Systems and methods for producing hyperpolarized materials

Abstract

The present disclosure describes hyperpolarized materials for nuclear magnetic resonance, magnetic resonance imaging, or similar applications. The present disclosure describes methods for producing hyperpolarized materials for nuclear magnetic resonance, magnetic resonance imaging, or similar applications. The present disclosure describes precursor compounds for producing hyperpolarized materials for nuclear magnetic resonance, magnetic resonance imaging, or similar applications.

Description

[0001] Cross - reference to related applications

[0002] This application claims the priority of U.S. Provisional Application No. 63 / 375,392, filed on September 13, 2022, entitled "SYSTEMS AND METHODS FOR GENERATION OF HYPERPOLARIZED MATERIALS", which is incorporated herein by reference in its entirety for all purposes. Technical field

[0003] The disclosed embodiments generally relate to the generation and purification of hyperpolarized materials for nuclear magnetic resonance, magnetic resonance imaging, or similar applications. Background art

[0004] Parahydrogen - induced polarization (PHIP) is a method for polarizing metabolites for hyperpolarized (HP) magnetic resonance imaging (MRI), which has low cost and high throughput. Parahydrogen - induced polarization with side - arm hydrogenation (PHIP - SAH) can be used to polarize metabolites such as acetate molecules. However, existing PHIP - SAH polarization methods may not be suitable for pre - clinical or clinical HP MRI applications. Brief description of the drawings

[0005] The drawings, which are a part of this specification, illustrate several embodiments and, together with the description, are used to explain certain principles and features of the disclosed embodiments. Brief description of the drawings

[0006] Figure 1 Depicts a first exemplary process for generating a polarized bio - relevant contrast agent according to various embodiments.

[0007] Figure 2 Depicts a second exemplary process for generating a polarized bio - relevant contrast agent according to various embodiments.

[0008] Figure 3 Depicts a third exemplary process for generating a polarized bio - relevant contrast agent according to various embodiments.

[0009] Figure 4 Depicts a fourth exemplary process for generating a polarized bio - relevant contrast agent according to various embodiments.

[0010] Figure 5 Shows exemplary singlet lifetimes of (Z) - 4 - ((2 - oxopropanoyl)oxy)but - 2 - enoic acid tert - butyl ester, (Z) - 4 - ((2 - oxopropanoyl)oxy)but - 2 - enoic acid tert - butyl ester - D1, and (Z) - 4 - ((2 - oxopropanoyl)oxy)but - 2 - enoic acid tert - butyl ester - D2 according to various embodiments.

[0011] Figure 6A Shows exemplary polarization transfer pulse sequences according to various embodiments, the exemplary polarization transfer pulse sequences being used to transfer protons associated with secondary hydrogens in tert-butyl (Z)-4-((2-oxopropanoyl)oxy)but-2-enoate, tert-butyl (Z)-4-((2-oxopropanoyl)oxy)but-2-enoate-D1, and tert-butyl (Z)-4-((2-oxopropanoyl)oxy)but-2-enoate-D2 to the natural abundance 13 C nuclei present in tert-butyl (Z)-4-((2-oxopropanoyl)oxy)but-2-enoate, tert-butyl (Z)-4-((2-oxopropanoyl)oxy)but-2-enoate-D1, and tert-butyl (Z)-4-((2-oxopropanoyl)oxy)but-2-enoate-D2.

[0012] Figure 6B Shows the use according to various embodiments Figure 6A of various first time periods t in the polarization transfer pulse sequence 扫掠 to achieve exemplary 13 C polarization of tert-butyl (Z)-4-((2-oxopropanoyl)oxy)but-2-enoate-D2.

[0013] Figure 7A Shows according to various embodiments the exemplary Figure 6A and 6B C polarization levels of tert-butyl (Z)-4-((2-oxopropanoyl)oxy)but-2-enoate-D2 after the polarization transfer procedure. 13 C polarization levels.

[0014] Figure 8 Shows exemplary 13 C polarization levels of deuterated secondary hydrogenated ester side arm derivatives of lactate.

[0015] Figure 9 Shows exemplary 13 C polarization levels of deuterated secondary hydrogenated derivatives of ethyl 2-oxoglutarate.

[0016] Figure 10 Shows exemplary 13 C polarization levels of deuterated secondary hydrogenated derivatives of Z-OMPD monomethyl ester. Detailed Description

[0017] Reference will now be made in detail to exemplary embodiments discussed with reference to the drawings. Unless otherwise defined, technical and / or scientific terms have the meaning as commonly understood by one of ordinary skill in the art. The disclosed embodiments are described in sufficient detail to enable those skilled in the art to practice the disclosed embodiments. It is to be understood that other embodiments may be utilized and changes may be made without departing from the scope of the disclosed embodiments. Thus, the materials, methods, and examples are illustrative only and not intended to be necessarily limiting.

[0018] Recent work in the fields of nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI) has demonstrated that, using various so-called hyperpolarization techniques, the NMR and MRI signals associated with various biorelevant contrast agents can be enhanced by many orders of magnitude. This signal enhancement allows for improved spectroscopic analysis of biorelevant contrast agents as they are metabolized by various tissues at different locations within the body. Analysis of the metabolic information determined by such spectroscopic imaging can allow for non-invasive determination of the health state of tissues within the body. For example, abnormal metabolism of a biorelevant contrast agent may signal a disease, such as cancer, at some location in the body.

[0019] Prior art for hyperpolarizing biorelevant contrast agents includes dissolution dynamic nuclear polarization (DNP), parahydrogen-induced polarization (PHIP), PHIP sidearm hydrogenation (PHIP-SAH), and signal amplification by reversible exchange (SABRE). In PHIP and PHIP-SAH, a precursor of the biorelevant contrast agent is reacted with parahydrogen to form a parahydrogenated derivative of the precursor. Then, the spin order is transferred from the protons added via the parahydrogenation reaction to the nucleus of interest (such as a carbon-13 nucleus) contained within the biorelevant contrast agent. In PHIP-SAH, the parahydrogenated derivative of the precursor is cleaved (e.g., hydrolyzed) to produce the hyperpolarized biorelevant contrast agent. The biorelevant contrast agent is then purified and used in NMR or MRI procedures. In PHIP-SAH, the precursor can comprise the biorelevant contrast agent coupled to a sidearm containing at least one unsaturated bond (e.g., at least one carbon-carbon double bond or at least one carbon-carbon triple bond) suitable for reacting with parahydrogen. However, prior precursors have used sidearms that may not allow for the production of biorelevant contrast agents with clinically relevant polarization, concentration, volume, or purity. Such behavior may be related to poor solubility of the precursor in organic solvents (where parahydrogen is highly soluble), poor yields in the reaction between the unsaturated bond and parahydrogen, and poor transfer of the spin order (e.g., from the sidearm to the nucleus of interest) or a variety of other factors. Thus, there is a need for novel PHIP-SAH precursors that produce hyperpolarized biorelevant contrast agents with clinically relevant polarization, concentration, volume, or purity.

[0020] The disclosed embodiments include systems and methods for generating bio - relevant contrast agents at clinically relevant polarizations, concentrations, volumes, and purities. The disclosed embodiments provide technical improvements in polarizing bio - relevant contrast agents in solution. These technical improvements support an increase in the concentration of bio - relevant contrast agents and the degree of bio - relevant polarization.

[0021] Hyperpolarization and parahydrogen

[0022] As used in this disclosure, hyperpolarization describes a condition in which the absolute value of the difference between the spin - state populations (e.g., nuclear spin states, proton spin states, etc.) in one state (e.g., spin - up) and the spin - state populations in another state (e.g., spin - down) exceeds the absolute value of the corresponding difference under thermal equilibrium.

[0023] Consistent with the disclosed embodiments, parahydrogen can be used as a polarization source. As described herein, parahydrogen is a form of molecular hydrogen in which the two proton spins are in a singlet state. The disclosed embodiments are not limited to a particular method of generating parahydrogen. Parahydrogen can be formed in gaseous or liquid form. In some embodiments, gaseous parahydrogen is produced by flowing hydrogen gas at low temperature through a chamber having a catalyst (e.g., iron oxide or another suitable catalyst). The hydrogen gas can contain both parahydrogen and orthohydrogen. The low temperature can bring the hydrogen gas to thermodynamic equilibrium within the chamber, thereby increasing the parahydrogen population.

[0024] The disclosed embodiments are not limited to a particular location for parahydrogen generation or use. Parahydrogen can be generated at a first location and subsequently transported to a second location for use. In some embodiments, the first location is a chamber, which can be part of a container, bottle, holder, or other zone capable of containing a gas or liquid. Such a chamber can be maintained at a suitable pressure or temperature. In some embodiments, the first location is a physical location, such as a room, laboratory, specific storage facility, hospital, or other location where parahydrogen can be generated.

[0025] The disclosed embodiments are not limited to a particular method of transporting parahydrogen. The generated parahydrogen can be transported in a chamber, which can be different from the chamber in which parahydrogen is generated. The chamber for transporting parahydrogen gas can be maintained at a suitable pressure or temperature and can be transported by a vehicle or a person. Transporting parahydrogen can involve moving parahydrogen from one container to a different container. Transporting parahydrogen can involve moving parahydrogen within the same location, such as from one part of a room to another part of the room. Transporting parahydrogen can involve moving parahydrogen from one room in a building to a different room in the same building or to a nearby building. Transporting parahydrogen can involve moving parahydrogen to a different location in another part of the same city or to a different city. Transporting parahydrogen can involve bringing parahydrogen near a polarizer, NMR device, or MRI device. Transporting parahydrogen can involve packaging or transporting parahydrogen in a suitable container.

[0026] In some embodiments, the population difference between two spin states is the difference between the populations of the two spin states divided by the total population of the two spin states. The population difference can be expressed as a fractional population difference or a percentage population difference. In some embodiments, the fractional population difference is at least about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or more, at most about 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1 or less, or within a range defined by any two of the foregoing values.

[0027] Hydrogen can exhibit a population difference between proton spin states that greatly exceeds the population difference between proton spin states at thermal equilibrium. Para-hydrogen can have a large population difference between the singlet spin state and any triplet spin state. In the case of Iz1Iz2, for example, there is a large population difference between the spin states |↑>|↓> and the spin state |↑>|↑>. The population difference of the proton spin states can be at least about 0.1 (e.g., a 10% difference in spin states - 55% of the para-hydrogen molecules in the sample are in the singlet state and 45% are in the triplet state), 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or more, at most about 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1 or less, or within a range defined by any two of the foregoing values.

[0028] Biologically relevant contrast agents

[0029] The disclosed embodiments include systems and methods for generating and utilizing bio-related contrast agents at clinically relevant polarizations, concentrations, volumes, or purities. In some embodiments, the method is for preparing NMR materials. In some embodiments, the NMR materials are suitable for NMR or MRI operations. In some embodiments, the NMR materials increase the NMR or MRI signal and the signal-to-noise ratio (SNR). In some embodiments, the NMR materials are suitable for solution NMR spectroscopy. In some embodiments, the NMR materials are chemical compounds. In some embodiments, the NMR materials are metabolites (e.g., molecules with biological relevance such as amino acids, sugars, their derivatives, etc.), such as metabolites suitable for NMR metabolomics applications. In some embodiments, the NMR materials are suitable for in vitro probing of the metabolism of cell cultures or other biological tissues. In some embodiments, the NMR materials are used in NMR probes to study transient effects where high signal enhancement due to hyperpolarization is required, such as proton exchange between water and biomolecules. In some embodiments, the NMR materials are small molecules or metabolites suitable for injection into cells, tissues, or organisms for detection in MRI scans. In some embodiments, the NMR materials are introduced into a chamber for further analysis by NMR or MRI operations. In some embodiments, the NMR materials are enriched with one or more deuteriums (2 H), or carbon-13 ( 13 C) atoms.

[0030] Consistent with the disclosed embodiments, the NMR material can include a bio-related contrast agent. In some embodiments, the bio-related contrast agent can be suitable for NMR or MRI operations. In some embodiments, the bio-related contrast agent can increase the NMR or MRI signal or signal-to-noise ratio (SNR). In some embodiments, the bio-related contrast agent can be suitable for solution NMR spectroscopy. In some embodiments, the bio-related contrast agent can be a metabolite (e.g., a biologically relevant molecule such as an amino acid, sugar, its derivatives, etc.), such as a metabolite suitable for NMR metabolomics applications. In some embodiments, the bio-related contrast agent is used for perfusion imaging or contrast-enhanced imaging in MRI scans. In some embodiments, the bio-related contrast agent is suitable for in vitro detection of the metabolism of cell cultures or other biological tissues. In some embodiments, the bio-related contrast agent is used for in vitro detection of the metabolism of cell cultures or other biological tissues. In some embodiments, the bio-related contrast agent is used in an NMR probe to study transient effects, where high signal enhancement due to hyperpolarization is required, such as proton exchange between water and biomolecules. In some embodiments, the bio-related contrast agent is a small molecule or metabolite suitable for injection into cells, tissues, or organisms for detection in MRI scans. In some embodiments, the bio-related contrast agent is introduced into a chamber for further analysis by NMR or MRI operations. In some embodiments, the bio-related contrast agent is enriched with one or more 2 H or 13 C atoms.

[0031] In some embodiments, the bio-related contrast agent comprises pyruvate, lactate, α-ketoglutarate, bicarbonate, fumarate, urea, dehydroascorbate, glutamate, glutamine, acetate, dihydroxyacetone, acetoacetate, glucose, ascorbate, ketoisocaproate, alanine, fructose, imidazole, nicotinamide, nitroimidazole, pyrazinamide, isoniazid, the conjugate acid of any of the above natural and unnatural amino acids, its esters, or an 2 H, 13 C or nitrogen-15 ( 15 N)-enriched version. In some embodiments, the bio-related contrast agent comprises pyruvate, lactate, α-ketoglutarate. In some embodiments, the bio-related contrast agent comprises pyruvate. In some embodiments, the bio-related contrast agent comprises lactate. In some embodiments, the bio-related contrast agent comprises α-ketoglutarate (e.g., ethyl α-ketoglutarate).

[0032] In some embodiments, a biorelevant contrast agent comprises at least one non-hydrogen nuclear spin. In some embodiments, the non-hydrogen nucleus comprises at least one spin-1 / 2 atom. In some embodiments, the non-hydrogen nuclear spin comprises 13 C or 15 N. In some embodiments, the biorelevant contrast agent is at least partially isotopically labeled with non-hydrogen nuclear spins. In some embodiments, the biorelevant contrast agent is at least partially enriched in non-hydrogen nuclear spins as compared to an analogue of the biorelevant contrast agent that characterizes non-hydrogen nuclear spins at their natural abundance. In some embodiments, the biorelevant contrast agent is enriched to characterize the non-hydrogen nuclear spins at an abundance of at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, up to about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less, or within a range defined by any two of the foregoing values.

[0033] In some embodiments, the non-hydrogen nuclear spin replaces an NMR-inactive (i.e., spin-0) nucleus (e.g., 12 C or a quadrupolar (i.e., spin > 1 / 2) nucleus) (e.g., nitrogen-14, 14 N) of an analogue of the biorelevant contrast agent that characterizes non-hydrogen nuclear spins at their natural abundance. For example, an analogue of pyruvate that characterizes 13 C may include about 98.9% 12 C and about 1.1% 13 C at any one of the C* in the structure H3C-C*(=O)-C*OOH. As a biorelevant contrast agent, pyruvate may alternatively be enriched with 13 C isotopes such that one or both of the C* contain 13 C at any of the abundances described herein. As used herein, *C and C* describe carbon that may be either 12 C or 13 C carbon isotopes. Again, an analogue of urea that characterizes 15 N may include about 99.6% 14 N and about 0.4% 15N. As a biologically relevant contrast agent, urea can alternatively be enriched with 15 N isotopes such that one or both N* contain 15 N at any abundance described herein. As used herein, *N and N* describe nitrogen that can be 14 N or 15 N nitrogen isotopes.

[0034] Precursors of biologically relevant contrast agents

[0035] In some embodiments, the present disclosure describes a precursor (i.e., a precursor compound) comprising a biologically relevant contrast agent and a side arm. In some embodiments, the biologically relevant contrast agent is covalently attached to the side arm. In some embodiments, the biologically relevant contrast agent is attached to the side arm via a transfer moiety, such as a PHIP transfer moiety, which is part of the side arm.

[0036] In some embodiments, the present disclosure describes a precursor (i.e., a precursor compound) comprising an acyl derivative (i.e., R-C(=O)-) of a biologically relevant contrast agent and a side arm. As used herein, the term "acyl derivative of a biologically relevant contrast agent" refers to a covalently bonded derivative of a biologically relevant contrast agent in which the terminal acid moiety [R-C(=O)OH)] of the unbound biologically relevant contrast agent is altered to an acyl group and a covalent bond [R-C(=O)-)] in the bound biologically relevant contrast agent. In some embodiments, the acyl derivative of the biologically relevant contrast agent is covalently linked to the side arm. In some embodiments, the acyl derivative of the biologically relevant contrast agent is attached to the side arm via a transfer moiety, such as a PHIP transfer moiety, which is part of the side arm.

[0037] The side arm can be parahydrogenated (e.g., by mixing the precursor and parahydrogen). In some embodiments, the hydrogenation produces an Iz1Iz2 level, a lower energy state between |↑>|↓>, |↓>|↑> or singlet spin order on two hydrogen spins, depending on whether the hydrogenation is carried out at low or high magnetic field.

[0038] In some embodiments, the precursor is selected such that after hydrogenation and any other optional chemical reactions, the bio - relevant contrast agent is suitable for hyperpolarized NMR or MRI applications. In some embodiments, additional chemical reactions after hydrogenation can be used to separate the bio - relevant contrast agent from the precursor. Such additional chemical reactions can include, for example, cleaving the side arm of the precursor by hydrolysis. For example, the bio - relevant contrast agent can be a metabolite molecule, such that the precursor can be a derivative of the metabolite molecule, where the derivative has a general chemical structure of formula Ia or formula Ib. The bio - relevant contrast agent can be polarized using the PHIP - SAH method (i.e., secondary hydrogenation of the side arm and subsequent polarization transfer to the bio - relevant contrast agent). After hydrogenation and polarization transfer, the linking bond (e.g., an ester bond) in the precursor can be hydrolyzed to produce the polarized bio - relevant contrast agent and a separate side - arm element.

[0039] As used herein, hydrolysis is defined as the cleavage of a molecule in the presence of added water elements through a nucleophilic substitution reaction. Hydrolysis can also be carried out under anhydrous conditions in the presence of hydroxide ions.

