -1.7μT, -1.8μT, -1.9μT, -2μT, or smaller, or within the range defined by any two of the aforementioned values. The upper limits are at least approximately 0.1μT, 0.2μT, 0.3μT, 0.4μT, 0.5μT, 0.6μT, 0.7μT, 0.8μT, 0.9μT, 1μT, 1.1μT, 1.2μT, 1.3μT, 1.4μT, 1.5μT, 1.6μT, 1.7μT, 1.8μT, 1.9μT, 2μT, or greater, with a maximum of approximately 2μT and 1.9μT. , 1.8μT, 1.7μT, 1.6μT, 1.5μT, 1.4μT, 1.3μT, 1.2μT, 1μT, 1μT, 0.9μT, 0.8μT, 0.7μT, 0.6μT, 0.5μT, 0.4μT, 0.3μT, 0.2μT, 0.1μT, or smaller, or within the range defined by any two of the aforementioned values.In some embodiments, the modulation duration may be at least about 100 milliseconds, 200 milliseconds, 300 milliseconds, 400 milliseconds, 500 milliseconds, 600 milliseconds, 700 milliseconds, 800 milliseconds, 900 milliseconds, 1 second, 2 seconds, 3 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 20 seconds, 30 seconds, or longer, and at most about 30 seconds, 20 seconds, 10 seconds, 9 seconds, 8 seconds, 7 seconds, 6 seconds, 5 seconds, 4 seconds, 3 seconds, 2 seconds, 1 second, 900 milliseconds, 800 milliseconds, 700 milliseconds, 600 milliseconds, 500 milliseconds, 400 milliseconds, 300 milliseconds, 200 milliseconds, 100 milliseconds, or shorter, or within the range defined by any two of the aforementioned values. In some embodiments, the amplitude change rate varies along the amplitude profile. In some embodiments, a constant adiabatic sweep is calculated by selecting a specific subgroup of level pseudocrossings in the spin system. In some embodiments, magnetic amplitude modulation includes a combination of non-adiabatic jumps, monotonic amplitude modulation, and rate of change sign reversal. In some embodiments, the precursor may be selected or designed such that, following hydrogenation and other potential chemical reactions, one of the products is a target molecule usable in hyperpolarized NMR or MRI applications.

[0098] Hydrolysis, purification, and separation procedures This disclosure presents methods and systems for producing compositions (e.g., clinical dose compositions) containing a hyperpolarized molecule of interest (or a pharmaceutically acceptable salt thereof) in a solvent. In some embodiments, the molecule of interest is produced by additional chemical reactions and / or processing steps after hydrogenation and polarization transfer according to this disclosure. Such additional chemical reactions and / or processing steps may include, but are not limited to, (i) catalytic filtration and collection (e.g., filtration and collection of rhodium atoms and / or iridium atoms), (ii) cleaving the side arms of the precursor molecule of the molecule of interest by hydrolysis, for example, using an aqueous sodium hydroxide solution (e.g., cleaving the precursor) to form the molecule of interest and the side arms, (iii) washing the solution with an organic solvent to separate any resulting aqueous mixture phase from the organic mixture phase, (iv) evaporative extraction of volatile organic compounds from the aqueous mixture (e.g., using nitrogen gas bubbling), and (v) additional filtration / purification / concentration / finishing steps known in the art.

[0099] The volume of the solution containing the target molecule (e.g., after cleavage) and / or the concentration of the target molecule produced may depend on the volume of the solution used for polarization transfer and the concentration of the precursor in that solution. Exemplary ranges of solution volume and precursor concentration are described herein. In more specific examples, solutions of at least about 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 20 mL, 30 mL, 40 mL, 50 mL, 60 mL, 70 mL, 80 mL, 90 mL, 100 mL or larger, up to about 100 mL, 90 mL, 80 mL, 70 mL, 60 mL, 50 mL, 40 mL, 30 mL, 20 mL, 10 mL, 9 mL, 8 mL, 7 mL, 6 mL, 5 mL, 4 mL, 3 mL, 2 mL, 1 mL or smaller, or within the range defined by any two of the aforementioned values, may be produced. In some embodiments, the solution may contain at least about 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 200 mM, 300 mM, 400 mM, 500 mM or larger, up to about 500 mM, 400 mM, 300 mM, 200 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM or smaller, or an amount of the target molecule within the range defined by any two of the aforementioned values.

[0100] In some embodiments, the disclosure describes multi-step liquid-to-liquid separation and purification procedures for generating a dose (e.g., a clinical dose) of an administration composition containing the molecule of interest (e.g., the hyperpolarizing molecule of interest or a pharmaceutically acceptable salt thereof).

