阅读说明：本技术 一种稳定同位素标记的硝基咪唑类化合物及其制备和应用方法 (Stable isotope labeled nitroimidazole compound and preparation and application methods thereof ) 是由 阮英恒 周静洋 杨晓云 杨晓军 于 2021-06-11 设计创作，主要内容包括：本发明提供一种稳定同位素标记的硝基咪唑类化合物及其制备和应用方法,其中所述硝基咪唑类化合物的化学结构通式为：本发明的有益效果是：以氘代硝基咪唑类化合物作为造影剂,该造影剂可用于肿瘤乏氧组织的显像。(The invention provides a stable isotope labeled nitroimidazole compound and preparation and application methods thereof, wherein the chemical structure general formula of the nitroimidazole compound is as follows: the invention has the beneficial effects that: the deuterated nitroimidazole compound is used as a contrast agent, and the contrast agent can be used for imaging tumor hypoxic tissues.)

1. A stable isotope labeled nitroimidazole compound, wherein the chemical structure general formula of the deuterated nitroimidazole compound is as follows:

wherein

R₁And/or R₂Independently of one another, hydrogen or deuterium or halogen, wherein none or one or more hydrogen atoms are replaced by deuterium;

R₃、R₄、R₅、R₆and/or R₇Independently of one another, hydrogen or deuterium, branched or straight-chain C₁-C₁₀Alkoxy, branched or straight chain C₁-C₁₀Alkyl, substituted or unsubstituted C₆-C₁₀Monocyclic or bicyclic aryl, substituted or unsubstituted alkyl-C₆-C₁₀Monocyclic or bicyclic aryl, substituted or unsubstituted C₅-C₁₀Monocyclic or bicyclic heteroaryl, substituted or unsubstituted alkyl-C₆-C₁₀Monocyclic or bicyclic heteroaryl or hydroxy, wherein none or one or more hydrogen atoms are replaced by deuterium;

wherein R is₁～R₇Wherein at least one hydrogen atom is replaced by deuterium, and to diastereomeric and/or enantiomeric mixtures and enantiomerically pure compounds and pharmaceutical salts thereof.

2. The nitroimidazole compound of claim 1, wherein the nitroimidazole compound has the chemical formula:

different deuteration positions and deuteration numbers of different substituent groups.

3. A method for using stable isotope labeled nitroimidazoles, characterized in that the deuterated nitroimidazoles according to any one of claims 1 to 2 are used as contrast agents for magnetic resonance imaging.

4. The method of use according to claim 3, wherein the deuterated nitroimidazole compound is used as a tumor hypoxia imaging agent.

5. The method of use according to claim 3, wherein said contrast agent further comprises at least one pharmaceutically acceptable excipient.

6. The method of use of claim 5, wherein the excipient is one of a carrier, a bulking agent, or a solvent.

7. A preparation method of stable isotope labeled nitroimidazole compounds is characterized by comprising the following steps:

carrying out a condensation reaction on a compound with a structure shown in a formula I-2 and a compound with a structure shown in a formula I-3 to obtain a deuterated nitroimidazole intermediate compound, and carrying out an upper protection reaction, a fluoro reaction or a deprotection reaction to obtain a target deuterated nitroimidazole compound;

wherein R is₁And/or R₂Independently of one another, hydrogen or deuterium or halogen, wherein none or one or more hydrogen atoms are replaced by deuterium;

wherein R is₃、R₄、R₅、R₆And/or R₇Independently of one another, hydrogen or deuterium, branched or straight-chain C₁-C₁₀Alkoxy, branched or straight chain C₁-C₁₀Alkyl, substituted or unsubstituted C₆-C₁₀Monocyclic or bicyclic arylSubstituted or unsubstituted alkyl-C₆-C₁₀Monocyclic or bicyclic aryl, substituted or unsubstituted C₅-C₁₀Monocyclic or bicyclic heteroaryl, substituted or unsubstituted alkyl-C₆-C₁₀Monocyclic or bicyclic heteroaryl or hydroxy, wherein none or one or more hydrogen atoms are replaced by deuterium;

wherein R is₈And/or R₉Independently of one another, hydroxy or halogen;

wherein R is₁₀is-C (CH)₃)₂-CHPh, -CHO-branched or straight C₁-C₁₀Alkyl, -CHO-substituted or unsubstituted C₆-C₁₀Monocyclic or bicyclic aryl, -Si-branched or straight-chain C₁-C₁₀Alkyl or-Si-substituted or unsubstituted C₆-C₁₀Monocyclic or bicyclic aryl;

wherein R is₁₁Is hydrogen or deuterium or an amino protecting group;

wherein R is₁～R₇Wherein at least one hydrogen atom is replaced by deuterium, and to diastereomeric and/or enantiomeric mixtures and enantiomerically pure compounds and pharmaceutical salts thereof.

