Injection device for preparation and delivery of MRI probes

Abstract

A sterile MRI probe injection device is disclosed for preparing and administering a hyperpolarized MRI probe to a patient in need thereof. The device includes one or more reaction chambers in which the hyperpolarized MRI probe is prepared, separated from the reaction mixture, and concentrated to prepare a solution of a suitable concentration for administration to a patient. Also disclosed is a method for using the device to prepare and administer a hyperpolarized MRI probe to a patient in need thereof for diagnosing the stage of a disease or adverse condition or for monitoring the progress of a treatment for a patient having a disease or adverse condition.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This patent application claims the benefit of U.S. Provisional Patent Application No. 63 / 328,556, filed April 7, 2022, which is incorporated by reference in its entirety.

[0002] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with Government support and the Government has certain rights in this invention. [Background technology]

[0003] Background technology Nuclear magnetic resonance spectroscopy (NMR) and magnetic resonance imaging (MRI) methods are powerful tools that are widely used in biomedical, chemical, and materials science applications. These methods rely on the population differences in the energy levels of nuclear spins (called polarization) that arise after the application of a strong magnetic field. Spins aligned with or opposite to the applied magnetic field produce a net polarization, which is detected. Unfortunately, nuclear polarization at thermal equilibrium (i.e., normal conditions) is inherently poor, which generally remains a limitation in the range of sensitivity and capabilities of magnetic resonance (Gunther, NMR Spectrosc. Basic Princ. Concepts Appl. Chem., 13-28 (2013)).

[0004] To overcome this problem and enable NMR / MRI signal enhancement by orders of magnitude, hyperpolarization techniques have been developed. The most widely used hyperpolarization techniques are electron (dynamic nuclear polarization, DNP) (Hausser et al., Adv. Magn. Opt. Reson., 3, 79-139 (1968); Abagam et al., Reports Prog. Phys., 41, 395-467 (1978); and Ardenkjaer-Larsen et al., Proc. Natl. Acad. Sci. USA, 100, 10158-10163 (2003)), photon (spin-exchange optical pumping) (Bhaskar et al., Phys. Rev. Lett., 49, 25 (1982); Ebert et al., Lancet, 347, 1297 (1996); Albert et al., Nature, 370, 199-201 (1994); and Schroder et al., Nature, 370, 199-201 (1995)). al., Science, 314, 446-449 (2006)); Eisenschmid et al., J. Am. Chem. Soc., 109, 8089 (1987); Haake et al., J. Am. Chem. Soc., 118, 8688 (1996); Goldman et al., CR Phys., 6, 575 (2005); and Chekmenev et al., J. Am. Chem. Soc., 130, 4212 (2008)) Hyperpolarized magnetic resonance (MR) is a novel molecular imaging method for monitoring metabolic, enzymatic transformations, or biochemical pathways that were previously inaccessible by MR.

[0005] Current hyperpolarized imaging using dissolution DNP and superconducting MRI scanners is extremely powerful because of its unique ability to track chemical changes in vivo. However, DNP-based experiments are relatively laborious, time-consuming, and expensive.

[0006] The PHIP approach and its subcategory SABRE (Signal Amplification by Reversible Exchange) are able to transfer 100% pure singlet spin order of para-hydrogen (para-H2) to target molecules. PHIP is a traditional hydrogenation method that relies on a catalytic hydrogenation reaction in which a precursor in the form of a hydrogen acceptor is reduced and polarized by para-hydrogen. In contrast, reversible exchange using SABRE leaves the hyperpolarized drug chemically unchanged. Also, since it is not limited to one para-H2 molecule per molecule, it is possible to achieve extraordinary levels of hyperpolarization through multiple spin transfer steps. This effect has been shown to be highly effective in a wide range of biologically relevant molecules. 1 H, 13 C. 19 F, 31 P and 15 N and / or 29 It has also been shown to transfer polarization to nuclei such as Si nuclei (Barskiy et al., ChemPhysChem, 18, 1493-1498 (2017); Theis et al., J. Am. Chem. Soc., 137, 1404-1407 (2015); Shchepin et al., ChemPhysChem, 18, 1961-1965 (2017); Zhivonitko et al., Chem. Commun., 51, 2506-2509 (2015); Iali et al., Angew. Chemie - Int. Ed., 58, 10271-10275 (2019); and Gemeinhardt et al., Angew. Chemie Int. Ed., 59, 418-423 (2019)).

[0007] The high polarization rate, short signal build-up time, low cost and scalability make SABRE a promising modality for studying in vivo metabolism using magnetic resonance spectroscopy techniques.

[0008] Currently available hyperpolarized contrast agents are accompanied by a spin-transfer catalyst component that contains the heavy metal, e.g., transition metal atom, necessary to enable polarization transfer from para-H2 to the substrate. Toxicity concerns arise when administering hyperpolarized contrast agents in vivo, since potentially toxic heavy-metal-based complexes (e.g., catalysts are typically Ir-based organometallic compounds) are present in solution along with the hyperpolarized contrast agent.

[0009] Another obstacle to the successful implementation of the SABRE process is the low solubility of parahydrogen in water compared to its solubility in alcoholic solvents (H2 solubility in water is about 1.6 mg / L). SABRE hyperpolarization is primarily activated in organic solvents, preferably methanol, which is not compatible with in vivo administration.

[0010] The above indicates that a need exists for improved SABRE catalysts that can be easily separated from a hyperpolarized substrate such that the hyperpolarized substrate is free of heavy metals. Additionally, a need exists for methods of separating the hyperpolarized substrate from the SABRE catalyst and / or hyperpolarized SABRE catalyst complexes that contain heavy metals. Also, a need exists for methods of administering the hyperpolarized substrate in a solvent medium suitable for in vivo administration.

[0011] Since the lifespan of hyperpolarized MRI probes is very short, there is an unmet need for an apparatus that can generate hyperpolarized MRI probes in situ and administer them to patients in need. Apparatuses have been proposed in the art to meet this need (Maslana et al., J. Autom. Methods Manag. Chem., 22(6), 187-194 (2000); Kainz et al., “Synthesis of perfluoroalkyl-substituted aryl bromides and their purification over fluorous reverse phase silica,” Synthesis (10), 1425-1427 (1998)); however, the proposed apparatus may be ineffective due to the disclosed method of hyperpolarization and / or separation of the hyperpolarized probe from the reaction mixture. Previous attempts to automate and design apparatus for generating hyperpolarized MRI probes or other molecules of interest have suffered from several limitations. To be efficiently hyperpolarized, the polarity of the Ir catalyst and the solubility of the hyperpolarized MRI probe in methanol or other solvents are often comparable. Attempts to use extraction between the aqueous and organic layers do not always allow separation of the hyperpolarized MRI probe from the catalyst, as the activated catalyst is somewhat soluble in water (Iali et al., “Achieving High Levels of NMR-Hyperpolarization in Aqueous Media with Minimal Catalyst Contamination using SABRE,” Chem. - A. Eur. J., 23, 10491-10495 (2017)). The Ir catalyst is attached to silica gel to allow filtration, but at the expense of reduced hyperpolarization levels.Ir catalysts have been captured by solid-supported thiols with mixed success and failure (Barskiy et al., “Rapid Catalyst Capture Enables Metal-Free Parahydrogen-Based Hyperpolarized Contrast Agents,” J. Phys. Chem. Lett. 9(11), 2721-2724 (2018); Kidd et al., “Facile Removal of Homogenous SABRE Catalysts for Purifying Hyperpolarized Metronidazole, a Potential Hypoxia Sensor,” J. Phys. Chem. 122, 16848-16852 (2018)). Previous devices also lacked clinical capability due to inadequate purity and safety controls. Furthermore, devices that disclose venting spent gases to a fume hood are not practical in clinical settings where such devices would be useful.

[0012] The above indicates that a need exists for improved SABRE catalysts that can be easily separated from a hyperpolarized substrate such that the hyperpolarized substrate is free of heavy metals. Additionally, a need exists for methods of separating the hyperpolarized substrate from SABRE catalysts and / or hyperpolarized SABRE catalyst complexes that contain heavy metals. Also, a need exists for methods of administering the hyperpolarized substrate in a solvent medium suitable for in vivo administration.

[0013] The present invention provides such a SABRE catalyst and method. These and other advantages of the present invention, as well as additional inventive features, will become apparent from the description of the invention provided herein. Summary of the Invention

[0014] BRIEF SUMMARY OF THE INVETION The present disclosure provides an MRI probe injection device that satisfies the unmet needs highlighted above. According to a first aspect of the present disclosure, there is provided an MRI probe injection device, comprising: (i) one or more reaction chambers, each reaction chamber containing: (a) a structure configured to attenuate a magnetic field within the reaction chamber from an external source, the structure being such that the strength of the magnetic field within the reaction chamber from the external source is below a threshold; (b) one or more inlet ports; (c) a coil configured to generate an electromagnetic field within the reaction chamber; (d) one or more temperature control devices; and (e) one or more exit ports; (ii) one or more MRI probe separators configured to receive a reaction mixture containing the perfluorinated SABRE catalyst, a solvent, and a hyperpolarized MRI probe from the one or more reaction chambers and to extract the hyperpolarized MRI probe from the reaction mixture; and (iii) one or more MRI probe collectors configured to form a solution containing a desired concentration of hyperpolarized MRI probes.

[0015] According to an embodiment of the first aspect, the structure is a mu-metal shield that attenuates magnetic fields from external sources to have a strength of 10 nT or less in one or more reaction chambers.

[0016] According to an embodiment of the first aspect, the one or more inlet ports include one or more gas ports and one or more liquid ports.

[0017] According to an embodiment of the first aspect, each reaction chamber is adapted to withstand an atmospheric pressure of at least 10 bar.

[0018] According to an embodiment of the first aspect, the magnetic field within the reaction chamber induced by the coil is between 0 and 200 mT.

[0019] According to a first embodiment, the one or more temperature control devices include non-magnetic heating and / or cooling elements configured to maintain a temperature within the reaction chamber between -25°C and 100°C.

[0020] According to an embodiment of the first aspect, the one or more temperature control devices are configured to cycle the temperature within the reaction chamber between at least two different temperatures between -25°C and 100°C over a period of time.

[0021] According to an embodiment of the first aspect, one or more reaction chambers are equipped to perform hyperpolarization on a reaction mixture comprising a perfluorinated SABRE catalyst, a solvent, and a substrate to be hyperpolarized into a hyperpolarized MRI probe.

[0022] According to an embodiment of the first aspect, the solvent is a mono- or bi-phase system comprising water, methanol, ethanol, a fluorous solvent, or a mixture thereof.

[0023] According to an embodiment of the first aspect, the one or more MRI probe separators are configured to separate the hyperpolarized MRI probe from the perfluorinated SABRE catalyst by one of: filtration; extraction; or column chromatography.

[0024] According to an embodiment of the first aspect, the MRI probe injection device further comprises a gas trap, a gas leak detector, and / or an oxygen level monitor.

[0025] According to an embodiment of the first aspect, the device further comprises a processor configured to execute instructions causing the processor to: control a flow rate of gas and / or liquid through the device, monitor safety indicators of the device and / or environment, administer a desired amount of hyperpolarized MRI probe to a patient, or calculate a decay rate of the hyperpolarized MRI probe as a function of the flow rate of said gas and / or liquid.

[0026] According to an embodiment of the first aspect, the one or more MRI probe collectors include one or more driers.

[0027] According to an embodiment of the first aspect, the one or more reaction chambers comprise at least two reaction chambers configured to be operable in series or in parallel.

[0028] According to an embodiment of the first aspect, the components of the MRI probe injection device are made of non-magnetic material or plastic.

[0029] According to a second aspect of the present disclosure, there is provided a method of administering a hyperpolarized MRI probe to a patient in need thereof, the method comprising: (i) providing an MRI probe injection device according to a first aspect; (ii) providing a reaction mixture comprising a perfluorinated SABRE catalyst comprising a d-block element and a perfluorinated ligand, a solvent, a co-ligand, and a substrate to be hyperpolarized by an MRI probe into one or more reaction chambers; (iii) agitating the reaction mixture, the agitation being provided by bubbling parahydrogen gas or a mixture of parahydrogen and nitrogen gas through the reaction mixture; (iv) applying a magnetic field suitable for hyperpolarizing the perfluorinated SABRE catalyst and the substrate, hyperpolarizing the substrate onto the hyperpolarized MRI probe; (v) separating the hyperpolarized MRI probe from the reaction mixture by at least one of filtration, extraction, or column chromatography to obtain a solution containing the hyperpolarized MRI probe; (vi) concentrating the hyperpolarized MRI probe present in the solution obtained in step (v) to obtain a concentrate and reconstituting the concentrate into a solution of the hyperpolarized MRI probe of a desired concentration for administration to a patient; (vii) analyzing at least one of the purity or concentration of the hyperpolarized MRI probe present in the solution; and (viii) administering a hyperpolarized MRI probe to the patient.

[0030] According to an embodiment of the second aspect, the solvent is selected from perfluorohexane / diethyl ether mixtures, mixtures of methoxynonafluorobutane and ethyl acetate with non-polar solvents, mixtures of perfluorohexane and ether, mixtures of perfluorobutyl methyl ether and ethyl acetate, ether, fluorocarbon derivatives of THF FC 75, decafluoromethoxytrifluoromethylpentane, hexafluoropropanol, nonafluorobutyl methyl ether, perfluoromethylcyclohexane, perfluoroalkanes, perfluorohexane, and methoxynonafluorobutane.

[0031] According to an embodiment of the second aspect, the substrate is 1- 13 C-Ketoglutaric acid, 1- 13 C-5- 12 C-Ketoglutaric acid, 1- 13 C-pyruvic acid, 1- 13 CN-acetylcysteine, 15 N2-isoniazid (or pyridyl-4-carbobis- 15 N2-hydrazide), 13 C2, 15 N3-metronidazole, 15 N2-1-aminoisoquinoline (1-AIQ), deuterated versions thereof, and salts thereof.

[0032] According to an embodiment of the second aspect, the perfluorinated ligand has the formula (I): [L m -(NHC)-(YZ) q or a salt thereof, wherein: each L is independently selected from hydrogen, adamantyl, a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; NHC is a 4- to 7-membered N-heterocyclic carbenyi group, and NHC is bonded to a d-block element via a carbene; each Y is independently selected from a bond or a spacer group; each Z is a perfluorinated tag; m is an integer from 1 to 4, and q is an integer from 1 to 3.

[0033] According to an embodiment of the second aspect, the perfluorinated tag is one of the following: perfluorinated C, which contains only carbon and fluorine atoms. 3-60 Group: Perfluorinated C containing only carbon and fluorine atoms 3-40 group; or perfluorinated C 3-20 Base.

[0034] According to an embodiment of the second aspect, the perfluorinated ligand is:

[0035] [ka]

[0036] or a salt thereof,

[0037] [ka]

[0038] is a single or double bond, and

[0039] [ka]

[0040] represents a bond to a d-block element via a carbene. [Brief description of the drawings]

[0041] [Figure 1] FIG. 1 shows a reaction chamber and separator of a hyperpolarized MRI probe according to one embodiment of the present disclosure. [Diagram 2] FIG. 2 shows the internal structure of the reaction chamber according to one embodiment of the present disclosure. [Diagram 3] FIG. 3 illustrates a schematic of certain components and their arrangement of an MRI probe injection device, according to one embodiment of the present disclosure. [Figure 4]FIG. 4 illustrates an MRI probe injection apparatus with multiple reaction chambers and separation of hyperpolarized probes from the reaction mixture from each reaction chamber according to one embodiment of the present disclosure. [Diagram 5] FIG. 5 shows some of the steps involved in a method of preparing an MRI probe for administration to a patient, according to one embodiment of the present disclosure. [Figure 6] FIG. 6 illustrates a fluorous solid-phase extraction (F-SPE) process according to one embodiment of the present disclosure, in which an organic fraction containing the hyperpolarized probe is separated from the fluorous fraction on a fluorous silica gel using a fluorophobic solvent, i.e., by a fluorophilic pass, the hyperpolarized probe is eluted into the fluorous fraction, and the catalyst is subsequently recovered from the fluorous silica gel by a fluorophilic pass that includes passing an aqueous / organic solvent, such as methanol, ethanol, ethyl acetate, acetonitrile, or THF, over the silica gel. [Figure 7] FIG. 7 illustrates the substrate polarization process and extraction of the probe as shown in FIG. 6 by a fluorophilic path according to one embodiment of the present disclosure. [Figure 8] Figure 8 shows a "reverse" F-SPE process in which the probe is separated from the reaction mixture by a combination of fluorophilic and fluorophobic processes. Thus, the reaction mixture is contacted with standard silica gel with a fluorophilic solvent and the probe is retained on the silica gel. The probe is then eluted with a standard organic solvent or recovered from the silica gel by a fluorophobic path. [Figure 9] FIG. 9 illustrates the partitioning of the SABRE catalyst and hyperpolarized substrate between two immiscible phases (a fluorous solvent fraction and water or another water-immiscible solvent such as a chlorinated solvent and water or other hydrophilic solvent) according to one embodiment of the present disclosure, which allows the principles of phase transfer catalysis to be employed in combination with parahydrogen to generate high levels of hyperpolarization in the aqueous phase without catalyst contamination. [Figure 10] FIG. 10 shows the hyperpolarization of a fluorinated SABRE catalyst that includes a metal coordinated to a co-ligand and a substrate that includes a half-spin nucleus to form a metal-free hyperpolarized substrate according to one embodiment of the present disclosure.

[0042] Detailed Description of the Invention The present disclosure provides an MRI probe injection device for manufacturing and administering a hyperpolarized MRI probe (i.e., a hyperpolarized substrate) to a patient. The MRI probe injection device includes one or more reaction chambers, one or more MRI probe separators, and one or more MRI probe collectors. The MRI probe injection device may also include a hyperpolarized MRI probe administrator for administering the hyperpolarized MRI probe to the patient. Each reaction chamber may also include: (a) a structure configured to attenuate a magnetic field within the reaction chamber from an external source, the structure being such that the strength of the magnetic field within the reaction chamber from the external source is below a threshold; (b) one or more inlet ports; (c) a coil configured to generate an electromagnetic field within the reaction chamber; (d) one or more temperature control devices; and (e) One or more exit ports.

[0043] The present disclosure provides a method of administering a hyperpolarized MRI probe to a subject (e.g., a human or animal), the method including: (i) providing an MRI probe injection device according to a first aspect; (ii) providing a reaction mixture comprising a perfluorinated SABRE catalyst comprising a d-block element and a perfluorinated ligand, a solvent, a co-ligand, and a substrate to be hyperpolarized by an MRI probe into one or more reaction chambers; (iii) agitating the reaction mixture, the agitation being provided by bubbling parahydrogen gas or a mixture of parahydrogen and nitrogen gas through the reaction mixture; (iv) applying a magnetic field suitable for hyperpolarizing the perfluorinated SABRE catalyst and the substrate, hyperpolarizing the substrate onto the hyperpolarized MRI probe; (v) separating the hyperpolarized MRI probe from the reaction mixture by at least one of filtration, extraction, or column chromatography to obtain a solution containing the hyperpolarized MRI probe; (vi) concentrating the hyperpolarized MRI probe present in the solution obtained in step (v) to obtain a concentrate and reconstituting the concentrate into a solution of the hyperpolarized MRI probe of a desired concentration for administration to a patient; (vii) analyzing at least one of the purity and concentration of the hyperpolarized MRI probe present in the solution; and (viii) administering a hyperpolarized MRI probe to the patient.

[0044] The present disclosure also provides a perfluorinated SABRE catalyst introduced into a reaction chamber of at least one MRI probe injection device, the perfluorinated SABRE catalyst comprising a d-block element and a perfluorinated ligand, the perfluorinated ligand having formula (I):

[0045] [ka]

[0046] or a salt thereof, each L is independently selected from hydrogen, adamantyl, a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; NHC is a 4- to 7-membered N-heterocyclic carbenyi group, and NHC is bonded to a d-block element via a carbene; each Y is independently selected from a bond or a spacer group; each Z is a perfluorinated tag; m is an integer from 1 to 4 (e.g., 1, 2, 3, or 4); and q is an integer from 1 to 3 (e.g., 1, 2, or 3).

