Deuterium-labeled probe for diagnosing and monitoring ad using magnetic resonance deuterium imaging application technology

Abstract

Provided is a deuterium-labeled probe for diagnosing and monitoring AD using a magnetic resonance deuterium imaging application technology. The present invention provides use of a deuterium-labeled molecular probe or a composition comprising a deuterium-labeled molecular probe in the preparation of a kit or system, wherein the kit or system is used for accurately diagnosing Alzheimer's disease or dynamically monitoring progression of Alzheimer's disease in a subject, the deuterium-labeled molecular probe is selected from deuterium-labeled glucose, a deuterium-labeled acetate, or a deuterium-labeled amino acid, and the Alzheimer's disease is early-stage, intermediate-stage, or late-stage Alzheimer's disease. The present invention achieves early and accurate detection of Alzheimer's disease.

Description

Deuterium-labeled probes are used in magnetic resonance deuterium imaging techniques to diagnose and monitor Alzheimer's disease (AD).

[0001] This application claims priority to Chinese invention patent application No. 202510095910.0, filed on January 21, 2025, entitled "Diagnosis and Monitoring of AD Using Deuterium-Labeled Probes with Magnetic Resonance Deuterium Imaging". Technical Field

[0002] This invention relates to the field of magnetic resonance imaging application technology, specifically to the use of deuterium-labeled probes in magnetic resonance deuterium imaging application technology for the diagnosis and monitoring of AD. Background Technology

[0003] Alzheimer's disease (AD) is a neurodegenerative disease with an extremely high rate of death and disability. Due to its long course, severe impact, and large number of patients, Alzheimer's disease is one of the most burdensome socioeconomic diseases worldwide. Currently, there is still no cure for Alzheimer's disease; early detection and intervention are the most effective means of slowing its progression in clinical practice.

[0004] Clinically, the diagnosis of Alzheimer's disease (AD) mainly relies on cognitive scale tests and neuroimaging examinations. However, structural abnormalities such as brain atrophy and functional cognitive impairment appear late, and cognitive assessments are highly subjective and have low accuracy, leading to a large number of AD patients being diagnosed late or not at all. As typical pathological features of AD, molecular markers such as β-amyloid protein (Aβ) and neurofibrillary tangles (tau protein) have been suggested for the early clinical diagnosis of AD. Aβ and tau protein can be detected through cerebrospinal fluid (CSF) puncture or PET imaging. However, in 2021, the Alzheimer's Disease International Working Group published an article in Lancet Neurology pointing out that abnormalities in molecular markers such as Aβ are not necessarily related to functional cognitive impairment and AD.

[0005] Mounting evidence suggests that abnormalities in brain molecular metabolism can appear as early as 20 years before the onset of clinical symptoms in Alzheimer's disease (AD). Pathological changes resulting from abnormalities in brain metabolic pathways (such as those for glucose, acetate, and amino acids) and metabolic flux characteristics (such as reaction rates and clearance rates) may be decisive factors driving AD progression. For example, in the early stages of the disease, oxidative damage to the glucose glycolysis and tricarboxylic acid (TCA) cycle metabolic pathways in the brain leads to a decrease in aerobic glycolysis levels in neurons, resulting in excessive accumulation of Aβ protein and hyperphosphorylation of tau protein, ultimately causing synaptic dysfunction and impaired neurocognitive function. Furthermore, changes in metabolic flux characteristics such as the rate of acetate consumption in microglia associated with neuroinflammation and the rate of neurotransmitter circulation in neurons are also considered major contributing factors to AD progression. A 2020 article in JAMA pointed out that energy metabolism, represented by glucose, is taking center stage in AD research. Therefore, in-depth research into the pathogenesis of AD from the perspective of brain energy metabolism pathways and metabolic flux characteristics, to achieve early diagnosis and reflect disease progression, has become an inevitable trend.