8. The method of claim 7, comprising the following synthetic steps:

under the protection of inert gas, cooling the system to-30-10 ℃, adding boron sodium deuteride into a deuterated methanol solution of 2, 3-epoxypropionic acid ethyl ester, carrying out heat preservation reaction at-30-10 ℃, quenching the reaction by using deuterium water, and carrying out post-treatment to obtain 2, 3-epoxy-1, 1-d 2-propanol;

adding 2, 3-epoxy-1, 1-d 2-propanol, 2-nitro-1H-imidazole and alkali into a solvent, stirring for reaction, and performing post-treatment to obtain 3- (2-nitro-1H-imidazol-1-yl) propane-3, 3-d2-1, 2-diol;

cooling the system to-15 ℃, adding paratoluensulfonyl chloride into a solution of 3- (2-nitro-1H-imidazol-1-yl) propane-3, 3-d2-1, 2-diol, keeping the temperature at 10-30 ℃ for reaction, adding a solution of imidazole, adding a solution of tert-butyldimethylchlorosilane, keeping the temperature at 10-30 ℃ for reaction, and performing aftertreatment to obtain 2- (tert-butyldimethylsilyloxy) -3- (2-nitro-1H-imidazol-1-yl) propyl-3, 3-d 2-4-methylbenzenesulfonate;

adding 2- (tert-butyldimethylsilyloxy) -3- (2-nitro-1H-imidazol-1-yl) propyl-3, 3-d 2-4-methylbenzenesulfonate into a solvent, adding a tetrabutylammonium fluoride solution, carrying out heat preservation reaction at 50-90 ℃, removing the solvent, adding hydrochloric acid for hydrolysis, and carrying out post-treatment to obtain the 1-fluoro-3- (2-nitro-1H-imidazol-1-yl) propane-3, 3-d 2-2-ol.

9. The method according to claim 8, wherein the base is one of sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide, cesium hydroxide, sodium methoxide, sodium ethoxide, sodium acetate, sodium tert-butoxide, potassium tert-butoxide, and aluminum tert-butoxide.

10. The method according to claim 8, wherein the solvent is one of methanol, ethanol, propanol, isopropanol, N-butanol, t-butanol, acetonitrile, N-dimethylformamide, and 1, 4-dioxane.

Technical Field

The invention relates to the field of nitroimidazole compound application, and particularly relates to a stable isotope labeled nitroimidazole compound and preparation and application methods thereof.

Background

The increasing use of hydrogen and its isotopes in the pharmaceutical industry, for example, research in magnetic resonance molecular imaging has resulted in two promising methods of imaging isotopes of glucose metabolism: hyperpolarization (HP)¹³C Magnetic Resonance Imaging (MRI) and deuterometabolic imaging (DMI). Although it is used for¹³There are many possible pathways for C hyperpolarization, but HP 13C MRI most often relies on¹³Dynamic nuclear polarization pre-polarization of the C-labeled substrate followed by rapid dissolution produced a large signal enhancement four to five orders of magnitude higher than Boltzmann polarization at clinically viable MRI field strengths, enabling target metabolic studies by spectral imaging. However, the magnetization lifetime of 13C is short, requiring compact studies and fast, carefully calibrated MRI scans. In contrast, a DMI that is not hyperpolarized takes advantage of the relatively large magnetic moment provided by Boltzmann polarization.

Positron Emission Tomography (PET) is the most advanced medical imaging technology at present, can realize high-resolution imaging of cell metabolism and functions, and perform noninvasive, three-dimensional and dynamic research on physiological and biochemical processes of a human body on a molecular level, and PET is applied to diagnosis of tumors including tumors, early diagnosis and identification of benign and malignant identification, staging, typing, relapse and metastasis of malignant tumors, selection of treatment schemes, monitoring of chemotherapeutic effects, observation of tumor change processes and detection of healed conditions. PET examination differs from other examinations in that it relies on positron drugs (PET drugs) that specifically concentrate on the target organ for diagnostic and evaluation purposes. Positron drugs currently used in PET examinations include fluorodopamine, which is a type of positron radionuclide that is susceptible to secondary damage upon multiple examinations on patients, and whose half-life is significantly improved (109.8min) compared to other imaging labeled nuclides such as oxygen-15 (O-15), nitrogen 13(N-13), and carbon 11(C-11), but still cannot be followed for the entire metabolic process.