[0047] The perfluorinated SABRE catalyst comprises a d-block element, such as, for example, Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Y, Zr, Nb, Mo, Tc, Ru, Rh, Pd, Ag, Cd, La, Hf, Ta, W, Re, Os, Ir, Pt, Au, and / or Hg. In some embodiments, the d-block element is a transition metal, such as, for example, Co, Rh, Ir, Ru, Pd, Pt, or Mt. In certain embodiments, the perfluorinated SABRE catalyst comprises an element from group 9 of the periodic table, i.e., Co, Rh, Ir, or Mt. In a preferred embodiment, the perfluorinated SABRE catalyst comprises Ir or Co. For example, the perfluorinated SABRE catalyst can be prepared from [Ir(COD)(IMes)(Cl)].

[0048] The perfluorinated SABRE catalyst has the formula (I):

[0049] [ka]

[0050] or a salt thereof, each L is independently selected from hydrogen, adamantyl, a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; NHC is a 4- to 7-membered N-heterocyclic carbenylic group, which is bonded to a d-block element via a carbene. each Y is independently selected from a bond or a spacer group; each Z is a perfluorinated tag, m is an integer from 1 to 4 (e.g., 1, 2, 3, or 4), and q is an integer from 1 to 3 (e.g., 1, 2, or 3).

[0051] According to some embodiments, NHC comprises an azolyl moiety, i.e., a 5-membered heterocyclic group having a nitrogen atom and at least one other heteroatom selected from nitrogen, sulfur, and oxygen.Thus, in some embodiments, NHC is a 5-membered N-heterocyclic carbenyi group.For example, the 5-membered N-heterocyclic carbenyi group can be imidazole-based, imidazoline-based, or thiazole-based.In other words, the 5-membered N-heterocyclic carbenyi group can be the carbene resulting from the treatment of a perfluorinated ligand having an imidazole, imidazoline, or thiazole core.

[0052] In some embodiments, the NHC is a 4,5-disubstituted, 1,3-disubstituted or 1,3,4,5-tetrasubstituted imidazole or imidazoline-based 5-membered N-heterocyclic carbenyi group. For example, the NHC can be 4,5-disubstituted imidazolidinyl, 1,3-disubstituted imidazolidinyl, 1,3,4,5-tetrasubstituted imidazolidinyl, 4,5-disubstituted 2,3-dihydro-imidazolyl, 1,3-disubstituted 2,3-dihydro-imidazolyl, or 1,3,4,5-tetrasubstituted 2,3-dihydro-imidazolyl. Examples of imidazolylidinyl moieties include N,N'-di-(2,4,6-trimethylphenyl)-imidazolylidinyl moiety, N,N'-di-(2,6-diisopropylphenyl)-imidazolidinyl moiety, N,N'-di-(2,6-dicyclohexyl)-imidazolidinyl moiety, N,N'-di-(2,6-t-butyl)-imidazolidinyl moiety, and N,N'-di-(1-adamantyl)-imidazolidinyl moiety.

[0053] In some embodiments, the perfluorinated ligand has formula (Ia) or (Ib):

[0054] [ka]

[0055] or a salt thereof, wherein each L is independently hydrogen, adamantyl, a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; each Y is independently a bond or a spacer group; each Z is independently a perfluorinated tag;

[0056] [ka]

[0057] is a single or double bond, and

[0058] [ka]

[0059] represents a bond to a d-block element via a carbene.

[0060] In some embodiments, the perfluorinated ligand has formula (Ic) or (Id):

[0061] [ka]

[0062] or a salt thereof, wherein each L is independently hydrogen, adamantyl, a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; a is 4 to 20 (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20); b=2a+1 or b=a-1, each n is independently an integer from 0 to 4 (e.g., 0, 1, 2, 3, or 4);

[0063] [ka]

[0064] is a single or double bond, and

[0065] [ka]

[0066] represents a bond to a d-block element via a carbene. In some embodiments of Formula (Ic) or (Id), a is 4 to 10 (e.g., 4, 5, 6, 7, 8, 9, or 10).

[0067] In some embodiments, the perfluorinated ligand has formula (Ie) or (If):

[0068] [ka]

[0069] or a salt thereof, each L is independently hydrogen, adamantyl, a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; each Ar is independently a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; Each G is independently bonded, C 1-6 Alkyl, C 1-6 Alkenyl or C 1-6 is heteroalkyl, a is 4 to 20 (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20); b=2a+1 or b=a-1,

[0070] [ka]

[0071] is a single or double bond, and

[0072] [ka]

[0073] represents a bond to a d-block element via a carbene. In some embodiments of formula (Ie) or (If), a is 4 to 10 (e.g., 4, 5, 6, 7, 8, 9, or 10).

[0074] In any of formulas (I) and (Ia) through (If), each L is independently selected from hydrogen, adamantyl, a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group.

[0075] As used herein, "substituted or unsubstituted aromatic" refers to substituted or unsubstituted aromatic rings having 5 to 60 ring carbon atoms (e.g., C 1-6 As used herein, "substituted or unsubstituted heteroaromatic" refers to a substituted (e.g., C) or unsubstituted aromatic ring, such as phenyl, naphthyl, phenanthryl, and anthracenyl, having one to two heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon. 1-6 In some embodiments, the term refers to a stable bicyclic or tricyclic system containing at least one 5- to 7-membered aromatic ring containing one to three, and in some embodiments one to two, alkyl-substituted or unsubstituted aromatic rings or heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon. Monocyclic heteroaryl groups typically have 5 to 7 ring atoms, and according to some embodiments, bicyclic heteroaryl groups are 9 to 10 membered heteroaryl groups, i.e., groups containing 9 or 10 ring atoms in which one 5 to 7 membered aromatic ring is fused to a second aromatic or non-aromatic ring. If the total number of S atoms and O atoms in a heteroaryl group is more than 1, these heteroatoms are not adjacent to each other. It is preferred that the total number of S atoms and O atoms in a heteroaryl group is 2 or less. It is particularly preferred that the total number of S atoms and O atoms in an aromatic heterocycle is 1 or less. Heteroaromatic groups include, but are not limited to, oxazolyl, piperazinyl, pyranyl, pyrazinyl, pyrazolopyrimidinyl, pyrazolyl, pyridinyl, pyridyl, pyrimidinyl, pyrrolyl, quinolinyl, tetrazolyl, thiazolyl, thienylpyrazolyl, thiophenyl, triazolyl, benzofrijoxazolyl, benzofuranyl, benzothiazolyl, benzolhiophenyl, benzoxadiazolyl, dihydrobenzodioxinyl, furanyl, imidazolyl, indolyl, isothiazolyl, and isoxazolyl.

[0076] In some embodiments, each L is independently selected from hydrogen, adamantyl, 2-methylphenyl, 3-methylphenyl, 4-methylphenyl, 2,4-dimethylphenyl, 2,5-dimethylphenyl, 2,6-dimethylphenyl, 3,5-dimethylphenyl, 2,4,6-trimethylphenyl, 2-ethylphenyl, 3-ethylphenyl, 4-ethylphenyl, 2,4-diethylphenyl, 2,5-diethylphenyl, 2,6-diethylphenyl, 3,5-diethylphenyl, 2,4,6-triethylphenyl, 2-npropylphenyl, 3-n Propylphenyl, 4-npropylphenyl, 2,4-di-npropylphenyl, 2,5-di-npropylphenyl, 2,6-di-npropylphenyl, 3,5-di-npropylphenyl, 2,4,6-tri-npropylphenyl, 2-isopropylphenyl, 3-isopropylphenyl, 4-isopropylphenyl, 2,4-di-isopropylphenyl, 2,5-di-isopropylphenyl, 2,6-di-isopropylphenyl, 3,5-di-isopropylphenyl, 2,4,6-tri-isopropylphenyl, 2-isobutylphenyl, 3-isobutylphenyl, 4-isobutylphenyl, 2,4-di-isobutylphenyl, 2,5-di-isobutylphenyl, 2,6-di-isobutylphenyl, 3,5-di-isobutylphenyl, 2,4,6-tri-isobutylphenyl, 2-sec-butylphenyl, 3-sec-butylphenyl, 4-sec-butylphenyl, 2,4-di-sec-butylphenyl, 2,5-di-sec-butylphenyl, 2,6-di-sec-butylphenyl, 3,5-di-sec-butylphenyl, 2,4,6-tri-sec-butylphenyl, 2-t-butylphenyl tert-butylphenyl, 3-tert-butylphenyl, 4-tert-butylphenyl, 2,4-di-tert-butylphenyl, 2,5-di-tert-butylphenyl, 2,6-di-tert-butylphenyl, 3,5-di-tert-butylphenyl, 2,4,6-tri-tert-butylphenyl, 2-cyclohexylphenyl, 3-cyclohexylphenyl, 4-cyclohexylphenyl, 2,4-di-cyclohexylphenyl, 2,5-di-cyclohexylphenyl, 2,6-di-cyclohexylphenyl, 3,5-di-cyclohexylphenyl, or 2,4,6-tri-cyclohexylphenyl.In certain embodiments of Formulas (I) and (Ia) through (If), each L is independently hydrogen or 2,4,6-trimethylphenyl.

[0077] In any of formulas (I) and (Ia)-(If), each Y is independently a bond or a spacer group. For example, Y can be a bond, a substituted or unsubstituted C 1-10 Alkyl groups, substituted or unsubstituted C 2-10 Alkenyl groups, substituted or unsubstituted C 2-10 Alkynyl group, substituted or unsubstituted C 1-10 Heteroalkyl groups, substituted or unsubstituted C 3-6 Cycloalkyl groups, substituted or unsubstituted C 3-6 It can be a heterocycloalkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, a substituted or unsubstituted alkaryl group, a substituted or unsubstituted arylalkyl group, or a linear or branched alkyleneoxy group (e.g., polyethylene oxide, polypropylene oxide, or a combination thereof).

[0078] In some embodiments, each Y is independently a bond, a substituted or unsubstituted C 1-10 Alkyl groups, substituted or unsubstituted C 2-10 Alkenyl groups, substituted or unsubstituted C 2-10 Alkynyl group, substituted or unsubstituted C 1-10 Heteroalkyl groups, substituted or unsubstituted C 3-6 Cycloalkyl groups, substituted or unsubstituted C 3-6 In certain embodiments, each Y is independently a bond, a substituted or unsubstituted C alkyl group, a substituted or unsubstituted C aryl ... 1-10 Alkyl groups, substituted or unsubstituted C 2-10In a preferred embodiment, each Y is independently a bond, a substituted or unsubstituted C alkyl group, a substituted or unsubstituted C aryl group, a substituted or unsubstituted C heteroaryl group, a substituted or unsubstituted alkaryl group, or a substituted or unsubstituted arylalkyl group. 1-10 Alkyl groups, substituted or unsubstituted C 2-10 It is an alkenyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, a substituted or unsubstituted alkaryl group, or a substituted or unsubstituted arylalkyl group.

[0079] The perfluorinated ligand comprises a perfluorinated tag. For example, in any of formulas (I) and (Ia)-(If), each Z is a perfluorinated tag. The perfluorinated tag can be any perfluorinated group, such as, for example, a perfluorinated alkyl (e.g., linear or branched), aryl, alkyarl, or arylalkyl group having up to 60 carbon atoms. In some embodiments, the perfluorinated tag is a perfluorinated C group containing only carbon and fluorine atoms. 3-60 In certain embodiments, the perfluorinated tag is a perfluorinated C group that contains only carbon and fluorine atoms. 3-40 In another embodiment, the perfluorinated tag is a perfluorinated C 3-20 For example, perfluorinated tags are C4F9 groups, C5F 11 Base, C6F 13 Base, C7F 15 Base, C8F 17 Base, C9F 19 group, C 10 F 21 group, C6F5 group, C4F7 group, C5F9 group, C6F 11 Base, C7F 13 Base, C8F 15 Base, C9F 17 Groups and C 10 F 19 groups, each of which may be a linear or branched alkyl, aryl, alkyarl, or arylalkyl group.

[0080] In one embodiment, Z is a linear or branched perfluoroalkyl chain having a chain length of up to 60 or more carbon atoms, e.g., the perfluoroalkyl chain has a chain length of 3 to 60, particularly 3 to 40, more particularly 3 to 20, and even more particularly 3 to 10 or more carbon atoms. For example, the perfluoroalkyl chain can be C4F9, C6F 13 , C7F 15 , C8F 17 , C9F 19 , and C 10 F 21 Preferably, the group consisting of C6F 13 , C8F 17 , and C 10 F 21 (each of which may be linear or branched and combinations thereof, where each of which may be linear or branched).

[0081] In one embodiment, the perfluorinated ligand has formula (Ig):

[0082] [ka]

[0083] or a salt thereof, a is 4 to 20 (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20); b=2a+1 or b=a-1,

[0084] [ka]

[0085] is a single or double bond, and

[0086] [ka]

[0087] represents a bond to a d-block element via a carbene. In some embodiments of Formula (Ig), a is 4 to 10 (e.g., 4, 5, 6, 7, 8, 9, or 10).

[0088] In one embodiment, the perfluorinated ligand has formula (Ih):

[0089] [ka]

[0090] or a salt thereof, wherein a is 4 to 20 (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20); b=2a+1 or b=a-1,

[0091] [ka]

[0092] is a single or double bond, and

[0093] [ka]

[0094] represents a bond to a d-block element via a carbene. In some embodiments of Formula (Ig), a is 4 to 10 (e.g., 4, 5, 6, 7, 8, 9, or 10).

[0095] Examples of perfluorinated ligands include:

[0096] [ka]

[0097] or a salt thereof, wherein

[0098] [ka]

[0099] is a single or double bond, and

[0100] [ka]

[0101] represents a bond to a d-block element via a carbene.

[0102] As used herein, the symbols

[0103] [ka]

[0104] represents a single bond or a double bond. In some embodiments,

[0105] [ka]

[0106] is a single bond. In other embodiments,

[0107] [ka]

[0108] is a double bond.

[0109] [ka]

[0110] In embodiments where is a single bond,

[0111] [ka]

[0112] The orientation of the two substituents resulting from may be of any suitable stereochemistry, i.e., cis or trans. In a preferred embodiment,

[0113] [ka]

[0114] is a single bond,

[0115] [ka]

[0116] The stereochemistry of the substituents resulting from is trans.

[0117] As used herein, the symbols

[0118] [ka]

[0119] represents a bond to a d-block element via a carbene. In other words,

[0120] [ka]

[0121] represents the binding of the catalyst to the metal.

[0122] In some embodiments, the perfluorinated SABRE catalyst further comprises an additional ligand. For example, the perfluorinated SABRE catalyst may further comprise an additional ligand selected from a phosphine ligand, a carbene ligand, an imidazole ligand, a pincer chelate ligand and a compound comprising a sulfoxide group. In certain embodiments, the perfluorinated SABRE catalyst comprises one or more phosphine ligands. Examples of phosphine ligands include, but are not limited to:

[0123] [ka]

[0124] In some embodiments, the perfluorinated SABRE catalyst comprises a pincer chelate ligand. Generally, when the perfluorinated SABRE catalyst comprises a phosphine ligand or a pincer chelate ligand, the perfluorinated SABRE catalyst is in pre-catalyst form. In some embodiments, the perfluorinated SABRE catalyst comprises a ligand that is a compound that comprises a sulfoxide group. Examples of compounds that comprise a sulfoxide group can be selected from the group consisting of dimethyl sulfoxide (DMSO), phenyl trifluoromethyl sulfoxide, phenyl methyl sulfoxide, phenyl chloromethyl sulfoxide, diphenyl sulfoxide, dibenzoyl sulfoxide and dibutyl sulfoxide. Generally, when the perfluorinated SABRE catalyst comprises a compound that comprises a sulfoxide group, the perfluorinated SABRE catalyst is in active form. As used herein, "perfluorinated SABRE catalyst" can refer to an active perfluorinated SABRE catalyst or a perfluorinated SABRE pre-catalyst.

[0125] The active perfluorinated SABRE catalyst can be prepared by any suitable method. In general, the active perfluorinated SABRE catalyst is prepared by combining the perfluorinated SABRE pre-catalyst with the substrate, para-hydrogen, and optionally a co-ligand in a solvent to form a mixture containing the active perfluorinated SABRE catalyst. In some embodiments, the active perfluorinated SABRE catalyst is prepared by combining the perfluorinated SABRE pre-catalyst with the substrate, para-hydrogen, and a co-ligand in a solvent to form a mixture containing the active perfluorinated SABRE catalyst. The co-ligand, when included in the preparation of the active perfluorinated SABRE catalyst, can be combined with the perfluorinated SABRE pre-catalyst in any order and by any suitable means. For example, the co-ligand, when included in the preparation of the active SABRE catalyst, can be provided first to interact with the transferred pre-catalyst to promote the formation of the active perfluorinated SABRE catalyst. Alternatively, the co-ligand, when included in the preparation of the active perfluorinated SABRE catalyst, can be added together with the substrate to promote the formation of the active perfluorinated SABRE catalyst. In some embodiments, the co-ligand, substrate, and para-hydrogen are essentially combined simultaneously with the perfluorinated SABRE pre-catalyst in a solvent to promote the formation of an active perfluorinated SABRE catalyst. In other embodiments, a substrate is first provided that interacts with the perfluorinated SABRE pre-catalyst to promote the formation of an active SABRE catalyst. In some embodiments, the co-ligand and substrate are combined with the perfluorinated SABRE pre-catalyst in a solvent, and para-hydrogen is added (e.g., bubbled) to the resulting mixture. In other embodiments, the substrate is combined with the perfluorinated SABRE pre-catalyst in a solvent, and para-hydrogen is added (e.g., bubbled) to the resulting mixture. In some embodiments, the active perfluorinated SABRE catalyst is prepared by combining the perfluorinated SABRE pre-catalyst with the co-ligand in addition to the substrate and para-hydrogen.

[0126] In some embodiments, the active perfluorinated SABRE catalyst has the formula [Ir(H)2(F-IMes)(η2 -SUBSTRATE)(Co-ligand)] or [Ir(H)2(F-IMes)(η 1 -SUBSTRATE)(Co-ligand)2] wherein: SUBSTRATE is a target substrate to be hyperpolarized by the transfer of parahydrogen of pure singlet spin order by a spin transfer catalyst, preferably an atom with a 1 / 2 spin nucleus, e.g. 1 H, 13 C. 15 N, 19 F, 31 P and / or 29 The target substrate is enriched with Si. Without wishing to be bound by theory, the co-ligand interacts with the spin-transfer pre-catalyst to form an activated polarization transfer catalyst, facilitating polarization transfer to the target substrate. F-IMes refers to perfluorinated forms of N-heterocyclic carbenyI (NHC) ligands, such as 1,3-bis(2,4,6-trimethylphenyl)-1,3-dihydro-2H-imidazol-2-ylidene groups.

[0127] In some embodiments, a perfluorinated SABRE catalyst (e.g., a perfluorinated SABRE precatalyst) can be prepared by a method comprising reacting a perfluorinated compound with a base to form a carbene and reacting the carbene with [(d-block element)(COD)Cl]2, where COD represents cyclooctadienyl. In certain embodiments, the method comprises reacting a perfluorinated compound with a base to form a carbene and reacting the carbene with [Ir(COD)Cl]2. For example, a perfluorinated SABRE catalyst of the formula:

[0128] [ka]

[0129] The perfluorinated SABRE catalyst has the formula:

[0130] [ka]

[0131] can be prepared by reacting the carbene with [Ir(COD)Cl].

[0132] Examples of perfluorinated SABRE catalysts (e.g., perfluorinated SABRE precatalysts) include:

[0133] [ka]

[0134] [ka]

[0135] [ka]

[0136] [ka]

[0137] or

[0138] [ka]

[0139] or a salt thereof.

[0140] As used herein, the terms "hyperpolarized substrate" and "hyperpolarized MRI probe" are used interchangeably to refer to the desired hyperpolarized compound.

[0141] As described herein, methods of preparing a hyperpolarized substrate can include hyperpolarizing a mixture that includes a perfluorinated SABRE catalyst (e.g., an active perfluorinated SABRE catalyst) by exposing the mixture to a magnetic field or radio frequency excitation to transfer polarization from parahydrogen to the substrate to form the hyperpolarized substrate. Initially, the hyperpolarized substrate is complexed with the hyperpolarized perfluorinated SABRE catalyst; however, one of skill in the art will understand that the hyperpolarized substrate can be replaced with another substrate molecule and the process repeated to generate a free hyperpolarized substrate bolus.

[0142] The transfer of polarization from the parahydrogen to the substrate to form the hyperpolarized substrate can occur under any suitable magnetic field or radio frequency excitation. For example, the transfer of polarization from the parahydrogen to the substrate can occur in a magnetic field below the Earth's magnetic field. The appropriate level of magnetic field or radio frequency excitation required to transfer the polarization from the parahydrogen to the substrate to form the hyperpolarized substrate will be readily apparent to one of skill in the art.