[0101] In some embodiments (i.e., the PHIP-SAH procedure), after the polarization step, side arms are cleaved from the molecular precursor of interest (e.g., via hydrolysis using an aqueous mixture) to produce the hyperpolarized molecule and unbound side arms of interest. In some embodiments, following the polarization step, the side arms are cleaved by mixing the solution (containing a first organic solvent and the polarized product, e.g., the hyperpolarized molecule of interest or a pharmaceutically acceptable salt thereof) with a hydrolyzing agent such as a base in an aqueous solution (e.g., sodium hydroxide). In some embodiments, the first organic solvent and the aqueous mixture (e.g., water) produce a biphasic solution. In some embodiments, the first organic solvent and the aqueous mixture (e.g., water) produce a biphasic solution in which a portion of the organic solvent is retained in the aqueous mixture. In some embodiments, the first organic solvent and the aqueous mixture (e.g., water) produce a partial mixture.

[0102] Clinically relevant purity In accordance with the embodiments, methods, and systems disclosed herein, the hyperpolarizing molecules of interest can be separated from other substances in the original solution (e.g., catalyst, solvent, reaction product, etc.). For example, the majority of the hydrogenation catalyst present in the original solution can be removed from the administration composition. In some embodiments, the administered composition contains a hydrogenation catalyst in amounts up to about 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, or less, at least about 0.001%, 0. It may hold hydrogenation catalyst in amounts of 0.02%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, or greater, or an amount of hydrogenation catalyst within the range defined by any two of the aforementioned values.Similarly, the administered composition may contain up to approximately 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, or less of cleavage by-products (e.g., side arms or other residues of cleavage), at least approximately 0. It may retain cleavage by-products in amounts of 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, or greater, or an amount of cleavage by-products within the range defined by any two of the aforementioned values.

[0103] In some embodiments, the methods and systems described herein produce a dose composition in which the concentration of hyperpolarized bio-associated contrast is at least about 100 mM, 150 mM, 200 mM, 250 mM, 300 mM, 350 mM, 400 mM, 450 mM, 500 mM or greater, and at most about 500 mM, 450 mM, 400 mM, 350 mM, 300 mM, 250 mM, 200 mM, 150 mM, 100 mM or less, or within the range defined by any two of the aforementioned values.

[0104] In some embodiments, the methods and systems described herein produce a dose composition in which the polarization of the hyperpolarized bio-associated contrast is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 40%, 45%, 50%, or greater, and at most about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, or within the range defined by any two of the aforementioned values. For example, in some embodiments, the methods and systems described herein allow the polarization of hyperpolarized bio-associated contrast to be 10%~15%, 10%~20%, 10%~25%, 10%~30%, 10%~35%, 10%~40%, 10%~45%, 10%~50%, 15%~20%, 15%~25%, 15%~30%, 15%~35%, 15%~40%, 15%~45%, 15%~50%, 20%~25% A dosage composition is produced that is %, 20%~30%, 20%~35%, 20%~40%, 20%~45%, 20%~50%, 25%~30%, 25%~35%, 25%~40%, 25%~45%, 25%~50%, 30%~35%, 30%~40%, 30%~45%, 30%~50%, 35%~40%, 35%~45%, 35%~50%, 40%~45%, 40%~50%, or 45%~50%.

[0105] In some embodiments, the methods and systems described herein have a catalyst concentration, precursor, or cleavage byproduct concentration of up to approximately 1 μM, 900 nanomolar (nM), 800 nM, 700 nM, 600 nM, 500 nM, 400 nM, 300 nM, 200 nM, 100 nM, 90 nM, 80 nM, 70 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 9 nM, 8 nM, 7 nM, 6 nM, 5 nM, 4 nM, 3 nM, 2 nM, 1 nM, or so The method produces a dosage composition that is smaller than the above, and may be within the range defined by at least about 1 nM, 2 nM, 3 nM, 4 nM, 5 nM, 6 nM, 7 nM, 8 nM, 9 nM, 10 nM, 20 nM, 30 nM, 40 nM, 50 nM, 60 nM, 70 nM, 80 nM, 90 nM, 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, 600 nM, 700 nM, 800 nM, 900 nM, 1 μM, or greater than or within the range defined by any two of the preceding values. The methods and systems described herein produce a dosage composition in which the purity of the hyperpolarized bio-associated contrast is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater, and at most about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, or less, or within the range defined by any two of the preceding values. In some embodiments, the fraction of at least the hyperpolarized compound is separated from the cleaved side arms, or other reaction by-products, if present.