[0040] Consistent with the disclosed embodiments, precursors in the general chemical form presented in formula Ia or formula Ib can be used as precursors for PHIP - SAH. After hydrogenation of such precursors, two 1 H spins with spin order are close (e.g., only three, four, or five bonds) to the target carbon or nitrogen on the metabolite, where the target carbon or nitrogen can be 13 C - enriched or 15 N - enriched as described herein. In some embodiments, achieving 13 C or 15 N spins and the 1High J-couplings between at least one of the H spins. In some embodiments, a J-coupling of at least about 0.1 Hertz (Hz), 0.2 Hz, 0.3 Hz, 0.4 Hz, 0.5 Hz, 0.6 Hz, 0.7 Hz, 0.8 Hz, 0.9 Hz, 1 Hz, 2 Hz, 3 Hz, 4 Hz, 5 Hz, 6 Hz, 7 Hz, 8 Hz, 9 Hz, 10 Hz or more, up to about 10 Hz, 9 Hz, 8 Hz, 7 Hz, 6 Hz, 5 Hz, 4 Hz, 3 Hz, 2 Hz, 1 Hz, 0.9 Hz, 0.8 Hz, 0.7 Hz, 0.6 Hz, 0.5 Hz, 0.4 Hz, 0.3 Hz, 0.2 Hz, 0.1 Hz or less, or a J-coupling within a range defined by any two of the foregoing values is achieved. For example, in some embodiments, the J-coupling is between 1 Hz and 2 Hz, between 1 Hz and 3 Hz, between 1 Hz and 4 Hz, between 1 Hz and 5 Hz, between 1 Hz and 6 Hz, between 1 Hz and 7 Hz, between 1 Hz and 8 Hz, between 1 Hz and 9 Hz, between 1 Hz and 10 Hz, between 2 Hz and 3 Hz, between 2 Hz and 4 Hz, between 2 Hz and 5 Hz, between 2 Hz and 6 Hz, between 2 Hz and 7 Hz, between 2 Hz and 8 Hz, between 2 Hz and 9 Hz, between 2 Hz and 10 Hz, between 3 Hz and 4 Hz, between 3 Hz and 5 Hz, between 3 Hz and 6 Hz, between 3 Hz and 7 Hz, between 3 Hz and 8 Hz, between 3 Hz and 9 Hz, between 3 Hz and 10 Hz, between 4 Hz and 5 Hz, between 4 Hz and 6 Hz, between 4 Hz and 7 Hz, between 4 Hz and 8 Hz, between 4 Hz and 9 Hz, between 4 Hz and 10 Hz, between 5 Hz and 6 Hz, between 5 Hz and 7 Hz, between 5 Hz and 8 Hz, between 5 Hz and 9 Hz, between 5 Hz and 10 Hz, between 6 Hz and 7 Hz, between 6 Hz and 8 Hz, between 6 Hz and 9 Hz, between 6 Hz and 10 Hz, between 7 Hz and 8 Hz, between 7 Hz and 9 Hz, between 7 Hz and 10 Hz, between 8 Hz and 9 Hz, between 8 Hz and 10 Hz or between 9 Hz and 10 Hz. Such J-couplings can achieve 13 Effective polarization of the C spins.

[0041] Novel precursors are disclosed herein, including compounds of formulas Ia, Ib, IIa, IIb, IIIa, IIIb, and IVa, their tautomers, deuterated derivatives of those compounds and their tautomers, their salts, and at one or more sites within the molecule 13 C or 15N-enriched derivatives (which derivatives may in turn be hyperpolarized), and progeny of precursors given by general formulas Ia, Ib, IIa, IIb, IIIa, IIIb, and IVa.

[0042] Precursors of formula Ia and formula Ib

[0043] In some embodiments, the precursor comprises a compound of formula Ia. Formula Ia encompasses the following structures:

[0044]

[0045] and includes its tautomers, deuterated derivatives of those compounds and their tautomers, pharmaceutically acceptable salts thereof, and at one or more sites 13 C or 15 N-enriched derivatives. In some embodiments, Z of formula Ia describes: (i) a carbon-carbon double bond (-C═C-) that is fully substituted to include 2 H (deuterium, also known as D) (i.e., -CD═CD-) or (ii) a carbon-carbon triple bond (-C≡C-). In some embodiments, R1 of formula Ia comprises a parahydrogen induced polarization (PHIP) transfer moiety as described herein. In some embodiments, R2 of formula Ia comprises an optionally substituted hydrocarbon, alkoxy, primary amine, secondary amine, or tertiary amine as described herein. In some embodiments, R3 of formula Ia comprises a bio-related contrast agent as described herein. In formula Ia, all of the portions to the right of the R3-R1 bond (i.e., -R1-Z-(C═O)-R2) may be collectively referred to as the side arm.

[0046] In some embodiments, the precursor comprises a compound of formula Ib. Formula Ib encompasses the following structures:

[0047]

[0048] and includes its tautomers, deuterated derivatives of those compounds and their tautomers, pharmaceutically acceptable salts thereof, and at one or more sites 13C-enriched derivatives. In some embodiments, Z in formula Ib represents an ethynyl (-C≡C-) group, a fully deuterated propargyl (-CD2-C≡CD2-) group, a fully deuterated vinyl (-CD=CD-) group, a fully deuterated allyl (-CD2-CD=CD-) group, or a fully deuterated but-3-enyl (-CD2-CD2-CD=CD-) group. In some embodiments, R2 in formula Ib comprises an optionally substituted hydrocarbyl group, alkyl group, cycloalkyl group, aryl group, carboxyl group, ketone group, or alkoxy group as described herein. In some embodiments, R3 in formula Ib comprises an acyl derivative of a biologically relevant contrast agent as described herein. In formula Ib, all moieties to the right of the R3 group (i.e., -S-Z-R2) may be collectively referred to as the side arm.

[0049] In some embodiments, the compound of formula Ia or formula Ib has a solubility in water of at least about 1 millimolar (mM), 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 150 mM, 200 mM, 250 mM, 300 mM, 350 mM, 400 mM, 450 mM, 500 mM, 550 mM, 600 mM, 650 mM, 700 mM, 750 mM, 800 mM, 850 mM, 900 mM, 950 mM, 1,000 mM or more, up to about 1,000 mM, 950 mM, 900 mM, 850 mM, 800 mM, 750 mM, 700 mM, 650 mM, 600 mM, 550 mM, 500 mM, 450 mM, 400 mM, 350 mM, 300 mM, 250 mM, 200 mM, 150 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, 9 mM, 8 mM, 7 mM, 6 mM, 5 mM, 4 mM, 3 mM, 2 mM, 1 mM or less, or has a solubility in water within a range defined by any two of the foregoing values.

[0050] In some embodiments, the solubility of a compound of formula Ia or formula Ib in an organic solvent (e.g., acetone, ethanol, chloroform, toluene) is at least about 1 millimolar (mM), 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 150 mM, 200 mM, 250 mM, 300 mM, 350 mM, 400 mM, 450 mM, 500 mM, 550 mM, 600 mM, 650 mM, 700 mM, 750 mM, 800 mM, 850 mM, 900 mM, 950 mM, 1,000 mM or more, up to about 1,000 mM, 950 mM, 900 mM, 850 mM, 800 mM, 750 mM, 700 mM, 650 mM, 600 mM, 550 mM, 500 mM, 450 mM, 400 mM, 350 mM, 300 mM, 250 mM, 200 mM, 150 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, 9 mM, 8 mM, 7 mM, 6 mM, 5 mM, 4 mM, 3 mM, 2 mM, 1 mM or less, or the solubility in the organic solvent is within a range defined by any two of the foregoing values.

[0051] Parahydrogenated precursors of formula IIa and formula IIb

[0052] In some embodiments, a compound of formula Ia is deuterated (i.e., modified by addition of deuterium protons across Z via a hydrogenation reaction between a compound of formula Ia and a deuterium molecule), as described herein. In some embodiments, deuteration of a compound of formula Ia yields a compound of formula IIa. Formula IIa encompasses the following structures:

[0053]

[0054] And includes its tautomers, deuterated derivatives of those compounds and their tautomers, pharmaceutically acceptable salts thereof, and 13 C or 15 N-enriched derivatives at one or more sites. In some embodiments, Z' of formula IIa is: (i) fully substituted to include 2The secondary hydrogenated carbon-carbon single bond (-CH*-CH*-) of H (deuterium, also known as D), i.e., -CDH*-CDH*-, or (ii) the secondary hydrogenated carbon-carbon double bond (-CH*=CH*-). In some embodiments, H* represents hydrogen having a spin order derived from secondary hydrogen (i.e., a hydrogen atom or proton added across a carbon-carbon double bond or carbon-carbon triple bond Z via a hydrogenation reaction between a compound of Formula Ia and secondary hydrogen, as described herein). In some embodiments, H* represents hydrogen having a spin order derived from secondary hydrogen (e.g., prior to polarization transfer). In some embodiments, R1 of Formula IIa comprises a PHIP transfer moiety, as described herein. In some embodiments, R2 of Formula IIa comprises an optionally substituted hydrocarbon, alkoxy, primary amine, secondary amine, or tertiary amine, as described herein. In some embodiments, R3 of Formula IIa comprises a bio-related contrast agent, as described herein. In Formula IIa, all moieties to the right of the R3-R1 bond (i.e., -R1-Z'-(C=O)-R2) may be collectively referred to as the secondary hydrogenated side arm.

[0055] In some embodiments, a compound of Formula Ib is secondary hydrogenated (i.e., modified by adding a secondary hydrogen proton across Z via a hydrogenation reaction between Formula Ib and a secondary hydrogen molecule), as described herein. In some embodiments, the secondary hydrogenation of a compound of Formula Ib yields a compound of Formula IIb. Formula IIb encompasses the following structures:

[0056]

[0057] And includes its tautomers, deuterated derivatives of those compounds and their tautomers, its pharmaceutically acceptable salts, and at one or more sites 13C-enriched derivatives. In some embodiments, Z' in Formula IIb represents a secondary vinyl (-CH*=CH*-) group, a fully deuterated secondary vinyl prop-2-enyl (-CD2-CH*=CH*-) group, a fully deuterated secondary ethyl (-CDH*-CDH*-) group, a fully deuterated secondary propyl (-CD2-CDH*-CDH*-) group, or a fully deuterated secondary butyl (-CD2-CD2-CH*=CH*-) group. In some embodiments, H* represents a hydrogen having a spin order derived from secondary hydrogen (i.e., a hydrogen atom or proton added across a carbon-carbon double bond or carbon-carbon triple bond Z by a hydrogenation reaction between a compound of Formula Ib and secondary hydrogen, as described herein). In some embodiments, H* represents a hydrogen having a spin order derived from secondary hydrogen (e.g., prior to polarization transfer). In some embodiments, R2 in Formula IIb comprises an optionally substituted hydrocarbyl group, alkyl group, cycloalkyl group, aryl group, carboxyl group, ketone group, or alkoxy group, as described herein. In some embodiments, R3 in Formula IIb comprises an acyl derivative of a biorelevant contrast agent, as described herein. In Formula IIb, all moieties to the right of the R3 group (i.e., S-Z'R2) may be collectively referred to as a secondary hydrogenated side arm.

[0058] In some embodiments, the compound of Formula IIa or Formula IIb has a solubility in water of at least about 1 millimolar (mM), 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 150 mM, 200 mM, 250 mM, 300 mM, 350 mM, 400 mM, 450 mM, 500 mM, 550 mM, 600 mM, 650 mM, 700 mM, 750 mM, 800 mM, 850 mM, 900 mM, 950 mM, 1,000 mM or more, up to about 1,000 mM, 950 mM, 900 mM, 850 mM, 800 mM, 750 mM, 700 mM, 650 mM, 600 mM, 550 mM, 500 mM, 450 mM, 400 mM, 350 mM, 300 mM, 250 mM, 200 mM, 150 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, 9 mM, 8 mM, 7 mM, 6 mM, 5 mM, 4 mM, 3 mM, 2 mM, 1 mM or less, or has a solubility in water within a range defined by any two of the foregoing values.

[0059] In some embodiments, the solubility of the compound of Formula IIa or Formula IIb in an organic solvent (e.g., acetone, ethanol, chloroform, toluene) is at least about 1 millimole (mM), 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 150 mM, 200 mM, 250 mM, 300 mM, 350 mM, 400 mM, 450 mM, 500 mM, 550 mM, 600 mM, 650 mM, 700 mM, 750 mM, 800 mM, 850 mM, 900 mM, 950 mM, 1,000 mM or more, up to about 1,000 mM, 950 mM, 900 mM, 850 mM, 800 mM, 750 mM, 700 mM, 650 mM, 600 mM, 550 mM, 500 mM, 450 mM, 400 mM, 350 mM, 300 mM, 250 mM, 200 mM, 150 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, 9 mM, 8 mM, 7 mM, 6 mM, 5 mM, 4 mM, 3 mM, 2 mM, 1 mM or less, or the solubility in the organic solvent is within the range defined by any two of the foregoing values.

[0060] In some embodiments, when the composition of Formula Ia or Formula IIb reacts with secondary hydrogen, the chemical yield (e.g., the chemical yield of the compound of Formula IIa or Formula IIb) is at least about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more, up to about 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30% or less, or within the range defined by any two of the foregoing values. For example, in some embodiments, when the composition of Formula Ia or Formula Ib reacts with secondary hydrogen, the chemical yield is between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 30% and 55%, between 30% and 60%, between 30% and 65%, between 30% and 70%, between 30% and 75%, between 30% and 80%, between 30% and 85%, between 30% and 90%, between 30% and 95%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 35% and 55%, between 35% and 60%, between 35% and 65%, between 35% and 70%, between 35% and 75%, between 35% and 80%, between 35% and 85%, between 35% and 90%, between 35% and 95%, between 40% and 45%, between 40% and 50%, between 40% and 55%, between 40% and 60%, between 40% and 65%, between 40% and 70%, between 40% and 75%, between 40% and 80%, between 40% and 85%, between 40% and 90%, between 40% and 95%, between 45% and 50%, between 45% and 55%, between 45% and 60%, between 45% and 65%, between 45% and 70%, between 45% and 75%, between 45% and 80%, between 45% and 85%, between 45% and 90%, between 45% and 95%, between 50% and 55%, between 50% and 60%, between 50% and 65%, between 50% and 70%, between 50% and 75%, between 50% and 80%, between 50% and 85%, between 50% and 90%, between 50% and 95%, between 55% and 60%, between 55% and 65%, between 55% and 70%, between 55% and 75%, between 55% and 80%, between 55% and 85%, between 55% and 90%, between 55% and 95%,Between 60% and 65%, between 60% and 70%, between 60% and 75%, between 60% and 80%, between 60% and 85%, between 60% and 90%, between 60% and 95%, between 65% and 70%, between 65% and 75%, between 65% and 80%, between 65% and 85%, between 65% and 90%, between 65% and 95%, between 70% and 75%, between 70% and 80%, between 70% and 85%, between 70% and 90%, between 70% and 95%, between 75% and 80%, between 75% and 85%, between 75% and 90%, between 75% and 95%, between 80% and 85%, between 80% and 90%, between 80% and 95%, between 85% and 90%, between 85% and 95% or between 90% and 95%.

[0061] Cleaved precursors of formula IIIa and formula IIIb

[0062] In some embodiments, the compound of formula IIa is cleaved (e.g., hydrolyzed) as described herein. In some embodiments, the compound of formula IIa is cleaved (e.g., hydrolyzed) as described herein to provide a side arm compound and a corresponding biologically relevant contrast agent. In some embodiments, cleavage of the compound of formula IIa produces a compound of formula IIIa and a corresponding biologically relevant contrast agent as described herein. Formula IIIa encompasses the following structures:

[0063]

[0064] And includes its tautomers, deuterated derivatives of those compounds and their tautomers, pharmaceutically acceptable salts thereof and at one or more sites 13 C or 15 N-enriched derivatives. In some embodiments, Z” of formula IIIa is: (i) a secondary hydrocarbyl-carbon single bond (-CH*-CH*-) that is fully substituted to include 2 H (deuterium, also known as D) (i.e., -CDH*-CDH*-), or (ii) a secondary hydrocarbyl-carbon double bond (-CH*=CH*-). In some embodiments, R1' of formula IIIa comprises a PHIP transfer moiety as described herein. In some embodiments, R2 of formula IIIa comprises an optionally substituted hydrocarbon, alkoxy, primary amine, secondary amine or tertiary amine as described herein. In formula IIIa, all moieties R1-Z”-(C=O)-R2 can be collectively referred to as a cleaved side arm or a hydrolyzed side arm.

[0065] In some embodiments, the compound of formula IIb is cleaved (e.g., hydrolyzed) as described herein. In some embodiments, the compound of formula IIb is cleaved (e.g., hydrolyzed) as described herein to provide a side arm compound and a corresponding biorelevant contrast agent. In some embodiments, cleavage of the compound of formula IIb yields a compound of formula IIIb and a corresponding biorelevant contrast agent, as described herein. Formula IIIb encompasses the following structures:

[0066]

[0067] and includes its tautomers, deuterated derivatives of those compounds and their tautomers, pharmaceutically acceptable salts thereof, and 13 C-enriched derivatives at one or more sites. In some embodiments, Z” in formula IIIb represents a secondary vinyl (-CH*=CH*-) group, a fully deuterated secondary vinyl prop-2-enyl (-CD2-CH*=CH*-) group, a fully deuterated secondary vinyl ethyl (-CDH*-CDH*-) group, a fully deuterated secondary vinyl propyl (-CD2-CDH*-CDH*-) group, or a fully deuterated secondary vinyl butyl (-CD2-CD2-CH*=CH*-) group. R2 in formula IIIb comprises an optionally substituted hydrocarbyl, alkyl, cycloalkyl, aryl, carboxyl, keto, or alkoxy group, as described herein. In formula IIIb, all moieties H-S-Z'-R2 can be collectively referred to as a cleaved side arm or a hydrolyzed side arm.

[0068] In some embodiments, the solubility of the compound of formula IIIa or formula IIIb in water is at least about 1 millimole (mM), 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 150 mM, 200 mM, 250 mM, 300 mM, 350 mM, 400 mM, 450 mM, 500 mM, 550 mM, 600 mM, 650 mM, 700 mM, 750 mM, 800 mM, 850 mM, 900 mM, 950 mM, 1,000 mM or more, up to about 1,000 mM, 950 mM, 900 mM, 850 mM, 800 mM, 750 mM, 700 mM, 650 mM, 600 mM, 550 mM, 500 mM, 450 mM, 400 mM, 350 mM, 300 mM, 250 mM, 200 mM, 150 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, 9 mM, 8 mM, 7 mM, 6 mM, 5 mM, 4 mM, 3 mM, 2 mM, 1 mM or less, or the solubility in water is within the range defined by any two of the foregoing values.

[0069] In some embodiments, the solubility of the compound of formula IIIa or formula IIIb in an organic solvent (e.g., acetone, ethanol, chloroform, toluene) is at least about 1 millimole (mM), 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 150 mM, 200 mM, 250 mM, 300 mM, 350 mM, 400 mM, 450 mM, 500 mM, 550 mM, 600 mM, 650 mM, 700 mM, 750 mM, 800 mM, 850 mM, 900 mM, 950 mM, 1,000 mM or more, up to about 1,000 mM, 950 mM, 900 mM, 850 mM, 800 mM, 750 mM, 700 mM, 650 mM, 600 mM, 550 mM, 500 mM, 450 mM, 400 mM, 350 mM, 300 mM, 250 mM, 200 mM, 150 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, 9 mM, 8 mM, 7 mM, 6 mM, 5 mM, 4 mM, 3 mM, 2 mM, 1 mM or less, or the solubility in the organic solvent is within the range defined by any two of the foregoing values.

[0070] Side arms of formula IVa

[0071] In some embodiments, a biorelevant contrast agent is conjugated with a side arm (such as a side arm compound of Formula IVa) to form a precursor compound, such as a compound of Formula Ia, as described herein. Formula IVa encompasses the following structures:

[0072]

[0073] and includes its tautomers, deuterated derivatives of those compounds and their tautomers, pharmaceutically acceptable salts thereof, and 13 C or 15 N-enriched derivatives at one or more sites. In some embodiments, Z of Formula IVa describes: (i) a carbon-carbon double bond (-C═C-) that is fully substituted to include 2 H (deuterium, also known as D) (i.e., -CD═CD-), or (ii) a carbon-carbon triple bond (-C≡C-). In some embodiments, R1 of Formula IVa contains a parahydrogen-induced polarization (PHIP) transfer moiety, as described herein. In some embodiments, R2 of Formula IVa contains a solubilizing moiety, as described herein. In some embodiments, R2 of Formula IVa contains an optionally substituted hydrocarbon, alkoxy, primary amine, secondary amine, or tertiary amine. In some embodiments, conjugation of a compound of Formula IVa with a biorelevant contrast agent yields a compound of Formula Ia, as described herein.