Tumor hypoxia is a phenomenon commonly existing in the occurrence and development processes of solid tumors, and is a state that the available oxygen is reduced or the partial pressure of oxygen is reduced to be below a critical value due to the abnormal microenvironment in vivo, so that a series of physiological functions of organs, tissues and cells are limited or even terminated. Tumor cells undergo a series of corresponding biological changes in order to adapt to hypoxic environments. Hypoxia-inducible factor (HIF-1) is an oxygen-dependent transcription activator, which consists of two subunits of HIF-1 alpha and HIF-1 beta, HIF-1 alpha is degraded quickly only in normal oxygenation state, but its protein stability is increased in hypoxia state, and forms heterodimer with HIF-1 beta, and is an important regulator in a series of self-adaptive reactions after oxygen partial pressure in body organs and tissues is changed, and it can regulate transcription and translation of a series of downstream genes by combining with hypoxia response element, at present, 60 more downstream target genes are directly regulated by HIF-1 alpha, including Vascular Endothelial Growth Factor (VEGF), Erythropoietin (EPO), glycolytic enzyme, Carbonic Anhydrase (CA), Epidermal Growth Factor (EGF), P-glycoprotein, etc. which are involved in the regulation of the pH value of the tumor, energy metabolism,

Angiogenesis and remodeling, cell proliferation and apoptosis, invasion and metastasis, malignant phenotype and other tumors have important functions in the evolution process of the tumors, and resist radiotherapy and chemotherapy, resulting in local recurrence, distant metastasis and other bad prognosis. Due to the specific expression characteristics in malignant cells, the HIF-1 alpha gene has been studied.

Hypoxia exists in various disease states, and the degree of hypoxia of tissues is known to have a certain value for clinic. The accurate determination of the hypoxic condition in the tumor not only creates conditions for basic research of tumor hypoxic cells, but also provides reliable basis for clinical judgment of prognosis of tumor treatment and implementation of changing growth environment of the hypoxic cells in the tumor to improve treatment effect. At present, the invasive examination method for detecting hypoxic oxygen in clinic is mainly an oxygen electrode method. The oxygen electrode is considered as the "gold standard" for direct detection of the tumor oxygen partial pressure, which is obtained by measurement calculation of a 300 μm electrode inserted inside the tumor. This approach can predict the therapeutic response and potential metastatic potential of the tumor and is used by some institutions for routine examination of the primary focus. Direct measurements of cervical cancer, breast cancer, renal cell carcinoma, melanoma, soft tissue sarcoma and head and neck tumors using oxygen electrode methods have been used to demonstrate neoplastic refractory oxygen. However, the oxygen electrode method has certain limitations, for example, there is no accepted standard for determining the critical value of the oxygen deficiency at present; oxygen distribution in the tumor needs to be measured at multiple points; the oxygen electrode has the technical limitation, and certain errors exist in the measurement process; the oxygen electrode method is an invasive detection method. These limit the application of the oxygen electrode method.

Imaging detection methods have been used in clinical settings because of their advantages of being non-invasive, accurate, and highly repeatable, including color doppler ultrasound, magnetic resonance spectroscopy, and nuclear medicine imaging. Positron Emission Tomography (PET) is carried out by using radionuclide-labeled hypoxic tissue imaging agent, so that hypoxic oxygen can be repeatedly detected for many times in a positioning and quantitative manner. Commonly used hypoxia imaging agents include nitroimidazole hypoxia imaging agents and non-nitroimidazole hypoxia imaging agents. The FMISO is the nitroimidazole hypoxia imaging agent which is the earliest and most widely used in clinical research, is the most common technology for hypoxic detection research at present due to high stability and specificity, can be used for diagnosing and judging the stage of tumors, can judge the effect of radiotherapy and chemotherapy, and can adjust and optimize a radiotherapy and chemotherapy scheme in time. The imaging agent of the non-nitroimidazole has a certain sensitivity to hypoxic tissues, but has higher uptake rate in intestinal tract and liver, and is not suitable for the examination of abdominal tumors.