[0106] transportation

[0107] In line with the disclosed embodiments, the polarization transfer and use of the molecule in question may occur at different locations. In some embodiments, the administered composition is transported to a different location. The disclosed embodiments are not necessarily limited to any specific transport distance or duration. Instead, the maximum distance or duration may be determined based on the molecule in question, its initial degree or polarization, the desired final degree of polarization, and the transport conditions. In some embodiments, the administered composition is transported at least 1 meter in a suitable transport device.

[0108] In accordance with the disclosed embodiments, the transport device may be configured to transport a sample of a precursor or target molecule. The transport device may be arranged and configured to transport one or more samples (e.g., one or more dosing compositions) simultaneously. The transport device may include a transport chamber configured to receive one or more samples. The transport device may be configured to maintain the transport chamber within a predetermined temperature range and a predetermined magnetic field strength. The transport device may be configured to maintain one or more samples in a magnetic field of at least approximately 10G, 20G, 30G, 40G, 50G, 60G, 70G, 80G, 90G, 100G, 200G, 300G, 400G, 500G, 600G, 700G, 800G, 900G, 1,000G or greater, and at most approximately 1,000G, 900G, 800G, 700G, 600G, 500G, 400G, 300G, 200G, 100G, 90G, 80G, 70G, 60G, 50G, 40G, 30G, 20G, 10G or less, or within the range defined by any two of the aforementioned values.

[0109] Permanent magnets or electromagnets included in the transport device can provide a magnetic field. In some embodiments, the permanent magnets or electromagnets are shielded to reduce the strength of the magnetic field outside the transport device. The transport device may also be configured to include a cooling system. The cooling system may be configured to maintain the sample at a predetermined temperature or within a predetermined temperature range during transport. For example, the cooling system may be configured to maintain the sample at a temperature below 270K, below 80K, or below 4K. In some embodiments, the transport device is configured to maintain the sample at approximately liquid nitrogen temperature. The transport device may include insulation between the cooling system and the outside of the transport device to minimize heat exchange with the external environment. In some embodiments, the cooling system is configured to maintain the sample temperature using a cold gas flow. In some embodiments, the cooling system is configured to maintain the sample temperature using a coolant. In some embodiments, the transport device includes a dewar to provide cooling for the sample. To distribute hyperpolarized samples over long distances, containers may be transported by standard transport vehicles such as airplanes, trains, trucks, cars, and ships.

[0110] In some embodiments, the administration composition containing the hyperpolarizing molecule of interest is transported within a transport device. In some embodiments, the relaxation time of the hyperpolarizing molecule of interest within the transport device is at least about 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, or longer, and at most about 10 hours, 9 hours, 8 hours, 7 hours, 6 hours, 5 hours, 4 hours, 3 hours, 2 hours, 1 hour, 50 minutes, 40 minutes, 30 minutes, 20 minutes, 10 minutes, 9 minutes, 8 minutes, 7 minutes, 6 minutes, 5 minutes, 4 minutes, 3 minutes, 2 minutes, 1 minute, or shorter, or within the range defined by any two of the aforementioned values. [Examples]

[0111] Figure 5 shows an example related to hyperpolarized pyruvate in H2O and D2O solvents at pH 7. 13This shows the CT1 relaxation time, as shown in Figure 5. 13 The C T1 relaxation time generally remains relatively high when experiencing magnetic fields up to 1T. At higher magnetic fields, 13 The C T1 relaxation time is dramatically reduced. Therefore, by performing the hyperpolarization procedure and / or purification procedure with a magnetic field of up to 1 T, hyperpolarization is achieved. 13 The 13C NMR / MRI signal decays relatively slowly during hyperpolarization and / or purification procedures, allowing pyruvate to retain strong polarization until it is used in the NMR / MRI experiment. Furthermore, 13 The C T1 relaxation time is significantly longer in D2O than in H2O. Therefore, hyperpolarization can be achieved by performing hyperpolarization and / or purification procedures in D2O. 13 The 13C NMR / MRI signal decays relatively slowly during hyperpolarization and / or purification procedures, allowing pyruvate to retain strong polarization until it is used in the NMR / MRI experiment.

[0112] Figure 6 shows an example relating to hyperpolarized pyruvate in D2O solvent at two different pH values ​​(5.5 and 7) under various magnetic fields. 13 This shows the CT1 relaxation time, as shown in Figure 6. 13 The C T1 relaxation time generally remains relatively high when experiencing a magnetic field of up to 1 T at pH 7. At pH 5.5, 13 The C T1 relaxation time reaches its peak at approximately 100 mT and decreases rapidly at lower and higher magnetic fields. Therefore, hyperpolarization can be achieved by performing hyperpolarization and / or purification procedures at a magnetic field of up to 1 T at pH 7, or at a magnetic field of approximately 100 mT at pH 5.5. 13 The 13C NMR / MRI signal decays relatively slowly during hyperpolarization and / or purification procedures, allowing pyruvate to retain strong polarization until it is used in the NMR / MRI experiment.