[0143] In some embodiments, the method includes replenishing para-hydrogen into the mixture during the step of hyperpolarizing the mixture including the perfluorinated SABRE catalyst (e.g., an active perfluorinated SABRE catalyst) by exposing the mixture to a magnetic field or radio frequency excitation to transfer polarization from the para-hydrogen to the substrate to form a hyperpolarized substrate. In other words, in some embodiments, the method includes bubbling para-hydrogen into the mixture including the perfluorinated SABRE catalyst (e.g., an active perfluorinated SABRE catalyst) by exposing the mixture to a magnetic field or radio frequency excitation to transfer polarization from the para-hydrogen to the substrate to form a hyperpolarized substrate during the step of hyperpolarizing the mixture including the perfluorinated SABRE catalyst (e.g., an active perfluorinated SABRE catalyst).

[0144] The method of preparing a hyperpolarized substrate may include providing a co-ligand for interacting with the perfluorinated SABRE catalyst to promote the formation of an active perfluorinated SABRE catalyst. The co-ligand may be any suitable compound that includes one or more sulfoxide groups, thioester groups, phosphine groups, amine groups, CO groups, isonitrile groups, nitrogen-containing heterocyclic groups, or combinations thereof. In some embodiments, the co-ligand is a compound that includes a sulfoxide group. Examples of compounds that include a sulfoxide group may be selected from the group consisting of DMSO, phenylmethyl sulfoxide, phenylchloromethyl sulfoxide, diphenyl sulfoxide, dibenzoyl sulfoxide, phenyltrifluoromethyl sulfoxide, and dibutyl sulfoxide. In certain embodiments, the co-ligand is dimethyl sulfoxide or phenyltrifluoromethyl sulfoxide.

[0145] In some embodiments of the method for preparing a hyperpolarized substrate, the magnetic field is an electromagnetic field. For example, the strength of the electromagnetic field can range from 0 to 200 millitesla (mT). In some embodiments, the electromagnetic field can be at least partially supplied by one or more permanent magnets in addition to or instead of the coil. In some embodiments, the electromagnetic field can be an alternating magnetic field supplied at a frequency adapted to the particular nucleus. The alternating magnetic field can change direction (i.e., alternate between positive and negative relative to the positive direction). In some embodiments, the frequency can be a radio frequency, preferably between 50 and 500 MHz, although other frequencies outside this range are considered to be within the scope of the present disclosure.

[0146] A perfluorinated SABRE catalyst (e.g., an active perfluorinated SABRE catalyst) is combined with a substrate comprising parahydrogen and 1 / 2 spin nuclei or nuclei in a solvent to obtain a reaction mixture. The solvent can be any suitable solvent capable of forming a heterogeneous or homogeneous mixture. In some embodiments, the solvent comprises water, methanol, ethanol, a fluorous solvent, or a mixture thereof. For example, the solvent can be ethanolic or methanolic, i.e., comprising at least a combination of ethanol or methanol and water. In certain embodiments, the solvent comprises a fluorous solvent. In other embodiments, the solvent is deuterated, such that the deuterated solvent can be prepared without deuterium-hydrogen exchange (i.e., with limited deuterium-hydrogen exchange).

[0147] The fluorous solvent can be any organic solvent that contains at least one compound having a fluorine atom. Without wishing to be bound by a particular theory, it is believed that the fluorous solvent increases the solubility of the perfluorinated SABRE catalyst. In some embodiments, the solvent (e.g., the fluorous solvent) is selected from a perfluorohexane / diethyl ether mixture, a mixture of methoxynonafluorobutane and ethyl acetate with a non-polar solvent, a mixture of perfluorohexane and ether, a mixture of perfluorobutyl methyl ether and ethyl acetate, ether, a fluorocarbon derivative of THF FC 75, decafluoromethoxytrifluoromethylpentane, hexafluoropropanol, nonafluorobutyl methyl ether, perfluoromethylcyclohexane, perfluoroalkane, perfluorohexane, and methoxynonafluorobutane.

[0148] In some embodiments, the method of preparing a hyperpolarized substrate further includes isolating the hyperpolarized substrate. The hyperpolarized substrate can be isolated by any suitable method. For example, the hyperpolarized substrate can be isolated by extraction, filtration, column chromatography, distillation, crystallization, or a combination thereof.

[0149] In some embodiments, the hyperpolarized substrate is isolated by treating the reaction mixture with a solid phase adsorbent to adsorb the perfluorinated SABRE catalyst and recovering a liquid containing the hyperpolarized substrate, where the liquid is free (i.e., undetectable) or substantially free (e.g., less than 100 ppm, less than 50 ppm, less than 10 ppm, less than 5 ppm, or less than 1 ppm) of the perfluorinated SABRE catalyst. See, e.g., FIG. 10. The solid phase adsorbent can be any suitable adsorbent capable of preferentially adsorbing the perfluorinated SABRE catalyst over the hyperpolarized substrate. For example, the solid phase adsorbent can be a fluorous solid phase adsorbent, a reversed phase adsorbent (e.g., C18 adsorbent, etc.), and a polyethylene-based filter (e.g., ultra-high molecular weight polyethylene). See, e.g., FIGS. 6 and 7. In certain embodiments, the method of preparing the hyperpolarized substrate further includes passing a fluorophobic solvent over the adsorbent and recovering an eluate comprising the hyperpolarized substrate, the eluate being free (i.e., undetectable) or substantially free (e.g., less than 100 ppm, less than 50 ppm, less than 10 ppm, less than 5 ppm, or less than 1 ppm) of perfluorinated SABRE catalyst. The fluorophobic solvent can be any suitable solvent capable of preferentially washing the hyperpolarized substrate off the solid-phase adsorbent relative to the perfluorinated SABRE catalyst. For example, the fluorophobic solvent can include water and one or more of methanol, ethanol, acetonitrile, and dimethylformamide. Alternatively, or additionally, the method of preparing the hyperpolarized substrate can further include passing a fluorophilic solvent (e.g., an organic solvent selected from methanol, ethanol, acetonitrile, THF, ethyl acetate, chlorinated solvents (e.g., chlorinated alkanes such as methylene chloride, chloroform, and ethylene dichloride), and combinations thereof) over the adsorbent to recover, for example, the perfluorinated SABRE catalyst. Examples of fluorophilic solvent systems include perfluorohexane / diethyl ether mixtures, mixtures of methoxynonafluorobutane and ethyl acetate, mixtures of perfluorohexane and diethyl ether, mixtures of perfluorobutyl methyl ether and ethyl acetate, or diethyl ether.

[0150] In some embodiments, the hyperpolarized substrate is isolated by treating the reaction mixture with a solid phase adsorbent to adsorb the hyperpolarized substrate and recovering a liquid comprising the perfluorinated SABRE catalyst, wherein the liquid is free of (i.e., not detectable) or substantially free of (e.g., less than 100 ppm, less than 50 ppm, less than 10 ppm, less than 5 ppm, or less than 1 ppm) the hyperpolarized substrate. The solid phase adsorbent can be any suitable adsorbent capable of preferentially adsorbing the hyperpolarized substrate over the perfluorinated SABRE catalyst. For example, the solid phase adsorbent can be a normal phase adsorbent, such as, for example, silica, alumina, and the like. See, for example, FIG. 8. In certain embodiments, the method of preparing the hyperpolarized substrate further comprises passing a fluorophilic solvent over the adsorbent and recovering an eluate comprising the perfluorinated SABRE catalyst, wherein the eluate is free of (i.e., not detectable) or substantially free of (e.g., less than 100 ppm, less than 50 ppm, less than 10 ppm, less than 5 ppm, or less than 1 ppm) the hyperpolarized substrate. The fluorophilic solvent can be any suitable solvent that can preferentially wash the perfluorinated SABRE catalyst from the solid-phase adsorbent relative to the hyperpolarized substrate. For example, the fluorophobic solvent can include a perfluorohexane / diethyl ether mixture, a mixture of methoxynonafluorobutane and ethyl acetate, a mixture of perfluorohexane and diethyl ether, a mixture of perfluorobutyl methyl ether and ethyl acetate, or diethyl ether. Alternatively, or additionally, the method of preparing the hyperpolarized substrate can further include passing a fluorophobic solvent (e.g., a solvent comprising water, methanol, ethanol, acetonitrile, dimethylformamide, or a combination thereof) over the adsorbent, e.g., to recover the hyperpolarized substrate.

[0151] In any of the embodiments disclosed herein, the perfluorinated SABRE catalyst and / or hyperpolarized substrate may be present in a monophasic or biphasic mixture. The monophasic or biphasic mixture may include any combination of the solvents described herein. For example, the biphasic mixture may include a polar solvent (e.g., water, methanol, and ethanol) in combination with a nonpolar solvent (e.g., an organic solvent or a fluorous solvent). In embodiments in which the perfluorinated SABRE catalyst and / or hyperpolarized substrate are present in a biphasic solvent, the perfluorinated SABRE catalyst and / or hyperpolarized substrate may be isolated by liquid / liquid extraction. See, for example, FIG. 9. Thus, in some embodiments, the hyperpolarized substrate is isolated by liquid / liquid extraction, such as, for example, partitioning the perfluorinated SABRE catalyst and hyperpolarized substrate between a methanol mixture and a fluorous solvent, or partitioning the perfluorinated SABRE catalyst and hyperpolarized substrate between a methanol mixture and an organic solvent.

[0152] In some embodiments, the hyperpolarized substrate is isolated by precipitating the perfluorinated SABRE catalyst and filtering the precipitated perfluorinated SABRE catalyst from the hyperpolarized substrate. Typically, the perfluorinated SABRE catalyst is precipitated by adding a solvent in which the perfluorinated SABRE catalyst is insoluble (e.g., hexane, pentane, water, ethanol, etc.). In certain embodiments, the perfluorinated SABRE catalyst is precipitated by adding water.

[0153] The perfluorinated SABRE catalyst and / or hyperpolarized substrate can be dried or concentrated (e.g., under reduced pressure, using a desiccant, heating, or a combination thereof). Alternatively, or in addition, the perfluorinated SABRE catalyst and / or hyperpolarized substrate can be diluted or reconstituted with a solvent (e.g., water) to provide a desired concentration. For example, the perfluorinated SABRE catalyst can be isolated and reused as a hyperpolarized catalyst. Similarly, the hyperpolarized substrate can be dried or concentrated to remove the organic solvent and reconstituted in water for administration to a subject.

[0154] The substrate can be a 1 / 2 spin nucleus or any compound that contains a nucleus. For example, the substrate can be 1 H, 13 C. 15 N, 19 F, 31 P, 29 In some embodiments, the substrate may further comprise at least one of: 2 Thus, the methods described herein include the step of determining the identity of a target substrate. 1 H, 13 C. 15 N, 19 F, 31 P and / or 29 This can be used to enhance the signal of the Si response. In general, the spin polarization transfer described here is based on the SABRE effect; however, the method can be extended to parahydrogen induced polarization (PHIP).

[0155] In some embodiments, the substrate is selected from ketoglutarate, pyruvate, N-acetylcysteine, and salts or esters thereof. 13 C-Ketoglutaric acid, 1- 13 C-5- 12 C-Ketoglutaric acid, 1- 13 C-pyruvic acid, 1- 13 CN-acetylcysteine, 15 N2-isoniazid (or pyridyl-4-carbobis- 15 N2-hydrazide), 13 C2, 15 N3-metronidazole, 15 N2-1-aminoisoquinoline (1-AIQ), deuterated versions thereof, and salts thereof.

[0156] In some embodiments, the substrate has formula (II):

[0157] [ka]

[0158] (each R1 is independently selected from hydrogen, deuterium, a cation, C1-C6 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C1-C6 alkyl, (heterocycloalkyl)C1-C6 alkyl, (heteroaryl)C1-C6 alkyl, and (aryl)C1-C6 alkyl; and Xa, Xb, Xc, and Xd are each independently hydrogen or deuterium, with the proviso that at least one of Xa, Xb, Xc, and Xd is deuterium, or a pharma- ceutically acceptable salt thereof.

[0159] Each R1 may be independently selected from hydrogen, deuterium, a cation, C1-C6 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C1-C6 alkyl, (heterocycloalkyl)C1-C6 alkyl, (heteroaryl)C1-C6 alkyl, and (aryl)C1-C6 alkyl. In some embodiments, each R1 is independently selected from C1-C6 alkyl, for example, each R1 may be methyl, ethyl, propyl (e.g., isopropyl or n-propyl), butyl (e.g., isobutyl, n-butyl, tert-butyl or sec-butyl), pentyl or hexyl. In some embodiments, each R1 is independently selected from hydrogen, deuterium, and a cation. In embodiments where R1 is a cation, it will be readily understood by one of skill in the art that the compound of formula (II) is a salt (e.g., a pharma- ceutically acceptable salt) in which the negative charge on the oxygen is balanced by a cation. In certain embodiments, each R1 is independently selected from a cation or a C1-C6 alkyl.

[0160] The present disclosure further provides a hyperpolarized substrate or a pharma- ceutical composition comprising a hyperpolarized substrate or a pharma- ceutical acceptable salt obtained from any of the methods described herein, or a pharmaceutical composition comprising the hyperpolarized substrate or a pharma- ceutical acceptable salt and a pharma- ceutical acceptable carrier. In other words, the present invention provides imaging media (e.g., aqueous imaging compositions) with enhanced sensitivity to water-soluble compounds containing hyperpolarized nuclei, which imaging media are particularly well suited for nuclear magnetic resonance (NMR) spectroscopy and / or magnetic resonance imaging (MRI).

[0161] The present disclosure further provides a method for obtaining a magnetic resonance image of a tissue of a subject having or suspected of having cancer or an adverse vascular condition, comprising administering to the subject a hyperpolarized substrate or pharmaceutical composition thereof described herein and imaging the subject by magnetic resonance imaging. In one embodiment, the subject has cancer, such as a cancer selected from breast cancer, colon cancer, rectal cancer, bladder cancer, endometrial cancer, kidney cancer, lung cancer, melanoma, non-Hodgkin's lymphoma, pancreatic cancer, prostate cancer, and thyroid cancer. In other embodiments, the subject has an adverse vascular condition, such as a vascular condition selected from myocardial infarction, stroke, and pulmonary disease (e.g., COPD, pulmonary fibrosis, long-term COVID-19 symptoms, and combinations thereof).

[0162] In some embodiments, the disclosure provides a method of diagnosing or monitoring a patient with cancer or suspected of having cancer, the method comprising administering a hyperpolarized substrate or pharmaceutical composition as described above and administering a hyperpolarized 13 The present invention includes diagnosing or monitoring a patient by C-MRI. For example, the hyperpolarized substrate can be used in a method for diagnosing or monitoring a patient with cancer or a patient suspected of having cancer. In a particular embodiment, the method or use includes identifying a mutation that causes cancer. In a particular embodiment, the method or use identifies an IDH1 mutation as the cause of cancer. In other words, the method or use can be used to identify whether a patient has a tumor, for example, an IDH1 mutation.

[0163] The figures are now described to illustrate various aspects of the disclosed MRI probe injection device. Features of any one aspect of the present disclosure are not intended to limit the scope of other aspects of the present disclosure, and it will be understood that features from multiple aspects disclosed herein may be combined, unless otherwise contradicted by context.

[0164] FIG. 1 illustrates a reaction chamber 110 and a separator 120 of a hyperpolarized MRI probe injection apparatus according to one embodiment of the present disclosure. As shown in FIG. 1, the reaction chamber 110 includes multiple inlet ports 112, 114, and multiple outlet ports 116, 118. According to one embodiment, the reaction chamber 110 includes a first inlet port 112 for receiving a liquid input (e.g., a solvent, a catalyst, a substrate, etc.) and a second inlet port 114 for receiving a gas input (e.g., parahydrogen gas). The reaction chamber 110 also includes a first outlet port 116 for exhausting gas from the reaction chamber 110, and a second outlet port 118 for drawing a reaction mixture from the reaction chamber 110 to the separator 120.

[0165] According to one embodiment, the first inlet port 112 can introduce a solution including one or more solvents, a catalyst, and a substrate into the reaction chamber. The solvent can include ethanol, a mixture of ethanol and a fluorous solvent, methanol, or a mixture of methanol and a fluorous solvent, for example, the one or more solvents include ethanol and water. The substrate can include 13 C. 15 N, 19 F, 31 P and 29The reaction chamber 110 may include an element having a 1 / 2 spin nucleus selected from the group consisting of Si, as well as deuterated versions thereof. A second inlet port 114 may introduce parahydrogen gas into the reaction chamber. Although not shown, one or more additional inlet ports may be included in the reaction chamber 110 for introducing other liquids or gases into the reaction chamber. Alternatively, two or more components may be mixed in a separate vessel to form an intermediate solution, which is then introduced into the reaction chamber 110 through one of the inlet ports 112, 114.

[0166] According to one embodiment, a first outlet port 116 allows for the evacuation of gas from the reaction chamber 110. A continuous flow of parahydrogen gas can be connected to the inlet port 114, which is exhausted through the outlet port 116. The outlet port 116 may include a valve incorporated therein or connected in series therewith to allow a desired pressure to be maintained within the reaction chamber 110. According to one embodiment, the reaction chamber 110 is configured to withstand atmospheric pressures of up to 10 bar or more.

[0167] According to one embodiment, the line connected to the outlet port 116 may be connected to a gas trap 122 and / or a storage tank 124 to ensure that gas exhausted from the reaction chamber 110 is not vented to the atmosphere in order to implement safety protocols. Although not explicitly shown, a compressor, valve, gauge, or other typical air system components may be connected to the exhausted gas line to safely store gas exhausted from the reaction chamber 110.

[0168] According to one embodiment, the second exit port 118 allows for the withdrawal of the reaction mixture from the reaction chamber 110 after the reaction to produce the hyperpolarized MRI probe is complete. The second exit port 118 may include a valve incorporated therein or connected in series with it such that a vacuum may be created in the separator 120 and / or collection vessel 130 that causes liquid to be drawn up a tube in the reaction chamber 110 and into the separator. This valve is closed to create the vacuum while the reaction is taking place and is opened to withdraw the reaction mixture from the reaction chamber 110 once the reaction is complete.

[0169] The separator 120 is configured to receive the reaction mixture containing the perfluorocarbon hyperpolarized transfer SABRE catalyst, the solvent, and the hyperpolarized MRI probe from the reaction chamber and separate the hyperpolarized MRI probe from the catalyst. According to one embodiment, the separator 120 can include a filter medium, which can include an adsorbent, such as fluorous silica gel. The separator 120 can be configured to dilute the reaction mixture with water and / or ethanol before filtering the diluted mixture through the filter medium. The catalyst is absorbed into the fluorous silica gel and separated from the hyperpolarized probe in the solvent. According to another embodiment, the filter medium can be omitted and the mixture can be separated by stirring the mixture and extracting the ethanol layer from the other layer containing the fluorous solvent-catalyst mixture. Optionally, the reaction mixture can be diluted with water before stirring and extraction, such that the water / ethanol layer is extracted from the column. Alternatively, the above method can be adapted by adding a step of concentrating and redissolving using water when methanol is used as the solvent. Finally, the separated reaction mixture 132 containing the hyperpolarized MRI probe is collected in a collection vessel 130, which can be, for example, a vacuum flask or other device.

[0170] It will be appreciated that when extracted from the low magnetic field in the reaction chamber 110, the hyperpolarized MRI probe should be maintained in a high magnetic field to maintain polarization during separation, concentration, reconstitution, and administration. The high magnetic field may be provided by one or more permanent magnets or electromagnets (e.g., coils) in proximity to the separator 120 and collection vessel 130. In some cases, the high magnetic field may be provided by a coil of an MRI device located in proximity to the MRI probe injection device. The strength of the high magnetic field may be determined based on the molecule to be hyperpolarized.

[0171] Although not explicitly shown in FIG. 1 , reaction chamber 110 may include a number of internal components, such as a temperature control device and / or coils (e.g., solenoids) for generating an internal magnetic field within reaction chamber 110, which are connected to an external controller via interface 126. The controller is configured to execute software for controlling the temperature within the reaction chamber, controlling the flow rates of gases and liquids through the device, monitoring the safety of the device and / or the environment, administering a desired amount of hyperpolarized probe to a patient, and / or calculating the decay rate of the hyperpolarized probe as a function of gas and / or liquid flow rate. Interface 126 may include electrical connections for providing control signals to the internal components and / or for receiving signals from one or more sensors (e.g., temperature sensors, gaussmeters, pressure gauges, etc.) connected to or disposed within reaction chamber 110.