[0074] PHIP transfer moiety

[0075] In some embodiments, the compositions of the present disclosure contain a PHIP transfer moiety. In some embodiments, the compositions of the present disclosure contain a PHIP transfer moiety between a Z, Z' or Z'' moiety and a sulfur atom of Formula Ib, Formula IIb or Formula IIIb. In some embodiments, the PHIP transfer moiety described herein contains a chemical moiety that is configured to permit or enhance polarization transfer from one or more parahydrogenated protons H* (e.g., H* in the side arm) to one or more non-hydrogen nuclear spins of a biorelevant contrast agent (such as one or more 13 C or 15 N atoms of the biorelevant contrast agent, as described herein). In some embodiments, the PHIP transfer moiety permits or enhances polarization transfer from a parahydrogen proton H* in the side arm of a compound of Formula IIa or Formula IIb to a non-hydrogen nuclear spin of the corresponding biorelevant contrast agent of the compound of Formula IIa or Formula IIb. In some embodiments, after a parahydrogenation reaction between parahydrogen and Formula Ia or Formula Ib, the PHIP transfer moiety permits or enhances polarization transfer from a parahydrogen proton H* in the side arm of the compound of Formula IIa or Formula IIb to a non-hydrogen nuclear spin of the corresponding biorelevant contrast agent of the compound of Formula IIa or Formula IIb.

[0076] In some embodiments, the PHIP transfer moiety comprises a fully deuterated C1 hydrocarbon (i.e., -CD2-) or a fully deuterated C2 hydrocarbon (i.e., -CD2-CD2-).

[0077] In some embodiments, the PHIP transfer moiety comprises a chemical moiety of the form *CR4R5 or any fully deuterated version thereof. In some embodiments, *C is 12 a C carbon isotope. In some embodiments, R4 and R5 are each independently selected from: 2 H, a fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbon, a fully deuterated C6 aryl, a fully deuterated benzyl, a fully deuterated phenyl, a fully deuterated heteroaryl and a fully deuterated haloalkyl.

[0078] In some embodiments, the PHIP transfer moiety comprises a chemical moiety of the form *CR6R7–*CR8R9 or any fully deuterated version thereof. In some embodiments, *C is 12 a C carbon isotope. In some embodiments, R6, R7, R8 and R9 are each independently selected from: 2 H, a fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbon, a fully deuterated C6 aryl, a fully deuterated benzyl, a fully deuterated phenyl, a fully deuterated heteroaryl and a fully deuterated haloalkyl.

[0079] In some embodiments, the PHIP transfer moiety comprises a chemical moiety of the form *CH2, *CH2–*CH2 or any fully deuterated version thereof. In some embodiments, *C is 12 a C carbon isotope.

[0080] In some embodiments, the compositions described herein include a first J-coupling J 12 between the spin-1 / 2 atoms described herein and the non-hydrogen nuclear spins described herein. In some embodiments, the compositions described herein contain a second J-coupling J 13 between the spin-1 / 2 atoms described herein and the secondary hydrogen protons H* described herein. In some embodiments, the compositions described herein include a third J-coupling J 23 between the non-hydrogen nuclear spins described herein and the secondary hydrogen protons H* described herein. In some embodiments, J 12 and / or J 13 is greater than J 23 . In such cases, the PHIP transfer moiety can permit or enhance polarization transfer.

[0081] In some embodiments, the J-coupling between one or both of the *H nuclear spins induced by the PHIP transfer portion and a non-hydrogen nuclear spin is at least about 0.1 Hz, 0.2 Hz, 0.3 Hz, 0.4 Hz, 0.5 Hz, 0.6 Hz, 0.7 Hz, 0.8 Hz, 0.9 Hz, 1 Hz or more, at most about 1 Hz, 0.9 Hz, 0.8 Hz, 0.7 Hz, 0.6 Hz, 0.5 Hz, 0.4 Hz, 0.3 Hz, 0.2 Hz, 0.1 Hz or less, or the J-coupling between the non-hydrogen nuclear spin is within a range defined by any two of the foregoing values. For example, in some embodiments, the J-coupling is between 0.1 Hz and 0.2 Hz, between 0.1 Hz and 0.3 Hz, between 0.1 Hz and 0.4 Hz, between 0.1 Hz and 0.5 Hz, between 0.1 Hz and 0.6 Hz, between 0.1 Hz and 0.7 Hz, between 0.1 Hz and 0.8 Hz, between 0.1 Hz and 0.9 Hz, between 0.1 Hz and 1 Hz, between 0.2 Hz and 0.3 Hz, between 0.2 Hz and 0.4 Hz, between 0.2 Hz and 0.5 Hz, between 0.2 Hz and 0.6 Hz, between 0.2 Hz and 0.7 Hz, between 0.2 Hz and 0.8 Hz, between 0.2 Hz and 0.9 Hz, between 0.2 Hz and 1 Hz, between 0.3 Hz and 0.4 Hz, between 0.3 Hz and 0.5 Hz, between 0.3 Hz and 0.6 Hz, between 0.3 Hz and 0.7 Hz, between 0.3 Hz and 0.8 Hz, between 0.3 Hz and 0.9 Hz, between 0.3 Hz and 1 Hz, between 0.4 Hz and 0.5 Hz, between 0.4 Hz and 0.6 Hz, between 0.4 Hz and 0.7 Hz, between 0.4 Hz and 0.8 Hz, between 0.4 Hz and 0.9 Hz, between 0.4 Hz and 1 Hz, between 0.5 Hz and 0.6 Hz, between 0.5 Hz and 0.7 Hz, between 0.5 Hz and 0.8 Hz, between 0.5 Hz and 0.9 Hz, between 0.5 Hz and 1 Hz, between 0.6 Hz and 0.7 Hz, between 0.6 Hz and 0.8 Hz, between 0.6 Hz and 0.9 Hz, between 0.6 Hz and 1 Hz, between 0.7 Hz and 0.8 Hz, between 0.7 Hz and 0.9 Hz, between 0.7 Hz and 1 Hz, between 0.8 Hz and 0.9 Hz, between 0.8 Hz and 1 Hz or between 0.9 Hz and 1 Hz.

[0082] Deuterated compounds

[0083] In certain embodiments, the use of deuterated Z, Z', or Z'' groups or PHIP transfer moieties increases the period of time that the spin order associated with the secondary hydrogen singlet (referred to herein as "singlet lifetime") persists. In certain embodiments, the singlet lifetime associated with a compound containing a deuterated Z, Z', or Z'' group or PHIP transfer moiety is increased compared to a similar compound containing a Z, Z', or Z'' group or PHIP transfer moiety, where the PHIP transfer moiety contains nuclei such as 1 H, 13 C, 19 F, 31 P, etc. or other nuclei coupled to H* hydrogen atoms having a spin order derived from the secondary hydrogen described herein. In certain embodiments, the singlet lifetime associated with a compound containing a deuterated Z, Z', or Z'' group or PHIP transfer moiety described herein is at least about 10 seconds, 15 seconds, 20 seconds, 25 seconds, 30 seconds, 35 seconds, 40 seconds, 45 seconds, 50 seconds, 55 seconds, 60 seconds, 65 seconds, 70 seconds, 75 seconds, 80 seconds, 85 seconds, 90 seconds, 95 seconds, 100 seconds, 105 seconds, 110 seconds, 115 seconds, 120 seconds or longer, up to about 120 seconds, 115 seconds, 110 seconds, 105 seconds, 100 seconds, 95 seconds, 90 seconds, 85 seconds, 80 seconds, 75 seconds, 70 seconds, 65 seconds, 60 seconds, 55 seconds, 50 seconds, 45 seconds, 40 seconds, 35 seconds, 30 seconds, 25 seconds, 20 seconds, 15 seconds, 10 seconds or shorter, or within a range defined by any two of the foregoing values.

[0084] In certain embodiments, the Z, Z', or Z'' group or PHIP transfer moiety contains at most 2, 1, or 0 nuclei such as 1 H, 13 C, 19 F, 31 P, etc. or other nuclei coupled to H* hydrogen atoms having a spin order derived from the secondary hydrogen described herein. In certain embodiments, the Z, Z', or Z'' group or PHIP transfer moiety contains at most 2, 1, or 0 1 H nuclei.

[0085] As used herein, the phrase "fully deuterated" means the replacement of 2 H (deuterium, also referred to as D) for 1 H (proton) at every site of the chemical moiety. Thus, for example, a fully deuterated straight-chain alkyl hydrocarbon moiety has the form (-CD2) n -CD3, where for a fully deuterated straight-chain C1-C10 alkyl hydrocarbon, n ranges from 0 to 9. Similarly, a fully deuterated phenyl moiety has the form -C6D5, a fully deuterated benzyl moiety has the form -CD2-C6D5, etc.

[0086] R2 group

[0087] In some embodiments, the R2 group described herein comprises an optionally substituted hydrocarbon, alkoxy, primary amine, secondary amine, or tertiary amine. In some embodiments, the R2 group described herein comprises an optionally substituted hydrocarbon, alkoxy, primary amine, secondary amine, or tertiary amine that functions as a solubilizing moiety. In some embodiments, the R2 group described herein comprises a solubilizing moiety. In some embodiments, the solubilizing moiety comprises any chemical moiety configured to permit or enhance the solubility of a compound, such as any one of the compounds of formulae Ia, Ib, IIa, IIb, IIIa, IIIb, and / or IVa, in a solution in which a secondary hydrogenation reaction or a cleavage (e.g., hydrolysis) reaction occurs. In some embodiments, an enhancement in the solubility of a variant of a compound of formulae Ia, Ib, IIa, IIb, IIIa, IIIb, and / or IVa that uses one or more protons in place of the R2 group is measured. In some embodiments, an enhancement in the solubility of a variant of a compound of formulae Ia, Ib, IIa, IIb, IIIa, IIIb, and / or IVa that uses methyl as the R2 group is measured.

[0088] In some embodiments, the solubilizing moiety comprises a hydrophobic moiety or a lipophilic moiety. In some embodiments, the solubilizing moiety comprises an organic solubilizing moiety. For example, in some embodiments, the solubilizing moiety comprises a hydrophobic moiety, a lipophilic moiety, or an organic solubilizing moiety. In some embodiments, the solubilizing moiety comprises a hydrophilic moiety or a lipophobic moiety.

[0089] In some embodiments, the R2 group comprises or is selected from: methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, hydroxy, methoxy, ethoxy, propoxy, isopropoxy, propionate, butoxy, tert-butoxy, sec-butoxy, ester, phenyl, substituted phenyl, primary amino, secondary amino, tertiary amino, primary amide, secondary amide, and tertiary amide. In some embodiments, the substituted phenyl is selected from fluorobenzene, chlorobenzene, bromobenzene, iodobenzene, toluene, cumene, ethylbenzene, styrene, o-xylene, m-xylene, p-xylene, phenol, benzoic acid, benzaldehyde, acetophenone, methyl benzoate, anisole, aniline, nitrobenzene, benzonitrile, benzamide, benzenesulfonic acid, naphthalene, and anthracene.

[0090] R3 group

[0091] In some embodiments, the R3 group described herein comprises a biorelevant contrast agent. In some embodiments, the biorelevant contrast agent has the formula R4C(=O)X-. In some embodiments, R4 is selected from straight-chain, branched-chain, or cyclic C1-C10 alkyl, wherein one or more C atoms are optionally substituted with CO, COOH, CH2COOH, CONH2, OH, amino (NR'R''), one or more halogen atoms, one or more haloalkyls, or one or more carbocycles, wherein the carbocycle is optionally substituted with one or more aliphatic or aromatic rings, and the one or more aliphatic or aromatic rings are optionally substituted with one or more functional groups. In some embodiments, X is selected from NR''' and O. In some embodiments, R', R'', and R''' are each independently selected from 1 H, 2 H, 3 H, and an amino protecting group, which is optionally selected from trifluoroacetyl, acetyl, benzoyl, benzyloxycarbonyl, tert-butyl carbonate, and benzyl. In some embodiments, the R3 group comprises any of the biorelevant contrast agents described herein.

[0092] In some embodiments, the R3 group described herein comprises an acyl derivative of a biorelevant contrast agent. In some embodiments, the biorelevant contrast agent has the formula R9C(=O)O-. In some embodiments, R9 is selected from straight-chain, branched-chain, or cyclic C1-C10 alkyl, wherein one or more C atoms are optionally substituted with CO, COOH, CH2COOH, CONH2, OH, amino (NR'R''), one or more halogen atoms, one or more haloalkyls, or one or more carbocycles, wherein the carbocycle is optionally substituted with one or more aliphatic or aromatic rings, and the one or more aliphatic or aromatic rings are optionally substituted with one or more functional groups. In some embodiments, R' and R'' are each independently selected from 1 H, 2 H, 3 H, and an amino protecting group, which is optionally selected from trifluoroacetyl, acetyl, benzoyl, benzyloxycarbonyl, tert-butyl carbonate, and benzyl. In some embodiments, the R3 group comprises an acyl derivative of any of the biorelevant contrast agents described herein.

[0093] In some embodiments, the R3 group comprises at least one non-hydrogen nuclear spin. In some embodiments, the non-hydrogen nucleus comprises at least one spin-1 / 2 atom. In some embodiments, the non-hydrogen nuclear spin comprises 13 C or 15N. In some embodiments, the R3 group is at least partially isotopically labeled with non-hydrogen nuclear spins. In some embodiments, the R3 group is at least partially enriched in non-hydrogen nuclear spins as compared to an analog of the R3 group that characterizes non-hydrogen nuclear spins at their natural abundance. In some embodiments, the R3 group is enriched to characterize the non-hydrogen nuclear spins at an abundance of at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, up to about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less, or within a range defined by any two of the foregoing values.

[0094] In some embodiments, the non-hydrogen nuclear spin replaces an NMR-inactive (i.e., spin-0) nucleus (e.g., 12 C or a quadrupolar (i.e., spin > 1 / 2) nucleus) (e.g., 14 N) of an analog of the R3 group that characterizes non-hydrogen nuclear spins at their natural abundance, as described herein. In some embodiments, the non-hydrogen nuclear spin is no more than about 1 or 2 chemical bonds away from the carbonyl (C=O) carbon in the R3 group.

[0095] Parahydrogenation

[0096] In some embodiments, the non-hydrogen nuclear spin replaces an NMR-inactive (i.e., spin-0) nucleus (e.g., 12 C or a quadrupolar (i.e., spin > 1 / 2) nucleus) (e.g., 14 N) of an analog of the R3 group that characterizes non-hydrogen nuclear spins at their natural abundance, as described herein. In some embodiments, the non-hydrogen nuclear spin is no more than about 1 or 2 chemical bonds away from the carbonyl (C=O) carbon in the R3 group.

[0097] Consistent with the disclosed embodiments, the precursor of a biorelevant contrast agent (such as a compound of formula Ia or formula Ib, as described herein) can be parahydrogenated by combining a precursor, parahydrogen, and a hydrogenation catalyst. The disclosed embodiments are not limited to a particular method of producing a parahydrogenated precursor. In some embodiments, the precursor is added to a mixture containing parahydrogen. In some embodiments, parahydrogen gas is added to a solution containing the precursor (for example, the parahydrogen gas can be bubbled into such a solution). During the process of hydrogenating the precursor, parahydrogen can produce a Iz1Iz2 level, preferentially populating the lower energy state between the |↑>|↓>, |↓>|↑>, or singlet spin order on the two hydrogen spins in the precursor.

[0098] The precursor can have an unsaturated bond (such as an unsaturated carbon-carbon double bond or an unsaturated carbon-carbon triple bond) that can be hydrogenated by parahydrogen gas. After the combination of the precursor and parahydrogen, at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more of the precursor, at most about 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10% or less of the precursor, or a percentage of the precursor within a range defined by any two of the foregoing values can be hydrogenated.

[0099] In some embodiments, the population difference of the parahydrogen precursor in the para-hydrogen proton spin state is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 40%, 45%, 50% or more, at most about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less, or a population difference within a range defined by any two of the foregoing values. For example, in some embodiments, the population difference is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, between 20% and 50%, between 25% and 30%, between 25% and 35%, between 25% and 40%, between 25% and 45%, between 25% and 50%, between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 40% and 45%, between 40% and 50% or between 45% and 50%. In some embodiments, the population difference is between spin states including para-hydrogen protons and other nuclear spins, such as the spins of additional protons on the compound. In some embodiments, the parahydrogen precursor includes a side arm, and the para-hydrogen spin can be located on the side arm.

[0100] In some embodiments, the concentration of the hydrogenation catalyst during hydrogenation is at least about 0.1 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM or more, up to about 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, 9 mM, 8 mM, 7 mM, 6 mM, 5 mM, 4 mM, 3 mM, 2 mM, 1 mM, 0.9 mM, 0.8 mM, 0.7 mM, 0.6 mM, 0.5 mM, 0.4 mM, 0.3 mM, 0.2 mM, 0.1 mM or less, or within a range defined by any two of the foregoing values.

[0101] The disclosed embodiments can include methods for generating hyperpolarized bio-related contrast agents implemented by the disclosed systems. The disclosed methods can include mixing (e.g., via a mixing mechanism) a solution comprising a precursor of a bio-related contrast agent and a hydrogenation catalyst. The mixing mechanism can be a device for introducing, holding, and facilitating the blending, mixture, or solution of two or more materials. In some embodiments, the mixing mechanism is disposed within a chamber, and mixing occurs inside the chamber. In some embodiments, the solution is mixed at a location remote from the chamber. The volume of the solution can be at least about 1 milliliter (ml), 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, 10 ml, 20 ml, 30 ml, 40 ml, 50 ml, 60 ml, 70 ml, 80 ml, 90 ml, 100 ml or more, up to about 100 ml, 90 ml, 80 ml, 70 ml, 60 ml, 50 ml, 40 ml, 30 ml, 20 ml, 10 ml, 9 ml, 8 ml, 7 ml, 6 ml, 5 ml, 4 ml, 3 ml, 2 ml, 1 ml or less, or within a volume range defined by any two of the foregoing values.

[0102] In some embodiments, the mixing mechanism is a gas-liquid exchange mechanism. For example, the gas-liquid exchange mechanism can be a bubbler or a diffusion system. In some embodiments, the mixing mechanism comprises a membrane adapted to allow the diffusion of molecular hydrogen. In some embodiments, mixing can be performed using an injection chamber, wherein the solution is injected into a chamber filled with pressurized parahydrogen.

[0103] In some embodiments, the catalyst is a molecular, complex, or particulate system for catalytic hydrogenation. In some embodiments, the catalyst comprises a homogeneous metal catalyst, such as a rhodium complex or a ruthenium complex. The rhodium complex can be used for the coordination and activation of precursor molecules and secondary hydrogen. In some embodiments, a heterogeneous metal catalyst is linked to nanoparticles.

[0104] Various embodiments of the present disclosure describe introducing a solution comprising a precursor of a bio-related contrast agent and a hydrogenation catalyst into a chamber configured to hold the solution during polarization transfer. In some embodiments, the solution is mixed in the chamber. In some embodiments, the solution is hydrogenated in the chamber. In some embodiments, the chamber is within a magnetic shield (e.g., a mu-metal shield). The magnetic shield can reduce the influence of the Earth's magnetic field (or other external magnetic fields), thereby allowing the amplitude of a low-level magnetic field applied to the solution to be modulated. Thus, placing the solution in the chamber can include placing the solution within the magnetic shield.

[0105] As described herein, in some embodiments, secondary hydrogenation occurs before polarization transfer (e.g., before modulating the amplitude of the magnetic field applied to the solution, etc.). In some embodiments, secondary hydrogenation occurs during polarization transfer. For example, during modulation of the magnetic field amplitude, secondary hydrogen can be combined with the solution (e.g., flowed through or bubbled through the solution).

[0106] In some embodiments, secondary hydrogen gas is combined with the solution in the hydrogenation chamber under pressure. The pressure can be at least about 10 bar, 15 bar, 20 bar, 30 bar, 50 bar or more, up to about 50 bar, 30 bar, 20 bar, 15 bar, 10 bar or less, or within a range defined by any two of the foregoing values. In some embodiments, secondary hydrogen is combined with the solution in a metal chamber capable of withstanding pressure. The secondary hydrogen can be combined with the solution at an interval of time (or the dissolution of the secondary hydrogen can occur in less than an interval of time). The interval of time can be up to about 90 seconds, 60 seconds, 30 seconds, 20 seconds, 10 seconds, 9 seconds, 8 seconds, 7 seconds, 6 seconds, 5 seconds, 4 seconds, 3 seconds, 2 seconds, 1 second or less, at least about 1 second, 2 seconds, 3 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 20 seconds, 30 seconds, 60 seconds, 90 seconds or more, or within a range defined by any two of the foregoing values. In some embodiments, hydrogenation occurs or takes place within the interval of time.