[0113] Figure 7 shows illustrative examples related to hyperpolarized pyruvate in D2O solvent in the absence and presence of EDTA under various magnetic fields. 13 This shows the CT1 relaxation time, as shown in Figure 3. 13The C T1 relaxation time generally peaks at a magnetic field of approximately 1 T, both in the absence and in the presence of EDTA. However, 13 The CT1 relaxation time is typically significantly higher in the presence of EDTA than in the absence of EDTA. Furthermore, 13 The C T1 relaxation time is relatively constant at lower magnetic fields in the presence of EDTA, while it drops sharply at lower magnetic fields in the absence of EDTA. Therefore, by performing the hyperpolarization procedure and / or purification procedure in the presence of EDTA, hyperpolarization can be achieved. 13 The 13C NMR / MRI signal decays relatively slowly during hyperpolarization and / or purification procedures, allowing pyruvate to retain strong polarization until it is used in the NMR / MRI experiment.

[0114] List of embodiments

[0115] Embodiment 1. A method, (a) To obtain a solution containing the target molecule containing an overpolarizable nucleus or a derivative of the target molecule containing an overpolarizable nucleus, (b) Applying a first magnetic field of up to 1 Tesla (T) to the target molecule or the derivative of the target molecule, (c) Subjecting the target molecule or a derivative of the target molecule to a nuclear spin hyperpolarization procedure to generate a target hyperpolarizable molecule, thereby imparting a first nuclear spin polarization to the hyperpolarizable nucleus, (d) Applying a second magnetic field of up to 1T to the hyperpolarizing molecule of the target, wherein the solution containing the hyperpolarizing molecule is characterized by a pH of 5 to 9, and the hyperpolarizable nucleus has a spin lattice (T1) relaxation time of more than 60 seconds (s) in the solution with respect to the second magnetic field and the pH. (e) Subjecting the target hyperpolarization molecule to a purification procedure to produce a purified target hyperpolarization molecule, thereby imparting a second nuclear spin polarization to the hyperpolarizable nucleus, (f) The administration of the target hyperpolarization molecule, (g) A method comprising performing a magnetic resonance spectroscopy (MRS) procedure on the subject.

[0116] Embodiment 2. The first magnetic field has a maximum of 900 millitesla (mT), 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, The method according to Embodiment 1, wherein the power is 900 microtesla (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, or 1 μT.

[0117] Embodiment 3. The second magnetic field has a maximum of 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, 9 The method according to Embodiment 1 or 2, wherein the μT is 00μT, 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, or 1μT.

[0118] Embodiment 4. The method according to any one of Embodiments 1 to 3, wherein the nuclear spin hyperpolarization procedure includes signal amplification by a para-hydrogen-induced polarization (PHIP) procedure, a PHIP-side-arm hydrolysis (PHIP-SAH) procedure, a PHIP nuclear over-Hausser effect system (PHIPNOESYS) procedure, or a reversible exchange (SABRE) procedure.

[0119] Embodiment 5. The method according to any one of Embodiments 1 to 4, wherein Embodiments 5(a) to 5(g) are carried out collectively in a maximum of 60 seconds, 50 seconds, 40 seconds, 30 seconds, 20 seconds, or 10 seconds.

[0120] The method according to any one of Embodiments 1 to 5, wherein Embodiment 6(a) to (g) are carried out collectively for at least 10 seconds, 20 seconds, 30 seconds, 40 seconds, 50 seconds, or 60 seconds.

[0121] Embodiment 7. The method according to any one of Embodiments 1 to 6, wherein the second nuclear spin polarization is 90%, 80%, 70%, 60%, or 50% or more of the first nuclear spin polarization.

[0122] Embodiment 8. The method according to any one of Embodiments 1 to 7, wherein the second nuclear spin polarization is at least 10%, 20%, 30%, 40%, or 50%.

[0123] Embodiment 9. The method according to any one of Embodiments 1 to 7, wherein the hyperpolarizable nucleus is characterized by a second magnetic field and a T1 relaxation time of at least 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, 190 seconds, or 200 seconds in a solution at pH 5-9, 5-8, 5-7, 5-6, 6-9, 6-8, 6-7, 7-9, 7-8, or 8-9.