[0172] Some components of the device can be made from non-magnetic materials or plastics, for example, various connectors, sensors, valves, gauges, etc. can be made from brass or plastic to reduce their effect on the magnetic field induced in the reaction chamber.

[0173] FIG. 2 illustrates the internal components of the reaction chamber 110 according to one embodiment of the present disclosure. The reaction chamber 110 can include one or more layers 202, 204, 206 of metals shaped to form a chamber configured to contain liquids and / or gases at a volume and pressure suitable for carrying out the disclosed reactions therein. As shown in FIG. 2, the reaction chamber 110 can include three metal layers forming a substantially cylindrical shape. It will be understood that the reaction chamber is not limited to a cylindrical shape and other desired shapes are contemplated for use as the reaction chamber. According to one embodiment, the layers 202, 204, 206 include a ferromagnetic alloy. According to some embodiments, one or more layers 202, 204, 206 may include a nickel-iron (Ni-Fe) alloy, such as Invar, Permalloy, or Mumetal. At least one layer 202, 204, 206 is configured to shield the reaction chamber from magnetic fields associated with external sources, such as the Earth's magnetic field or other electrical components that may emit EM radiation. According to one embodiment, layers 202, 204, 206 include a mu-metal shield that attenuates magnetic fields from external sources to have a strength below a threshold within one or more reaction chambers. According to one embodiment, the threshold is 10 nT. According to some embodiments, different layers can include different materials. For example, outer layer 202 can include a ferromagnetic alloy used to shield the chamber interior from external magnetic fields, while inner layer 206 can include a separate material that does not react with solvents or chemicals introduced into the reaction chamber.

[0174] According to one embodiment, the reaction chamber 110 also includes a coil 210 that is used to generate an electromagnetic field within the reaction chamber in response to an electric current being passed therethrough. According to some embodiments, the coil is a highly homogeneous solenoid. The electric current can provide a constant or alternating magnetic field of 0-200 millitesla (mT) and / or a radio frequency of 0 to 500 MHz.

[0175] According to one embodiment, the reaction chamber 110 also includes a temperature control device 220, which can include both heating and cooling elements. The temperature control device may include a non-magnetic (i.e., non-ferrous) material. According to one embodiment, the cooling element includes a liquid nitrogen bath outside at least the inner layer 206 of the reaction chamber such that at least one surface of the layer 206 can be in contact with liquid nitrogen. The liquid nitrogen evaporates to draw heat from the reaction chamber through the surface and provide cooling. The resulting nitrogen gas can be vented to the atmosphere and / or into a gas trap 122. The heating element can be a resistive heating element, such as a convection coil, proximate to the surface of the layer 206 or inserted into the reaction chamber through a port in the reaction chamber. In operation, the temperature control device can be connected to a temperature sensor / probe and configured to operate the heating and cooling elements to regulate the temperature of the reaction chamber 110. According to one embodiment, the temperature control device 220 is configured to maintain the temperature of the reaction chamber between -25°C and 100°C. According to other embodiments, the temperature of the reaction chamber can be cycled between two or more different temperatures up to 10-20 times within a period of 1 to 5 minutes. For example, during operation, liquid nitrogen can be dispensed into the bath to cool the reaction chamber to a desired low temperature. Once the reaction chamber 110 has reached a low temperature and / or the liquid nitrogen has boiled into nitrogen gas, the heating element can be turned on to heat the reaction chamber 110 to a desired high temperature. This cycle can be repeated multiple times during the reaction.

[0176] 3 is a schematic diagram illustrating components and their arrangement of an MRI probe injection device according to one embodiment of the present disclosure. The MRI probe injection device includes at least one reaction chamber 110, a separator 120, a collection and measurement facility 320, and an MRI probe administrator device 330. The reaction chamber 110, the collection and measurement facility 320, and the MRI probe administrator device 330 are connected to a controller 310. The injection device also includes a dryer or concentrator 322 and an analyzer 332 for analyzing at least one of the purity or concentration of the hyperpolarized MRI probe in the solution.

[0177] According to one embodiment, the controller 310 includes software and / or hardware for operating the MRI probe injection device. The controller 310 can include a computing device including at least one processor, a memory, and one or more input / output devices. The software stored in the memory can be executed by the processor to control the functions of the MRI probe injection device. According to one embodiment, the controller 310 includes a processor configured to execute instructions that cause the processor to: control the flow rate of gas and / or liquid through the device, monitor safety indicators of the device and / or environment, administer a desired amount of hyperpolarized MRI probe to the patient, and / or calculate the decay rate of the hyperpolarized MRI probe as a function of the flow rate of the gas and / or liquid.

[0178] Although not explicitly shown, the controller 310 may receive input from one or more sensors that provide feedback to control the operation of the MRI probe injection device. For example, the sensors may include a temperature sensor to monitor the temperature of the reaction chamber and control the temperature control device (i.e., heating and / or cooling elements). The sensors may also include a pressure transducer, flow meter, or other type of sensor to monitor the flow rate of liquids and / or gases entering or leaving the reaction chamber 110. The sensors may also include a gas leak detector and / or an oxygen level monitor to maintain safety protocols in operating the device. For example, the gas leak detector may monitor the ambient environment around the device to monitor the level of hydrogen gas that may be leaking from the reaction chamber and / or any associated gas lines or storage tanks. The oxygen level monitor may detect the oxygen level in the room to prevent accidental asphyxiation due to nitrogen gas leaks. It will be understood that any other necessary sensors, such as encoders, limit switches, proximity sensors, optical sensors, etc., may be included in the device to monitor one or more operating characteristics or the state of any components or actuators incorporated therein.

[0179] The controller 310 may also include an interface that allows for human input, for example, via a touch screen display, a keypad or keyboard, switches, buttons, etc. This interface allows a technician to manually operate the device. Alternatively, the interface may include a wired or wireless interface that allows the controller to communicate with an external terminal (e.g., a computer device, a server device, a tablet device, etc.) that is used to control the operation of the MRI probe injection device. For example, the controller 310 may include a network interface that allows the device to be connected to other terminals via a wired or wireless interface. The terminal may display a graphical user interface that allows the technician to interact with various menus to control the operation of the device.

[0180] As shown in FIG. 3, in one embodiment, the MRI probe injection device includes a collection and measurement facility 320. In a clinical setting, the mixture containing the hyperpolarized MRI probe can be analyzed and further processed to facilitate automated generation and administration of the hyperpolarized MRI probe to a patient (e.g., a human or an animal) during treatment. According to one embodiment, the collection and measurement facility 320 can include a dryer 322 configured to concentrate the solution or emulsion containing the hyperpolarized MRI probe received from the one or more MRI probe separators 120 to form a concentrate. The dryer 322 can include a spray dryer and / or an evaporative dryer. In one embodiment, the one or more evaporative dryers form one or more azeotropes with the inert solvent, resulting in a lower temperature of the evaporative dryer.

[0181] For example, the vessel may be equipped with a spray dryer with a nozzle configured to vaporize the solvent and deposit the hyperpolarized MRI probe on the vessel wall. The solvent vapor may be exhausted to a gas trap and / or a storage tank. The solid hyperpolarized MRI probe may be dissolved in a buffer solution (e.g., saline or a mixture of saline and ethanol) to form a solution containing a desired concentration of the hyperpolarized MRI probe. The collection and measurement facility 320 may also include an analyzer 324 and / or other measurement tools used to analyze the reaction mixture before and / or after concentration and dilution to determine the effectiveness of the reaction in the reaction chamber and / or adjust operational parameters to create a buffer solution with a desired concentration of the hyperpolarized MRI probe. For example, after analyzing the buffer solution, the controller 310 may use the measured concentration of the hyperpolarized MRI probe in the buffer solution to dispense additional buffer solution to reach the target concentration. In one embodiment, the analyzer 324 may include a liquid chromatography / mass spectrometer (LC / MS) and / or a nuclear magnetic resonance (NMR) spectrometer.

[0182] The administrator 330 may include any equipment necessary for metering and delivery of the hyperpolarized MRI probe to a patient. The administrator 330 is configured to administer a dose of a solution containing a desired concentration of the hyperpolarized MRI probe to an animal or patient in need thereof. According to one embodiment, the administrator 330 may include, but is not limited to, a peristaltic pump, fluid lines, and the like. In one embodiment, the administrator 330 may include a power injector common to other MRI or X-ray contrast agents. In another embodiment, the administrator 330 may include an aerosol delivery attachment for pulmonary administration.

[0183] Again, it will be appreciated that the separator 120, collection and measurement equipment 320, and administrator 330 should be maintained in a high magnetic field during extraction and preparation of the hyperpolarized MRI probe prior to delivery to the patient. The magnetic field can be provided by one or more permanent magnets or electromagnets, which can be contained in close proximity to the device and / or can be provided by a separate device, such as an MRI machine.

[0184] FIG. 4 illustrates an MRI injection device with multiple reaction chambers and the separation of hyperpolarized probes from the reaction mixture from each reaction chamber according to one embodiment of the present disclosure. As shown in FIG. 4, the MRI injection device 400 may include two or more reaction chambers 402, 404, 406 to facilitate multiple reactions to different hyperpolarized MRI probes substantially simultaneously and / or to increase the total amount of a single hyperpolarized MRI probe through multiple independent reactions performed in parallel. For example, if it takes 10 minutes to produce a single dose of hyperpolarized MRI probe from a particular reaction chamber, multiple reaction chambers can be used to produce separate doses every 5 minutes or 3.3 minutes, for example, using a two or three chamber system. Each reaction chamber 402, 404, 406 can be associated with a separate and distinct separator 120, collection and measurement equipment 320, and / or administrator 330, as described above. Alternatively, multiple reaction chambers can share a single separator 120, collection and measurement equipment 320, and / or administrator 330.

[0185] It will be appreciated that in some cases, one or more reaction chambers are configured to be operated in series. For example, a reaction mixture generated based on a reaction carried out in a first reaction chamber can be provided to a second reaction chamber, where a second reaction can take place. The reaction can be the same reaction or a different reaction (e.g., by introducing a different gas or chemical compound into the second reaction chamber, etc.).

[0186] 5 illustrates some of the steps involved in a method of preparing a hyperpolarized MRI probe for administration to a patient, according to one embodiment of the present disclosure. At step 502, a reaction mixture is provided to one or more chambers of an MRI probe injection device. Again, the reaction mixture can include a perfluorinated SABRE catalyst including a d-block element and a perfluorinated ligand, a solvent, a co-ligand, and a substrate to be hyperpolarized into the MRI probe.

[0187] According to one aspect of the present disclosure, the substrate is 1- 13 C-Ketoglutaric acid, 1- 13 C-5- 12 C-Ketoglutaric acid, 1- 13 C-pyruvic acid, 1- 13 CN-acetylcysteine, 15 N2-isoniazid (or pyridyl-4-carbobis- 15 N2-hydrazide), 13 C2, 15 N3-metronidazole, 15 It may be selected from one of N2-1-aminoisoquinoline (1-AIQ), deuterated versions thereof, and salts thereof.

[0188] According to one aspect of the present disclosure, the perfluorinated ligand has the formula (I): [L m -(NHC)-(YZ) q or a salt thereof. In formula (I), each L is independently selected from hydrogen, adamantyl, a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; NHC is a 4- to 7-membered N-heterocyclic carbenyi group, where NHC is attached to a d-block element via a carbene; each Y is independently selected from a bond or a spacer group; each Z is a perfluorinated tag; m is an integer from 1 to 4; and q is an integer from 1 to 3. The perfluorinated tag is one of the following: a perfluorinated C that contains only carbon and fluorine atoms; 3-60 Group: Perfluorinated C containing only carbon and fluorine atoms 3-40 group; or perfluorinated C 3-20 Base.

[0189] According to one aspect of the present disclosure, the perfluorinated ligand is:

[0190] [ka]

[0191] or a salt thereof,

[0192] [ka]

[0193] is a single or double bond, and

[0194] [ka]

[0195] represents a bond to a d-block element via a carbene.

[0196] At 504, the reaction mixture is agitated via injection of parahydrogen gas or a mixture of parahydrogen gas and nitrogen gas. For example, the inlet port 114 connected to the parahydrogen gas can be connected to a tube descending to the bottom of the reaction chamber 110 such that the gas entering the reaction chamber is bubbled through the solution containing the solvent, catalyst, and substrate. According to one embodiment, the coil 210 and / or the temperature controller 220 can be controlled to maintain a target temperature of the reaction chamber 110 or to cycle the temperature between two or more temperatures to promote the reaction to produce a hyperpolarized MRI probe. According to one embodiment, a magnetic field is applied within the reaction chamber using a coil 210 suitable for hyperpolarizing the perfluorinated SABRE catalyst and substrate to hyperpolarize the substrate into a hyperpolarized MRI probe.

[0197] In step 506, a solution containing the hyperpolarized MRI probe is extracted from the reaction chamber. The solution may be separated into at least two portions by separating the hyperpolarized MRI probe from the reaction mixture by at least one of filtration, extraction, or column chromatography to obtain a solution containing the hyperpolarized MRI probe. According to one embodiment, the solution is filtered through a separator 120 to separate the catalyst from the hyperpolarized MRI probe dissolved in a solvent comprising ethanol, a mixture of ethanol and water, methanol, or a mixture of ethanol and methanol. According to one embodiment of the present disclosure, the solvent may be selected from one of a perfluorohexane / diethyl ether mixture, a mixture of methoxynonafluorobutane and ethyl acetate with a non-polar solvent, a mixture of perfluorohexane and ether, a mixture of perfluorobutyl methyl ether and ethyl acetate, ether, a fluorocarbon derivative of THF FC 75, decafluoromethoxytrifluoromethylpentane, hexafluoropropanol, nonafluorobutyl methyl ether, perfluoromethylcyclohexane, perfluoroalkanes, perfluorohexane, and methoxynonafluorobutane.

[0198] In step 508, the solution is concentrated to obtain a concentrate. To evaporate the solvent and increase the concentration of the hyperpolarized MRI probe in the solution, the solution can be dried using one or more dryers. In some cases, all of the solvent is removed, leaving a powder, solid residue, or viscous liquid containing the hyperpolarized MRI probe on the surface of the dryer.

[0199] In step 510, a buffer solution is added to the concentrate to obtain a solution of the desired concentration. According to one embodiment, saline or a solution of saline and ethanol can be added to the concentrate to create a buffer solution with a desired concentration of hyperpolarized MRI probe.

[0200] In step 512, the purity and / or concentration of the hyperpolarized MRI probe in the buffer is analyzed. According to some embodiments, the analysis can be performed using at least one of a liquid chromatograph / mass spectrometer or an NMR spectrometer. According to some embodiments, the analysis results can be used to further dilute the solution with additional buffer to adjust the concentration of the hyperpolarized MRI probe in the solution.

[0201] A hyperpolarized MRI probe can be administered to a subject (e.g., a patient or an animal) in step 514. The hyperpolarized MRI probe can be administered at a desired dose and / or at desired intervals using the administrator 330.

[0202] Again, it will be appreciated that step 504 occurs in a low magnetic field inside the reaction chamber, and steps 506-512 occur in a high magnetic field (e.g., orders of magnitude greater than the low magnetic field inside the reaction chamber) provided by one or more permanent magnets and / or electromagnets outside the reaction chamber.

[0203] In one embodiment, the hyperpolarized MRI probe can be separated from the reaction mixture in an MRI separator by a fluorophobic path through a filter medium, as shown in FIG. 6 and further described in Example 8.

[0204] FIG. 7 shows one embodiment of hyperpolarization, probe separation, and recovery of the SABRE catalyst.

[0205] In another embodiment, as shown in FIG. 8 and further described in Example 9, the hyperpolarized MRI probe can be separated from the reaction mixture in an MRI separator by a fluorophobic path preceded by a fluorophilic path.

[0206] In another embodiment, FIG. 9 illustrates the process of probe separation and injection into a subject (eg, an animal or patient).

[0207] Aspects including embodiments of the present disclosure may be beneficial alone or in combination with one or more other aspects or embodiments. Without limiting the foregoing, certain non-limiting aspects of the present disclosure, numbered 1-47, are provided below. As will be apparent to one of skill in the art upon reading this disclosure, each of the individually numbered aspects may be used or combined with any of the preceding or succeeding individually numbered aspects. This is intended to provide support for all combinations of such aspects, and is not limited to the combinations of aspects explicitly provided below:

[0208] (1) In aspect (1), an MRI probe injection device is provided that includes: (i) one or more reaction chambers, each reaction chamber containing: (a) a structure configured to attenuate a magnetic field within the reaction chamber from an external source, where the strength of the magnetic field within the reaction chamber from the external source is below a threshold; (b) one or more inlet ports; (c) a coil configured to generate an electromagnetic field within the reaction chamber; (d) one or more temperature control devices; and (e) one or more exit ports; (ii) one or more MRI probe separators configured to receive a reaction mixture containing the perfluorinated SABRE catalyst, a solvent, and a hyperpolarized MRI probe from the one or more reaction chambers and to extract the hyperpolarized MRI probe from the reaction mixture; and (iii) one or more MRI probe collectors configured to form a solution containing a desired concentration of hyperpolarized MRI probes.

[0209] (2) In embodiment (2), structure (a) is a mu-metal shield that attenuates magnetic fields from external sources to have a strength of 10 nT or less in one or more reaction chambers.

[0210] (3) In embodiment (3), the one or more inlet ports include one or more gas ports and one or more liquid ports.

[0211] (4) In embodiment (4), each reaction chamber is configured to withstand an atmospheric pressure of at least 10 bar.

[0212] (5) In embodiment (5), the magnetic field within the reaction chamber induced by the coil is between 0 and 200 millitesla.

[0213] (6) In embodiment (6), the one or more temperature control devices include non-magnetic heating and / or cooling elements configured to provide temperature cycling of the reaction chamber.

[0214] (7) In embodiment (7), the one or more temperature control devices provide cooling with nitrogen gas by evaporation of liquid nitrogen and heating with convection coils.

[0215] (8) In embodiment (8), the one or more temperature control devices are configured to provide temperature cycling or maintain a temperature of each reaction chamber between -25°C and 100°C.

[0216] (9) In embodiment (9), the one or more reaction chambers are equipped to perform hyperpolarization on a reaction mixture that includes a perfluorinated SABRE catalyst, a solvent, and a substrate to be hyperpolarized into a hyperpolarized MRI probe.

[0217] (10) In embodiment (10), the solvent is a one- or two-phase system comprising water, methanol, ethanol, a fluorous solvent, or a mixture thereof.

[0218] (11) In embodiment (11), the one or more MRI probe separators are configured to separate the hyperpolarized MRI probe from the perfluorinated SABRE catalyst by one of: filtration; extraction; or column chromatography.

[0219] (12) In embodiment (12), the MRI probe injection device further comprises a gas trap, a gas leak detector, and / or an oxygen level monitor.

[0220] (13) In embodiment (13), the MRI probe injection device further includes a processor configured to execute instructions that cause the processor to: control a flow rate of gas and / or liquid through the device, monitor safety indicators of the device and / or environment, administer a desired amount of hyperpolarized MRI probe to a patient, or calculate a decay rate of the hyperpolarized MRI probe as a function of the flow rate of the gas and / or liquid.

[0221] (14) In embodiment (14), the one or more MRI probe concentrators include one or more dryers.

[0222] (15) In embodiment (15), the one or more dryers include an evaporative dryer that forms one or more azeotropes with the inert solvent, resulting in a lower temperature of the evaporative dryer.

[0223] (16) In embodiment (16), the one or more reaction chambers include at least two reaction chambers configured to be operable in series or parallel.

[0224] (17) In embodiment (17), the components of the MRI probe injection device are made of a non-magnetic material or plastic.

[0225] (18) In embodiment (18), there is provided a method of administering a hyperpolarized MRI probe to a patient in need thereof, the method comprising: (i) providing an MRI probe injection device; (ii) providing a reaction mixture comprising a perfluorinated SABRE catalyst comprising a d-block element and a perfluorinated ligand, a solvent, a co-ligand, and a substrate to be hyperpolarized by an MRI probe into one or more reaction chambers; (iii) agitating the reaction mixture, the agitation being provided by bubbling parahydrogen gas or a mixture of parahydrogen and nitrogen gas through the reaction mixture; (iv) applying a magnetic field suitable for hyperpolarizing the perfluorinated SABRE catalyst and the substrate, hyperpolarizing the substrate onto the hyperpolarized MRI probe; (v) separating the hyperpolarized MRI probe from the reaction mixture by at least one of filtration, extraction, or column chromatography to obtain a solution containing the hyperpolarized MRI probe; (vi) concentrating the hyperpolarized MRI probe present in the solution obtained in step (v) to obtain a concentrate and reconstituting the concentrate into a solution of the hyperpolarized MRI probe of a desired concentration for administration to a patient; (vii) analyzing at least one of the purity or concentration of the hyperpolarized MRI probe present in the solution; and (viii) administering a hyperpolarized MRI probe to the patient.