[0107] Polarization transfer using radiofrequency waveforms

[0108] In some embodiments, prior to polarization transfer, the concentration of the precursor or target molecule in solution is at least about 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 200 mM, 300 mM, 400 mM, 500 mM, 600 mM, 700 mM, 800 mM, 900 mM, 1,000 mM or more, up to about 1,000 mM, 900 mM, 800 mM, 700 mM, 600 mM, 500 mM, 400 mM, 300 mM, 200 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM 20 mM, 10 mM or less, or within a range defined by any two of the foregoing values. The volume of the solution can be at least about 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, 10 ml, 20 ml, 30 ml, 40 ml, 50 ml, 60 ml, 70 ml, 80 ml, 90 ml, 100 ml, 200 ml, 300 ml, 400 ml, 500 ml, 600 ml, 700 ml, 800 ml, 900 ml, 1,000 ml, 2,000 ml or more, up to about 2,000 ml, 1,000 ml, 900 ml, 800 ml, 700 ml, 600 ml, 500 ml, 400 ml, 300 ml, 200 ml, 100 ml, 90 ml, 80 ml, 70 ml, 60 ml, 50 ml, 40 ml, 30 ml, 20 ml, 10 ml, 9 ml, 8 ml, 7 ml, 6 ml, 5 ml, 4 ml, 3 ml, 2 ml, 1 ml or less, or within a range defined by any two of the foregoing values.

[0109] Various embodiments of the present disclosure describe the application of polarization transfer magnetic perturbations, which are intended to generate a magnetic field around a solution (e.g., around a solution containing Formula IIa or Formula IIb as described herein). In some embodiments, the strength of the magnetic field is at least about 0.1 gauss (G), 0.2 G, 0.3 G, 0.4 G, 0.5 G, 0.6 G, 0.7 G, 0.8 G, 0.9 G, 1 G, 2 G, 3 G, 4 G, 5 G, 6 G, 7 G, 8 G, 9 G, 10 G, 20 G, 30 G, 40 G, 50 G, 60 G, 70 G, 80 G, 90 G, 100 G, 200 G, 300 G, 400 G, 500 G, 600 G, 700 G, 800 G, 900 G, 1,000 G, 2,000 G, 3,000 G, 4,000 G, 5,000 G, 6,000 G, 7,000 G, 8,000 G, 9,000 G, 10,000 G, 20,000 G, 30,000 G, 40,000 G, 50,000 G, 60,000 G, 70,000 G, 80,000 G, 90,000 G, 100,000 G, 200,000 G or more, up to about 200,000 G, 100,000 G, 90,000 G, 80,000 G, 70,000 G, 60,000 G, 50,000 G, 40,000 G, 30,000 G, 20,000 G, 10,000 G, 9,000 G, 8,000 G, 7,000 G, 6,000 G, 5,000 G, 4,000 G, 3,000 G, 2,000 G, 1,000 G, 900 G, 800 G, 700 G, 600 G, 500 G, 400 G, 300 G, 200 G, 100 G, 90 G, 80 G, 70 G, 60 G, 50 G, 40 G, 30 G, 20 G, 10 G, 9 G, 8 G, 7 G, 6 G, 5 G, 4 G, 3 G, 2 G, 1 G, 0.9 G, 0.8 G, 0.7 G, 0.6 G, 0.5 G, 0.4 G, 0.3 G, 0.2 G, 0.1 G or less, or within a range defined by any two of the foregoing values. In some embodiments, the strength of the magnetic field around the solution is from 0.1 G to 200,000 G. The magnetic perturbation can be generated by an electromagnet or a permanent magnet. The magnetic field can be pulsed or applied to the sample as a continuous wave (CW). The magnetic perturbation can be static or vary with time.

[0110] The signal generator can be configured to generate one or more radio frequency (RF) waveforms, and the one or more RF waveforms can be applied to a sample to transfer polarization. The signal generator can include a plurality of computing units, processors, controllers, associated memories, PCs, computer services, or any device capable of performing computational operations using inputs and generating outputs. In some embodiments, the RF coil can radiate or 'apply' a pulse sequence, including a first RF waveform. In some embodiments, the RF coil can have one or more channels. A channel can be a path for an RF signal. At least one channel can be provided for each different type of NMR spectroscopy. In some embodiments, at least one channel is for 1 H, and at least one channel is for 2 H, 13 C, 15 N, 19 F, and 31 P, any one of them. For example, the first RF waveform can be applied to the 1 H channel of one or more radio frequency coils (RF coils) placed around the sample. In some embodiments, a second RF waveform is applied to the 13 C channel of the RF coil. In some embodiments, 1 the RF waveforms on the H channel and 13 the C channel are configured to apply polarization transfer sequences, such as PH-INEPT, Goldman's sequence, S2M, S2hM, SLIC, ADAPT, or ESOTERIC.

[0111] In some embodiments, the RF waveforms are configured to support polarization transfer even in the presence of a large proton full width at half maximum (FWHM). Such RF waveforms can include pulse sequences that can contain dozens to hundreds of RF pulses. The sequence can be configured such that the pulses prevent the adverse effects of magnetic field inhomogeneity on polarization transfer.

[0112] In some embodiments, the pulse sequence for polarization is configured to transfer spin order from two non-equivalent 1 H-hydrogenated spins, for example, when the chemical shift difference is greater than the J-coupling between them. For example, ESOTHERIC can be a pulse sequence suitable for polarization transfer in this state.

[0113] In some embodiments, the pulse sequence is configured to transfer from equivalent 1H hydrogen spin transfer spin order, for example, when the chemical shift difference is less than the J-coupling between them. Such pulse sequences can be used in magnetic fields having the following intensities: at least about 0.01 millitesla (mT), 0.02 mT, 0.03 mT, 0.04 mT, 0.05 mT, 0.06 mT, 0.07 mT, 0.08 mT, 0.09 mT, 0.1 mT, 0.2 mT, 0.3 mT, 0.4 mT, 0.5 mT, 0.6 mT, 0.7 mT, 0.8 mT, 0.9 mT, 1 mT, 2 mT, 3 mT, 4 mT, 5 mT, 6 mT, 7 mT, 8 mT, 9 mT, 10 mT, 20 mT, 30 mT, 40 mT, 50 mT, 60 mT, 70 mT, 80 mT, 90 mT, 100 mT, 200 mT, 300 mT, 400 mT, 500 mT, 600 mT, 700 mT, 800 mT, 900 mT, 1,000 mT, 2,000 mT, 3,000 mT, 4,000 mT, 5,000 mT, 6,000 mT or more, up to about 6,000 mT, 5,000 mT, 4,000 mT, 3,000 mT, 2,000 mT, 1,000 mT, 900 mT, 800 mT, 700 mT, 600 mT, 500 mT, 400 mT, 300 mT, 200 mT, 100 mT, 90 mT, 80 mT, 70 mT, 60 mT, 50 mT, 40 mT, 30 mT, 20 mT, 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 0.9 mT, 0.8 mT, 0.7 mT, 0.6 mT, 0.5 mT, 0.4 mT, 0.3 mT, 0.2 mT, 0.1 mT, 0.09 mT, 0.08 mT, 0.07 mT, 0.06 mT, 0.05 mT, 0.04 mT, 0.03 mT, 0.02 mT, 0.01 mT or less, or in a range defined by any two of the foregoing values. Examples of such sequences can be the Goldman sequence (M. Goldman, H. Jóhannesson, C.R. Phys. 2005, 6, 575–581, the literature on the pulse sequence configuration for transfer spin order is incorporated herein by reference), the singlet-to-heteronuclear magnetization (S2hM) sequence or other sequences used in singlet NMR (such as ADAPT, SLIC, etc.).

[0114] In some embodiments, the magnetic shield is configured to maintain the magnetic field applied to the solution at at least about 0 mG, 0.1 mG, 0.2 mG, 0.3 mG, 0.4 mG, 0.5 mG, 0.6 mG, 0.7 mG, 0.8 mG, 0.9 mG, 1 mG, 2 mG, 3 mG, 4 mG, 5 mG, 6 mG, 7 mG, 8 mG, 9 mG, 10 mG, 20 mG, 30 mG, 40 mG, 50 mG, 60 mG, 70 mG, 80 mG, 90 mG, 100 mG or more, up to about 100 mG, 90 mG, 80 mG, 70 mG, 60 mG, 50 mG, 40 mG, 30 mG, 20 mG, 10 mG, 9 mG, 8 mG, 7 mG, 6 mG, 5 mG, 4 mG, 3 mG, 2 mG, 1 mG, 0.9 mG, 0.8 mG, 0.7 mG, 0.6 mG, 0.5 mG, 0.4 mG, 0.3 mG, 0.2 mG, 0.1 mG or less, or a magnetic field within a range defined by any two of the foregoing values. During application of the polarization waveform to one or more radio frequency coils, the magnetic shield can maintain the magnetic field strength within the polarization chamber at such an amplitude.

[0115] Consistent with the disclosed embodiments, an RF waveform can be applied to a solution containing a parahydrogen precursor.

[0116] Polarization transfer using magnetic field modulation

[0117] In some embodiments, polarization transfer magnetic perturbations are performed in a magnetic shield (such as a μ - shield, etc.) to achieve a uniform low magnetic field. The magnetic shield is capable of transferring polarization to 13 C nuclear spins at a magnetic field in the microtesla (μT) range below the Earth's magnetic field. The low magnetic field can be at least about 0 mG, 0.1 mG, 0.2 mG, 0.3 mG, 0.4 mG, 0.5 mG, 0.6 mG, 0.7 mG, 0.8 mG, 0.9 mG, 1 mG, 2 mG, 3 mG, 4 mG, 5 mG, 6 mG, 7 mG, 8 mG, 9 mG, 10 mG, 20 mG, 30 mG, 40 mG, 50 mG, 60 mG, 70 mG, 80 mG, 90 mG, 100 mG or more, up to about 100 mG, 90 mG, 80 mG, 70 mG, 60 mG, 50 mG, 40 mG, 30 mG, 20 mG, 10 mG, 9 mG, 8 mG, 7 mG, 6 mG, 5 mG, 4 mG, 3 mG, 2 mG, 1 mG, 0.9 mG, 0.8 mG, 0.7 mG, 0.6 mG, 0.5 mG, 0.4 mG, 0.3 mG, 0.2 mG, 0.1 mG or less, or within a range defined by any two of the foregoing values.

[0118] In such fields, by using proton spin and other spin species of interest, including 2 H,13 C, 15 N, 19 F, and 31 P) to transfer polarization by avoiding level anti-crossing (LAC). In some embodiments, the magnetic field can be adjusted to a specific magnetic field strength of the LAC, such as that performed in the SABRE-SHEATH experiment. In various embodiments, to achieve robust polarization transfer in a larger volume of sample, the magnetic field strength can be time-modulated. For example, the magnetic field strength can be swept through the LAC condition. Alternatively or additionally, the sample can be physically moved inside the magnetic field. Such modulation can relax the constraints on magnetic field homogeneity and on magnetic field offset. Thus, robust polarization transfer can be performed with a larger volume and higher efficiency. Furthermore, relaxing the constraints on magnetic field homogeneity and on magnetic field offset can allow the use of less complex, less precise, or less expensive polarization systems.

[0119] The lower limit of the magnetic field modulation can be at least about -10 μT, -9 μT, -8 μT, -7 μT, -6 μT, -5 μT, -4 μT, -3 μT, -2 μT, -1.9 μT, -1.8 μT, -1.7 μT, -1.6 μT, -1.5 μT, -1.4 μT, -1.3 μT, -1.2 μT, -1.1 μT, -1 μT, -0.9 μT, -0.8 μT, -0.7 μT, -0.6 μT, -0.5 μT, -0.4 μT, -0.3 μT, -0.2 μT, -0.1 μT or more, up to about -0.1 μT, -0.2 μT, -0.3 μT, -0.4 μT, -0.5 μT, -0.6 μT, -0.7 μT, -0.8 μT, -0.9 μT, -1 μT, -1.1 μT, -1.2 μT, -1.3 μT, -1.4 μT, -1.5 μT, -1.6 μT, -1.7 μT, -1.8 μT, -1.9 μT, -2 μT, -3 μT, -4 μT, -5 μT, -6 μT, -7 μT, -8 μT, -9 μT, -10 μT or less, or within the range defined by any two of the foregoing values. The upper limit of the modulation can be at least about 0.1 μT, 0.2 μT, 0.3 μT, 0.4 μT, 0.5 μT, 0.6 μT, 0.7 μT, 0.8 μT, 0.9 μT, 1 μT, 1.1 μT, 1.2 μT, 1.3 μT, 1.4 μT, 1.5 μT, 1.6 μT, 1.7 μT, 1.8 μT, 1.9 μT, 2 μT, 3 μT, 4 μT, 5 μT, 6 μT, 7 μT, 8 μT, 9 μT, 10 μT or more, up to about 10 μT, 9 μT, 8 μT, 7 μT, 6 μT, 5 μT, 4 μT, 3 μT, 2 μT, 1.9 μT, 1.8 μT, 1.7 μT, 1.6 μT, 1.5 μT, 1.4 μT, 1.3 μT, 1.2 μT, 1.1 μT, 1 μT, 0.9 μT, 0.8 μT, 0.7 μT, 0.6 μT, 0.5 μT, 0.4 μT, 0.3 μT, 0.2 μT, 0.1 μT or less, or within the range defined by any two of the foregoing values.

[0120] The magnetic field can have such an amplitude over a volume of at least about 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, 10 ml, 20 ml, 30 ml, 40 ml, 50 ml, 60 ml, 70 ml, 80 ml, 90 ml, 100 ml, 200 ml, 300 ml, 400 ml, 500 ml, 600 ml, 700 ml, 800 ml, 900 ml, 1,000 ml, 2,000 ml or more, up to about 2,000 ml, 1,000 ml, 900 ml, 800 ml, 700 ml, 600 ml, 500 ml, 400 ml, 300 ml, 200 ml, 100 ml, 90 ml, 80 ml, 70 ml, 60 ml, 50 ml, 40 ml, 30 ml, 20 ml, 10 ml, 9 ml, 8 ml, 7 ml, 6 ml, 5 ml, 4 ml, 3 ml, 2 ml, 1 ml or less, or a volume within a range defined by any two of the foregoing values. The modulation can be carried out over a period of time. The period of time can be at least about 100 milliseconds (ms), 200 milliseconds, 300 milliseconds, 400 milliseconds, 500 milliseconds, 600 milliseconds, 700 milliseconds, 800 milliseconds, 900 milliseconds, 1 second (s), 2 seconds, 3 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 20 seconds, 30 seconds or longer, up to about 30 seconds, 20 seconds, 10 seconds, 9 seconds, 8 seconds, 7 seconds, 6 seconds, 5 seconds, 4 seconds, 3 seconds, 2 seconds, 1 second, 900 milliseconds, 800 milliseconds, 700 milliseconds, 600 milliseconds, 500 milliseconds, 400 milliseconds, 300 milliseconds, 200 milliseconds, 100 milliseconds or shorter, or within a range defined by any two of the foregoing values.

[0121] Thus, the rate of change of the amplitude of the magnetic field can be at least about 0.01 μT / second, 0.015 μT / second, 0.02 μT / second, 0.025 μT / second, 0.03 μT / second, 0.035 μT / second, 0.04 μT / second, 0.045 μT / second, 0.05 μT / second, 0.055 μT / second, 0.06 μT / second, 0.065 μT / second, 0.07 μT / second, 0.075 μT / second, 0.08 μT / second, 0.085 μT / second, 0.09 μT / second, 0.095 μT / second, 0.1 μT / second, 0.15 μT / second, 0.2 μT / second, 0.25 μT / second, 0.3 μT / second, 0.35 μT / second, 0.4 μT / second, 0.45 μT / second, 0.5 μT / second, 0.55 μT / second, 0.6 μT / second, 0.65 μT / second, 0.7 μT / second, 0.75 μT / second, 0.8 μT / second, 0.85 μT / second, 0.9 μT / second, 0.95 μT / second, 1 μT / second or greater, and at most about 1 μT / second, 0.95 μT / second, 0.9 μT / second, 0.85 μT / second, 0.8 μT / second, 0.75 μT / second, 0.7 μT / second, 0.65 μT / second, 0.6 μT / second, 0.55 μT / second, 0.5 μT / second, 0.45 μT / second, 0.4 μT / second, 0.35 μT / second, 0.3 μT / second, 0.25 μT / second, 0.2 μT / second, 0.15 μT / second, 0.1 μT / second, 0.095 μT / second, 0.09 μT / second, 0.08 μT / second, 0.075 μT / second, 0.07 μT / second, 0.065 μT / second, 0.06 μT / second, 0.055 μT / second, 0.05 μT / second, 0.045 μT / second, 0.04 μT / second, 0.035 μT / second, 0.03 μT / second, 0.025 μT / second, 0.02 μT / second, 0.015 μT / second, 0.01 μT / second or less, or within a range defined by any two of the foregoing values. The upper limit of the rate of change of the amplitude of the magnetic field can be determined by the capabilities of the device used to perform the sweep.

[0122] In some embodiments, when the magnetic field is within the upper and lower limits disclosed above, the spatial deviation of the magnetic field in volume during modulation is less than about half (or one quarter, or one eighth, or one tenth) of the magnitude of the magnetic field. For example, when the magnetic field strength is less than 2 μT (or greater than -2 μT), then the spatial deviation of the magnetic field in volume during modulation can be less than 1 μT. As another example, when the magnetic field strength is less than 10 μT (or greater than -10 μT), then the spatial deviation of the magnetic field in volume during modulation can be less than 5 μT. For example, the spatial deviation can be measured by performing at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500 or more spatial random samplings or spatially uniform distribution measurements of the magnetic field within the volume and calculating the standard deviation of the sampled magnetic field measurement results. Such uniformity can be achieved, for example, in a large uniform magnetic shield by having a large penetration solenoid passing through the magnetic shield or by using a large Helmholtz coil with a large uniform region for generating magnetic field amplitude modulation. In some embodiments, the modulation is a sweep of the magnetic field. In some embodiments, the magnetic field amplitude modulation includes non-adiabatic jumps, monotonic amplitude changes, or a combination thereof.

[0123] In some embodiments, after the polarization transfer step, the non-hydrogen nuclear spins of the bio-related contrast agent (such as the 13 C or 15N) having a nuclear spin polarization of at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% or more, and at most about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less, or a polarization within a range defined by any two of the foregoing values. For example, in some embodiments, after the polarization transfer step, the nuclear spin polarization of the non-hydrogen nuclei of the biorelevant contrast agent is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, between 20% and 50%, between 25% and 30%, between 25% and 35%, between 25% and 40%, between 25% and 45%, between 25% and 50%, between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 40% and 45%, between 40% and 50% or between 45% and 50%.

[0124] In some embodiments, this polarization is achieved for the following solution volumes: at least about 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, 10 ml, 20 ml, 30 ml, 40 ml, 50 ml, 60 ml, 70 ml, 80 ml, 90 ml, 100 ml, 200 ml, 300 ml, 400 ml, 500 ml or more, and at most about 500 ml, 400 ml, 300 ml, 200 ml, 100 ml, 90 ml, 80 ml, 70 ml, 60 ml, 50 ml, 40 ml, 30 ml, 20 ml, 10 ml, 9 ml, 8 ml, 7 ml, 6 ml, 5 ml, 4 ml, 3 ml, 2 ml, 1 ml or less, or a volume within a range defined by any two of the foregoing values.

[0125] In some embodiments, after polarization transfer, a portion of the population difference of the para-hydrogen proton spin states has been transferred to the polarization of the target (e.g., 13 C or 15 N) nuclear spins of a bio-related contrast agent. This portion can be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% or more, up to about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less, or within a range defined by any two of the foregoing values. For example, in some embodiments, this portion is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, between 20% and 50%, between 25% and 30%, between 25% and 35%, between 25% and 40%, between 25% and 45%, between 25% and 50%, between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 40% and 45%, between 40% and 50% or between 45% and 50%.