FMISO is a nitroimidazole compound that enters cells via passive diffusion through the cell membrane, the lipophilicity of its ligand determining its ability to enter cells. The nitro groups of the ligands are reduced by the action of intracellular associated enzymes to produce free anions which, in normal cells, are peroxidized and diffuse out of the cell, whereas in hypoxic cells the intermediate is further reduced and the product binds to the intracellular components and is thus retained in the cell. FMISO accumulates only in hypoxic cells rich in functional nitroreductase enzymes, and thus deuterium-forming tracers that emit positrons on FMISO tags can be used for imaging of hypoxic tissues.

Hypoxia plays a crucial role in the pathological mechanism of tumors, and is directly related to the curative effect and prognosis of patients. The deuterated FMISO is used for noninvasive evaluation of hypoxic PET tracer, provides effective basis for definite diagnosis, optimal treatment scheme formulation and prognosis evaluation at different stages of tumor, has great potential advantages in clinic, and can be widely applied in the future. Therefore, the prior art is still in need of improvement and development.

Disclosure of Invention

In view of the defects of the prior art, the invention aims to provide a stable isotope labeled nitroimidazole compound and a preparation and application method thereof, and aims to solve the problems that in the prior art, a positive electron radionuclide is used as a contrast agent, secondary damage is easily caused to a patient during multiple detections, and the tracking of the whole metabolic process cannot be performed.

The technical scheme of the invention is as follows:

a stable isotope labeled nitroimidazole compound, wherein the chemical structure general formula of the nitroimidazole compound is as follows:

wherein

R₁And/or R₂Independently of one another, hydrogen or deuterium or halogen, wherein none or one or more of the hydrogen atoms are replaced by deuterium;

R₃、R₄、R₅、R₆and/or R₇Independently of one another, hydrogen or deuterium, branched or straight-chain C₁-C₁₀Alkoxy, branched or straight chain C₁-C₁₀Alkyl, substituted or unsubstituted C₆-C₁₀Monocyclic or bicyclic aryl, substituted or unsubstituted alkyl-C₆-C₁₀Monocyclic or bicyclic aryl, substituted or unsubstituted C₅-C₁₀Single ringOr bicyclic heteroaryl, substituted or unsubstituted alkyl-C₆-C₁₀Monocyclic or bicyclic heteroaryl or hydroxy, wherein none or one or more hydrogen atoms are replaced by deuterium;

wherein R is₁～R₇In which at least one hydrogen atom is replaced by deuterium, and to possible diastereomeric and/or enantiomeric mixtures and enantiomerically pure compounds and pharmaceutical salts thereof.

The chemical structural formula of the stable isotope labeled nitroimidazole compound is as follows:

or one of the array combinations between different deuterated positions and deuterated numbers of different substituent groups.

A preparation method of stable isotope labeled nitroimidazole compounds comprises the following steps:

under the protection of inert gas, cooling the system to-30-10 ℃, adding boron sodium deuteride into a deuterated methanol solution of 2, 3-epoxypropionic acid ethyl ester, carrying out heat preservation reaction at-30-10 ℃, quenching the reaction by using deuterium water, and carrying out post-treatment to obtain 2, 3-epoxy-1, 1-d 2-propanol;

adding 2, 3-epoxy-1, 1-d 2-propanol, 2-nitro-1H-imidazole and a base into a solvent, wherein the base is sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide, cesium hydroxide, sodium methoxide, sodium ethoxide, sodium acetate, sodium tert-butoxide, potassium tert-butoxide, aluminum tert-butoxide and the like, and the solvent is methanol, ethanol, propanol, isopropanol, N-butanol, tert-butanol, acetonitrile, N-dimethylformamide, 1, 4-dioxane and the like, stirring for reaction, and carrying out aftertreatment to obtain 3- (2-nitro-1H-imidazol-1-yl) propane-3, 3-d2-1, 2-diol;