[0226] (19) In embodiment (19), the solvent comprises water, methanol, ethanol, a fluorous solvent, or a mixture thereof.

[0227] (20) In embodiment (20), the solvent is selected from perfluorohexane / diethyl ether mixtures, mixtures of methoxynonafluorobutane and ethyl acetate with a non-polar solvent, mixtures of perfluorohexane and ether, mixtures of perfluorobutyl methyl ether and ethyl acetate, ether, fluorocarbon derivatives of THF FC 75, decafluoromethoxytrifluoromethylpentane, hexafluoropropanol, nonafluorobutyl methyl ether, perfluoromethylcyclohexane, perfluoroalkanes, perfluorohexane, and methoxynonafluorobutane.

[0228] (21) In embodiment (21), the substrate is 1 H, 13 C. 15 N, 19 F, 31 P, 29 Si, or combinations thereof.

[0229] (22) In embodiment (22), the substrate further comprises 2 Includes D.

[0230] (23) In embodiment (23), the substrate is 1- 13 C-Ketoglutaric acid, 1- 13 C-5- 12 C-Ketoglutaric acid, 1- 13 C-pyruvic acid, 1- 13 CN-acetylcysteine, 15 N2-isoniazid (or pyridyl-4-carbobis- 15 N2-hydrazide), 13 C2, 15 N3-metronidazole, 15 N2-1-aminoisoquinoline (1-AIQ), deuterated versions thereof and salts thereof.

[0231] (24) In embodiment (24), the substrate has the formula (II):

[0232] [ka]

[0233] wherein each R1 is independently selected from hydrogen, deuterium, a cation, C1-C6 alkyl, C3-C7 cycloalkyl, (C3-C7 cycloalkyl)C1-C6 alkyl, (heterocycloalkyl)C1-C6 alkyl, (heteroaryl)C1-C6 alkyl, and (aryl)C1-C6 alkyl; and Xa, Xb, Xc, and Xd are each independently hydrogen or deuterium, with the proviso that at least one of Xa, Xb, Xc, and Xd is deuterium, or a pharma- ceutically acceptable salt thereof.

[0234] (25) In embodiment (25), the perfluorinated ligand is represented by the formula (I): [L m -(NHC)-(YZ) qwherein each L is independently selected from hydrogen, adamantyl, a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; NHC is a 4- to 7-membered N-heterocyclic carbenyi group, which is attached to a d-block element via a carbene; each Y is independently selected from a bond or a spacer group; each Z is a perfluorinated tag; m is an integer from 1 to 4; and q is an integer from 1 to 3, or a salt thereof.

[0235] (26) In embodiment (26), NHC is a 5-membered N-heterocyclic carbene group.

[0236] (27) In embodiment (27), the five-membered N-heterocyclic carbene group is an imidazole-based, imidazoline-based, or thiazole-based.

[0237] (28) In embodiment (28), the NHC is a 4,5-disubstituted, 1,3-disubstituted, or 1,3,4,5-tetrasubstituted imidazole- or imidazoline-based five-membered N-heterocyclic carbene group.

[0238] (29) In embodiment (29), the NHC is a 4,5-disubstituted imidazolidinyl, a 1,3-disubstituted imidazolidinyl, a 1,3,4,5-tetrasubstituted imidazolidinyl, a 4,5-disubstituted 2,3-dihydro-imidazolyl, a 1,3-disubstituted 2,3-dihydro-imidazolyl, or a 1,3,4,5-tetrasubstituted 2,3-dihydro-imidazolyl.

[0239] (30) In embodiment (30), the perfluorinated ligand has the formula (Ia) or (Ib):

[0240] [ka]

[0241] each L is independently hydrogen, adamantyl, a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; each Y is independently a bond or a spacer group; each Z is independently a perfluorinated tag;

[0242] [ka]

[0243] is a single or double bond, and

[0244] [ka]

[0245] represents a bond to a d-block element via a carbene) or a salt thereof.

[0246] (31) In embodiment (31), the perfluorinated ligand has the formula (Ic) or (Id):

[0247] [ka]

[0248] wherein each L is independently hydrogen, adamantyl, a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; a is 4 to 20, b=2a+1 or b=a-1; and each n is independently an integer from 0 to 4;

[0249] [ka]

[0250] is a single or double bond, and

[0251] [ka]

[0252] represents a bond to a d-block element via a carbene) or a salt thereof.

[0253] (32) In embodiment (32), the perfluorinated ligand has the formula (Ie) or (If):

[0254] [ka]

[0255] wherein each L is independently hydrogen, adamantyl, a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; each Ar is independently a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; each G is independently a bond, C 1-6 Alkyl, C 1-6 Alkenyl or C 1-6 heteroalkyl, a is 4 to 20, and b=2a+1 or b=a-1;

[0256] [ka]

[0257] is a single or double bond, and

[0258] [ka]

[0259] represents a bond to a d-block element via a carbene) or a salt thereof.

[0260] (33) In embodiment (33), each L is independently hydrogen, adamantyl, 2-methylphenyl, 3-methylphenyl, 4-methylphenyl, 2,4-dimethylphenyl, 2,5-dimethylphenyl, 2,6-dimethylphenyl, 3,5-dimethylphenyl, 2,4,6-trimethylphenyl, 2-ethylphenyl, 3-ethylphenyl, 4-ethylphenyl, 2,4-diethylphenyl, 2,5-diethylphenyl, 2,6-diethylphenyl, 3,5-diethylphenyl, 2,4,6-triethylphenyl, 2-npropylphenyl, 3-n Propylphenyl, 4-npropylphenyl, 2,4-di-npropylphenyl, 2,5-di-npropylphenyl, 2,6-di-npropylphenyl, 3,5-di-npropylphenyl, 2,4,6-tri-npropylphenyl, 2-isopropylphenyl, 3-isopropylphenyl, 4-isopropylphenyl, 2,4-di-isopropylphenyl, 2,5-di-isopropylphenyl, 2,6-di-isopropylphenyl, 3,5-di-isopropylphenyl, 2,4,6-tri-isopropylphenyl, 2-isobutylphenyl, 3-isobutylphenyl, 4-isobutylphenyl, 2,4-di-isobutylphenyl, 2,5-di-isobutylphenyl, 2,6-di-isobutylphenyl, 3,5-di-isobutylphenyl, 2,4,6-tri-isobutylphenyl, 2-sec-butylphenyl, 3-sec-butylphenyl, 4-sec-butylphenyl, 2,4-di-sec-butylphenyl, 2,5-di-sec-butylphenyl, 2,6-di-sec-butylphenyl, 3,5-di-sec-butylphenyl, 2,4,6-tri-sec-butylphenyl, 2-t-butylphenyl tert-butylphenyl, 3-tert-butylphenyl, 4-tert-butylphenyl, 2,4-di-tert-butylphenyl, 2,5-di-tert-butylphenyl, 2,6-di-tert-butylphenyl, 3,5-di-tert-butylphenyl, 2,4,6-tri-tert-butylphenyl, 2-cyclohexylphenyl, 3-cyclohexylphenyl, 4-cyclohexylphenyl, 2,4-di-cyclohexylphenyl, 2,5-di-cyclohexylphenyl, 2,6-di-cyclohexylphenyl, 3,5-di-cyclohexylphenyl, or 2,4,6-tri-cyclohexylphenyl.

[0261] (34) In embodiment (34), each L is independently hydrogen or 2,4,6-trimethylphenyl.

[0262] (35) In embodiment (35), each Y is independently a bond, a substituted or unsubstituted C 1-10 Alkyl groups, substituted or unsubstituted C 2-10 Alkenyl groups, substituted or unsubstituted C 2-10 Alkynyl group, substituted or unsubstituted C 1-10 Heteroalkyl groups, substituted or unsubstituted C 3-6 Cycloalkyl groups, substituted or unsubstituted C 3-6 It is a heterocycloalkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, a substituted or unsubstituted alkaryl group, a substituted or unsubstituted arylalkyl group, or a straight-chain or branched alkyleneoxy group.

[0263] (36) In embodiment (36), the perfluorinated tag is a perfluorinated C 3-60 It is based on

[0264] (37) In embodiment (37), the perfluorinated tag is a perfluorinated C 3-40 It is based on

[0265] (38) In embodiment (38), the perfluorinated tag is perfluorinated C 3-20 It is based on

[0266] (39) In embodiment (39), the perfluorinated tag is a C4F9 group, C5F 11 Base, C6F 13 Base, C7F 15 Base, C8F 17 Base, C9F 19 group, C 10 F 21 group, C6F5 group, C4F7 group, C5F9 group, C6F 11 Base, C7F 13 Base, C8F 15 Base, C9F 17 Groups and C 10 F19 is selected from the group.

[0267] (40) In embodiment (40), the perfluorinated ligand is:

[0268] [ka]

[0269] (In the formula,

[0270] [ka]

[0271] is a single or double bond, and

[0272] [ka]

[0273] represents a bond to a d-block element via a carbene) or one of its salts.

[0274] (41) In embodiment (41), the d-block element is a transition metal.

[0275] (42) In embodiment (42), the d-block element is one of Co, Rh, Ir, Ru, Pd, Pt, or Mt.

[0276] (43) In embodiment (43), the method further comprises using the hyperpolarized MRI probe to diagnose a stage of a disease or monitor a progress of treatment of a patient having the disease.

[0277] (44) In embodiment (44), the disease is cancer or an adverse vascular condition.

[0278] (45) In embodiment (45), the cancer is selected from breast cancer, colon cancer, rectal cancer, bladder cancer, endometrial cancer, renal cancer, lung cancer, melanoma, non-Hodgkin's lymphoma, pancreatic cancer, prostate cancer, and thyroid cancer.

[0279] (46) In embodiment (46), the adverse vascular condition is selected from myocardial infarction, stroke, and pulmonary disease.

[0280] (47) In embodiment (47), the pulmonary disease is selected from COPD, pulmonary fibrosis, long-term COVID-19 symptoms, or a combination thereof.

[0281] The following examples further illustrate various aspects of the disclosure but, of course, should not be construed as limiting its scope. EXAMPLES

[0282] Example 1 This example illustrates 1H,1H,2H,2H-perfluorooctyl-N,N'-bis(2,4,6-trimethylphenyl)-9,10-diamine:

[0283] [ka]

[0284] (x=6 and y=13)

[0285] A 1.7 M solution of tert-butyllithium in pentane (5 mL, 9 mmol, 8 equiv.) was added to a solution of 1H,1H,2H,2H-perfluorooctyl iodide (2 g, 4 mmol, 4 equiv.) in dry Et2O (60 mL) at -78 °C. The mixture was stirred at -78 °C for 20 min, after which solid N,N'-dimesitylethanediimine (0.30 g, 1.03 mmol, 1 equiv.) was added portionwise. The reaction mixture was stirred for 4 h. The reaction was slowly warmed to -30 °C and quenched with a saturated solution of ammonium chloride (0.6 mL).

[0286] Water (20 mL) was added, the organic layer was separated, and the aqueous layer was extracted with diethyl ether (3×15 mL). The combined organic layers were dried over anhydrous MgSO4 and concentrated. The crude products were purified by flash column chromatography (4:1 hexane / dichloromethane (DCM)) to give the diamines, diimines, and threodiamines (0.6 g, 60% yield, white solid). 1 H NMR (400 MHz, CDCl3) δ 1.64-1.82 (m, 2H, CH2CHHCH(NHAr)), 2.00 (s, 12H, o-CH3), 2.07-2.27 (m, 4H,CHHCHHCH(NHAr)), 2.33 (s, 6H, p-CH3), 2.67-2.91 (m, 2H, CHHCH2CH(NHAr)), 2.95 (br s, 2H, NH), 3.23 (br d, 3 J H-H = 10 Hz, 2H, CH ), 6.82 (s, 4H, Ar-CH ) ppm. 19 F NMR (282.23 MHz, CDCl3) δ -83.8 (t, 3 J F-F =10 Hz, 6F, CF3), -114.4 (m, 4F, CF2CH2), -121.9 (m, 4F, CF2CF2CH2), -122.8 (m, 4F, CF3CF2CF2CF2), -123.4 (m, 4F, CF3CF2CF2), -126.1 (m, 4F,CF3CF2) ppm. 13 C NMR (400 MHz, CDCl3) δ 18.3 (o -CH3), 20.3 (p -CH3), 21.7 (m, CF2CH2CH2), 29.2 (t, 2 J F-C = 22.2 Hz, CF2CH2), 57.1 (CH), 107-115 (m, 8 CF2), 117.4 (qt, 1 J F-C = 288.3 Hz, 2 J F-C = 33.3 Hz, CF3), 118.7 (tt, 1 JF-C = 255.1 Hz, 2MS (ESI), m / z (%): 990 [M+H] + (100).

[0287] Example 2 This example illustrates the synthesis of trans-4,5-bis(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)-1,3-bis(2,4,6-trimethylphenyl)-4,5-dihydroimidazolium tetrafluoroborate:

[0288] [ka]

[0289] The synthesis of

[0290] A mixture of 1H,1H,2H,2H-perfluorooctyl-N,N'-bis(2,4,6-trimethylphenyl)-9,10-diamine from Example 1 (0.250 g, 0.25 mmol), ammonium tetrafluoroborate (~10% molar excess) (0.030 g, 0.25 mmol), and triethyl orthoformate (0.5 mL) was heated to 125 °C and stirred for 15 h. After cooling to room temperature, the solution was evaporated and the solid was triturated with diethyl ether (6 x 3 mL). The residue was redissolved in acetone, filtered, and concentrated to give dihydroimidazolium tetrafluoroborate (0.2 g, 70% yield, yellowish solid). 1 H NMR (400 MHz, acetone-d6) δ 2.15-2.70 (m, 8H, CF2CH2CH2), 2.34 (s, 6H, p-CH3), 2.51 (s, 6H, o-CH3), 2.94 (br s, 2H, NH), 5.16 (m, 2H, CH2CH2CH), 7.17 (s, 4H, Ar-CH), 9.06 (s, 1H, N-CH=N) ppm. 19F NMR (400 MHz, acetone-d6) δ -81.6 (t, 4 J F-F =10 Hz, 6F, CF3), -115.1 (m, 4F, CF2CF2CH2), -122.5 (m, 4F, CF2CF2CH2), -123.6 (m, 4F, CF3CF2CF2CF2), -124.5 (m, 4F, CF3CF2CF2), -126.9 (m, 4F, CF3CF2CF2), -150.9 (4F, BF4) ppm. 13 C NMR (400 MHz, acetone-d6) δ 17.4 (o -CH3), δ 17.7 (o -CH3), 19.9 (p -CH3), 24.4 (m, CF2CH2CH2), 26.2 (t, 2 J F-C = 22.2 Hz, CF2CH2), 67.9 (CH), 105-118 (m, 8 CF2), 118 (qt, 1 J F-C = 288.3 Hz, 2 J F-C = 33.3 Hz, CF3), 118.9 (tt, 1 JF-C = 254 Hz, 2 MS (ESI) m / z (%): 999 [M+H] + (100).

[0291] Example 3 This example illustrates the synthesis of trans-4,5-bis(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluoroctyl)-1,3 bis(2,4,6-trimethylphenyl)-4,5-dihydroimidazolium chloride:

[0292] [ka]

[0293] The synthesis of

[0294] Dihydroimidazolidium tetrafluoroborate salt (0.5 g, 0.46 mmol) was dissolved in MeOH (1.5 mL) and passed through a short column of Amberlite 400 ion exchange resin. The column was washed with MeOH until no spots were visible on TLC under UV. The solvent was removed and the resulting yellowish solid was dried under vacuum pump to give the product (0.48 g, 99%). 1 H NMR (400 MHz, acetone-d6) δ 2.15-2.70 (m, 8H, CF2CH2CH2), 2.34 (s, 6H, p-CH3), 2.51 (s, 6H, o-CH3), 2.94 (br s, 2H, NH), 5.16 (m, 2H, CH2CH2CH), 7.17 (s, 4H, Ar-CH), 9.06 (s, 1H, N-CH=N) ppm. 19 F NMR (400 MHz, acetone-d6) δ -81.8 (t, 4 J F-F =10 Hz, 6F, CF3), -115.1 (m, 4F, CF2CF2CH2), -122.5 (m, 4F, CF2CF2CH2), -123.6 (m, 4F, CF3CF2CF2CF2), -124.5 (m, 4F, CF3CF2CF2), -127 (m, 4F,CF3CF2CF2) ppm. 13 C NMR (400 MHz, acetone-d6) δ 17.4 (o -CH3), δ 17.7 (o -CH3), 19.9 (p -CH3), 24.4 (m, CF2CH2CH2), 26.2 (t, 2 J F-C = 22.2 Hz, CF2CH2), 67.9 (CH), 105-118 (m, 8 CF2), 118 (qt, 1 J F-C = 288.3 Hz, 2 J F-C= 33.3 Hz, CF3), 118.9 (tt, 1 JF-C = 254 Hz, 2 MS (ESI) m / z (%): 999 [M+H] + (100).

[0295] Example 4 This example illustrates a method for the synthesis of a transition metal-containing fluorinated SABRE catalyst in accordance with one embodiment of the present invention.

[0296] [ka]

[0297] In a glove box at room temperature, potassium tert-butoxide (112 mg, 1.00 mmol, 2.5 eq.) was added to a stirred solution of trans-4,5-bis(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)-1,3-bis(2,4,6-trimethylphenyl)-4,5-dihydroimidazolium chloride (320 mg, 0.88 mmol, 2.2 eq.) in tetrahydrofuran (10 mL) from Example 3. The resulting suspension was stirred for 30 min. A solution of [Ir(COD)Cl]2 (268 mg, 0.40 mmol, 1.0 eq.) was added and the resulting solution was stirred at room temperature overnight (Cowley et al., J. Am. Chem. Soc., 133, 6134-6137 (2011)). The solvent was removed under reduced pressure to give the crude product, which was dried under vacuum overnight. The sample was dissolved in hexane and added to a 60 mL filter packed with SiO2 gel in hexane. The crude solution was absorbed on top of the silica gel, and then hexane was added to elute the compound, yielding 104 mg (40% yield). 1H NMR (400 MHz, CDCl3) δ 1.04, 1.18, 1.49, 1.76, 2.13, 2.22, 2.25, 2.4, 2.48, 2.66, 2.94, 3.73, 4.01, 4.2, 6.84, 6.91 6.97 ppm. 19 F NMR (400 MHz, CDCl3) δ-81.08, -114.33, -122.05, -123.07, -124.02, -126.39 ppm. 13 C NMR (100 MHz, CDCl3) δ 18.7, 20.95, 21.07, 21.34, 26.45, 27.21, 29.83, 30.74, 31.42, 49.93, 55.12, 68.11, 68.47, 83.26, 86.69, 108.32, 111.0, 112.95, 115.85, 117.75, 118.69, 129.02, 129.3, 130.36, 130.55, 134.71, 134.8, 135.32, 136.33, 137.92, 138.28, 138.45, 138.5, 206.72 ppm. MS (ESI) m / z (%): 1299 [M+H] + (100).

[0298] Example 5 This example is in accordance with one embodiment of the disclosure, as shown in Scheme 1, 13 C] A method for hyperpolarizing pyruvate is shown. Scheme 1. [1- 13 C] Hyperpolarization of pyruvate.

[0299] [ka]

[0300] (Wherein, co-ligand = phenyltrifluoromethylsulfoxide.)

[0301] In the above reaction scheme, the co-ligand approach developed by Duckett and co-workers (Iali et al., Angew. Chemie - Int. Ed., 58, 10271-10275 (2019)) was used to obtain [1- 13 Hyperpolarization of [C]pyruvate is 13 The alignment was performed using the SHield Enabled Alignment Transfer to Heteronuclei SABRE (SABRE-SHEATH) (Theis et al., J. Am. Chem. Soc., 137, 1404-1407 (2015) and Truong et al., J. Phys. Chem. C, 119, 8786-8797 (2015)) adjusted for the C nucleus (Barskiy et al., ChemPhysChem, 18, 1493-1498 (2017)). 13 [C]-sodium pyruvate and deuterated methanol-d4 solvent were purchased from Sigma-Aldrich and used without further purification. The [IrCl(COD)(F-IMes)]SABRE catalyst used in this example was prepared according to Example 4. The active catalyst used herein was prepared in 0.6 mL of methanol-d4 in a 5 mm NMR tube with a fixed ratio of substrate, Ir(F-IMes)SABRE catalyst from Example 4, and phenyl trifluoromethyl sulfoxide (PTFSO).