[0124] Embodiment 10. The method according to any one of Embodiments 1 to 9, wherein the molecule of interest comprises a carboxylate, a carbon-13 labeled carboxylate, a partially or completely deuterated carboxylate, or a carbon-13 labeled and partially or completely deuterated carboxylate.

[0125] Embodiment 11. The method according to any one of Embodiments 1 to 10, wherein the target molecule, a derivative of the target molecule, or the target hyperpolarizing molecule is dissolved in a solution.

[0126] Embodiment 12. The method according to any one of Embodiments 1 to 11, wherein the solution is characterized by a pH of 5-9, 5-8, 5-7, 6-9, 6-8, 6-7, 7-9, 7-8, or 8-9.

[0127] Embodiment 13. The method according to any one of Embodiments 1 to 12, wherein the solution further comprises a scavenger selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2',2'',2'''-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid (DOTA), 1,7,10,16-tetraoxa-4,13-diazacyclooctadecane (Kryptofix® 22), 1,4,7,10,13,16-hexaazacyclooctadecane (hexacyclene), and crown ethers.

[0128] Embodiment 14. The method according to Embodiment 13, wherein the solution contains a scavenger at a concentration of at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%.

[0129] Embodiment 15. The method according to Embodiment 13 or 14, wherein the solution contains a scavenger at a maximum concentration of about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%.

[0130] Embodiment 16. The method according to any one of Embodiments 1 to 15, wherein the solution comprises a deuterated water (D2O) solvent.

[0131] Embodiment 17. A hyperpolarizable nucleus is carbon-13 ( 13 C) The method according to any one of embodiments 1 to 16, including a nucleus.

[0132] Embodiment 18. A method, (a) Obtaining a target molecule or a derivative of a target molecule dissolved in a solution characterized by a pH of 5 to 9, wherein the target molecule or derivative of the target molecule contains a hyperpolarizable nucleus, and the hyperpolarizable nucleus is characterized by a spin lattice (T1) relaxation time of at least 60 seconds (s) in the solution at a magnetic field of up to 1 Tesla (T) and a pH of 5 to 9. (b) subjecting the target molecule or a derivative of the target molecule to a nuclear spin hyperpolarization procedure to generate a target hyperpolarizable molecule, thereby imparting a first nuclear spin polarization to the hyperpolarizable nucleus, (c) Subjecting the target hyperpolarization molecule to a purification procedure to produce a purified target hyperpolarization molecule, thereby imparting a second nuclear spin polarization to the hyperpolarizable nucleus, (d) The administration of the target hyperpolarization molecule, (e) A method comprising performing a magnetic resonance spectroscopy (MRS) procedure on the subject.

[0133] Embodiment 19. The method of Embodiment 18, further comprising, before (b), a maximum of 1T, 900 millitesla (mT), 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, A method comprising applying a first magnetic field of 900 microtesla (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, or 1 μT to a target molecule or a derivative of a target molecule.

[0134] Embodiment 20. The method according to Embodiment 18 or 19, further comprising, before (c), a maximum of 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, 2m A method comprising applying a second magnetic field of T, 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, or 1 μT to a target hyperpolarized molecule.

[0135] Embodiment 21. The method according to any one of Embodiments 18 to 20, wherein the nuclear spin hyperpolarization procedure includes signal amplification by a para-hydrogen-induced polarization (PHIP) procedure, a PHIP-sidearm hydrolysis (PHIP-SAH) procedure, a PHIP nuclear over-Hausser effect system (PHIPNOESYS) procedure, or a reversible exchange (SABRE) procedure.

[0136] The method according to any one of Embodiments 18 to 21, wherein Embodiment 22(a) to (e) are carried out collectively in a maximum of 60 seconds, 50 seconds, 40 seconds, 30 seconds, 20 seconds, or 10 seconds.

[0137] The method according to any one of Embodiments 18 to 22, wherein Embodiment 23.(a) to (e) are carried out collectively for at least 10 seconds, 20 seconds, 30 seconds, 40 seconds, 50 seconds, or 60 seconds.

[0138] Embodiment 24. The method according to any one of Embodiments 18 to 23, wherein the second nuclear spin polarization is 90%, 80%, 70%, 60%, or 50% or more of the first nuclear spin polarization.

[0139] Embodiment 25. The method according to any one of Embodiments 18 to 24, wherein the second nuclear spin polarization is at least 10%, 20%, 30%, 40%, or 50%.

[0140] Embodiment 26. The method according to any one of Embodiments 18 to 25, wherein the hyperpolarizable nucleus is characterized by a T1 relaxation time of at least 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, 190 seconds, or 200 seconds in solution, with a pH of 5-9, 5-8, 5-7, 5-6, 6-9, 6-8, 6-7, 7-9, 7-8, or 8-9.