cooling the system to-15 ℃, adding paratoluensulfonyl chloride into a solution of 3- (2-nitro-1H-imidazol-1-yl) propane-3, 3-d2-1, 2-diol, keeping the temperature at 10-30 ℃ for reaction, adding a solution of imidazole, adding a solution of tert-butyldimethylchlorosilane, keeping the temperature at 10-30 ℃ for reaction, and performing aftertreatment to obtain 2- (tert-butyldimethylsilyloxy) -3- (2-nitro-1H-imidazol-1-yl) propyl-3, 3-d 2-4-methylbenzenesulfonate;

adding 2- (tert-butyldimethylsilyloxy) -3- (2-nitro-1H-imidazol-1-yl) propyl-3, 3-d 2-4-methylbenzenesulfonate into a solvent, adding a tetrabutylammonium fluoride solution, carrying out heat preservation reaction at 50-90 ℃, removing the solvent, adding hydrochloric acid for hydrolysis, and carrying out post-treatment to obtain the 1-fluoro-3- (2-nitro-1H-imidazol-1-yl) propane-3, 3-d 2-2-ol.

An application method of a stable isotope labeled nitroimidazole compound, wherein the nitroimidazole compound is used as a contrast agent for magnetic resonance imaging.

The application method of the nitroimidazole compound, wherein the nitroimidazole compound is used as a tumor hypoxia imaging agent.

The application method of the nitroimidazole compound is characterized in that the contrast agent further comprises at least one pharmaceutically acceptable excipient.

The application method of the nitroimidazole compound is characterized in that the excipient is one of a carrier, a filling agent or a solvent.

The invention has the beneficial effects that: the deuterated nitroimidazole compound is used as the contrast agent, and the contrast agent can be used for imaging tumor hypoxic tissues. The deuterated nitroimidazole compound used as the contrast agent can stably exist in vivo, and cannot cause damage to human bodies after multiple detections; the method has the advantages of simple and practical method, high precision, reliable result and capability of analyzing the metabolic condition in a quantitative positioning manner.

Drawings

FIG. 1 is a chemical reaction diagram of a preparation method of nitroimidazole compounds according to an embodiment of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments of the present invention without making any creative effort, shall fall within the protection scope of the present invention.

A stable isotope labeled nitroimidazole compound (FMISO), which has a chemical structural formula as follows:

wherein:

R₁and/or R₂Independently of one another, hydrogen or deuterium or halogen, wherein none or one or more of the hydrogen atoms are replaced by deuterium;

R₃、R₄、R₅、R₆and/or R₇Independently of one another, hydrogen or deuterium, branched or straight-chain C₁-C₁₀Alkoxy, branched or straight chain C₁-C₁₀Alkyl, substituted or unsubstituted C₆-C₁₀Monocyclic or bicyclic aryl, substituted or unsubstituted alkyl-C₆-C₁₀Monocyclic or bicyclic aryl, substituted or unsubstituted C₅-C₁₀Monocyclic or bicyclic heteroaryl, substituted or unsubstituted alkyl-C₆-C₁₀Monocyclic or bicyclic heteroaryl or hydroxy, wherein none or one or more hydrogen atoms are replaced by deuterium;

wherein R is₁、R₇In which at least one hydrogen atom is replaced by deuterium, and to possible diastereomers and/or mixtures of enantiomers, as well as enantiomerically pure compounds and pharmaceutical salts thereof.

The chemical structural formula of the stable isotope labeled nitroimidazole compound is as follows:

or one of the array combinations between different deuterated positions and deuterated numbers of different substituent groups.

In some embodiments, there is also provided a method for preparing deuterated nitroimidazoles (FMISO) compounds, wherein the reaction process is as shown in fig. 1, and specifically comprises the following steps:

step 1, compound B: adding boron sodium deuteride (3.97g,94.82mmol) into a stirred methanol-d 4(100mL) solution of the compound A (10.0g,86.20mmol) at the temperature of 10 ℃ below zero under the protection of nitrogen, preserving the temperature for 1h, adding deuterium water (3mL) to quench the reaction, evaporating the methanol-d 4 at the temperature of 40 ℃ under reduced pressure, adding methyl tert-butyl ether to extract for 3 times, combining organic phases, drying with anhydrous sodium sulfate, and concentrating under reduced pressure to obtain 5.05g of oily compound B, wherein the yield is 77.0%;

step 2, compound D: adding compound B (4.74g,62.28mmol), compound C (3.52g,31.14mmol) and potassium carbonate (0.36g) into ethanol (200mL), stopping heating after the exothermic reaction starts, continuing stirring for 40min, concentrating under reduced pressure, and purifying the residue by silica gel column chromatography to obtain 2.61g of compound D with a yield of 44.3%;