[0126] In some embodiments, the magnetic field modulation includes non-adiabatic jumps of the magnetic field. Non-adiabatic jumps can be made to a magnetic field in which an avoided crossing of energy levels including proton spins and non-proton spins occurs. Given the J-coupling between nuclear spins in a given system, this value can be determined by analytical calculations or by plotting the energy levels of the Hamiltonian for different magnetic fields and identifying the LAC. In some embodiments, the duration of the magnetic field amplitude being in the LAC condition is at most about 5 seconds, 4 seconds, 3 seconds, 2 seconds, 1 second, 0.9 seconds, 0.8 seconds, 0.7 seconds, 0.6 seconds, 0.5 seconds, 0.4 seconds, 0.3 seconds, 0.2 seconds, 0.1 seconds or less, at least about 0.1 seconds, 0.2 seconds, 0.3 seconds, 0.4 seconds, 0.5 seconds, 0.6 seconds, 0.7 seconds, 0.8 seconds, 0.9 seconds, 1 second, 2 seconds, 3 seconds, 4 seconds, 5 seconds or more or within a range defined by any two of the foregoing values.

[0127] In some embodiments, the modulation of the magnitude of the magnetic field includes changing the magnetic field magnitude monotonically (or monotonically over a finite number of intervals - such as one to ten increasing intervals and / or one to ten decreasing intervals). In some embodiments, the modulation of the magnitude of the magnetic field comprises linearly changing the magnitude of the magnetic field. The initial magnetic field magnitude, the ending magnetic field magnitude, and the total duration of the sweep can be optimized for the target molecule. In some embodiments, the magnetic field magnitude during the sweep is within a lower limit and an upper limit. The lower limit can be at least about -2 μT, -1.9 μT, -1.8 μT, -1.7 μT, -1.6 μT, -1.5 μT, -1.4 μT, -1.3 μT, -1.2 μT, -1.1 μT, -1 μT, -0.9 μT, -0.8 μT, -0.7 μT, -0.6 μT, -0.5 μT, -0.4 μT, -0.3 μT, -0.2 μT, -0.1 μT or more, up to about -0.1 μT, -0.2 μT, -0.3 μT, -0.4 μT, -0.5 μT, -0.6 μT, -0.7 μT, -0.8 μT, -0.9 μT, -1 μT, -1.1 μT, -1.2 μT, -1.3 μT, -1.4 μT, -1.5 μT, -1.6 μT, -1.7 μT, -1.8 μT, -1.9 μT, -2 μT or less, or within a range defined by any two of the foregoing values. The upper limit can be at least about 0.1 μT, 0.2 μT, 0.3 μT, 0.4 μT, 0.5 μT, 0.6 μT, 0.7 μT, 0.8 μT, 0.9 μT, 1 μT, 1.1 μT, 1.2 μT, 1.3 μT, 1.4 μT, 1.5 μT, 1.6 μT, 1.7 μT, 1.8 μT, 1.9 μT, 2 μT or more, up to about 2 μT, 1.9 μT, 1.8 μT, 1.7 μT, 1.6 μT, 1.5 μT, 1.4 μT, 1.3 μT, 1.2 μT, 1.1 μT, 1 μT, 0.9 μT, 0.8 μT, 0.7 μT, 0.6 μT, 0.5 μT, 0.4 μT, 0.3 μT, 0.2 μT, 0.1 μT or less, or within a range defined by any two of the foregoing values. In some embodiments, the duration of the modulation can be at least about 100 milliseconds, 200 milliseconds, 300 milliseconds, 400 milliseconds, 500 milliseconds, 600 milliseconds, 700 milliseconds, 800 milliseconds, 900 milliseconds, 1 second, 2 seconds, 3 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 20 seconds, 30 seconds or longer, up to about 30 seconds, 20 seconds, 10 seconds, 9 seconds, 8 seconds, 7 seconds, 6 seconds, 5 seconds, 4 seconds, 3 seconds, 2 seconds, 1 second, 900 milliseconds, 800 milliseconds, 700 milliseconds, 600 milliseconds, 500 milliseconds, 400 milliseconds, 300 milliseconds, 200 milliseconds, 100 milliseconds or less, or within a range defined by any two of the foregoing values. In some embodiments, the rate of change of the magnitude varies along the magnitude profile.In some embodiments, a constant adiabatic sweep is calculated by selecting a certain subset of the level-avoided crossings of a spin system. In some embodiments, the magnetic amplitude modulation includes a combination of non-adiabatic jumps, monotonic amplitude modulation, and sign changes in the rate of change. In some embodiments, precursors can be selected or designed such that, after hydrogenation and other potential chemical reactions, one of the products is a biologically relevant contrast agent useful for hyperpolarized NMR or MRI applications.

[0128] Hydrolysis, purification and separation

[0129] The present disclosure provides methods and systems for producing a composition (e.g., a clinical dose composition) comprising a hyperpolarized biologically relevant contrast agent (or a pharmaceutically acceptable salt thereof) in a solvent. In some embodiments, according to the present disclosure, the biologically relevant contrast agent is produced by additional chemical reactions and / or processing steps after hydrogenation and polarization transfer. Such additional chemical reactions and / or processing steps can include, but are not limited to: (i) catalyst filtration and removal (e.g., filtration to remove rhodium atoms and / or iridium atoms); (ii) cleavage of the side arm of a biologically relevant contrast agent precursor molecule (e.g., cleavage of a compound of formula IIa or formula IIb as described herein) to form a biologically relevant contrast agent and a side arm (e.g., a compound of formula IIIa or formula IIIb as described herein), e.g., by hydrolysis with an aqueous sodium hydroxide solution; (iii) washing the solution with an organic solvent and separating any resulting aqueous and organic phases; (iv) evaporation of volatile organic compounds extracted from the aqueous mixture (e.g., using nitrogen bubbling); and (v) additional filtration / purification / concentration / conditioning steps known in the art.

[0130] The volume of the solution comprising the bio-related contrast agent (e.g., after lysis) and / or the concentration of the bio-related contrast agent produced can depend on the volume of the solution used for polarization transfer and the concentration of the precursor in the solution. Exemplary ranges of solution volume and precursor concentration are described herein. As additional specific examples, a solution of at least about 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, 10 ml, 20 ml, 30 ml, 40 ml, 50 ml, 60 ml, 70 ml, 80 ml, 90 ml, 100 ml or more, at most about 100 ml, 90 ml, 80 ml, 70 ml, 60 ml, 50 ml, 40 ml, 30 ml, 20 ml, 10 ml, 9 ml, 8 ml, 7 ml, 6 ml, 5 ml, 4 ml, 3 ml, 2 ml, 1 ml or less, or an amount of solution within the range defined by any two of the foregoing values can be produced. In some embodiments, the solution can comprise at least about 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 200 mM, 300 mM, 400 mM, 500 mM or more of the bio-related contrast agent, at most about 500 mM, 400 mM, 300 mM, 200 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM or less of the bio-related contrast agent, or an amount of the bio-related contrast agent within the range defined by any two of the foregoing values.

[0131] In some embodiments, the present disclosure describes a multi-step liquid-liquid separation and purification process for producing a dosage (e.g., a clinical dosage) of a dosage composition comprising a bio-related contrast agent (e.g., a hyperpolarized bio-related contrast agent or a pharmaceutically acceptable salt thereof).

[0132] In some embodiments (i.e., for the PHIP-SAH procedure), the polarization step is followed by cleavage (e.g., via hydrolysis with an aqueous mixture) of the side arm from the target molecule precursor (e.g., a bio-related contrast agent precursor) to produce the target molecule (e.g., a bio-related contrast agent) and the unbound side arm (e.g., a compound of formula IIIa as described herein). In some embodiments, the polarization step is followed by cleavage of the side arm by mixing a solution (which includes a first organic solvent and the polarized product, e.g., a hyperpolarized bio-related contrast agent or a pharmaceutically acceptable salt thereof) with a hydrolyzing agent such as a base (e.g., sodium hydroxide) in an aqueous solution. In some embodiments, the first organic solvent and the aqueous mixture (e.g., water) produce a two-phase solution. In some embodiments, the first organic solvent and the aqueous mixture (e.g., water) produce a two-phase solution, wherein a portion of the organic solvent remains in the aqueous mixture. In some embodiments, the first organic solvent and the aqueous mixture (e.g., water) produce a partial mixture.

[0133] Clinically relevant purity

[0134] Consistent with the disclosed embodiments, the steps, methods, and systems described herein can separate hyperpolarized bio-related contrast agents from other substances in the original solution (e.g., catalysts, original solvents, reaction products, etc.). For example, most of the hydrogenation catalysts present in the original solution can be removed from the dosage composition. In some embodiments, the dosage composition can retain up to about 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001% or less of the hydrogenation catalyst, at least about 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1% or more of the hydrogenation catalyst, or an amount of the hydrogenation catalyst within a range defined by any two of the foregoing values. Similarly, the dosage composition can retain up to about 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001% or less of the cleavage by-products (e.g., side arms or other residues of cleavage), at least about 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1% or more of the cleavage by-products, or an amount of the cleavage by-products within a range defined by any two of the foregoing values.

[0135] In some embodiments, the methods and systems described herein produce a dose composition wherein the concentration of the hyperpolarized bio-related contrast agent is at least about 100 mM, 150 mM, 200 mM, 250 mM, 300 mM, 350 mM, 400 mM, 450 mM, 500 mM or more, up to about 500 mM, 450 mM, 400 mM, 350 mM, 300 mM, 250 mM, 200 mM, 150 mM, 100 mM or less, or within a range defined by any two of the foregoing values.

[0136] In some embodiments, the methods and systems described herein produce a dose composition wherein the polarization of the hyperpolarized bio-related contrast agent is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 40%, 45%, 50% or more, up to about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less, or a polarization within a range defined by any two of the foregoing values. For example, in some embodiments, the methods and systems described herein produce a dose composition wherein the polarization of the hyperpolarized bio-related contrast agent is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, between 20% and 50%, between 25% and 30%, between 25% and 35%, between 25% and 40%, between 25% and 45%, between 25% and 50%, between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 40% and 45%, between 40% and 50% or between 45% and 50%.

[0137] In some embodiments, the methods and systems described herein produce a dose composition, wherein the concentration of the catalyst, precursor, or cleavage by - product can each be at most about 1 μM, 900 nanomolar (nM), 800 nM, 700 nM, 600 nM, 500 nM, 400 nM, 300 nM, 200 nM, 100 nM, 90 nM, 80 nM, 70 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 9 nM, 8 nM, 7 nM, 6 nM, 5 nM, 4 nM, 3 nM, 2 nM, 1 nM or less, at least about 1 nM, 2 nM, 3 nM, 4 nM, 5 nM, 6 nM, 7 nM, 8 nM, 9 nM, 10 nM, 20 nM, 30 nM, 40 nM, 50 nM, 60 nM, 70 nM, 80 nM, 90 nM, 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, 600 nM, 700 nM, 800 nM, 900 nM, 1 μM or more, or within a range defined by any two of the foregoing values. The methods and systems described herein produce a dose composition, wherein the purity of the hyperpolarized bio - relevant contrast agent is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher, at most about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90% or lower, or within a range defined by any two of the foregoing values. In some embodiments, at least a portion of the hyperpolarized compound is separated from the cleaved side - arm or other reaction by - products (if such by - products are present).

[0138] Transportation

[0139] Consistent with the disclosed embodiments, polarization transfer and use of the bio - relevant contrast agent can occur at different locations. In some embodiments, the dose composition is transported to another location. In some embodiments, the dose composition is transported to another location. The disclosed embodiments are not necessarily limited to any particular transport distance or duration. Instead, the maximum distance or duration can be determined based on the target molecule, the degree of original or polarization, the desired final degree of polarization, and the transport conditions. In some embodiments, the dose composition is transported at least one meter in a suitable transport device.

[0140] Consistent with the disclosed embodiments, the transport device can be configured to transport samples of a precursor or a bio-related contrast agent. The transport device can be arranged and configured to transport one or more samples (e.g., one or more dose compositions) simultaneously. The transport device can include a transport chamber configured to receive one or more samples. The transport device can be configured to maintain the transport chamber within a predetermined temperature range and a predetermined magnetic field strength. The transport device can be configured to maintain one or more samples in a magnetic field of at least about 10 G, 20 G, 30 G, 40 G, 50 G, 60 G, 70 G, 80 G, 90 G, 100 G, 200 G, 300 G, 400 G, 500 G, 600 G, 700 G, 800 G, 900 G, 1,000 G or more, up to about 1,000 G, 900 G, 800 G, 700 G, 600 G, 500 G, 400 G, 300 G, 200 G, 100 G, 90 G, 80 G, 70 G, 60 G, 50 G, 40 G, 30 G, 20 G, 10 G or less, or in a magnetic field within a range defined by any two of the foregoing values.

[0141] A permanent magnet or an electromagnet included in the transport device can provide the magnetic field. In some embodiments, the permanent magnet or the electromagnet is shielded to reduce the magnetic field strength outside the transport device. The transport device can also include a cooling system. The cooling system can be configured to maintain the sample at a predetermined temperature or within a predetermined temperature range during transport. For example, the cooling system can be configured to maintain the sample at a temperature below 270 K, below 80 K, or below 4 K. In some embodiments, the transport device is configured to maintain the sample at approximately the temperature of liquid nitrogen. The transport device can include a thermal insulation layer between the cooling system and the exterior of the transport device to minimize heat exchange with the external environment. In some embodiments, the cooling system is configured to use a cold gas flow to maintain the temperature of the sample. In some embodiments, the cooling system is configured to use a liquid coolant to maintain the temperature of the sample. In some embodiments, the transport device includes a Dewar to provide cooling of the sample. To also distribute the hyperpolarized sample over large distances, the container can be transported by standard transport means such as airplanes, trains, trucks, cars, and ships.

[0142] In some embodiments, a dosage composition containing a hyperpolarized bio-related contrast agent is transported in a transport device. In some embodiments, the relaxation time of the hyperpolarized bio-related contrast agent in the transport device is at least about 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours or more, up to about 10 hours, 9 hours, 8 hours, 7 hours, 6 hours, 5 hours, 4 hours, 3 hours, 2 hours, 1 hour, 50 minutes, 40 minutes, 30 minutes, 20 minutes, 10 minutes, 9 minutes, 8 minutes, 7 minutes, 6 minutes, 5 minutes, 4 minutes, 3 minutes, 2 minutes, 1 minute or less, or a relaxation time within a range defined by any two of the foregoing values.

[0143] Generation of polarized biologically relevant contrast agents

[0144] Figure 1 Depicted is a first exemplary process 100 for generating a polarized bio-related contrast agent according to various embodiments. In some embodiments, at step 110, the first process 100 includes providing a composition that includes a compound of formula Ia. In some embodiments, the compound of formula Ia includes: a Z group that includes: (i) a carbon-carbon double bond (-C═C-) that is fully substituted with 2 H (deuterium, also known as D) (i.e., -CD═CD-); or (ii) a carbon-carbon triple bond (-C≡C-), as described herein; an R1 group that includes a PHIP transfer moiety as described herein; an R2 group that includes an optionally substituted hydrocarbon, alkoxy, primary amine, secondary amine, or tertiary amine or solubilizing moiety, as described herein; and an R3 group that includes a bio-related contrast agent, as described herein.

[0145] In some embodiments, at step 120, the double bond or triple bond in the compound of formula Ia is hydrogenated with secondary hydrogen to form a secondary hydrogenated derivative of the compound of formula Ia, wherein the secondary hydrogenated derivative is a compound having the structure of formula IIa. In some embodiments, the compound of formula IIa includes: Z', which is: (i) fully substituted to include 2The secondary-hydrogenated carbon-carbon single bond (-CH*-CH*-) of H (deuterium, also known as D), i.e., -CDH*-CDH*-; or (ii) the secondary-hydrogenated carbon-carbon double bond (-CH*=CH*-), where H* is a hydrogen having a spin order derived from secondary hydrogen; an R1 group that includes a PHIP transfer moiety as described herein; an R2 group that includes an optionally substituted hydrocarbon, alkoxy, primary amine, secondary amine, or tertiary amine, or a solubilizing moiety as described herein; and an R3 group that includes a biologically relevant contrast agent as described herein. In some embodiments, the compound of formula Ia is hydrogenated with secondary hydrogen using the hydrogenation process described herein.

[0146] In some embodiments, at step 130, as described herein, a polarization transfer waveform is applied to transfer the nuclear spin order from at least one H* in the side arm of the compound of formula IIa to any non-hydrogen nuclear spin in the biologically relevant contrast agent of the compound of formula IIa, thereby forming a derivative of the compound of formula IIa having a hyperpolarized biologically relevant contrast agent. In some embodiments, the nuclear spin order is transferred using any polarization transfer method described herein.

[0147] Figure 2 Depicted is a second exemplary process 200 for generating a polarized biologically relevant contrast agent according to various embodiments of the present disclosure. In some embodiments, at step 210, the second process includes providing a composition that includes a compound of formula IIa. In some embodiments, formula IIa includes: a Z' group that is: (i) fully substituted to include 2 The secondary-hydrogenated carbon-carbon single bond (-CH*-CH*-) of H (deuterium, also known as D), i.e., -CDH*-CDH*-; or (ii) the secondary-hydrogenated carbon-carbon double bond (-CH*=CH*-), as described herein, where H* is a hydrogen having a spin order derived from secondary hydrogen; an R1 group that includes a PHIP transfer moiety as described herein; an R2 group that includes an optionally substituted hydrocarbon, alkoxy, primary amine, secondary amine, or tertiary amine, or a solubilizing moiety as described herein; and an R3 group that includes a biologically relevant contrast agent as described herein.

[0148] In some embodiments, at step 220, as described herein, a polarization transfer waveform is applied to transfer the nuclear spin order from at least one H* in the side arm of the compound of formula IIa to any non-hydrogen nuclear spin in the biologically relevant contrast agent of the compound of formula IIa, thereby forming a derivative of the compound of formula IIa having a hyperpolarized biologically relevant contrast agent.

[0149] In some embodiments, at step 230, a derivative compound of formula IIa is hydrolyzed to form a composition that includes a hyperpolarized bio-related contrast agent and a separate side arm compound of formula IIIa. In some embodiments, the compound of formula IIIa includes: Z", which is: (i) a secondary hydrocarbyl-carbon single bond (-CH*-CH*-) that is fully substituted to include 2 H (deuterium, also known as D) (i.e., -CDH*-CDH*); or (ii) a secondary hydrocarbyl-carbon double bond (-CH*=CH*-), as described herein; an R1' group that includes a parahydrogen-induced polarization (PHIP) transfer moiety, as described herein; and an R2 group that includes an optionally substituted hydrocarbon, alkoxy, primary amine, secondary amine, or tertiary amine or a solubilizing moiety, as described herein.

[0150] In some embodiments, at step 240, the hyperpolarized bio-related contrast agent is washed one or more times with an organic solvent. In some embodiments, after the washing step, the non-hydrogen nuclear spins in the bio-related contrast agent have at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% or more, and at most about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less non-hydrogen spin polarization, or non-hydrogen spin polarization within a range defined by any two of the foregoing values.

[0151] In some embodiments, the first process or the second process includes one or more additional steps or operations. In some embodiments, the first process or the second process omits one or more steps or operations. In some embodiments, one or more steps or operations of the first process or the second process are combined. In some embodiments, all steps or operations of the first process or the second process are combined to produce a complete process for generating a hyperpolarized contrast agent from a precursor having the structure of formula Ia.