[0302] Parahydrogen was generated using a Gas-Delivery Manifold. Ultra-high purity hydrogen gas (Airgas) was delivered to a parahydrogen flow cryostat (Xeus technology LTD) to concentrate parahydrogen to approximately 50% at liquid nitrogen temperature (77 K) in the presence of a spin-exchange catalyst (Fe2O3). The p-H2 flow rate was delivered via PTFE tubing to a mass flow controller (MFC, Sierra Instruments SmartTrak 100 series) set at 90 scc / m, which was then delivered to a conventional 5 mm NMR tube (Norell) and bubbled through the sample. The entire pH2 line was pressurized to 100 psi.

[0303] The magnetic shielding conditions were as follows: Fields near or below ~1μT were achieved using an apparatus consisting of a solenoid coil mounted within a mu-metal shield (Magnetic Shield Corporation, model No. ZG-206). The shield was demagnetized when required using an internal home-made coil driven by a Variac. The solenoid was 41mm in diameter (40mm core), had a 20cm long winding of 220 turns of AWG20 (0.9mm) copper wire and a 220Ω resistor in series. The solenoid coil was powered by a commercial 1.5V battery and had a variable resistance decade box in series to give finer control of the internal magnetic field inside the shield. Typical values ​​of the magnetic field within the shield were between ±1.2μT, with the SABRE SHEATH experiments typically ranging between -0.7μT and +0.8μT in the sample area. This value was monitored during the SABRE experiment using a Lakeshore Cryotronics Gaussmeter (Model No. 475 DSP with HMMA-2512-VR Hall Probe).

[0304] MR experiments were performed using a 1T Magritek Spinsolve benchtop NMR spectrometer. All 13 C NMR spectra were obtained during the experiment. 1 Acquisition was performed with H decoupling turned off. For low-field NMR acquisitions, the time required to manually transfer the sample from the shielded region into the magnet was typically less than 5 s.

[0305] From the newly generated p-H2-derived hydride [1- 13 C]pyruvic acid 13 Efficient hyperpolarization transfer to C nuclear spins was observed in deuterated methanol using [IrCl(H)2(PTFSO)2(F-IMes)], [1- 13This was achieved by performing SABRE in a sub-microtesla magnetic field using SABRE in SHield Enables Alignment Transfer to Heteronuclei (SABRE-SHEATH) with a solution mixture of [C]pyruvic acid and p-H2. Figure 6 shows the top curve plus a single-scan HPT obtained for the hyperpolarized probe. 13 The lower curve depicts the 4M[1- 13 C]Single-scan thermal polarization from sodium acetate. 13 C signal. The enhancement is ε~9000 and the polarization is P( 13 C) About 1%.

[0306] All experiments were performed using 0.6 mL of CD3OD containing the fluorination catalyst, co-ligand (phenyltrifluoromethylsulfoxide), and [1- 13 The experiment was carried out using a solution containing [1-C]pyruvate. The ratio and concentration can be further adjusted to achieve better enhancement and polarization. The results were obtained using a solution containing 8 mM fluorinated catalyst, 16 mM phenyltrifluoromethylsulfoxide (PTFSO), and 30 mM [1-C]pyruvate in 0.6 mL of CD3OD. 13 C]pyruvic acid. Experiments were performed at room temperature, with a p-H2 flow rate of ~100 scc / m, and a p-H2 overpressure of 96 PSI. Parahydrogen used in this example was obtained from a low-cost 50% p-H2 generator. For each experiment, p-H2 bubbling was performed for ~1 min, the sample was quickly transferred to a 1T NMR spectrometer for detection, and the sample was returned to the mu-metal shield to continue p-H2 bubbling for the next experiment. 13 C signal enhancement is Eq.1:

[0307]

number

[0308] The area under the HP signal curve (AUC) was calculated using 13 C signal thermal signal standard (4M[1- 13Calculated by comparing with sodium acetate (C) where S HP and S REF is HP[1- 13 C]pyruvate and thermal signal criteria [1- 13 C] from acetic acid 13 C signal, C REF and C HP are the thermal signal standards [1- 13 C]acetic acid (4M) and HP[1- 13 C] is the concentration of pyruvate, A REF and A HP is the thermal signal criterion [1- 13 C] acetate and HP[1- 13 C] is the effective cross-sectional area of ​​the NMR tube for the pyruvate sample.

[0309] Example 6 This example is based on one embodiment of the present invention and is shown in Scheme 2, 13 C] A method for hyperpolarizing pyruvate is shown. Scheme 2. The perfluorinated SABRE catalyst of Example 4 was used with dimethylsulfoxide as a co-ligand to prepare [1- 13 C] Hyperpolarization of pyruvate.

[0310] [ka]

[0311] In the above reaction scheme, the co-ligand approach developed by Duckett and co-workers (Iali et al., Angew. Chemie - Int. Ed., 58, 10271-10275 (2019)) was used to prepare [1- 13 Hyperpolarization of [C]pyruvate is 13The alignment was performed using the SHield Enabled Alignment Transfer to Heteronuclei SABRE (SABRE-SHEATH) (Theis et al., J. Am. Chem. Soc., 137, 1404-1407 (2015) and Truong et al., J. Phys. Chem. C, 119, 8786-8797 (2015)) adjusted for the C nucleus (Barskiy et al., ChemPhysChem, 18, 1493-1498 (2017)). 13 Sodium [C]-pyruvate and deuterated methanol-d4 solvent were purchased from Sigma-Aldrich and used without further purification. The [IrCl(COD)(F-IMes)]SABRE catalyst used in this example was prepared according to Example 4. The active catalyst used herein is a catalyst that reacts with the substrate [1- 13 C]pyruvic acid, the Ir(F-IMes) SABRE catalyst from Example 4, and the co-ligand dimethylsulfoxide (DMSO) were prepared in a fixed ratio in 0.6 mL of methanol-d4 in a 5 mm NMR tube.

[0312] Parahydrogen concentrated to approximately 70% to 95% was used and delivered via PTFE tubing to a mass flow controller (MFC, Sierra Instruments SmartTrak 100 series) set at 50 to 120 scc / m and into a 5 mm medium-walled NMR tube (Norell) where it was bubbled through the sample. The entire pH2 line was pressurized to a value between 50 and 100 psi.

[0313] A polarized traveling magnetic field was established as follows. Fields near or below ~1 μT were achieved using an apparatus consisting of a solenoid coil mounted within a three-layer mu-metal shield (6 in. ID & 15 in. in length, part number ZG-206, Magnetic Shield Corp., Bensenville, IL). The magnetic field was generated using a custom-built solenoid coil and a triple independent channel DC power supply (KEITHLEY 2231A-30-3). The solenoid had a 41 mm diameter (40 mm core), 20 cm long winding of 220 turns of AWG20 (0.9 mm) copper wire and a 220 Ω resistor in series. The solenoid coil was driven in series with a variable resistance decade box to provide finer control of the magnetic field inside the shield. Typical in-shield magnetic field values ​​are between ±1.2 μT, and in the SABRE SHEATH experiment they were typically between -0.7 μT and +0.8 μT in the sample area.

[0314] MR experiments were performed using an 80 MHz Magritek Spinsolve benchtop NMR spectrometer. The following acquisition parameters were used: Spectral width (SW) = 5 kHz; Dwell time (DT)=150μs; Number of scans (ns) = 1, receivergain=16; Excitation pulse angle (a) = 90°; 13 C resonance frequency = 20.25232790MHz.

[0315] All 13 CNMR spectra were obtained during the experiment. 1 Acquisition was performed with H decoupling turned off. For low-field NMR acquisitions, the time required to manually transfer the sample from the shielded region into the magnet was typically less than 5 s.

[0316] From p-H2 derived iridium hydride [1- 13 C]pyruvic acid 13Hyperpolarization transfer to C nuclear spins was observed in 0.5 mL of deuterated or non-deuterated methanol using [IrCl(H)2(PTFSO)2(F-IMes)], [1- 13 This was achieved by performing SABRE in a sub-microtesla magnetic field using SABRE in SHield Enables Alignment Transfer to Heteronuclei (SABRE-SHEATH) with a solution mixture of [C]pyruvate, co-ligand (dimethyl sulfoxide) and p-H2.

[0317] 13 C signal enhancement is given by Eq.:

[0318]

number

[0319] The area under the HP signal curve (AUC) was calculated using 13 C signal heat signal standard (4M [1- 13 Calculated by comparing with sodium acetate (C) where S HP and S REF HP [1- 13 C]pyruvate and thermal signal criteria [1- 13 C] from acetic acid 13 C signal, C REF and C HP are the thermal signal standards [1- 13 C]acetic acid (4M) and HP[1- 13 C] is the concentration of pyruvate, A REF and A HP is the thermal signal criterion [1- 13 C] acetate and HP[1- 13 C] is the effective cross-sectional area of ​​the NMR tube for the pyruvate sample. 13 C polarization rate (%P 13 C ) is the signal enhancement (ε 13 C ) at 1.81T 13 C nuclear spin polarization (1.5681*10 -4%) to obtain Equation S2:

[0320]

number

[0321] was calculated according to

[0322] Activation of the fluorinated SABRE catalyst is shown in FIG. 13 C polarization, which takes less than 15 min and is performed by bubbling a flow rate of ~95% p-H2 at 90 standard cubic centimeters per minute (scc / m) at a p-H2 partial pressure of 8 atm, which results in the formation of complex 2, complex 3a, complex 3b, and pyruvate, as depicted in Figure 8, in agreement with the notation introduced by Duckett and coworkers (Iali et al., Angew. Chemie - Int. Ed., 58, 10271-10275 (2019)). Without wishing to be bound by any particular theory, it is believed that complex 3B is the primary SABRE active species.

[0323] Example 7 This example is shown by the change in parahydrogen pressure and flow rate [1- 13 We show the effect of the magnetic field, temperature, and the concentration of the fluorination catalyst and DMSO on the hyperpolarization of [1-C]pyruvic acid. 13 The relaxation dynamics of [C]pyruvic acid were also studied.

[0324] [1- 13 Hyperpolarization of [C]pyruvate was repeated using SABRE in SHield Enables Alignment Transfer to Heteronuclei (SABRE-SHEATH), as described above in Example 6, to study the effects of parahydrogen pressure and flow rate, as well as the effects of magnetic transfer field, temperature, and concentration of fluorination catalyst and DMSO.

[0325] p-H2 parameters such as pressure and flow rate were evaluated, and the polarization rate results are shown in Figures 9A and 9B. 13 NMR samples contained sodium C]pyruvate, 2.6 mM fluorinated SABRE catalyst, and 40 mM dimethyl sulfoxide (DMSO), with a mixed magnetic field of 0.4 μT and a temperature of 0 °C. As can be seen from the results shown in Figures 9A and 9B, as the flow rate and pressure of parahydrogen increased, the polarization ratio also increased.

[0326] Temperatures and magnetic fields in the microtesla range were evaluated, and the 13C polarization level and polarization transfer field at 0°C are shown in Figures 10A and 10B, respectively. 13 NMR samples contained in [C] sodium pyruvate, 2.6 mM fluorinated SABRE catalyst, and 40 mM dimethyl sulfoxide (DMSO), with p-H2 flow and pressure of 70 scc / m and 100 PSI. As is evident from the results shown in Figures 10A and 10B, the best polarization transfer occurs between temperatures of -20°C and ~5°C, and between mixed magnetic fields of 0.3 μT and 0.5 μT. The optimal temperature is -7.24°C, and the optimal mixed magnetic field is 0.4 μT.

[0327] Perfluorinated SABRE catalyst and DMSO concentrations were evaluated at 0° C., magnetic transfer field of 0.4 μT, p-H2 flow rate of 90 scc / m, and p-H2 pressure of 110 PSI, and the polarization ratios are shown in FIG. 11A and FIG. 11B. As can be seen from the results shown in FIG. 11A, as the perfluorinated SABRE catalyst concentration increases, the polarization ratio increases. However, the polarization ratio remains relatively constant at concentrations above 20 mM.

[0328] As is apparent from the results shown in Figures 12A and 12B, [1- 13 The relaxation kinetics of [C]-pyruvate is 13C Total build-up time (T b = 6.6 ± 3.0 s) is substantially shorter than the corresponding T1 value of 16.1 ± 0.9 s, which reduces the P 13CMoreover, the relaxation kinetics at geomagnetic and 1.8 T are nearly identical to that of the non-fluorinated SABRE catalyst, T1 = 28.9 ± 1.6 s and 66.5 ± 7 s, respectively.

[0329] p-H2 and [1- 13 The simultaneous exchange of [C]pyruvate 13 In this regard, Figure 13 shows the P of ~13.48% obtained by comparing the NMR signal intensity with a reference sample. 13C with a signal enhancement of ∼86,500-fold, ε 13 C-Hyperpolarized [1- 13 A representative spectrum of 20 mM [1-C]-pyruvic acid is shown. 13 C] NMR samples contained sodium pyruvate, 2.6 mM fluorinated SABRE catalyst, and 40 mM dimethyl sulfoxide (DMSO), the mixing field was 0.4 μT and the temperature was 0 °C, and the parahydrogen pressure and flow rate were 110 PSI and 90 scc / m, respectively.

[0330] The temperature is the [1- 13 C]pyruvate exchange rate. In a recent study, Adelabu et al. (ChemPhysChem, 23, e202100839 (2022)) showed that the monotonically disappearing free HP resonance at low temperatures is due to the slow exchange rate of complex 3b to the free state. At room temperature (e.g., 22 °C), the [1- 13 C]pyruvate exchanged with the polarization transfer complex more quickly, resulting in hyperpolarization of both the free and bound 3b species at the expected pyruvate:precatalyst ratio. To rapidly release HP-pyruvate from 3b, the HP solution was rapidly warmed and the sample was inserted into the NMR detector (TomHon et al., J. Am. Chem. Soc., 144(1) 282-287 (2022)). Figure 14 shows a variable temperature SABRE-SHEATH experiment using the saturated perfluorinated SABRE catalyst of Example 4. 25 mM [1- 13C] sodium pyruvate, 6 mM perfluorinated SABRE catalyst, and NMR samples contained in 47 mM dimethyl sulfoxide (DMSO) in deuterated methanol, the mixing field was 0.4 μT, and the parahydrogen pressure and flow rate were 110 PSI and 90 scc / m, respectively.

[0331] As Figure 14 shows, HP[1- 13 Even at low temperatures such as −10 °C, where most of the [C]pyruvic acid is in the free state, the rate of exchange of complex 3b to the free state is faster. 13 C By lowering the temperature of SABRE-SHEATH to 0℃, 13C The exchange is more favorable for the “free” HP[1- 13 C] to remain fast enough to preferentially build pyruvate.

[0332] Example 8 This example illustrates the process for the preparation of hyperpolarized [1- 13 1 shows an exemplary method for isolation of [C] sodium pyruvate.

[0333] [1- 13 [C]Sodium pyruvate was hyperpolarized as described in Examples 6 and 7. 13 SABRE in SHield Enables Alignment Transfer to Heteronuclei (SABRE-SHEATH) was used, which was tuned to the C nucleus, with dimethylsulfoxide (DMSO), [IrCl(COD)(F-IMes)] as the co-ligand, and parahydrogen concentrated to approximately 70% to 90% in deuterated methanol-d4 solvent. SABRE samples were prepared using 30 mM [1- 13 The solution was prepared in 0.5 mL CD3OD using [C] sodium pyruvate, 2.6 mM perfluorinated SABRE catalyst from Example 4, and 35 mM dimethyl sulfoxide (DMSO). The parahydrogen flow rate was set at 90 scc / m, pressurized to 8 bar, the mixing field was 0.4 μT, and the temperature was 0 °C.

[0334] After the hyperpolarization procedure was completed, the samples were rapidly removed from the 0.40 μT magnetic field, depressurized, and 20% by volume (125 μL) of DO was added to the solution to precipitate the perfluorinated SABRE catalyst. The resulting mixture was transferred to a 1 mL plastic syringe attached to a Luer-Lok filter (Waters Oasis Prime HLB Plus Light Cartridge (part number: 186008866)) and the aqueous solution was directed through the filter into a 5 mm NMR tube already installed in an adjacent 1.8 T benchtop NMR spectrometer. The entire procedure took approximately 1.15 to 1.30 min, with no significant difference in the HP concentration for any sample. 13 No C signal was observed. The relaxation studies above show that the [1- 13 C]-T1 relaxation times are substantially shorter (e.g., in the Earth's magnetic field, [1- 13 C]-T1=28.9±1.6s). 13 To observe the [C]-pyruvate signal, automation and high-speed solution transfer is required.

[0335] Example 9 This example illustrates the use of hyperpolarized [1- 13 1 shows an exemplary method for isolation of [C] sodium pyruvate.

[0336] The precipitation and redissolution of HP[1-13C]pyruvate took place in the same NMR tube in which hyperpolarization was performed (Schmidt et al., ACS Sensors, 7(11), 3430-3439 (2022)). The SABRE samples were prepared by precipitating and redissolving between 20 and 30 mM [1- 13A solution was prepared in CD3OD using [1-C] sodium pyruvate, 7.5 mM fluorinated SABRE catalyst, and 50 mM dimethyl sulfoxide (DMSO). 100 μL of this solution was transferred to a 5 mm NMR tube and exposed to SABRE-SHEATH hyperpolarization conditions with the same setup and optimized conditions described in Examples 6 and 7. The solution was placed in a three-layer mu metal tube with 3'' ID and 9'' depth to shield the external magnetic field combined with a custom-made solenoid to generate a static magnetic field B0 of 0.4 μT. The NMR tube was pressurized (110 psi, i.e., about 8 bar total pressure) while bubbling p-H2 through the solution at a flow rate of 90 scc / m to activate the catalyst and generate the [1- 13 C] sodium pyruvate solution 13 C hyperpolarized. Activation of the catalyst was carried out at ambient temperature and magnetic field for 15 min. To build up the polarization, the sample was placed in a static magnetic field (typically around 0.4 μT) and a water bath, adjusting the reaction temperature to 0 °C. After polarization, the NMR tube was quickly transferred into a 1.8 T NMR spectrometer and kept at room temperature. Precipitation of pyruvic acid was performed by adding 400 μL of ethyl acetate (EtOAc) to the HP solution after depressurization, redissolving it by adding 300 μL of DO and reconstituting the pyruvic acid in water.

[0337] The NMR spectrum was acquired immediately after reconstitution in water using a 1.8T benchtop NMR and is shown in the top spectrum of Figure 15. Additionally, the bottom spectrum of Figure 15 shows the NMR spectrum of 4M [1- 13 C]Single-scan thermal polarization from sodium acetate. 13 C signal is shown.

[0338] In addition, using a calibration curve of naturally occurring isotopic pyruvate, 13C-pyruvic acid concentrations were determined by LCMS. The aqueous pyruvic acid samples were further analyzed for iridium elemental content by inductively coupled plasma Multi-Element Scan (ICP-MS) after the reconstructed SABRE-SHEATH method. The iridium content was determined to be only about 150 ppb to about 300 ppm.

[0339] Example 10 This example illustrates the synthesis of 2,2,2-trifluoro-N-(4-iodo-2,6-dimethylphenyl)acetamide:

[0340] [ka]

[0341] The synthesis method of

[0342] To a stirred solution of 2,6-diisopropylaniline (4.92 mL, 40.00 mmol, 1.0 equiv) in diethyl ether (50 mL) was added iodine (11.17 g, 44.00 mmol, 1.1 equiv) and saturated aqueous sodium bicarbonate (30 mL). The solution was stirred at room temperature for 3 h and gas evolution was observed. Excess iodine was destroyed by addition of sodium thiosulfate (1.33 g, 8.40 mmol). The phases were separated and the aqueous phase was further extracted with diethyl ether (2×20 mL). The combined organic phase was washed with water (200 mL) and a saturated solution of sodium thiosulfate. The organic phase was evaporated to dryness in vacuo to give the product as a brown oil which slowly became solid under vacuum.