step 3, compound E: p-toluenesulfonyl chloride (2.45g,12.86mmol) was added dropwise to a solution of compound D (2.43g,12.86mmol) in anhydrous pyridine (40mL) at 0 deg.C, the mixture was stirred at room temperature for 6h, a solution of imidazole (3.50g,51.44mmol) in dichloromethane (40mL) was added, a solution of tert-butyldimethylchlorosilane (5.81g,38.58mmol) in dichloromethane (40mL) was added, the mixture was stirred at room temperature for 2h, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was chromatographically purified to give compound E as a white solid, 3.96g, yield 67.3%;

step 4, compound F: adding a compound E (3.87g,8.46mmol) into acetonitrile (80mL), adding a tetrahydrofuran solution (42mL,42 mmol) of TBAF, heating for reflux reaction for 6h, removing the solvent under reduced pressure, adding 1M hydrochloric acid into the obtained fluoro intermediate, heating for reflux hydrolysis, cooling to room temperature after the reaction is finished, adding a 2N sodium hydroxide solution for neutralization, extracting with ethyl acetate, drying with anhydrous sodium sulfate, concentrating under reduced pressure, and purifying the residue by silica gel column chromatography to obtain 173mg of a compound F, wherein the yield is 10.7%.

Compound F is one of the deuterated nitroimidazoles (FMISO) compounds.

In some embodiments, a method of use of deuterated nitroimidazoles (FMISO) compounds is also provided, wherein the deuterated nitroimidazoles (FMISO) compounds of the invention are used as contrast agents for magnetic resonance imaging. Further, deuterated nitroimidazoles (FMISO) compounds are used as tumor hypoxia imaging agents.

FMISO is a nitroimidazole compound that enters cells via passive diffusion through the cell membrane, the lipophilicity of its ligand determining its ability to enter cells. The nitro groups of the ligands are reduced by the action of intracellular associated enzymes to produce free anions which, in normal cells, are peroxidized and diffuse out of the cell, whereas in hypoxic cells the intermediate is further reduced and the product binds to the intracellular components and is thus retained in the cell. FMISO accumulates only in hypoxic cells rich in functional nitroreductase enzymes, and thus positron-emitting deuterium forming tracers labeling FMISO can be used for imaging hypoxic tissues.

This example uses deuterated nitroimidazoles (FMISO) as imaging agents for tumor hypoxia and uses magnetic resonance imaging to detect metabolic processes in tumors, thereby making a diagnosis. The embodiment using deuterated FMISO as the contrast agent has the following advantages: high content of deuterium atoms; accumulation in brain tissue for a reasonable period of time to a concentration sufficient for diagnosis; the toxicity is low and the intact agent is almost completely excreted from the body. This allows effective diagnosis using a dose that is not harmful to the human body.

In some embodiments, there is provided a method of using deuterated nitroimidazole (FMISO) compounds as tumor hypoxia imaging agents, comprising the steps of:

administering the contrast agent to a subject;

after a sufficient accumulation time after administration of the contrast agent, magnetic resonance imaging is performed at the frequency of deuterons to obtain²H-MR images;

obtained by analysis²The H-MR image finds an abnormally high intensity signal area, and diagnoses the tumor metabolic process based on the observed abnormal signal intensity of the deuterons.

In this embodiment, the signal intensity measured based on the contrast agent of this embodiment may be compared with typical signal intensities observed in corresponding tissues in healthy subjects, so as to compare the judgment difference, thereby making a judgment on the condition of the subject.

The detection method provided by the present embodiment is independent of any detrimental effects of ionizing radiation (typically, for example, for CT, PET, SPECT methods), which in turn increases the safety of the study and allows the study to be repeated more frequently. The present invention aims to obtain diagnostic information similar to Positron Emission Tomography (PET), but unlike the latter, it can eliminate the risks associated with ionizing radiation of radiopharmaceuticals.

In some embodiments, the contrast agent further comprises at least one pharmaceutically acceptable excipient, such as, for example, one of a carrier, a filler, or a solvent.

In some embodiments, the contrast agent is administered to the subject orally or parenterally.

In some specific embodiments, the contrast agent is administered to the subject at 0.60-0.75 grams of contrast agent per 1kg of body weight.