[0152] Figure 3Depicts a third exemplary process 300 for generating polarized bio-related contrast agents according to various embodiments. In the illustrated example, the process comprises providing, at step 310, a composition comprising a compound of formula Ib. In some embodiments, the compound of formula Ib comprises: a Z group comprising an ethynyl (-C≡C-) group, a fully deuterated prop-2-ynyl (-CD2-C≡C-) group, a fully deuterated vinyl (-CD=CD-) group, a fully deuterated prop-2-enyl (-CD2-CD=CD-) group, or a fully deuterated but-3-enyl (-CD2-CD2-CD=CD-) group, as described herein; an R2 group comprising an optionally substituted hydrocarbyl, alkyl, cycloalkyl, aryl, carboxyl, keto, or solubilizing moiety, as described herein; and an R3 group comprising an acyl derivative of a bio-related contrast agent, as described herein.

[0153] In some embodiments, at step 320, the double bond or triple bond in the compound of formula Ib is hydrogenated with parahydrogen to form a parahydrogenated derivative of the compound of formula Ib, wherein the parahydrogenated derivative is a compound having the structure of formula IIb. In some embodiments, the compound of formula IIb comprises a Z' group comprising a parahydrogenated vinyl (-CH*=CH*-) group, a fully deuterated parahydrogenated prop-2-enyl (-CD2-CH*=CH*-) group, a fully deuterated parahydrogenated ethane (-CDH*-CDH*-) group, a fully deuterated parahydrogenated propane (-CD2-CDH*-CDH*-) group, or a fully deuterated parahydrogenated butane (-CD2-CD2-CH*=CH*-) group, where H* is a hydrogen having a spin order derived from parahydrogen, as described herein; an R2 group comprising an optionally substituted hydrocarbyl, alkyl, cycloalkyl, aryl, carboxyl, keto, or alkoxy or solubilizing moiety, as described herein; and an R3 group comprising an acyl derivative of a bio-related contrast agent, as described herein. In some embodiments, the compound of formula Ib is hydrogenated with parahydrogen using a hydrogenation method described herein.

[0154] In some embodiments, at step 330, as described herein, a polarization transfer waveform is applied to transfer the nuclear spin order from at least one H* in the side arm of the compound of formula IIb to any non-hydrogen nuclear spin in the acyl derivative of the bio-related contrast agent of the compound of formula IIb, thereby forming a derivative of the compound of formula IIb having a hyperpolarized acyl derivative of the bio-related contrast agent. In some embodiments, the nuclear spin order is transferred using any polarization transfer method described herein.

[0155] Figure 4Depicts a fourth exemplary process 400 for generating polarized bio-related contrast agents in accordance with various embodiments of the present disclosure. In the illustrated example, the process includes providing, at step 410, a composition that includes a compound of Formula IIb. In some embodiments, Formula IIb includes a Z' group that includes a secondary vinyl (-CH*=CH*-) group, a fully deuterated secondary vinylidene (-CD2-CH*=CH*-) group, a fully deuterated secondary ethyl (-CDH*-CDH*-) group, a fully deuterated secondary propyl (-CD2-CDH*-CDH*-) group, or a fully deuterated butyl (-CD2-CD2-CH*=CH*-) group, as described herein, where H* is a hydrogen having a spin order derived from secondary hydrogen, as described herein; an R2 group that includes an optionally substituted hydrocarbon group, alkyl group, cycloalkyl group, aryl group, carboxyl group, ketone group, or alkoxy group or solubilizing moiety, as described herein; and an R3 group that includes an acyl derivative of a bio-related contrast agent, as described herein.

[0156] In some embodiments, at step 420, as described herein, a polarization transfer waveform is applied to transfer nuclear spin order from at least one H* in a side arm of the compound of Formula IIb to any non-hydrogen nuclear spin in the acyl derivative of the bio-related contrast agent of the compound of Formula IIb, thereby forming a derivative of the compound of Formula IIb having a hyperpolarized acyl derivative of the bio-related contrast agent.

[0157] In some embodiments, at step 430, the derivative compound of Formula IIb is hydrolyzed to form a composition that includes a hyperpolarized bio-related contrast agent and a separate side arm compound of Formula IIIb. In some embodiments, the compound of Formula IIIb includes a Z” group that includes a secondary vinyl (-CH*=CH*-) group, a fully deuterated secondary vinylidene (-CD2-CH*=CH*-) group, a fully deuterated secondary ethyl (-CDH*-CDH*-) group, a fully deuterated secondary propyl (-CD2-CDH*-CDH*-) group, or a fully deuterated secondary butyl (-CD2-CD2-CH*=CH*-) group, as described herein; and an R2 group that includes an optionally substituted hydrocarbon group, alkyl group, cycloalkyl group, aryl group, carboxyl group, ketone group, or alkoxy group or solubilizing moiety, as described herein.

[0158] In some embodiments, at step 440, the hyperpolarized bio-related contrast agent is washed one or more times with an organic solvent. In some embodiments, after the washing step, the non-hydrogen nuclear spins in the bio-related contrast agent have at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% or more, and at most about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less non-hydrogen spin polarization, or non-hydrogen spin polarization within a range defined by any two of the foregoing values.

[0159] In some embodiments, the third process or the fourth process includes one or more additional steps or operations. In some embodiments, the third process or the fourth process omits one or more steps or operations. In some embodiments, one or more steps or operations of the third process or the fourth process are combined. In some embodiments, all steps or operations of the third process or the fourth process are combined to produce a complete process for generating a hyperpolarized contrast agent from a precursor having the structure of formula Ib.

[0160] Examples

[0161] Example 1: Polarization transfer in non-deuterated, partially deuterated and fully deuterated pyruvate esters derivatives

[0162] To evaluate the effect of the deuteration level on the PHIP transfer moiety in pyruvate esters, the following ester side-arm derivatives of pyruvate esters were prepared: (1) tert-butyl 4-((2-oxopropanoyl)oxy)but-2-ynoate (CH2 ester), (2) tert-butyl 4-((2-oxopropanoyl)oxy)but-2-ynoate-D1 (CDH ester), and (3) tert-butyl 4-((2-oxopropanoyl)oxy)but-2-ynoate-D2 (CD2 ester). Compounds (1), (2), and (3) were prepared at natural abundance 13 C (about 1.1%). Compounds (1), (2), and (3) were reacted with parahydrogen. The reactions respectively produced parahydrogenated compounds: (4) (Z)-tert-butyl 4-((2-oxopropanoyl)oxy)but-2-enoate, (5) (Z)-tert-butyl 4-((2-oxopropanoyl)oxy)but-2-enoate-D1, and (6) (Z)-tert-butyl 4-((2-oxopropanoyl)oxy)but-2-enoate-D2. The singlet relaxation times T s . Figure 5 Exemplary singlet lifetimes of compounds (4), (5), and (6) are shown. As Figure 5As shown, compared to the non-deuterated PHIP transfer moiety (Compound (4)), full deuteration of the PHIP transfer moiety (Compound (6)) results in a singlet relaxation time T s that is increased by more than 8-fold.

[0163] Without limitation, it is believed that the increased singlet relaxation time T s is the most important factor in the performance improvement achieved by the fully deuterated compounds described herein. Without limitation, the increased singlet relaxation time T s increase can transfer the spin order associated with para-hydrogen to other nuclei (such as the 13 C or 15 N nuclei) described herein. Without limitation, such increased polarization transfer times allow for more efficient transfer of spin order to such nuclei.

[0164] Figure 6A An exemplary polarization transfer pulse sequence is shown that is used to transfer the spin order from the para-hydrogen associated protons in Compounds (4), (5), and (6) to the natural abundance 13 C nuclei present in Compounds (4), (5), and (6). Compounds (4), (5), and (6) are placed in a μ-metal magnetic shield and subjected to a magnetic field supplied by a solenoid coil. As Figure 6A shown, the magnitude B1 of the magnetic field is subjected to a first amplitude modulation by linearly changing the magnetic field strength from 1.0 μT to 1.3 μT. During the first amplitude modulation, the magnetic field B1 is linearly polarized and oscillates at the Larmor frequency of the target 13 C nuclei. The first amplitude modulation is performed within a first time period t 扫掠 . During the first amplitude modulation, magnetization in the target 扫掠 C is built up within the first time period t 13 .

[0165] After the first time period t 扫掠 , the magnitude B1 of the magnetic field is subjected to a second amplitude modulation by linearly changing the magnetic field strength from 1.3 μT to 0 μT. During the second amplitude modulation, the magnetic field B1 is linearly detuned from the Larmor frequency of the target 13 C nuclei.

[0166] Figure 6B Shows the Figure 6A polarization transfer pulse sequence t 扫掠 of Compound (6) achieved using various first time periods. As 13 shown, Figure 6B the 13 C polarization is between about 10 seconds and about 12 seconds t 扫掠reaches a maximum during the first time period between them.

[0167] Figure 7A shows the Figure 6A and 6B 13C polarization level of compound (6) after the polarization transfer procedure of 13 As Figure 7A shown, a 13C polarization level of up to 36% is achieved. 13 13C polarization level.

[0168] Figure 7B shows the highest achieved 13C polarization levels of compounds (4), (5), and (6). As 13 shown, Figure 7B the 13 13C polarization level increases with increasing deuteration.

[0169] Example 2: Polarization transfer in deuterated lactate esters derivatives

[0170] To evaluate the effect of the deuteration level on the PHIP transfer moiety in the lactate ester, compound (7), an ester side-arm derivative of the lactate ester, was prepared. Compound (7) was prepared at natural abundance 13C (ca. 1.1%). Compound (7) was hydrogenated in an NMR pressure tube with hydrogen enriched to >90% parahydrogen at a flow rate of 0.5 standard liters per minute at 10 bar pressure. 50 mM of compound (7) was reacted with parahydrogen in acetone-d6 in the presence of a 2.5 mM Rh(dppb)(COD)BF4 catalyst solution. The reaction produced the parahydrogenated derivative of compound (7). Polarization transfer was carried out using a transverse magnetic field at the Larmor frequency of 13C at a static field of 50 μT. The field was ramped from 1.8 μT to 2.2 μT in 9 seconds. 13 13C 13 13C Figure 8 shows the 13 13C polarization level of the parahydrogenated compound (7). As Figure 8 shown, a 13C polarization level of up to 31% is achieved. 13 13C polarization level.

[0171] Compound (7):

[0172] Example 3: Polarization transfer in deuterated monoethyl ketoglutarate derivatives

[0173] To evaluate the effect of the deuteration level on the PHIP transfer moiety in the monethyl ketoglutarate, compound (8), an ester side-arm derivative of the monethyl ketoglutarate, was prepared. Compound (8) was prepared at natural abundance 13C 13Prepared with C (about 1.1%). The compound (8) was hydrogenated in an NMR pressure tube with >90% parahydrogen enrichment at a flow rate of 0.5 standard liters per minute under a pressure of 10 bar using hydrogen. 50 mM of compound (8) was reacted with parahydrogen in acetone-d6 in the presence of a 2.5 mM Rh(dppb)(COD)BF4 catalyst solution. The reaction produced a parahydrogenated derivative of compound (8). Polarization transfer was carried out using a transverse magnetic field at the Larmor frequency of 13 C in a static field of 50 μT. The field was ramped from 1.8 μT to 2.2 μT in 9 seconds. Figure 8 Shows the 13 C polarization level of the parahydrogenated compound (8). As Figure 8 shown, a C polarization level of up to 31% was achieved. As 13 shown, a C polarization level of up to 21% was achieved. Figure 9 shown, a C polarization level of up to 21% was achieved. 13 C polarization level.

[0174] Compound (8):

[0175] Example 4: Polarization transfer in Z-OMPD monomethyl ester derivatives

[0176] To evaluate the effect of the deuteration level on the PHIP transfer moiety in Z-OMPD monomethyl ester, compound (9), an ester side-arm derivative of Z-OMPD monomethyl ester, was prepared. Compound (9) was prepared with natural abundance 13 C (about 1.1%). The compound (9) was hydrogenated in an NMR pressure tube with >90% parahydrogen enrichment at a flow rate of 0.5 standard liters per minute under a pressure of 10 bar using hydrogen. 220 mM of compound (9) was reacted with parahydrogen in acetone-d6 in the presence of a 2.5 mM Rh(dppb)(COD)BF4 catalyst solution. The reaction produced a parahydrogenated derivative of compound (9). Polarization transfer was carried out using a transverse magnetic field at the Larmor frequency of 13 C in a static field of 50 μT. The field was ramped from 1.8 μT to 2.2 μT in 9 seconds. Figure 8 Shows the 13 C polarization level of the parahydrogenated compound (9). Figure 10 Shows the 13 C polarization level of the parahydrogenated compound (9). As Figure 10 shown, a C polarization level of up to 24% was achieved. 13 C polarization level.

[0177] Compound (9):

[0178] Description of the examples

[0179] Example 1. A composition comprising a compound of formula Ia:

[0180]

[0181] Wherein:

[0182] Z comprises a carbon-carbon double bond (-CD=CD-) or a carbon-carbon triple bond (-C≡C-) that is fully substituted to include deuterium (D);

[0183] R1 comprises a parahydrogen-induced polarization (PHIP) transfer moiety;

[0184] R2 comprises an optionally substituted hydrocarbon or alkoxy group; and

[0185] R3 comprises a bio-related contrast agent that comprises a non-hydrogen nuclear spin.

[0186] Example 2. A composition comprising a compound of formula IIa:

[0187]

[0188] Wherein:

[0189] Z' is a secondary hydrogenated carbon-carbon single bond (-CDH*-CDH*-) or a secondary hydrogenated carbon-carbon double bond (-CH*=CH*-) that is fully substituted to include deuterium;

[0190] Wherein H* is a hydrogen having a spin order derived from parahydrogen;

[0191] R1 comprises a parahydrogen-induced polarization (PHIP) transfer moiety;

[0192] R2 comprises an optionally substituted hydrocarbon or alkoxy group; and

[0193] R3 comprises a bio-related contrast agent that comprises a non-hydrogen nuclear spin.

[0194] Example 3. A composition comprising: (i) a bio-related contrast agent that comprises a non-hydrogen nuclear spin; and (ii) a compound of formula IIIa:

[0195]

[0196] Wherein:

[0197] Z” is a secondary hydrogenated carbon-carbon single bond (-CDH*-CDH*-) or a secondary hydrogenated carbon-carbon double bond (-CH*=CH*-) that is fully substituted to include deuterium;

[0198] Wherein H* is a hydrogen having a spin order derived from parahydrogen;

[0199] R1' includes a secondary hydrogen induced polarization (PHIP) transfer moiety; and

[0200] R2 includes an optionally substituted hydrocarbon or alkoxy group.

[0201] Example 4. A composition comprising: (i) a hyperpolarized bio-related contrast agent, said hyperpolarized bio-related contrast agent comprising a non-hydrogen nuclear spin; and (ii) a compound of formula IVa:

[0202]

[0203] wherein:

[0204] Z includes a carbon-carbon double bond (-CD=CD-) or a carbon-carbon triple bond (-C≡C-) that is fully substituted to include deuterium;

[0205] R1' includes a secondary hydrogen induced polarization (PHIP) transfer moiety; and

[0206] R2 includes an optionally substituted hydrocarbon or alkoxy group.

[0207] Example 5. The composition according to any one of Examples 1 to 4, wherein the PHIP transfer moiety comprises a fully deuterated C1 hydrocarbon (-CD2-) or a fully deuterated C2 hydrocarbon (-CD2-CD2-).

[0208] Example 6. The composition according to any one of Examples 1 to 5, wherein:

[0209] the PHIP transfer moiety comprises *CR4R5, *CR4Y, *C=Y or any fully deuterated version thereof;

[0210] *C is 12 C or 13 a C carbon isotope;

[0211] R4 and R5 are each independently selected from: deuterium, fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl and fully deuterated haloalkyl; and

[0212] Y is selected from the group consisting of spin-1 / 2 atoms and spin-1 / 2 atoms covalently bonded to one or more chemical moieties selected from: fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, halogen or fully deuterated haloalkyl, or heteroatoms such as N, O, S, etc., said heteroatoms being optionally substituted by fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, halogen or fully deuterated haloalkyl.

[0213] Example 7. The composition according to any one of Examples 1 to 5, wherein:

[0214] The PHIP transfer moiety comprises *CR6R7–*CR8R9 or any fully deuterated version thereof;

[0215] *C is 12 C or 13 a C carbon isotope; and

[0216] R6, R7, R8 and R9 are each independently selected from: deuterium, fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl and fully deuterated haloalkyl.

[0217] Example 8. The composition according to any one of Examples 1 to 5, wherein:

[0218] The PHIP transfer moiety comprises *CH2, *CH2–*CH2, *CHY, *C=Y or any fully deuterated version thereof;

[0219] wherein H* is a hydrogen having a spin order derived from secondary hydrogen;

[0220] *C is 12 C or 13 a C carbon isotope; and

[0221] Y is selected from the group consisting of spin-1 / 2 atoms and spin-1 / 2 atoms covalently bonded to one or more chemical moieties selected from: fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, halogen or fully deuterated haloalkyl, or heteroatoms such as N, O, S, etc., said heteroatoms being optionally substituted by fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, halogen or fully deuterated haloalkyl.

[0222] Example 9. The composition according to Example 6 or 8, wherein the spin-1 / 2 atom is selected from: 1 H, 13 C, 15 N, 19 F or 31 P.

[0223] Example 10. The composition according to any one of Examples 1 to 9, wherein the PHIP transfer moiety comprises at least one atom having a J-coupling with the non-hydrogen nuclear spin of at least 0.1 Hertz (Hz).

[0224] Example 11. The composition according to any one of Examples 1 to 10, wherein Z comprises at least one atom having a J-coupling with the non-hydrogen nuclear spin of at least 0.1 Hertz (Hz).

[0225] Example 12. The composition according to any one of Examples 1 to 11, wherein R2 contains a solubilizing moiety.

[0226] Example 13. The composition according to any one of Examples 1 to 12, wherein R2 contains a hydrophobic and / or organophilic moiety.

[0227] Example 14. The composition according to Example 13, wherein R2 contains an organic solubilizing moiety.

[0228] Example 15. The composition according to any one of Examples 1 to 12, wherein R2 contains a hydrophilic and / or organophobic moiety.

[0229] Example 16. The composition according to any one of Examples 1 to 15, wherein R2 is selected from: methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, hydroxy, methanolyl, ethanolyl, n-propanolyl, isopropanolyl, propionate alcoholyl, n-butanolyl, sec-butanolyl, tert-butanolyl, isobutanolyl, methoxy, ethoxy, propoxy, isopropoxy, propionate, butoxy, tert-butoxy, sec-butoxy, ester group, phenyl, substituted phenyl, primary amine group, secondary amine group, tertiary amine group, primary amide group, secondary amide group and tertiary amide group.

[0230] Example 17. The composition according to any one of Examples 1 to 16, wherein the biorelevant contrast agent comprises a compound of the formula R 10 C(=O)X–; wherein R 10 is selected from straight-chain, branched-chain or cyclic C1-C10 alkyl, wherein one or more C atoms are optionally replaced by C=C, CO, COH, CNH2, COOH, CH2COOH, CONH2, OC(=O); and X is selected from NR 11, S, and O; where R 11 is selected from hydrogen and amino protecting groups, and is optionally selected from trifluoroacetyl, acetyl, benzoyl, benzyloxycarbonyl, tert-butyl carbonate, and benzyl.

[0231] Example 18. The composition according to any one of Examples 1 to 17, wherein the bio-related contrast agent is selected from: pyruvate, glutamate, glutamine, lactate, acetate, acetoacetate, ketoacetate, alanine, fructose, fumarate, bicarbonate, urea, dehydroascorbate, α-ketoglutarate, dihydroxyacetone, glucose, ascorbate, and their conjugate acids.

[0232] Example 19. The composition according to any one of Examples 1 to 18, wherein the solubility of the composition in water is less than 50 millimoles (mM).

[0233] Example 20. The composition according to any one of Examples 1 to 19, wherein the composition is reacted with secondary hydrogen such that the chemical yield of the secondary hydrogenated product is at least 30%.

[0234] Example 21. The composition according to any one of Examples 1 to 20, which is used in the process of parahydrogen induced polarization (PHIP).