[0141] Embodiment 27. The method according to any one of Embodiments 18 to 26, wherein the molecule of interest comprises a carboxylate, a carbon-13 labeled carboxylate, a partially or fully deuterated carboxylate, or a carbon-13 labeled and partially or fully deuterated carboxylate.

[0142] Embodiment 28. The method according to any one of Embodiments 18 to 27, wherein the solution is characterized by a pH of 5-8, 5-7, 6-9, 6-8, 6-7, 7-9, 7-8, or 8-9.

[0143] Embodiment 29. The method according to any one of Embodiments 18 to 28, wherein the solution further comprises a scavenger selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2',2'',2'''-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid (DOTA), 1,7,10,16-tetraoxa-4,13-diazacyclooctadecane (Kryptofix® 22), 1,4,7,10,13,16-hexaazacyclooctadecane (hexacyclene), and crown ethers.

[0144] Embodiment 30. The method according to Embodiment 29, wherein the solution contains a scavenger at a concentration of at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%.

[0145] Embodiment 31. The method according to Embodiment 29 or 30, wherein the solution contains a scavenger at a maximum concentration of about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%.

[0146] Embodiment 32. The method according to any one of Embodiments 18 to 31, wherein the solution comprises a deuterated water (D2O) solvent.

[0147] Embodiment 33. A hyperpolarizable nucleus is carbon-13 ( 13 C) The method according to any one of embodiments 18 to 32, including a nucleus.

[0148] Embodiment 34. A method, (a) To obtain a target molecule containing an overpolarizable nucleus or a derivative of a target molecule containing an overpolarizable nucleus, (b) Subjecting the target molecule or a derivative of the target molecule to a parahydrogen-involved nuclear spin hyperpolarization procedure to generate the target hyperpolarizable molecule, thereby imparting a first nuclear spin polarization to the hyperpolarizable nucleus, (c) Subjecting the target hyperpolarization molecule to a purification procedure to produce a purified target hyperpolarization molecule, thereby imparting a second nuclear spin polarization to the hyperpolarizable nucleus, wherein the second nuclear spin polarization is 50% or more of the first nuclear spin polarization. (d) The administration of the target hyperpolarization molecule, (e) A method comprising performing a magnetic resonance spectroscopy (MRS) procedure on the subject.

[0149] Embodiment 35. The method of Embodiment 34, further comprising, before (b), a maximum of 1 Tesla (T), 900 millitesla (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 A method comprising applying a first magnetic field of mT, 900 microtesla (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, or 1 μT to a target molecule or a derivative of a target molecule.

[0150] Embodiment 36. The method according to Embodiment 34 or 35, further, prior to (c), up to 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, A method comprising applying a second magnetic field of 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, or 1 μT to a target hyperpolarized molecule.

[0151] Embodiment 37. The method according to any one of Embodiments 34 to 36, wherein the nuclear spin hyperpolarization procedure includes signal amplification by a para-hydrogen-induced polarization (PHIP) procedure, a PHIP-side-arm hydrolysis (PHIP-SAH) procedure, a PHIP nuclear over-Hausser effect system (PHIPNOESYS) procedure, or a reversible exchange (SABRE) procedure.

[0152] The method according to any one of Embodiments 34 to 37, wherein Embodiment 38.(a) to (e) are carried out collectively for a maximum of 60 seconds, 50 seconds, 40 seconds, 30 seconds, 20 seconds, or 10 seconds.

[0153] The method according to any one of embodiments 34 to 38, wherein embodiments 39(a) to (e) are carried out collectively for at least 10 seconds, 20 seconds, 30 seconds, 40 seconds, 50 seconds, or 60 seconds.

[0154] Embodiment 40. The method according to any one of embodiments 34 to 39, wherein the second nuclear spin polarization is 90%, 80%, 70%, 60%, or 50% or more of the first nuclear spin polarization.

[0155] Embodiment 41. The method according to any one of Embodiments 34 to 40, wherein the second nuclear spin polarization is at least 10%, 20%, 30%, 40%, or 50%.

[0156] Embodiment 42. The method according to any one of Embodiments 34 to 41, wherein the hyperpolarizable nucleus is characterized by a T1 relaxation time of at least 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, 190 seconds, or 200 seconds in a solution at pH 5-9, 5-8, 5-7, 5-6, 6-9, 6-8, 6-7, 7-9, 7-8, or 8-9.

[0157] Embodiment 43. The method according to any one of Embodiments 34 to 42, wherein the target molecule comprises a carboxylate, a carbon-13 labeled carboxylate, a fully or partially deuterated carboxylate, or a carbon-13 labeled and partially deuterated carboxylate.