In some embodiments, the deuterated FMISO as a contrast agent can also be used for tracking related metabolites, which specifically comprises the steps of:

scanning the site to be examined by NMR spectrometer and forming a site before using deuterated FMISO¹H pre-use profile; standard nmr scanners, such as 3T MRI scanners or 7T MRI scanners, can be used without the risk of exposure to ionizing radiation, without the need for special equipment, and are easy to use, the sequences and imaging parameters of which can be selected as follows: make itUsing the PRESS sequence (TR/TE: 2500/16ms, spectrum width: 4kHz, 90 pulse bandwidth: 5400Hz, 180 pulse bandwidth: 2400Hz, point: 4006, vapro water suppression, average: 128).

After obtaining the pre-use profile, the subject is administered a deuterated FMISO which may be a composition in the form of a single dosage unit, i.e., the composition may be a single dose or one or more unit doses contained in a container, each dose containing the same ingredients, i.e., each dose contains the same weight²The H-tag substance can be directly or after being diluted to be used for a subject, or the composition can be made into different specifications, for example, different specifications containing the deuterated FMISO can be used for administering different specifications of compositions according to the condition of the subject, the compositions can be made into different forms of liquid or solid, different doses of the deuterated FMISO can be administered according to different subjects, for example, the deuterated FMISO is dissolved in 200 to 300 ml of water according to the dosage of 0.60 to 0.75 g/kg of body weight and the maximum of 60 g for use, the deuterated FMISO can be used in an injection mode according to different specifications of the deuterated FMISO, and the deuterated FMISO can also be a composition in any one form of powder, tablets, pills, capsules or liquid. By using²H-mark means using²H for one or more¹H atoms, thereby giving rise to corresponding metabolites¹Overall reduction of H MRS signal, thus through quantization²H substitution¹The reduction of the signal in the proton magnetic resonance spectrum generated by H enables the status of the deuterated FMISO transformation to be obtained, thus knowing the cell metabolic status. Rescanning the site to be examined of the subject by means of a nuclear magnetic resonance spectrometer under the same parameters for a given time and forming a site after the use of a deuterated FMISO¹H, a post-use map; the given time is preferably 20 to 90 minutes, so that the deuterated FMISO can be sufficiently transformed in vivo to obtain more accurate detection results and reflect metabolic conditions, and the measurement can be performed at intervals within the given time, for example, the map data at the time point can be acquired every five minutes or ten minutes. Due to the fact that²H has a lower NMR frequencyWill have a broad intrinsic broad peak in the vibration spectrum, minimizing the effect of magnetic field inhomogeneity on deuterium imaging, but allowing²The nmr spectrum of H contains only a few metabolite peaks,¹the sensitivity of H MRS is higher, and the H MRS can be independently detected²And the H MRS can not obtain the map spectrum of certain metabolites, so that a more accurate detection result is obtained, and the metabolic dynamic process of key metabolites can be detected in the same acquisition process while the steady-state metabolic information of several metabolites is provided.

The method obtains the concentrations of the designated metabolites before and after the deuterium-substituted FMISO by analyzing the data of the before-use map and the after-use map of the deuterium-substituted FMISO, and obtains the metabolic condition of the tissue according to the concentration change before and after the deuterium-substituted FMISO. By using¹The advantages of universality and easy implementation of H proton magnetic resonance spectrum detection and the spectral resolution of color generation can track the metabolites transferred by deuterated FMISO, the detection resolution and sensitivity are higher, the dynamic exchange of single metabolites can be detected, and the 1H proton magnetic resonance spectrum change can be detected under the condition of high spectral resolution²Metabolites which cannot be detected by the H proton magnetic resonance spectrum are obtained, so that the rate of in vivo metabolic cycle is obtained, steady state information and metabolic rate of a plurality of metabolites can be provided by one-time acquisition, and meanwhile, the deuterated FMISO used in the invention can be taken, so that the human body cannot be injured by multiple detections; for the¹The detection of the H proton magnetic resonance spectrum can also use a standard nuclear magnetic resonance instrument, does not need special equipment, has lower cost and uses the standard nuclear magnetic resonance instrument¹The conversion of deuterium marks can be directly monitored by H proton magnetic resonance spectrum acquisition hardware and signal processing, and the method is simple and practical, has high precision and reliable result, and can quantitatively position and analyze the metabolic condition.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.