[0235] Example 22. A method for preparing a hyperpolarized bio-related contrast agent or a pharmaceutically acceptable salt thereof, the method comprising:

[0236] (a) providing a composition comprising a compound of formula Ia:

[0237]

[0238] wherein:

[0239] Z comprises a carbon-carbon double bond (-CD=CD-) or a carbon-carbon triple bond (-C≡C-) that is fully substituted to include deuterium;

[0240] R1 comprises a parahydrogen induced polarization (PHIP) transfer moiety;

[0241] R2 comprises an optionally substituted hydrocarbon or alkoxy group; and

[0242] R3 comprises a bio-related contrast agent comprising a non-hydrogen nuclear spin;

[0243] (b) hydrogenating the double bond or the triple bond in the compound of formula Ia with parahydrogen to form a parahydrogenated derivative of the compound of formula Ia having the structure of formula IIa:

[0244]

[0245] Wherein:

[0246] Z' is a secondary hydrocarbyl-carbon single bond (-CDH*-CDH*-) or a secondary hydrocarbyl-carbon double bond (-CH*=CH*-) that is fully substituted to include deuterium;

[0247] wherein H* is a hydrogen having a spin order derived from secondary hydrogen;

[0248] R1 comprises a secondary hydrogen induced polarization (PHIP) transfer moiety;

[0249] R2 comprises an optionally substituted hydrocarbon or alkoxy; and

[0250] R3 comprises a bio-related contrast agent, the bio-related contrast agent comprising a non-hydrogen nuclear spin; and

[0251] (c) Applying a polarization transfer waveform to transfer the nuclear spin order from at least one H* in the compound of formula IIa to the non-hydrogen nuclear spin, thereby forming a derivative of formula IIa having a hyperpolarized bio-related contrast agent.

[0252] Example 23. A method for preparing a hyperpolarized bio-related contrast agent or a pharmaceutically acceptable salt thereof, the method comprising:

[0253] (a) Providing a composition, the composition comprising a compound of formula IIa:

[0254]

[0255] Wherein:

[0256] Z' is a secondary hydrocarbyl-carbon single bond (-CDH*-CDH*-) or a secondary hydrocarbyl-carbon double bond (-CH*=CH*-) that is fully substituted to include deuterium;

[0257] wherein H* is a hydrogen having a spin order derived from secondary hydrogen;

[0258] R1 comprises a secondary hydrogen induced polarization (PHIP) transfer moiety;

[0259] R2 comprises an optionally substituted hydrocarbon or alkoxy; and

[0260] R3 comprises a bio-related contrast agent, the bio-related contrast agent comprising a non-hydrogen nuclear spin; and

[0261] (b) Applying a polarization transfer waveform to transfer the nuclear spin order from at least one H* in the compound of formula IIa to the non-hydrogen nuclear spin, thereby forming a derivative of formula IIa having a hyperpolarized bio-related contrast agent.

[0262] Example 24. The method according to Example 22 or 23, further comprising hydrolyzing the derivative of formula IIa to provide a composition comprising: (i) a hyperpolarized bio-related contrast agent comprising a non-hydrogen nuclear spin; and (ii) a compound of formula IIIa:

[0263]

[0264] Wherein:

[0265] Z” is a secondary hydrocarbyl-carbon single bond (-CDH*-CDH*-) or a secondary hydrocarbyl-carbon double bond (-CH*=CH*-) that is fully substituted to include deuterium;

[0266] Where H* is a hydrogen having a spin order derived from secondary hydrogen;

[0267] R1' comprises a parahydrogen-induced polarization (PHIP) transfer moiety; and

[0268] R2 comprises an optionally substituted hydrocarbon or alkoxy group.

[0269] Example 25. The method according to Example 24, further comprising washing the hyperpolarized bio-related contrast agent one or more times with an organic solvent.

[0270] Example 26. The method according to Example 25, wherein after the washing step, the non-hydrogen nuclear spin polarization of the non-hydrogen nuclear spin is greater than 10%.

[0271] Example 27. The method according to any one of Examples 22 to 26, wherein the PHIP transfer moiety comprises a fully deuterated C1 hydrocarbon (-CD2-) or a fully deuterated C2 hydrocarbon (-CD2-CD2-).

[0272] Example 28. The method according to any one of Examples 22 to 26, wherein:

[0273] The PHIP transfer moiety comprises *CR4R5, *CR4Y, *C=Y or any fully deuterated version thereof;

[0274] *C is 12 C or 13 C carbon isotope;

[0275] R4 and R5 are each independently selected from: deuterium, fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl and fully deuterated haloalkyl; and

[0276] Y is selected from: spin-1 / 2 atoms and spin-1 / 2 atoms covalently bonded to one or more chemical moieties selected from: fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls, halogens or fully deuterated haloalkyls, or heteroatoms such as N, O, S, etc., which heteroatoms are optionally substituted with fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls, halogens or fully deuterated haloalkyls.

[0277] Example 29. The method according to any one of Examples 22 to 26, wherein:

[0278] The PHIP transfer moiety comprises *CR6R7–*CR8R9 or any fully deuterated version thereof;

[0279] *C is 12 C or 13 a C carbon isotope; and

[0280] R6, R7, R8 and R9 are each independently selected from the group consisting of: deuterium, fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls and fully deuterated haloalkyls.

[0281] Example 30. The method according to any one of Examples 22 to 26, wherein:

[0282] The PHIP transfer moiety comprises *CH2, *CH2–*CH2, *CHY, *C=Y or any fully deuterated version thereof;

[0283] *C is 12 C or 13 a C carbon isotope; and

[0284] Y is selected from: spin-1 / 2 atoms and spin-1 / 2 atoms covalently bonded to one or more chemical moieties selected from: fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls, halogens or fully deuterated haloalkyls, or heteroatoms such as N, O, S, etc., which heteroatoms are optionally substituted with fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls, halogens or fully deuterated haloalkyls.

[0285] Example 31. The method according to Example 28 or 30, wherein the spin-1 / 2 atom is selected from: 1 H, 13 C, 15 N, 19 F or 31 P.

[0286] Example 32. The method according to any one of Examples 22 to 31, wherein the PHIP transfer moiety comprises at least one atom having a J-coupling with the non-hydrogen nuclear spin of at least 0.1 Hertz (Hz).

[0287] Example 33. The method according to any one of Examples 22 to 32, wherein Z or Z' comprises at least one atom having a J-coupling with the non-hydrogen nuclear spin of at least 0.1 Hertz (Hz).

[0288] Example 34. The method according to any one of Examples 22 to 33, wherein R2 contains a solubilizing moiety.

[0289] Example 35. The method according to any one of Examples 22 to 34, wherein R2 contains a hydrophobic and / or organophilic moiety.

[0290] Example 36. The method according to Example 35, wherein R2 contains an organic solubilizing moiety.

[0291] Example 37. The method according to any one of Examples 22 to 33, wherein R2 contains a hydrophilic and / or organophobic moiety.

[0292] Example 38. The method according to any one of Examples 22 to 37, wherein R2 is selected from: methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, hydroxy, methanolyl, ethanolyl, n-propanolyl, isopropanolyl, propionate alcoholyl, n-butanolyl, sec-butanolyl, tert-butanolyl, isobutanolyl, methoxy, ethoxy, propoxy, isopropoxy, propionate, butoxy, tert-butoxy, sec-butoxy, ester group, phenyl, substituted phenyl, primary amine group, secondary amine group, tertiary amine group, primary amide group, secondary amide group and tertiary amide group.

[0293] Example 39. The method according to any one of Examples 22 to 38, wherein the bio-related contrast agent comprises a compound of the formula R 10 C(=O)X–; wherein R 10 is selected from straight-chain, branched-chain or cyclic C1-C10 alkyl, wherein one or more C atoms are optionally replaced by C=C, CO, COH, CNH2, COOH, CH2COOH, CONH2, OC(=O); and X is selected from NR 11, S, and O; wherein R 11 is selected from hydrogen and an amino protecting group, and is optionally selected from trifluoroacetyl, acetyl, benzoyl, benzyloxycarbonyl, tert-butyl carbonate, and benzyl.

[0294] Example 40. The method according to any one of Examples 22 to 39, wherein the bio-related contrast agent is selected from pyruvate, glutamate, glutamine, lactate, acetate, acetoacetate, ketoacetate, alanine, fructose, fumarate, bicarbonate, urea, dehydroascorbate, α-ketoglutarate, dihydroxyacetone, glucose, ascorbate, and their conjugate acids.

[0295] Example 41. A hyperpolarized bio-related contrast agent or a pharmaceutically acceptable salt thereof, produced by the method according to any one of Examples 22 to 40.

[0296] Example 42. A composition comprising a compound of formula Ib:

[0297]

[0298] wherein:

[0299] Z contains an ethynyl (-C≡C-) group, a fully deuterated prop-2-ynyl (-CD2-C≡CD2-) group, a fully deuterated but-3-ynyl (-CD2-CD2-C≡C-) group, a fully deuterated vinyl (-CD=CD-) group, a fully deuterated prop-2-enyl (-CD2-CD=CD-) group, or a fully deuterated but-3-enyl (-CD2-CD2-CD=CD-) group;

[0300] R1 contains an optionally substituted hydrocarbon group, alkyl group, cycloalkyl group, aryl group, carboxyl group, keto group, or alkoxy group; and

[0301] R2 contains an acyl derivative of a bio-related contrast agent, the bio-related contrast agent containing a non-hydrogen nuclear spin.

[0302] Example 43. A composition comprising a compound of formula IIb:

[0303]

[0304] wherein:

[0305] Z' comprises a vinylidene (-CH*=CH*-) group, a deuterated vinylidene prop-2-enyl (-CD2-CH*=CH*-) group, a deuterated vinylidene but-3-enyl (-CD2-CD2-CH*=CH*-) group, a deuterated ethylidene (-CDH*-CDH*-) group, a deuterated propylidene (-CD2-CDH*-CDH*-) group or a deuterated butylidene (-CD2-CD2-CDH*-CDH*-) group;

[0306] wherein H* is a hydrogen having a spin order derived from secondary hydrogen;

[0307] R1 comprises an optionally substituted hydrocarbon group, an alkyl group, a cycloalkyl group, an aryl group, a carboxyl group, a ketone group or an alkoxy group; and

[0308] R2 comprises an acyl derivative of a bio-related contrast agent, the bio-related contrast agent comprising a non-hydrogen nuclear spin.

[0309] Example 44. A composition comprising: (i) a bio-related contrast agent comprising a non-hydrogen nuclear spin; and (ii) a compound of formula IIIb:

[0310]

[0311] wherein:

[0312] Z” comprises a vinylidene (-CH*=CH*-) group, a deuterated vinylidene prop-2-enyl (-CD2-CH*=CH*-) group, a deuterated vinylidene but-3-enyl (-CD2-CD2-CH*=CH*-) group, a deuterated ethylidene (-CDH*-CDH*-) group, a deuterated propylidene (-CD2-CDH*-CDH*-) group or a deuterated butylidene (-CD2-CD2-CDH*-CDH*-) group;

[0313] wherein H* is a hydrogen having a spin order derived from secondary hydrogen; and

[0314] R1 comprises an optionally substituted hydrocarbon group, an alkyl group, a cycloalkyl group, an aryl group, a carboxyl group, a ketone group or an alkoxy group.

[0315] Example 45. The composition according to any one of Examples 42 to 44, wherein the composition further comprises a PHIP transfer moiety between the Z, Z' and Z” moieties and the sulfur atom, the PHIP transfer moiety comprising a fully deuterated C1 hydrocarbon (-CD2-) or a fully deuterated C2 hydrocarbon (-CD2-CD2-).

[0316] Example 46. The composition according to Example 45, wherein:

[0317] The PHIP transfer moiety comprises *CR3R4, *CR3Y, *C═Y or any fully deuterated version thereof;

[0318] *C is 12 C or 13 a C carbon isotope;

[0319] R3 and R4 are each independently selected from: deuterium, fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl and fully deuterated haloalkyl; and

[0320] Y is selected from the group consisting of: spin-1 / 2 atoms and spin-1 / 2 atoms covalently bonded to one or more chemical moieties selected from: fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, halogen or fully deuterated haloalkyl, or heteroatoms such as N, O, S, etc., said heteroatoms being optionally substituted by fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, halogen or fully deuterated haloalkyl.

[0321] Example 47. The composition according to Example 45, wherein:

[0322] The PHIP transfer moiety comprises *CR5R6–*CR7R8 or any fully deuterated version thereof;

[0323] *C is 12 C or 13 a C carbon isotope; and

[0324] R5, R6, R7 and R8 are each independently selected from: deuterium, fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl and fully deuterated haloalkyl.

[0325] Example 48. The composition according to Example 45, wherein:

[0326] The PHIP transfer moiety comprises *CH2, *CH2–*CH2, *CHY, *C═Y or any fully deuterated version thereof;

[0327] *C is 12 C or 13 a C carbon isotope; and

[0328] Y is selected from the group consisting of: spin-1 / 2 atoms and spin-1 / 2 atoms covalently bonded to one or more chemical moieties selected from the following: fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls, halogens or fully deuterated haloalkyls, or heteroatoms such as N, O, S, etc., said heteroatoms being optionally substituted by fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls, halogens or fully deuterated haloalkyls.

[0329] Example 49. The composition according to Example 46 or 48, wherein the spin-1 / 2 atom is selected from: 1 H, 13 C, 15 N, 19 F or 31 P.

[0330] Example 50. The composition according to any one of Examples 45 to 49, wherein the PHIP transfer moiety comprises at least one atom having a J-coupling with the non-hydrogen nuclear spin of at least 0.1 Hertz (Hz).

[0331] Example 51. The composition according to any one of Examples 42 to 50, wherein Z comprises at least one atom having a J-coupling with the non-hydrogen nuclear spin of at least 0.1 Hertz (Hz).

[0332] Example 52. The composition according to any one of Examples 42 to 51, wherein R1 comprises a solubilizing moiety.

[0333] Example 53. The composition according to any one of Examples 42 to 52, wherein R1 comprises a hydrophobic and / or organophilic moiety.

[0334] Example 54. The composition according to Example 53, wherein R1 comprises an organic solubilizing moiety.

[0335] Example 55. The composition according to any one of Examples 42 to 52, wherein R1 comprises a hydrophilic and / or organophobic moiety.

[0336] Example 56. The composition according to any one of Examples 42 to 55, wherein R1 is selected from: methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, hydroxy, methanol group, ethanol group, n-propanol group, isopropanol group, propionic acid alcohol group, n-butanol group, sec-butanol group, tert-butanol group, isobutanol group, methoxy, ethoxy, propoxy, isopropoxy, propionate group, butoxy, tert-butoxy, sec-butoxy, ester group, phenyl, substituted phenyl, primary amine group, secondary amine group, tertiary amine group, primary amide group, secondary amide group, tertiary amide group, and ketone group.

[0337] Example 57. The composition according to any one of Examples 42 to 56, wherein the bio-related contrast agent comprises a compound of the formula R9C(=O)O–; wherein R9 is selected from straight-chain, branched-chain or cyclic C1-C10 alkyl, and one or more C atoms are optionally replaced by C=C, CO, COH, CNH2, COOH, CH2COOH, CONH2 or OC(=O).

[0338] Example 58. The composition according to any one of Examples 42 to 57, wherein the bio-related contrast agent is selected from: pyruvate, glutamate, glutamine, lactate, acetate, acetoacetate, ketoacetate, alanine, fructose, fumarate, bicarbonate, and their conjugate acids.

[0339] Example 59. The composition according to any one of Examples 42 to 58, wherein the solubility of the composition in water is less than 50 millimoles (mM).

[0340] Example 60. The composition according to any one of Examples 42 to 59, wherein the composition is reacted with parahydrogen such that the chemical yield of the parahydrogenated product is at least 30%.

[0341] Example 61. The composition according to any one of Examples 42 to 60, which is used in a parahydrogen induced polarization (PHIP) process.

[0342] Example 62. A method for preparing a hyperpolarized bio-related contrast agent or a pharmaceutically acceptable salt thereof, the method comprising:

[0343] (a) providing a composition comprising a compound of formula Ib:

[0344]

[0345] wherein:

[0346] Z contains an ethynyl (-C≡C-) group, a fully deuterated prop-2-ynyl (-CD2-C≡C-) group, a fully deuterated but-3-ynyl (-CD2-CD2-C≡C-) group, a fully deuterated vinyl (-CD=CD-) group, a fully deuterated prop-2-enyl (-CD2-CD=CD-) group or a fully deuterated but-3-enyl (-CD2-CD2-CD=CD-) group;

[0347] R1 contains an optionally substituted hydrocarbon group, an alkyl group, a cycloalkyl group, an aryl group, a carboxyl group, a ketone group or an alkoxy group; and

[0348] R2 contains an acyl derivative of a bio-related contrast agent, the bio-related contrast agent containing a non-hydrogen nuclear spin;

[0349] (b) Hydrogenating the double bond or the triple bond in the compound of formula Ib with secondary hydrogen to form a secondary hydrogenated derivative of the compound of formula Ib, the secondary hydrogenated derivative having the structure of formula IIb:

[0350]

[0351] wherein:

[0352] Z' contains a secondary hydrogenated vinyl (-CH*=CH*-) group, a deuterated secondary hydrogenated prop-2-enyl (-CD2-CH*=CH*-) group, a deuterated secondary hydrogenated but-3-enyl (-CD2-CD2-CH*=CH*-) group, a deuterated secondary hydrogenated ethanyl (-CDH*-CDH*-) group, a deuterated secondary hydrogenated propanyl (-CD2-CDH*-CDH*-) group or a deuterated secondary hydrogenated butanyl (-CD2-CD2-CDH*-CDH*-) group;

[0353] wherein H* is hydrogen having a spin order derived from secondary hydrogen;

[0354] R1 contains an optionally substituted hydrocarbon group, an alkyl group, a cycloalkyl group, an aryl group, a carboxyl group, a ketone group or an alkoxy group; and

[0355] R2 contains an acyl derivative of a bio-related contrast agent, the bio-related contrast agent containing a non-hydrogen nuclear spin; and

[0356] (c) Applying a polarization transfer waveform to transfer the nuclear spin order from at least one H* in the compound of formula IIb to the non-hydrogen nuclear spin, thereby forming a derivative of the compound of formula IIb having a hyperpolarized acyl derivative of a bio-related contrast agent.

[0357] Example 63. A method for preparing a hyperpolarized bio-related contrast agent or a pharmaceutically acceptable salt thereof, the method comprising:

[0358] (a) Provide a composition comprising a compound of formula IIb:

[0359]

[0360] Wherein:

[0361] Z' comprises a secondary vinylidene (-CH*=CH*-) group, a deuterated secondary vinylidene (-CD2-CH*=CH*-) group, a deuterated secondary vinylidene (-CD2-CD2-CH*=CH*-) group, a deuterated secondary ethylidene (-CDH*-CDH*-) group, a deuterated secondary propylidene (-CD2-CDH*-CDH*-) group or a deuterated secondary butylidene (-CD2-CD2-CDH*-CDH*-) group;

[0362] Wherein H* is hydrogen having a spin order derived from secondary hydrogen;

[0363] R1 comprises an optionally substituted hydrocarbon group, an alkyl group, a cycloalkyl group, an aryl group, a carboxyl group, a ketone group or an alkoxy group; and

[0364] R2 comprises an acyl derivative of a biorelevant contrast agent comprising a non-hydrogen nuclear spin; and

[0365] (b) Apply a polarization transfer waveform to transfer nuclear spin order from at least one H* in the compound of formula IIb to the non-hydrogen nuclear spin, thereby forming a derivative of the compound of formula IIb having a hyperpolarized acyl derivative of a biorelevant contrast agent.