[0158] Embodiment 44. The method according to any one of Embodiments 34 to 43, wherein the target molecule or a derivative of the target molecule is dissolved in a solution.

[0159] Embodiment 45. The method according to Embodiment 44, wherein the solution is characterized by a pH of 5-9, 5-8, 5-7, 5-6, 6-9, 6-8, 6-7, 7-9, 7-8, or 8-9.

[0160] Embodiment 46. The method according to Embodiment 44 or 45, wherein the solution further comprises a scavenger selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2',2'',2'''-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid (DOTA), 1,7,10,16-tetraoxa-4,13-diazacyclooctadecane (Kryptofix® 22), 1,4,7,10,13,16-hexaazacyclooctadecane (hexacyclene), and crown ethers.

[0161] Embodiment 47. The method according to Embodiment 46, wherein the solution contains a scavenger at a concentration of at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%.

[0162] Embodiment 48. The method according to Embodiment 46 or 47, wherein the solution contains a scavenger at a maximum concentration of about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%.

[0163] Embodiment 49. The method according to any one of Embodiments 44 to 48, wherein the solution comprises a deuterated water (D2O) solvent.

[0164] Embodiment 50. A hyperpolarizable nucleus is carbon-13 ( 13 C) Any of Embodiments 34 to 49, including a nucleus. 1 The method used.

[0165] Embodiment 51. A composition, It is a solution, Solvent and, A hyperpolarizing molecule of the target dissolved in the solvent, wherein the hyperpolarizing molecule contains a nucleus capable of hyperpolarization, A method characterized in which the hyperpolarizable nucleus has a spin lattice (T1) relaxation time of at least 60 seconds (s) in the solution at a magnetic field of up to 1 Tesla (T) and a pH of 5 to 9.

[0166] Embodiment 52. The composition according to Embodiment 51, wherein the hyperpolarizable nuclei of the hyperpolarizable molecule in question have at least 10%, 20%, 30%, 40%, or 50% nuclear spin polarization.

[0167] Embodiment 53. The composition according to Embodiment 51 or 52, wherein the hyperpolarizable nucleus has a maximum of 1 T, 900 millitesla (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 microtesla (μT), 800 μT, 700 μT, 600 μT, 500 μT, 400 μT, 300 μT, 200 μT, 10 A composition characterized by a T1 relaxation time of at least 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, 190 seconds, or 200 seconds in a solution with a pH of 5-9, 5-8, 5-7, 5-6, 6-9, 6-8, 6-7, 7-9, 7-8, or 8-9 in a magnetic field of 0 μT, 90 μT, 80 μT, 70 μT, 60 μT, 50 μT, 4 μT, 3 μT, 2 μT, or 1 μT.

[0168] Embodiment 54. The composition according to any one of Embodiments 51 to 53, wherein the target hyperpolarizing molecule comprises a hyperpolarizing carboxylate, a hyperpolarizing carbon-13 labeled carboxylate, a hyperpolarizing partially or fully deuterated carboxylate, or a hyperpolarizing carbon-13 labeled and partially or fully deuterated carboxylate.

[0169] Embodiment 55. The composition according to any one of Embodiments 51 to 54, wherein the solution is characterized by a pH of 5-9, 5-8, 5-7, 6-9, 6-8, 6-7, 7-9, 7-8, or 8-9.

[0170] Embodiment 56. The composition according to any one of Embodiments 51 to 55, wherein the solution further comprises a scavenger selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2',2'',2'''-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid (DOTA), 1,7,10,16-tetraoxa-4,13-diazacyclooctadecane (Kryptofix® 22), 1,4,7,10,13,16-hexaazacyclooctadecane (hexacyclene), and crown ethers.

[0171] Embodiment 57. The composition according to Embodiment 56, wherein the solution contains a scavenger at a concentration of at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%.

[0172] Embodiment 58. The composition according to Embodiment 56 or 57, wherein the solution contains a scavenger at a maximum concentration of about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%.

[0173] Embodiment 59. The composition according to any one of Embodiments 51 to 58, wherein the solvent comprises deuterated water (D2O) solvent.

[0174] Embodiment 60. A hyperpolarizable nucleus is carbon-13 ( 13 C) A composition according to any one of embodiments 51 to 59, comprising a nucleus.