[0343] Hexane was added to dissolve the product, the solution was filtered through Celite and evaporated to dryness in vacuo. No further purification was required. 1 H NMR (400 MHz, CDCl3): δ = 7.14 (s, 2H), 4.45 (s, 2H), 2.04 (s, 6H).

[0344] Example 11 This example illustrates the synthesis of N-(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-tridecafluorooct-1-en-1-yl)phenyl)-2,2,2-trifluoroacetamide:

[0345] [ka]

[0346] The synthesis method of

[0347] In a glove box, palladium(II) acetate (0.16 g, 0.051 equiv, 0.71 mmol), sodium acetate (1.72 g, 1.50 equiv, 21.0 mmol), tricyclohexylphosphane (660 mg, 0.168 equiv, 2.35 mmol), tetrabutylammonium bromide (30 mg, 0.0067 equiv, 93 μmol) and 2,2,2-trifluoro-N-(4-iodo-2,6-dimethylphenyl)acetamide (4 g, 0.8 equiv, 0.01 mol) were added to a Schlenk flask containing 10 mL of dimethylformamide. The Schlenk flask was warmed to 90°C and 3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-Tridecafluorooct-1-ene (4.84 g, 1 equiv, 14.0 mmol) was added to the reaction mixture and further heated to 120°C. The solution was stirred for 19 h. After cooling, the mixture was filtered through Celite and washed with diethyl ether (50 mL). Water (50 mL) and diethyl ether (30 mL) were added, the organic phase was separated and the aqueous phase was extracted with Et2O (3x15 mL). The combined organic layers were dried over anhydrous MgSO4 and concentrated under reduced pressure to give the crude product. Purification by column chromatography (hexane / DCM 4:1) gave the fluoroamide N-(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-tridecafluorooct-1-en-1-yl)phenyl)-2,2,2-trifluoroacetamide which was further crystallized (5 g, 70%, white needles).

[0348] Example 12 This example illustrates the synthesis of N-(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-tridecafluorooctyl)phenyl)-2,2,2-trifluoroacetamide:

[0349] [ka]

[0350] The synthesis of

[0351] To a solution of N-(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-tridecafluorooct-1-en-1-yl)phenyl)-2,2,2-trifluoroacetamide (0.40 g, 0.70 mmol) in ethyl acetate (10 mL) in a glass autoclave was added 10% Pd / C (0.08 g, 0.07 mmol). The autoclave was evacuated and charged with hydrogen gas to 500 kPa and stirred at room temperature for 5 h. The mixture was filtered through Celite and washed with EtOAc (30 mL). The solvent was removed on a vacuum rotary evaporator to give N-(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-tridecafluorooctyl)phenyl)-2,2,2-trifluoroacetamide.

[0352] Example 13 This example illustrates the synthesis of 2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-tridecafluorooctyl)aniline:

[0353] [ka]

[0354] The synthesis of

[0355] To a solution of N-(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-tridecafluorooctyl)phenyl)-2,2,2-trifluoroacetamide (0.14 g, 250 μmol) in n-butanol (1.5 mL) was added sodium hydroxide (0.10 g, 250 mmol) and the reaction mixture was heated to 110° C. for 21 h. After cooling to room temperature, water (4 mL) and ethyl acetate (4 mL) were added, the organic phase was separated and washed with 1 M HCl solution (4 mL), saturated NaHCO3 solution (4 mL) and brine (4 mL). The aqueous phase was neutralized and extracted with ethyl acetate (3×4 mL). The organic layers were combined, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to give 2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-tridecafluorooctyl)aniline (0.50 g, 92%, brown crystals).

[0356] Example 14 This example illustrates the synthesis of N,N'-bis[2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-tridecafluorooctyl)phenyl]ethane-1,2-diimine:

[0357] [ka]

[0358] The synthesis of

[0359] To a solution of 2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-tridecafluorooctyl)aniline (0.5 g, 1 mmol) in ethanol (5 mL) was added a 40% aqueous solution of glyoxal (0.1 mL, 1 mmol) and a catalytic amount of formic acid (few drops). The reaction mixture was stirred at room temperature for 20 h during which a yellow precipitate formed. The solid was filtered and washed with cold ethanol (3X5 mL) to give neat N,N-bis(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-tridecafluorooctyl)phenyl)ethane-1,2-diimine (0.2 g, 20%, yellow powder).

[0360] Example 15 This example illustrates the synthesis of 1,3-bis(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-tridecafluorooctyl)phenyl)-1H-imidazol-3-ium-2-ide, chloride salt:

[0361] [ka]

[0362] The synthesis of

[0363] A flask was charged with N,N-bis(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,10,10,10-tridecafluorooctyl)phenyl)ethane-1,2-diimine (2.40 g, 2.08 mmol) and tetrahydrofuran (80 mL). The mixture was cooled to 0° C. and a suspension of paraformaldehyde (262 mg, 2.91 mmol) and conc. HCl (112 mg, 3.12 mmol) in dioxane (0.78 mL) was added slowly. The mixture was heated to reflux overnight. Purification was achieved by column chromatography. 1H NMR (400 MHz, CDCl3) δ 2.07, 2.55, 2.92, 7.37, 8.28, 9.65. MS (ESI) m / z (%): 1168.8 [M-TFA] + (100).

[0364] Example 16 This example illustrates the synthesis of a fluorinated SABRE catalyst in accordance with one embodiment of the present invention.

[0365] [ka]

[0366] To a stirred solution of 1,3-bis(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10-tridecafluorooctyl)phenyl)-1H-imidazol-3-ium-2-ide chloride (2.2 equiv) in tetrahydrofuran was added potassium tert-butoxide (2.5 eq.) at room temperature in a glove box. The resulting suspension was stirred for 30 min. A solution of [Ir(COD)Cl]2 (1.0 eq.) was added and the resulting solution was stirred at room temperature for 2 h. The solvent was removed under reduced pressure to give the crude product, which was dried under vacuum overnight. The sample was purified by flash chromatography DCM / Hexane (4:1) to give the fluorinated SABRE catalyst. MS (ESI) m / z (%): 1469 [M-Cl] + (100).

[0367] Example 17 This example demonstrates the hyperpolarization of sodium pyruvate using the SABRE catalyst, as shown in Scheme 3, in accordance with one embodiment of the present invention. Scheme 3. Using the perfluorinated SABRE catalyst of Example 16 with dimethylsulfoxide as a co-ligand [1- 13 C] Hyperpolarization of pyruvate.

[0368] [ka]

[0369] In the above reaction scheme, [IrCl(H)2(DMSO)2(F-IMes)], [1- 13 Using a solution mixture of [1-C]pyruvic acid) and p-H2, SABRE in SHield Enables Alignment Transfer to Heteronuclei (SABRE-SHEATH) was performed in a submicrotesla magnetic field to convert p-H2-derived hydrides to [1- 13 C]pyruvic acid 13 Efficient hyperpolarization transfer to the C nuclear spins was achieved.

[0370] The SABRE sample contained 40 mM [1- 13 The SABRE sample was prepared in CD3OD using sodium [C]pyruvate, 6.6 mM of the perfluorinated SABRE catalyst of Example 16, and 50 mM dimethyl sulfoxide (DMSO). The SABRE sample was exposed to SABRE-SHEATH hyperpolarized conditions with the same setup and optimized conditions described in Examples 6 and 7. The NMR tube was pressurized (110 psi, i.e., approximately 8 bar total pressure) while bubbling p-H2 through the solution at a flow rate of 90 scc / m to activate the catalyst and generate the [1- 13 C] sodium pyruvate solution 13 C hyperpolarized. Activation of the catalyst was carried out at ambient temperature and magnetic field for approximately 15 minutes. To build up the polarization, the sample was placed in a magnetic field (typically around 0.4 μT) and a 5°C water bath to adjust the reaction temperature. The spectrum was acquired immediately after manually transferring the sample to a 1.8T benchtop NMR after 5 seconds, and the results are shown in the top spectrum of Figure 16. Additionally, the bottom spectrum of Figure 16 shows the 4M[1- 13 C]Single-scan thermal polarization from sodium acetate. 13 C signal. As is evident from the results shown in Figure 16, the signal enhancement is ε~16900 and the polarization is P( 13 C) about 2.17%.

[0371] Example 18 This example illustrates the synthesis of N-(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-en-1-yl)phenyl)-2,2,2-trifluoroacetamide:

[0372] [ka]

[0373] The synthesis method of

[0374] In a glove box, palladium(II) acetate (0.16 g, 0.051 equiv, 0.71 mmol), sodium acetate (1.72 g, 1.50 equiv, 21.0 mmol), tricyclohexylphosphane (660 mg, 0.168 equiv, 2.35 mmol), tetrabutylammonium bromide (30 mg, 0.0067 equiv, 93 μmol) and 2,2,2-trifluoro-N-(4-iodo-2,6-dimethylphenyl)acetamide (4 g, 0.8 equiv, 0.01 mol) were added to a Schlenk flask containing 10 mL of dimethylformamide. The Schlenk flask was warmed to 90°C and 3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-ene (4.84 g, 1 equiv, 14.0 mmol) was added to the reaction mixture and further heated to 120°C. The solution was stirred for 19 h. After cooling, the mixture was filtered through Celite and washed with diethyl ether (50 mL). Water (50 mL) and diethyl ether (30 mL) were added, the organic phase was separated and the aqueous phase was extracted with Et2O (3x15 mL). The combined organic layers were dried over anhydrous MgSO4 and concentrated under reduced pressure to give the crude product. Purification by column chromatography (hexane / DCM 4:1) gave the fluoroamide N-(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-en-1-yl)phenyl)-2,2,2-trifluoroacetamide which was further crystallized (5 g, 70%, white needles).

[0375] Example 19 This example illustrates the synthesis of 2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)aniline:

[0376] [ka]

[0377] The synthesis of

[0378] To a solution of N-(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-en-1-yl)phenyl)-2,2,2-trifluoroacetamide (0.14 g, 250 μmol) in n-butanol (1.5 mL) was added sodium hydroxide (0.10 g, 250 mmol) and the reaction mixture was heated to 110° C. for 21 h. After cooling to room temperature, water (4 mL) and ethyl acetate (4 mL) were added, the organic phase was separated and washed with 1 M HCl solution (4 mL), saturated NaHCO3 solution (4 mL) and brine (4 mL). The aqueous phase was neutralized and extracted with ethyl acetate (3×4 mL). The organic layers were combined, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to give 2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)aniline (0.50 g, 92%, brown crystals).

[0379] Example 20 This example illustrates the synthesis of N,N'-bis[2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-en-1-yl)phenyl]ethane-1,2-diimine:

[0380] [ka]

[0381] The synthesis of

[0382] To a solution of 2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-en-1-yl)aniline (0.5 g, 1 mmol) in ethanol (5 mL) was added a 40% aqueous solution of glyoxal (0.1 mL, 1 mmol) and a catalytic amount of formic acid (few drops). The reaction mixture was stirred at room temperature for 20 h during which a yellow precipitate formed. The solid was filtered and washed with cold ethanol (3X5 mL) to give neat N,N-bis(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)phenyl)ethane-1,2-diimine (0.2 g, 20%, yellow powder).

[0383] Example 21 This example illustrates the synthesis of 1,3-bis(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-en-1-yl)-1H-imidazol-3-ium-2-ide, chloride salt:

[0384] [ka]

[0385] The synthesis of

[0386] A flask was charged with N,N-bis(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-en-1-yl)phenyl)ethane-1,2-diimine (2.40 g, 2.08 mmol) and tetrahydrofuran (80 mL). The mixture was cooled to 0° C. and a suspension of paraformaldehyde (262 mg, 2.91 mmol) and conc. HCl (112 mg, 3.12 mmol) in dioxane (0.78 mL) was added slowly. The mixture was heated to reflux overnight. Purification was achieved by column chromatography. 1 H NMR (400 MHz, CDCl3) δ 2.27, 6.71, 7.32, 7.36, 7.67, 8.16, 9.59 ppm. 19F NMR (400 MHz, CDCl3) δ-82.43, -112.52, -122. 59, -123.89, -124.17, -127.33 ppm. MS (ESI) m / z (%): 964.8 [M+H] + (100).

[0387] Example 22 This example illustrates the preparation of a fluorinated SABRE catalyst according to one embodiment of the present invention.

[0388] [ka]

[0389] The synthesis of

[0390] To a stirred solution of 1,3-bis(2,6-dimethyl-4-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-en-1-yl)phenyl)-1H-imidazol-3-ium-2-ide chloride (2.2 equiv) in tetrahydrofuran was added potassium tert-butoxide (2.5 eq.) at room temperature in a glove box. The resulting suspension was stirred for 30 min. A solution of [Ir(COD)Cl]2 (1.0 eq.) was added and the resulting solution was stirred at room temperature for 2 h. The solvent was removed under reduced pressure to give the crude product, which was dried under vacuum overnight. The sample was purified by flash chromatography DCM / Hexane (4:1) to give the fluorinated SABRE catalyst. MS (ESI) m / z (%): 1266 [M-Cl] + (100).

[0391] Example 23 This example demonstrates the hyperpolarization of sodium pyruvate using the SABRE catalyst, as shown in Scheme 4, in accordance with one embodiment of the present invention. Scheme 4. Using the perfluorinated SABRE catalyst shown with dimethyl sulfoxide as a co-ligand [1- 13 C] Hyperpolarization of pyruvate.

[0392] [ka]

[0393] In the above reaction scheme, [IrCl(H)2(DMSO)2(F-IMes)], [1- 13 Using a solution mixture of [C]pyruvic acid and p-H2, SABRE in SHield Enables Alignment Transfer to Heteronuclei (SABRE-SHEATH) was performed in a submicrotesla magnetic field to convert p-H2-derived hydrides to [1- 13 C]pyruvic acid 13 Efficient hyperpolarization transfer to the C nuclear spins was achieved.

[0394] The SABRE samples were prepared by adding 20 mM [1- 13 SABRE samples were prepared in CD3OD using sodium [C]pyruvate, 7.6 mM of the perfluorinated SABRE catalyst shown in Scheme 4, and 50 mM dimethyl sulfoxide (DMSO). The SABRE samples were exposed to SABRE-SHEATH hyperpolarized conditions using the same setup and optimized conditions described in Examples 6 and 7. The NMR tube was pressurized (110 psi, i.e., approximately 8 bar total pressure) while bubbling p-H2 through the solution at a flow rate of 90 scc / m to activate the catalyst and catalyze the [1- 13 C] sodium pyruvate solution 13 C hyperpolarized. Activation of the catalyst was carried out at ambient temperature and magnetic field for approximately 15 minutes. To build up the polarization, the sample was placed in a magnetic field (typically around 0.4 μT) and a 5°C water bath to adjust the reaction temperature. The spectrum was acquired immediately after manually transferring the sample to a 1.8T benchtop NMR after 5 seconds, and the results are shown in the top spectrum of Figure 17. Additionally, the bottom spectrum of Figure 17 shows the 4M[1- 13 C]Single-scan thermal polarization from sodium acetate. 13C signal. As is evident from the results shown in Figure 17, the signal enhancement is ε ~ 19000 and the polarization is P( 13 C) Approximately 4.91%.

[0395] Example 24 This example illustrates, in accordance with one embodiment of the present invention, the use of a fluorous mixture instead of deuterated methanol alone to obtain [1- 13 C] A method for hyperpolarizing pyruvate is shown.

[0396] The hyperpolarization procedure of Example 6 was repeated using a mixture of nonafluorobutyl methyl ether (NFBME) and deuterated methanol instead of deuterated methanol alone.

[0397] The activation of the fluorinated SABRE catalyst is shown in FIG. 13 C polarization, which takes less than 25 minutes and is performed by bubbling ~95% p-H at a flow rate of 90 standard cubic centimeters per minute (scc / m) at a p-H partial pressure of 8 atm. NMR samples were prepared by mixing 0.4 μT and 0 °C with approximately 25 mM [1- 13 C] sodium pyruvate, 7.4 mM of the perfluorinated SABRE catalyst of Example 4, and 40 mM dimethyl sulfoxide (DMSO).

[0398] Example 25 This example illustrates the effect of varying the pressure and flow rate of parahydrogen and the magnetic transfer field on the [1- 13 We also show the effect of [1-C]pyruvic acid on hyperpolarization. 13 The relaxation dynamics of [C]pyruvic acid were also studied.

[0399] [1- 13Hyperpolarization of [C]pyruvate was repeated using SABRE in SHield Enables Alignment Transfer to Heteronuclei (SABRE-SHEATH), as described above in Example 24, to study the effects of parahydrogen flow rate, as well as the effect of a magnetic transfer field.

[0400] The p-H2 flow rate was evaluated and the polarization results are shown in Figure 19. The NMR sample contained approximately 22 mM [1- 13 C] sodium pyruvate, 7.4 mM of the perfluorinated SABRE catalyst of Example 4, and 45 mM dimethylsulfoxide (DMSO), with a mixing field of 0.4 μT and a temperature of 0° C. As can be seen from the results shown in FIG 19, as the flow rate of parahydrogen increased, the polarization rate also increased.

[0401] Evaluate the magnetic field in the microtesla range at 0℃ 13 The C polarization transfer field is shown in Figure 20. The NMR sample was prepared by adding 22 mM [1- 13 The mixture contained 7.4 mM of the perfluorinated SABRE catalyst of Example 4, and 46 mM of dimethylsulfoxide (DMSO). The p-H2 flow rate and pressure were 50 scc / m and 110 PSI, respectively. As can be seen from the results shown in Figure 20, the best polarization transfer occurs between a mixing field of 0.3 μT and 0.5 μT. The optimal mixing field is 0.4 μT.

[0402] As is apparent from the results shown in Figures 21A and 21B, [1- 13 The relaxation kinetics of [C]-pyruvate is 13C Total build-up time (T b = 3.0 ± 0.8 s) is substantially shorter than the corresponding T1 value of 11.3 ± 1.3 s, which reduces the P 13CMoreover, the relaxation kinetics at geomagnetic and 1.8 T are nearly identical to that of the non-fluorinated SABRE catalyst, T1 = 9.0 ± 1.9 s and 16.0 ± 1.4 s, respectively.

[0403] p-H2 and [1- 13 The simultaneous exchange of [C]pyruvate 13 In this regard, the upper spectrum of Figure 22 shows the results of the measurement of the reference sample (4M[1- 13 C]Single-scan thermal polarization from sodium acetate. 13 The P of ~6.02% obtained by comparing the NMR signal intensity with that of the C signal (the bottom spectrum in Figure 22). 13C with a signal enhancement of ∼38,600-fold, corresponding to 13 C-Hyperpolarized [1- 13 A representative spectrum of [1-C]-pyruvic acid is shown. The NMR sample was prepared from 23 mM [1-C]-pyruvic acid in 0.2 mL NFBME and 0.2 mL MeOD. 13 C] sodium pyruvate, 7.4 mM of the perfluorinated SABRE catalyst of Example 4, and 46 mM dimethylsulfoxide (DMSO). The mixing field was 0.4 μT and the temperature was 0° C., and the p-H2 flow rate and pressure were 90 ssc / m and 110 PSI, respectively.

[0404] The temperature is the [1- 13 C]pyruvate exchange rate. In a recent study, Adelabu et al. (ChemPhysChem, 23, e202100839 (2022)) showed that the monotonically disappearing free HP resonance at low temperatures is due to a slow exchange rate to the free state of complex 3b. At room temperature (e.g., 22 °C), the [1- 13C]pyruvate exchanged with the polarization transfer complex more quickly, resulting in hyperpolarization of both the free and bound 3b species at the expected pyruvate:precatalyst ratio. To rapidly release HP-pyruvate from 3b, the HP solution was rapidly warmed and the sample was inserted into the NMR detector (TomHon et al., J. Am. Chem. Soc., 144(1) 282-287(2022)). Figure 23 shows a variable temperature SABRE-SHEATH experiment using the saturated perfluorinated SABRE catalyst of Example 4. The NMR sample (in nonafluorobutyl methyl ether (NFBME) and deuterated methanol) was heated to 1000 K with 23 mM [1- 13 C] sodium pyruvate, 7.4 mM of the perfluorinated SABRE catalyst of Example 4, and 46 mM dimethyl sulfoxide (DMSO), with a mixing field of 0.4 μT and parahydrogen pressure and flow rate of 110 PSI and 90 scc / m, respectively.

[0405] As Figure 23 shows, HP[1- 13 Even at low temperatures such as −10 °C, where most of the [C]pyruvic acid is in the free state, the rate of exchange of complex 3b to the free state is faster. 13 C By lowering the temperature of SABRE-SHEATH to 0℃, 13C The exchange is more favorable for the “free” HP[1- 13 C] to remain fast enough to preferentially build pyruvate.