[0366] Example 64. The method according to Example 62 or 63, further comprising hydrolyzing the derivative of formula IIb to provide a composition comprising: (i) a hyperpolarized biorelevant contrast agent comprising a non-hydrogen nuclear spin; and (ii) a compound of formula IIIb:

[0367]

[0368] Wherein:

[0369] Z” comprises a secondary vinylidene (-CH*=CH*-) group, a deuterated secondary vinylidene (-CD2-CH*=CH*-) group, a deuterated secondary vinylidene (-CD2-CD2-CH*=CH*-) group, a deuterated secondary ethylidene (-CDH*-CDH*-) group, a deuterated secondary propylidene (-CD2-CDH*-CDH*-) group or a deuterated secondary butylidene (-CD2-CD2-CDH*-CDH*-) group;

[0370] wherein H* is hydrogen having a spin order derived from secondary hydrogen; and

[0371] R1 comprises an optionally substituted hydrocarbyl group, alkyl group, cycloalkyl group, aryl group, carboxyl group, ketone group or alkoxy group.

[0372] Example 65. The method according to Example 64, further comprising washing the hyperpolarized bio-related contrast agent one or more times with an organic solvent.

[0373] Example 66. The method according to Example 65, wherein after the washing step, the non-hydrogen nuclear spin polarization of the non-hydrogen nuclei is higher than 10%.

[0374] Example 67. The method according to any one of Examples 62 to 66, wherein the composition further comprises a PHIP transfer moiety between the Z, Z' or Z'' moiety and the sulfur atom, the PHIP transfer moiety comprising a fully deuterated C1 hydrocarbon (-CD2-) or a fully deuterated C2 hydrocarbon (-CD2-CD2-).

[0375] Example 68. The method according to Example 67, wherein:

[0376] the PHIP transfer moiety comprises *CR3R4, *CR3Y, *C=Y or any fully deuterated version thereof;

[0377] *C is 12 C or 13 a C carbon isotope;

[0378] R3 and R4 are each independently selected from: deuterium, fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl and fully deuterated haloalkyl; and

[0379] Y is selected from: spin-1 / 2 atoms and spin-1 / 2 atoms covalently bonded to one or more chemical moieties selected from: fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, halogen or fully deuterated haloalkyl, or heteroatoms such as N, O, S, etc., the heteroatoms optionally being substituted by fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, halogen or fully deuterated haloalkyl.

[0380] Example 69. The method according to Example 67, wherein:

[0381] The PHIP transfer moiety comprises *CR5R6–*CR7R8 or any fully deuterated version thereof;

[0382] *C is 12 C or 13 a C carbon isotope; and

[0383] R5, R6, R7, and R8 are each independently selected from the group consisting of deuterium, fully deuterated straight-chain, branched-chain, or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, and fully deuterated haloalkyl.

[0384] Example 70. The method according to Example 67, wherein:

[0385] The PHIP transfer moiety comprises *CH2, *CH2–*CH2, *CHY, *C=Y or any fully deuterated version thereof;

[0386] *C is 12 C or 13 a C carbon isotope; and

[0387] Y is selected from: spin-1 / 2 atoms and spin-1 / 2 atoms covalently bonded to one or more chemical moieties selected from: fully deuterated straight-chain, branched-chain, or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, halogen, or fully deuterated haloalkyl, or heteroatoms such as N, O, S, etc., said heteroatoms optionally substituted by fully deuterated straight-chain, branched-chain, or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, halogen, or fully deuterated haloalkyl.

[0388] Example 71. The method according to Example 68 or 70, wherein the spin-1 / 2 atom is selected from: 1 H, 13 C, 15 N, 19 F, or 31 P.

[0389] Example 72. The method according to any one of Examples 67 to 71, wherein the PHIP transfer moiety comprises at least one atom having a J-coupling to the non-hydrogen nuclear spin of at least 0.1 Hertz (Hz).

[0390] Example 73. The method according to any one of Examples 62 to 72, wherein Z or Z' comprises at least one atom having a J-coupling to the non-hydrogen nuclear spin of at least 0.1 Hertz (Hz).

[0391] Example 74. The method according to any one of Examples 62 to 73, wherein R1 comprises a solubilizing moiety.

[0392] Example 75. The method according to any one of Examples 62 to 74, wherein R1 comprises a hydrophobic and / or organophilic moiety.

[0393] Example 76. The method according to Example 75, wherein R1 comprises an organic solubilizing moiety.

[0394] Example 77. The method according to any one of Examples 62 to 73, wherein R1 comprises a hydrophilic and / or organophobic moiety.

[0395] Example 78. The method according to any one of Examples 62 to 77, wherein R1 is selected from: methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, hydroxy, methanolyl, ethanolyl, n-propanolyl, isopropanolyl, propionate alcoholyl, n-butanolyl, sec-butanolyl, tert-butanolyl, isobutanolyl, methoxy, ethoxy, propoxy, isopropoxy, propionate, butoxy, tert-butoxy, sec-butoxy, ester group, phenyl, substituted phenyl, primary amine group, secondary amine group, tertiary amine group, primary amide group, secondary amide group, tertiary amide group, and ketone group.

[0396] Example 79. The method according to any one of Examples 62 to 78, wherein the biorelevant contrast agent comprises a compound of the formula R9C(=O)O–; wherein R9 is selected from linear, branched or cyclic C1-C10 alkyl, wherein one or more C atoms are optionally replaced by C=C, CO, COH, CNH2, COOH, CH2COOH, CONH2, OC(=O).

[0397] Example 80. The method according to any one of Examples 62 to 79, wherein the biorelevant contrast agent is selected from: pyruvate, glutamate, glutamine, lactate, acetate, acetoacetate, ketoacetate, alanine, fructose, fumarate, bicarbonate, and their conjugate acids.

[0398] Example 81. A hyperpolarized biorelevant contrast agent or a pharmaceutically acceptable salt thereof, produced by the method according to any one of Examples 62 to 80.

Claims

1. A composition comprising a compound of formula Ia: Wherein: Z comprises a carbon-carbon double bond (-CD=CD-) or a carbon-carbon triple bond (-C≡C-) that is fully substituted to include deuterium (D); R1 comprises a parahydrogen-induced polarization (PHIP) transfer moiety; R2 comprises an optionally substituted hydrocarbon or alkoxy group; and R3 comprises a bio-related contrast agent comprising a non-hydrogen nuclear spin.

2. A composition comprising a compound of formula IIa: Wherein: Z' is a secondary-hydrogenated carbon-carbon single bond (-CDH*-CDH*-) or a secondary-hydrogenated carbon-carbon double bond (-CH*=CH*-) that is fully substituted to include deuterium; wherein H* is a hydrogen having a spin order derived from parahydrogen; R1 comprises a parahydrogen-induced polarization (PHIP) transfer moiety; R2 comprises an optionally substituted hydrocarbon or alkoxy group; and R3 comprises a bio-related contrast agent comprising a non-hydrogen nuclear spin.

3. A composition comprising: (i) a bio-related contrast agent comprising a non-hydrogen nuclear spin; and (ii) a compound of formula IIIa: Wherein: Z” is a secondary-hydrogenated carbon-carbon single bond (-CDH*-CDH*-) or a secondary-hydrogenated carbon-carbon double bond (-CH*=CH*-) that is fully substituted to include deuterium; wherein H* is a hydrogen having a spin order derived from parahydrogen; R1' comprises a parahydrogen-induced polarization (PHIP) transfer moiety; and R2 comprises an optionally substituted hydrocarbon or alkoxy group.

4. A composition comprising: (i) a hyperpolarized bio-related contrast agent comprising a non-hydrogen nuclear spin; and (ii) a compound of formula IVa: Wherein: Z comprises a carbon-carbon double bond (-CD=CD-) or a carbon-carbon triple bond (-C≡C-) that is fully substituted to include deuterium; R1' comprises a parahydrogen-induced polarization (PHIP) transfer moiety; and R2 comprises an optionally substituted hydrocarbon or alkoxy group.

5. The composition according to any one of claims 1 to 4, wherein the PHIP transfer moiety comprises a fully deuterated C1 hydrocarbon (-CD2-) or a fully deuterated C2 hydrocarbon (-CD2-CD2-).

6. The composition according to any one of claims 1 to 5, wherein: The PHIP transfer moiety comprises *CR4R5, *CR4Y, *C=Y or any fully deuterated version thereof; *C is 12 C or 13 C carbon isotope; R4 and R5 are each independently selected from: deuterium, fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl and fully deuterated haloalkyl; and Y is selected from the group consisting of spin-1 / 2 atoms and spin-1 / 2 atoms covalently bonded to one or more chemical moieties selected from: fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls, halogens or fully deuterated haloalkyls, or heteroatoms such as N, O, S, etc., which heteroatoms are optionally substituted with fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls, halogens or fully deuterated haloalkyls.

7. The composition according to any one of claims 1 to 5, wherein: the PHIP transfer moiety comprises *CR6R7–*CR8R9 or any fully deuterated version thereof; *C is 12 C or 13 C carbon isotope; and R6, R7, R8 and R9 are each independently selected from: deuterium, fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls and fully deuterated haloalkyls.

8. The composition according to any one of claims 1 to 5, wherein: the PHIP transfer moiety comprises *CH2, *CH2–*CH2, *CHY, *C=Y or any fully deuterated version thereof; *C is 12 C or 13 C carbon isotope; and Y is selected from the group consisting of spin-1 / 2 atoms and spin-1 / 2 atoms covalently bonded to one or more chemical moieties selected from: fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls, halogens or fully deuterated haloalkyls, or heteroatoms such as N, O, S, etc., which heteroatoms are optionally substituted with fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls, halogens or fully deuterated haloalkyls.

9. The composition according to claim 6 or 8, wherein the spin-1 / 2 atom is selected from: 1 H, 13 C 、15 N, 19 F or 31 P.

10. The composition according to any one of claims 1 to 9, wherein the PHIP transfer moiety includes at least one atom having a J-coupling with the non-hydrogen nuclear spin of at least 0.1 Hertz (Hz).

11. The composition according to any one of claims 1 to 10, wherein Z includes at least one atom having a J-coupling with the non-hydrogen nuclear spin of at least 0.1 Hertz (Hz).

12. The composition according to any one of claims 1 to 11, wherein R2 comprises a solubilizing moiety.

13. The composition according to any one of claims 1 to 12, wherein R2 comprises a hydrophobic and / or organophilic moiety.

14. The composition according to claim 13, wherein R2 comprises an organic solubilizing moiety.

15. The composition according to any one of claims 1 to 12, wherein R2 comprises a hydrophilic and / or organophobic moiety.

16. The composition according to any one of claims 1 to 15, wherein R2 is selected from: methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, hydroxy, methanolyl, ethanolyl, n-propanolyl, isopropanolyl, propionate alcohol group, n-butanolyl, sec-butanolyl, tert-butanolyl, isobutanolyl, methoxy, ethoxy, propoxy, isopropoxy, propionate group, butoxy, tert-butoxy, sec-butoxy, ester group, phenyl, substituted phenyl, primary amine group, secondary amine group, tertiary amine group, primary amide group, secondary amide group and tertiary amide group.

17. The composition according to any one of claims 1 to 16, wherein the bio-related contrast agent comprises a compound of the formula R 10 C(=O)X–; wherein R 10 is selected from straight-chain, branched-chain or cyclic C1-C10 alkyl, wherein one or more C atoms are optionally replaced by C=C, CO, COH, CNH2, COOH, CH2COOH, CONH2, OC(=O); and X is selected from NR 11 , S and O; wherein R 11 is selected from hydrogen and an amino protecting group, optionally selected from trifluoroacetyl, acetyl, benzoyl, benzyloxycarbonyl, tert-butyl carbonate and benzyl.

18. The composition according to any one of claims 1 to 17, wherein the bio-related contrast agent is selected from: pyruvate, glutamate, glutamine, lactate, acetate, acetoacetate, zymonate, alanine, fructose, fumarate, bicarbonate, urea, dehydroascorbate, α-ketoglutarate, dihydroxyacetone, glucose, ascorbate and its conjugate acid.

19. The composition according to any one of claims 1 to 18, wherein the solubility of the composition in water is less than 50 millimoles (mM).

20. The composition according to any one of claims 1 to 19, wherein the composition is reacted with parahydrogen such that the chemical yield of the parahydrogenated product is at least 30%.

21. The composition according to any one of claims 1 to 20, which is used in the parahydrogen-induced polarization (PHIP) process.

22. A method for preparing a hyperpolarized bio-related contrast agent or a pharmaceutically acceptable salt thereof, the method comprising: (a) providing a composition comprising a compound of formula Ia: wherein: Z comprises a carbon-carbon double bond (-CD=CD-) or a carbon-carbon triple bond (-C≡C-) that is fully substituted to include deuterium; R1 comprises a parahydrogen-induced polarization (PHIP) transfer moiety; R2 comprises an optionally substituted hydrocarbon or alkoxy; and R3 comprises a bio-related contrast agent comprising a non-hydrogen nuclear spin; (b) hydrogenating the double bond or the triple bond in the compound of formula Ia with parahydrogen to form a parahydrogenated derivative of the compound of formula Ia having the structure of formula IIa: wherein: Z' is a parahydrogenated carbon-carbon single bond (-CDH*-CDH*-) or a parahydrogenated carbon-carbon double bond (-CH*=CH*-) that is fully substituted to include deuterium; wherein H* is a hydrogen having a spin order derived from parahydrogen; R1 comprises a parahydrogen-induced polarization (PHIP) transfer moiety; R2 comprises an optionally substituted hydrocarbon or alkoxy; and R3 comprises a bio-related contrast agent comprising a non-hydrogen nuclear spin; and (c) applying a polarization transfer waveform to transfer the nuclear spin order from at least one H* in the compound of formula IIa to the non-hydrogen nuclear spin, thereby forming a derivative of formula IIa having a hyperpolarized bio-related contrast agent.

23. A method for preparing a hyperpolarized bio-related contrast agent or a pharmaceutically acceptable salt thereof, the method comprising: (a) providing a composition comprising a compound of formula IIa: wherein: Z' is a secondary hydrogenated carbon-carbon single bond (-CDH*-CDH*-) or a secondary hydrogenated carbon-carbon double bond (-CH*=CH*-) that is fully substituted to include deuterium; wherein H* is a hydrogen having a spin order derived from secondary hydrogen; R1 comprises a secondary hydrogen-induced polarization (PHIP) transfer moiety; R2 comprises an optionally substituted hydrocarbon or alkoxy group; and R3 comprises a bio-related contrast agent, the bio-related contrast agent comprising a non-hydrogen nuclear spin; and (b) applying a polarization transfer waveform to transfer the nuclear spin order from at least one H* in the compound of Formula IIa to the non-hydrogen nuclear spin, thereby forming a derivative of Formula IIa having a hyperpolarized bio-related contrast agent.

24. The method according to claim 22 or 23, further comprising hydrolyzing the derivative of Formula IIa to provide a composition comprising: (i) a hyperpolarized bio-related contrast agent, the hyperpolarized bio-related contrast agent comprising a non-hydrogen nuclear spin; and (ii) a compound of Formula IIIa: wherein: Z” is a secondary hydrogenated carbon-carbon single bond (-CDH*-CDH*-) or a secondary hydrogenated carbon-carbon double bond (-CH*=CH*-) that is fully substituted to include deuterium; wherein H* is a hydrogen having a spin order derived from secondary hydrogen; R1' comprises a secondary hydrogen-induced polarization (PHIP) transfer moiety; and R2 comprises an optionally substituted hydrocarbon or alkoxy group.

25. The method according to claim 24, further comprising washing the hyperpolarized bio-related contrast agent one or more times with an organic solvent.

26. The method according to claim 25, wherein after the washing step, the non-hydrogen nuclear spin polarization of the non-hydrogen nuclear spin is higher than 10%.

27. The method according to any one of claims 22 to 26, wherein the PHIP transfer moiety comprises a fully deuterated C1 hydrocarbon (-CD2-) or a fully deuterated C2 hydrocarbon (-CD2-CD2-).

28. The method according to any one of claims 22 to 26, wherein: the PHIP transfer moiety comprises *CR4R5, *CR4Y, *C=Y or any fully deuterated version thereof; *C is 12 C or 13 C carbon isotope; R4 and R5 are each independently selected from: deuterium, fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls and fully deuterated haloalkyls; and Y is selected from: spin-1 / 2 atoms and spin-1 / 2 atoms covalently bonded to one or more chemical moieties selected from: fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls, halogens or fully deuterated haloalkyls, or heteroatoms such as N, O, S, etc., the heteroatoms optionally being substituted by fully deuterated straight-chain, branched-chain or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryls, fully deuterated benzyls, fully deuterated phenyls, fully deuterated heteroaryls, halogens or fully deuterated haloalkyls.

29. The method according to any one of claims 22 to 26, wherein: The PHIP transfer moiety comprises *CR6R7–*CR8R9 or any fully deuterated version thereof; *C is 12 C or 13 C carbon isotope; and R6, R7, R8, and R9 are each independently selected from the group consisting of deuterium, fully deuterated straight-chain, branched-chain, or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, and fully deuterated haloalkyl.

30. The method according to any one of claims 22 to 26, wherein: The PHIP transfer moiety comprises *CH2, *CH2–*CH2, *CHY, *C=Y or any fully deuterated version thereof; *C is 12 C or 13 C carbon isotope; and Y is selected from: spin-1 / 2 atoms and spin-1 / 2 atoms covalently bonded to one or more chemical moieties selected from: fully deuterated straight-chain, branched-chain, or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, halogen, or fully deuterated haloalkyl, or heteroatoms such as N, O, S, etc., which heteroatoms are optionally substituted by fully deuterated straight-chain, branched-chain, or cyclic C1-C10 alkyl hydrocarbons, fully deuterated C6 aryl, fully deuterated benzyl, fully deuterated phenyl, fully deuterated heteroaryl, halogen, or fully deuterated haloalkyl.

31. The method according to claim 28 or 30, wherein the spin-1 / 2 atoms are selected from: 1 H, 13 C, 15 N, 19 F or 31 P.

32. The method according to any one of claims 22 to 31, wherein the PHIP transfer moiety comprises at least one atom having a J-coupling with the non-hydrogen nuclear spin of at least 0.1 Hertz (Hz).

33. The method according to any one of claims 22 to 32, wherein Z or Z' comprises at least one atom having a J-coupling with the non-hydrogen nuclear spin of at least 0.1 Hertz (Hz).

34. The method according to any one of claims 22 to 33, wherein R2 comprises a solubilizing moiety.

35. The method according to any one of claims 22 to 34, wherein R2 comprises a hydrophobic and / or organophilic moiety.

36. The method according to claim 35, wherein R2 comprises an organic solubilizing moiety.

37. The method according to any one of claims 22 to 33, wherein R2 comprises a hydrophilic and / or organophobic moiety.

38. The method according to any one of claims 22 to 37, wherein R2 is selected from: methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, hydroxy, methanolyl, ethanolyl, n-propanolyl, isopropanolyl, propionate alcoholyl, n-butanolyl, sec-butanolyl, tert-butanolyl, isobutanolyl, methoxy, ethoxy, propoxy, isopropoxy, propionate, butoxy, tert-butoxy, sec-butoxy, ester group, phenyl, substituted phenyl, primary amine group, secondary amine group, tertiary amine group, primary amide group, secondary amide group, and tertiary amide group.

39. The method according to any one of claims 22 to 38, wherein the bio-related contrast agent comprises a compound of the formula R 10 C(=O)X–; wherein R 10 is selected from linear, branched or cyclic C1-C10 alkyl, wherein one or more C atoms are optionally replaced by C=C, CO, COH, CNH2, COOH, CH2COOH, CONH2, OC(=O); and X is selected from NR 11 , S and O; wherein R 11 is selected from hydrogen and an amino protecting group, optionally selected from trifluoroacetyl, acetyl, benzoyl, benzyloxycarbonyl, tert-butyl carbonate and benzyl.

40. The method according to any one of claims 22 to 39, wherein the bio-related contrast agent is selected from pyruvate, glutamate, glutamine, lactate, acetate, acetoacetate, ketoacetate, alanine, fructose, fumarate, bicarbonate, urea, dehydroascorbate, α-ketoglutarate, dihydroxyacetone, glucose, ascorbate, and their conjugate acids.

41. A hyperpolarized bio-related contrast agent or a pharmaceutically acceptable salt thereof, produced by the method according to any one of claims 22 to 40.

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