Claims

1. It is a method, (a) Obtaining a target molecule or a derivative of a target molecule dissolved in a solution characterized by a pH of 5 to 9, wherein the target molecule or derivative of the target molecule contains a hyperpolarizable nucleus, and the hyperpolarizable nucleus is subjected to a spin lattice (T) in the solution at least 60 seconds (s) in a magnetic field of up to 1 Tesla (T) and a pH of 5 to 9. １ ) Characterized by the relaxation time, (b) Subjecting the target molecule or a derivative of the target molecule to a nuclear spin hyperpolarization procedure to generate a target hyperpolarizable molecule, thereby imparting a first nuclear spin polarization to the hyperpolarizable nucleus, (c) Subjecting the target hyperpolarization molecule to a purification procedure to produce a purified target hyperpolarization molecule, thereby imparting a second nuclear spin polarization to the hyperpolarizable nucleus, (d) Administering the hyperpolarizing molecules of the subject, (e) A method comprising performing a magnetic resonance spectroscopy (MRS) procedure on the subject.

2. The method according to claim 1, further comprising applying a first magnetic field of up to 1 T to the target molecule or the derivative of the target molecule before (b).

3. The method according to claim 1 or 2, further comprising applying a second magnetic field of up to 1 T to the hyperpolarized molecule of the target before (c).

4. The method according to any one of claims 1 to 3, wherein the nuclear spin hyperpolarization procedure includes signal amplification by a para-hydrogen-induced polarization (PHIP) procedure, a PHIP-sidearm hydrolysis (PHIP-SAH) procedure, a PHIP nuclear over-Hausser effect system (PHIPNOESSYS) procedure, or a reversible exchange (SABRE) procedure.

5. The method according to any one of claims 1 to 4, wherein (a) to (e) are carried out collectively in a maximum of 60 seconds.

6. The method according to any one of claims 1 to 5, wherein the second nuclear spin polarization is 50% or more of the first nuclear spin polarization.

7. The method according to any one of claims 1 to 6, wherein the second nuclear spin polarization is at least 10%.

8. The hyperpolarizable nucleus is subjected to at least 60 seconds of T in the solution at a pH of 5 to 9. １ The method according to any one of claims 1 to 7, characterized by a relaxation time.

9. The method according to any one of claims 1 to 8, wherein the target molecule includes a carboxylate, a carbon-13 labeled carboxylate, a partially or completely deuterated carboxylate, or a carbon-13 labeled and partially or completely deuterated carboxylate.

10. The method according to any one of claims 1 to 9, wherein the solution further comprises a scavenger selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2',2'',2'''-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid (DOTA), 1,7,10,16-tetraoxa-4,13-diazacyclooctadecane (Kryptofix® 22), 1,4,7,10,13,16-hexaazacyclooctadecane (hexacyclene), and crown ether.

11. The method according to claim 10, wherein the solution contains the scavenger in a concentration of at least 1%.

12. The method according to claim 10 or 11, wherein the solution contains the scavenger at a maximum concentration of 10%.

13. The aforementioned solution is deuterated water (D ２ O) The method according to any one of claims 1 to 12, comprising a solvent.

14. The aforementioned highly polarized nucleus is carbon-13 ( １３ C) The method according to any one of claims 1 to 13, comprising a nucleus.

15. A composition, It is a solution, Solvent and, A hyperpolarizing molecule of the target dissolved in the solvent, wherein the hyperpolarizing molecule contains a nucleus capable of hyperpolarization, The hyperpolarizable nuclei are subjected to a spin lattice (T) in the solution for at least 60 seconds (s) at a magnetic field of up to 1 Tesla (T) and a pH of 5 to 9. １ A method characterized by a relaxation period.

16. The composition according to claim 15, wherein the hyperpolar nuclei of the target molecule have at least 10% nuclear spin polarization.

17. The composition according to claim 15 or 16, wherein the target hyperpolarizing molecule includes a hyperpolarizing carboxylate, a hyperpolarizing carbon-13 labeled carboxylate, a hyperpolarizing partially or fully deuterated carboxylate, or a hyperpolarizing carbon-13 labeled and partially or fully deuterated carboxylate.

18. The composition according to any one of claims 15 to 17, wherein the solution further comprises a scavenger selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2',2'',2'''-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid (DOTA), 1,7,10,16-tetraoxa-4,13-diazacyclooctadecane (Kryptofix® 22), 1,4,7,10,13,16-hexaazacyclooctadecane (hexacyclene), and crown ether.

19. The composition according to claim 18, wherein the solution contains the scavenger at a concentration of at least 1%.

20. The composition according to claim 18 or 19, wherein the solution contains the scavenger at a maximum concentration of about 10%.

21. The solvent is deuterated water (D ２ O) The composition according to any one of claims 15 to 20, comprising a solvent.

22. The aforementioned hyperpolarizable nucleus is carbon-13 ( １３ C) The composition according to any one of claims 15 to 21, comprising a nucleus.

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