[0406] Example 26 This example demonstrates the use of hyperpolarized [1- 13 1 shows an exemplary method for isolation of [C] sodium pyruvate.

[0407] The hyperpolarization procedure of Example 6 was repeated using a mixture of nonafluorobutyl methyl ether (NFBME) and deuterated methanol instead of deuterated methanol alone. The SABRE sample contained 23 mM [1- 13C] sodium pyruvate, 7.4 mM of the perfluorinated SABRE catalyst of Example 4, and 46 mM dimethyl sulfoxide (DMSO) in 0.2 mL NFBME and 0.2 mL MeOD. The mixing field was 0.4 μT, and the parahydrogen pressure and flow rate were set at 110 PSI and 90 scc / m, respectively.

[0408] After the hyperpolarization procedure was completed, the sample was rapidly removed from the 0.40 μT magnetic field and transferred into the NMR spectrometer at room temperature at 1.8 T. The NMR sample was depressurized and the hyperpolarized [1- 13 400 μL of DO was added to the solution to move the sodium [C]pyruvate into the aqueous phase. The pyruvate signal in the aqueous phase was recovered, but upon separation an emulsion was formed, indicating the presence of both bound and free pyruvate. See Figure 24. In that regard, the upper spectrum in Figure 24 shows a ~10800-fold signal enhancement in ε. 13 C-Hyperpolarized [1- 13 A representative spectrum of [C]-pyruvate is shown, along with a reference sample (i.e., 4M[1- 13 C]Single-scan thermal polarization from sodium acetate. 13 The P value of ~1.68% obtained by comparing the NMR signal intensity with that of the C signal (lower spectrum in Figure 24). 13C Corresponds to.

[0409] [1- 13 The aqueous phase containing sodium [C]pyruvate was removed and tested for iridium content and pyruvate concentration. ICP-MS testing showed an iridium content of 637 ppb, and LCMS showed an iridium content of 1- 13 C] showed that approximately 50–75% of the sodium pyruvate concentration was recovered after filtration.

[0410] The fluorous mixture containing the perfluorinated SABRE catalyst in NFBME and MeOD was treated with additional [1- 13 After removing the aqueous phase, 23 mM [1-C]pyruvate was added to 0.2 mL of CD3OD. 13A solution containing [C] sodium pyruvate and 47 mM dimethyl sulfoxide was added to the fluorous mixture containing the 4-day-old perfluorinated SABRE catalyst. 13 Repeated hyperpolarization of sodium pyruvate [C] resulted in the formation of a [1- 13 The signal enhancement ε of [C]pyruvic acid showed a polarization of ∼3090 times and a P of ∼0.48%, obtained by comparing the NMR signal intensity with a reference sample. 13C Corresponds to [1- 13 Polarization of [C]pyruvic acid was repeatable and showed nearly identical polarization levels, demonstrating that the perfluorinated SABRE catalyst retained activity for at least 4 days after first use.

[0411] Example 27 This example illustrates the synthesis of 1,3-dimesityl-4,5-bis(2-(perfluorophenyl)propyl)-4,5-dihydro-1H-imidazol-3-ium, triflate salt:

[0412] [ka]

[0413] The synthesis of was depicted and prepared according to the synthetic sequence shown in FIG.

[0414] 4-Pentafluorophenylbutanal (3). To a solution of alcohol (2) (4.8 g, 20.0 mmol) in DCM (40 mL) was added Dess-Martin periodinane (16.96 g, 40.0 mmol) portionwise (5 min) at 0° C. with stirring under argon. The reaction was continued until the starting alcohol was consumed (2 h). The resulting solution was concentrated to about 10.0 mL, adsorbed onto 10.0 g of silica, and dried to a free-flowing powder. The silica containing the crude product was applied to a silica column (120.0 g) and eluted with 5% ethyl acetate in hexane to give the product as a colorless oil. Yield: 3.33 g (70%). 1 H NMR (CDCl3): 1H NMR (400 MHz, CDCl3) δ 9.71 (t, J = 1.3 Hz, 1H), 2.69 (tt, J = 7.9, 1.8 Hz, 2H), 2.44 (td, J = 7.3, 1.2 Hz, 2H), 1.86 (p, J = 7.3 Hz, 2H). 13 C NMR (101 MHz, CDCl3) δ 200.89, 42.81, 21.49.

[0415] 1,2,3,4,5-Pentafluoro-6-(4-iodobutyl)benzene (4). An ice-cold solution of alcohol (2) (16.0 g, 66.67 mmol) was stirred in DCM (100 mL) under argon with imidazole (5.89 g, 86.67 mmol) and triphenylphosphine (20.96 g, 80.0 mmol). Iodine (20.32 g, 80.0 mmol) was added portionwise over 15 min at 0° C. with vigorous stirring until a slight yellow color persisted. The reaction mixture was diluted with hexane (300 mL) and filtered. The filtrate was washed with saturated sodium thiosulfate (3×50 mL), water (3×100 mL) and dried over sodium sulfate. The clear solution was filtered, concentrated to an oil and chromatographed on silica gel (220 g). Elution with hexane gave compound (4) as a colorless oil. Yield: 21.0g (90%). 1 H NMR (400 MHz, cdcl3) δ 3.20 (t, J = 6.8 Hz, 1H), 2.73 (tt, J = 7.5, 1.8 Hz, 1H), 1.86 (dq, J = 8.6, 6.8 Hz, 1H), 1.76 - 1.66 (m, 1H). 13 C NMR (101 MHz, CDCL3) δ 37.84, 30.01, 21.27, 5.39.

[0416] Iodo-(4-(pentafluorophenyl)butyl)triphenyl-λ 5-Phosphane (5). A solution of iodobutane (4) (21.0 g, 60.0 mmol) and triphenylphosphine (17.29 g, 66.0 mmol) was refluxed under argon for 24 h. The precipitated solid was filtered, washed with anhydrous ether and dried under high vacuum for 20 h. Yield: 33.78 g (92%). 1 H NMR (400 MHz, CDCl3) δ 7.73 (m, 15H), 3.92, 3.86 (m, 2H), 2.76, 2.74 (m, 2H), 2.07, 2.025 (m, 2H), 1.6 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 135.23, 135.20, 133.75, 133.65, 130.65, 130.53, 118.32, 117.46, 29.43, 29.27, 23.05, 22.55, 21.78, 21.74, 21.66.

[0417] Compound 6. To a solution of phosphonium salt (5) (6.79 g, 11 mmol) in anhydrous THF (40 mL) was added dropwise potassium tert-butoxide in THF (2 M, 12.5 mL, 25.0 mmol) and stirred under argon for 30 min. Aldehyde (3) (2.2 g, 9.24 mmol) in THF (5 mL) was added dropwise at room temperature and stirred for 3 h. The solution was concentrated and purified on flash silica (120 g). Elution with hexane afforded the product as a colorless oil as a mixture of cis / trans isomers (90:10). Yield: 3.2 g (72%). 1 H NMR (400 MHz, CDCl3) δ 5.41 (m, 2H), 2.72, 2.68 (m, 4H), 2.12, 2.10, 2.06 (m, 4H), 1.64 (m, 4H). 13 C NMR (101 MHz, CDCl3) δ 129.46, 29.12, 26.76, 21.94

[0418] Compound 7. Olefin (6) (3.6 g, 8.1 mmol) in acetone (20 mL) and water (0.5 mL) was cooled to 0 °C and dibromamine-T (2.93 g, 8.91 mmol, Org. Biomol. Chem., 8, 1424-1430 (2010)) was added with vigorous stirring. After addition, the solution was allowed to warm to room temperature and stirred until the starting material disappeared (30 min). The solution was quenched with solid sodium thiosulfate (2.0 g) and stirred until the color of bromine completely disappeared. The mixture was concentrated under reduced pressure and the residue was diluted with water (50 mL) and extracted with EtOAc (3x30 mL). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure. The residue was chromatographed on flash silica (80 g) eluting with 10% EtOAc in hexane to give bromohydrin (7) as a colorless oil in a mixture of diastereomers. Yield: 2.41 g (55%). 1 H NMR (400 MHz), CDCl3) δ 4.02 (m, 1H), 3.48 (m, 1H), 2.73 (4H, CH2-C6F5), 1.6 - 2.0 (m, 9H, -CH2- and OH)). 13 C NMR (101 MHz, CDCl3) δ 143.76, 138.35, 129.62, 126.82, 59.74, 57.12, 35.29, 34.50, 27.39, 25.48, 21.74, 21.34.

[0419] Compound 8. Bromohydrin (7) (2.2 g, 4.06 mmol) was dissolved in DCM (10.0 mL) and stirred with 5.0 g of molecular sieves. Dess-Martin reagent (3.44 g, 8.12 mmol) was added and stirred until the starting material was completely consumed (30 min). The entire mixture was adsorbed onto flash silica (20.0 g) and dried to a free-flowing powder. The silica containing the crude product was loaded onto a flash silica (80.0 g) column and eluted with 0 to 20% EtOAc in hexane over 20 min (80.0 mL / min; elution rate). Bromoketone (8) eluted as a colorless syrup with 5% EtOAc in hexane. Yield: 1.82 g (83%). 1H NMR (400 MHz, CDCl3) δ 4.25 (dd, J = 8.3, 6.0 Hz, 1H), 2.94 - 2.81 (m, 1H), 2.80 - 2.68 (m, 4H), 2.67 - 2.52 (m, 1H), 2.09 - 1.86 (m, 4H), 1.86 - 1.59 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 202.53, 52.19, 38.18, 32.31, 26.87, 23.21, 21.62, 21.45.

[0420] Compound 9. To a stirred solution of bromoketone (8) (1.8 g, 3.34 mmol) in anhydrous acetonitrile (10.0 mL) was added sodium bicarbonate (0.56 g, 6.68 mmol). Trimethylanilineformamidine (0.94 g, 3.34 mmol) was then added as a solid to the stirred solution and the reaction mixture was heated to 60° C. with exclusion of moisture. After consumption of the bromoketone (70 h), the reaction mixture was filtered, concentrated under reduced pressure and chromatographed on flash silica (80 g). Elution with 20% DCM in hexane gave the ketoamidine (10) as a colorless syrup. Yield: 1.2 g (49%). 1 H NMR δ (CDCl3) 6.91 9s, 1H), 6.81 (s, 1H), 6.71 (s, 1H), 6.68 (s, 2H), 4.53 (m, 1H), 3.08 (m, 1H), 2.57 (m, 2h), 2.43 (m, 2H), 2.35 (m, 2H), 2.36 (s, 3H), 2.2 (s, 3H), 2.13 (s, 3H), 2.01 (s, 3H), 1.92 (s, 6H), 1.8 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 208.19, 151.90, 138.19, 131.45, 129.47, 128.74, 64.33, 60.38, 41.49, 28.39, 25.50, 22.96, 20.56, 18.44, 18.41, 18.23, 14.18. MS: 739.1 [M+H].

[0421] Compound 11 (triflate salt). To an ice-cold solution of the ketoamidine (0.994 g, 1.35 mmol) in ethanol was added lithium borohydride (2 M in THF, 0.675 mL, 1.35 mmol) and stirred under argon at 0-5 °C for 24 h. The reaction mixture was poured into 50 mL of water and extracted with ethyl acetate (3x50 mL). The combined organic layers were dried (sodium sulfate), filtered and concentrated at room temperature to a paste. The above paste was dissolved in 5.0 mL of anhydrous benzene and cooled to 0 °C. DIPEA (0.523 g, 4.05 mmol) was added and stirred under argon. Then trifluoromethanesulfonic anhydride (1.104 g, 1.34 mmol) was added dropwise and stirring was continued for 1 h. LC / MS showed complete consumption of the peak corresponding to 741. The mixture was adsorbed onto 10.0 g of silica gel and dried to a free-flowing powder. The crude product on silica gel was loaded onto an empty loading column on top of an 80.0 g silica column and chromatographed. Elution with 4% MeOH in DCM gave the product as a brown paste, a mixture of cis / trans isomers. Yield: 0.338 g (33%). 1 H NMR (CDCl3) δ 8.82 (s, 1H), 6.93 (s, 4H), 4.15 (m, 2H), 2.61 (m, 4H), 2.25, 2.23, 2.21 (3S, 18H), 1.77 (m, 4H), 1.41 (m, 4H). 13 C NMR (101 MHz, CDCl3) δ 159.41, 141.00, 135.66, 134.43, 130.60, 129.96, 128.58, 68.76, 66.17, 32.48, 26.91, 24.65, 21.71, 20.97, 18.38, 18.11. MS: 723.0 [M+H].

[0422] All references cited in this specification, including publications, patent applications, and patents, are incorporated by reference herein to the same extent as if each reference was individually and specifically indicated to be incorporated by reference and was set forth in its entirety herein.

[0423] Use of the terms "a," "an," "the," and "at least one," and similar referents in the context of describing the present invention (particularly in the context of the claims below) are to be construed to include both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The term "at least one," when followed by a list of one or more items (e.g., "at least one of A and B"), is to be construed to mean one item selected from the listed items (A or B), or any combination of two or more of the listed items (A and B), unless otherwise indicated herein or clearly contradicted by context. The terms "comprising," "having," "including," and "containing" are to be construed as open-ended terms (i.e., meaning "including, but not limited to"), unless otherwise indicated. The recitation of ranges of values ​​herein is intended only as a shorthand way of referring to each value within the range individually, unless otherwise noted herein, and each value is incorporated herein as if it were individually set forth herein. All methods described herein can be performed in any suitable order, unless otherwise noted herein or clearly contradicted by context. The use of all examples or exemplary language (e.g., "etc.") provided herein is intended only to more clearly illustrate the invention, and does not limit the scope of the invention unless otherwise stated in the claims. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0424] Preferred embodiments of the present invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of these preferred embodiments will become apparent to those skilled in the art upon reading the foregoing description. The inventors expect that such variations will be adopted by those skilled in the art as appropriate, and the inventors do not intend to carry out the invention in ways other than as specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

Claims

1. MRI probe injection device including: (i) one or more reaction chambers, each reaction chamber containing: (a) a structure configured to attenuate a magnetic field within the reaction chamber from an external source, the structure being such that the strength of the magnetic field within the reaction chamber from the external source is below a threshold; (b) one or more inlet ports; (c) a coil configured to generate an electromagnetic field within the reaction chamber; (d) one or more temperature control devices; and (e) one or more exit ports; (ii) one or more MRI probe separators configured to receive a reaction mixture including the perfluorinated SABRE catalyst, the solvent, and the hyperpolarized MRI probe from the one or more reaction chambers and extract the hyperpolarized MRI probe from the reaction mixture; and (iii) one or more MRI probe collectors configured to form a solution containing a desired concentration of hyperpolarized MRI probes.

2. 2. The MRI probe implantation apparatus of claim 1, wherein the structure is a mu-metal shield that attenuates magnetic fields from external sources to have a strength of 10 nT or less in the one or more reaction chambers.

3. The MRI probe injection device of claim 1 , wherein the one or more inlet ports include one or more gas ports and one or more liquid ports.

4. 2. The MRI probe injection device of claim 1, wherein each reaction chamber is configured to withstand an atmospheric pressure of at least 10 bar.

5. 2. The MRI probe injection apparatus of claim 1, wherein the magnetic field within the reaction chamber induced by the coil is between 0 and 200 millitesla.

6. 2. The MRI probe implantation apparatus of claim 1, wherein the one or more temperature control devices include non-magnetic heating and / or cooling elements configured to maintain the temperature of the reaction chamber between -25°C and 100°C.

7. 10. The MRI probe injection device of claim 1, wherein the one or more reaction chambers are equipped to perform hyperpolarization on a reaction mixture comprising a perfluorinated SABRE catalyst, a solvent, and a substrate to be hyperpolarized into the hyperpolarized MRI probe.

8. 8. The MRI probe injection apparatus of claim 7, wherein the solvent is a one-phase or two-phase system comprising water, methanol, ethanol, a fluorous solvent, or a mixture thereof.

9. One or more MRI probe separators include: Filtration; Extraction; or Column Chromatography 2. The MRI probe injection device of claim 1, configured to separate the hyperpolarized MRI probe from the perfluorinated SABRE catalyst by one of:

10. 10. The MRI probe injection device of claim 1, further comprising a gas trap, a gas leak detector, and / or an oxygen level monitor.

11. 10. The MRI probe injection device of claim 1 further comprising a processor configured to execute instructions that cause the processor to: Controlling the flow rate of gas and / or liquid through the device; Monitoring equipment and / or environmental safety indicators; administering a desired amount of a hyperpolarized MRI probe to the patient; or Calculating the decay rate of the hyperpolarized MRI probe as a function of the flow rate of said gas and / or liquid.

12. The MRI probe injection device of claim 1 , wherein the one or more MRI probe collectors include one or more dryers.

13. The MRI probe injection device of claim 1 , wherein the one or more reaction chambers include at least two reaction chambers configured to be operable in series or parallel.

14. 10. The MRI probe injection device of claim 1, wherein the components of the device are made of non-magnetic materials or plastics.

15. 1. A method of administering a hyperpolarized MRI probe to a patient in need thereof, the method comprising: (i) providing an MRI probe injection apparatus according to claim 1; (ii) providing a reaction mixture comprising a perfluorinated SABRE catalyst comprising a d-block element and a perfluorinated ligand, a solvent, a co-ligand, and a substrate to be hyperpolarized by an MRI probe into one or more reaction chambers; (iii) agitating the reaction mixture, the agitation being provided by bubbling parahydrogen gas or a mixture of parahydrogen and nitrogen gas through the reaction mixture; (iv) applying a magnetic field suitable for hyperpolarizing the perfluorinated SABRE catalyst and the substrate to hyperpolarize the substrate into a hyperpolarized MRI probe; (v) separating the hyperpolarized MRI probe from the reaction mixture by at least one of filtration, extraction, or column chromatography to obtain a solution containing the hyperpolarized MRI probe; (vi) concentrating the hyperpolarized MRI probe present in the solution obtained in step (v) to obtain a concentrate and reconstituting the concentrate into a solution of the hyperpolarized MRI probe of the desired concentration for administration to a patient; (vii) analyzing at least one of the purity or concentration of the hyperpolarized MRI probe present in the solution; and (viii) administering a hyperpolarized MRI probe to the patient.

16. 16. The method of claim 15, wherein the solvent is selected from perfluorohexane / diethyl ether mixtures, mixtures of methoxynonafluorobutane and ethyl acetate with a non-polar solvent, mixtures of perfluorohexane and ethers, mixtures of perfluorobutyl methyl ether and ethyl acetate, ethers, fluorocarbon derivatives of THF FC 75, decafluoromethoxytrifluoromethylpentane, hexafluoropropanol, nonafluorobutyl methyl ether, perfluoromethylcyclohexane, perfluoroalkanes, perfluorohexane, and methoxynonafluorobutane.

17. The substrate is 1- １３ C-ketoglutaric acid, 1- １３ C-5- １２ C-ketoglutaric acid, 1- １３ C-pyruvic acid, 1- １３ C-N-acetylcysteine, １５ N ２ -Isoniazid (or pyridyl-4-carb-bis- １５ N ２ -hydrazide), １３ C ２ , １５ N ３ -metronidazole, １５ N ２ 16. The method of claim 15, wherein the compound is selected from the group consisting of 1-aminoisoquinoline (1-AIQ), deuterated versions thereof, and salts thereof.

18. The perfluorinated ligand is represented by the formula (I): [L ｍ -(NHC)-(Y-Z) ｑ or a salt thereof. wherein each L is independently selected from hydrogen, adamantyl, a substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group; NHC is a 4- to 7-membered N-heterocyclic carbenyi group, and NHC is bonded to a d-block element via a carbene; each Y is independently selected from a bond or a spacer group; each Z is a perfluorinated tag; m is an integer from 1 to 4, and q is an integer from 1 to 3.

19. 20. The method of claim 18, wherein the perfluorinated tag is one of the following: Perfluorinated C containing only carbon and fluorine atoms ３－６０ Base; Perfluorinated C containing only carbon and fluorine atoms ３－４０ Group; or Perfluorinated C ３－２０ Base.

20. The method of claim 15, wherein the perfluorinated ligand is selected from one of the following: 【Chemistry 1】 or a salt thereof, wherein: 【Chemistry 2】 is a single or double bond, and 【Chemistry 3】 represents a bond to a d-block element via a carbene.

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