[0006] However, current cutting-edge research on AD molecular metabolism is mainly based on animal models, and related methods are difficult to translate into clinical applications. The fundamental reason for this is that the means to reflect the metabolic pathways and metabolic flows of energy substances in the human brain are extremely limited. Currently, PET and magnetic resonance imaging (MRI) molecular metabolic imaging methods, which can only be used on humans, can only reflect brain metabolic activities in a partial way and cannot accurately characterize the metabolic pathways and metabolic flows of brain energy substances: (1) PET uses radioactive probes to bind to specific receptors, transport proteins or biological enzymes, and reflects the concentration of specific markers in different areas of the brain through the amount of probe uptake. However, PET probes do not directly participate in the molecular metabolic process of the brain, and cannot detect the downstream metabolites of the probes. Therefore, they can only indirectly reflect the molecular metabolic activities of the brain; (2) Magnetic resonance imaging can directly analyze the concentration of various neurotransmitters in brain regions through spectral technology, but this method can only describe the metabolic state of the brain at a certain moment, lacks specific reflection of specific metabolic pathways, and traditional methods based on hydrogen protons ( 1 Magnetic resonance spectroscopy (MRS) for Alzheimer's disease (AD) suffers from problems such as low accuracy and poor stability in quantifying metabolites, making it extremely difficult to promote and use clinically. Therefore, developing new methods that can reflect the metabolic pathways and metabolic flows of energy substances in the human brain has become an urgent need for the early diagnosis of AD.

[0007] Glucose is one of the most important energy sources for the human body. Its metabolic process is very complex, involving multiple organs and biochemical pathways to produce energy. Abnormal glucose metabolism can lead to various diseases, the most common being diabetes. However, multiple pieces of evidence show that the development of many other major diseases, such as malignant tumors, are closely related to abnormal glucose metabolism.

[0008] The metabolism of energy substances, represented by glucose, has entered the central stage of research on various major diseases. Therefore, it has become an inevitable trend to conduct in-depth research on the pathogenesis of major diseases from the perspective of glucose metabolic pathways and metabolic flux characteristics, and to achieve early diagnosis to reflect disease progression. Summary of the Invention

[0009] In normal cells, glucose is taken up by the cell and converted into pyruvate, which is then transported to the mitochondria to participate in the tricarboxylic acid cycle. However, in tumor cells, even under sufficient oxygen conditions, tumor cells have a higher glucose uptake rate than normal cells and are more inclined to supply energy to the cell through glycolysis, resulting in the production of large amounts of lactic acid, a phenomenon known as the Warburg effect.

[0010] The inventors discovered that in the very early stages of AD, the brain of patients suffers oxidative damage due to glucose glycolysis and the tricarboxylic acid cycle metabolic pathway. The decrease in the level of aerobic glycolysis in neuronal cells promotes the excessive accumulation of β-amyloid protein (Aβ) and leads to the hyperphosphorylation of tau protein, ultimately resulting in synaptic dysfunction and impaired neurocognitive function.

[0011] Traditional cognitive scales and structural imaging can only be used to detect patients in the middle and late stages of Alzheimer's disease (AD). Detection methods based on molecular markers such as Aβ and tau proteins can detect early abnormalities, but they lack the ability to predict disease progression, thus limiting their clinical value. To address the shortcomings of existing methods, this invention develops a magnetic resonance deuterium (2H) imaging method to explore the pathogenesis of AD from the perspective of human brain energy metabolism pathways and metabolic flux characteristics, enabling early diagnosis and reflecting disease progression. This method is expected to fill the gap in current AD diagnosis by lacking information on abnormal brain energy metabolism activities and provide a new approach to elucidating the pathogenesis of AD from the perspective of molecular metabolic abnormalities, achieving early AD diagnosis, and discovering new targets for AD treatment.

[0012] Accurate detection of 2H-labeled probes and their downstream metabolites is a prerequisite and foundation for visualizing the metabolic pathways of energy substances in the human brain and quantifying metabolic flux. However, the concentration of 2H-labeled metabolites in the human body is extremely low (3-4 orders of magnitude lower than that of 1H), and it changes dynamically with metabolic activity. Furthermore, the metabolic characteristics differ in different brain regions, thus placing extremely high demands on the sensitivity, temporal resolution, and spatial resolution of 2H detection. In imaging principles, detection sensitivity and high temporal and spatial resolution are inherently mutually restrictive, and traditional methods struggle to simultaneously achieve all three. This application presents an innovative method for achieving high temporal and spatial resolution detection of low-concentration 2H-labeled metabolites and accurate quantification of metabolic flux.

[0013] In one aspect, the use of deuterium-labeled molecular probes or compositions containing deuterium-labeled molecular probes in the preparation of kits or systems for the precise diagnosis of Alzheimer's disease in subjects or for the dynamic monitoring of Alzheimer's disease progression or for determining the risk of subjects developing Alzheimer's disease is provided, wherein the deuterium-labeled molecular probes are selected from deuterium-labeled glucose, deuterium-labeled acetate, or deuterium-labeled amino acids.

[0014] In one implementation, the deuterium-labeled molecular probe is administered orally or by injection.

[0015] In one implementation, the subject is a human or a mammal.

[0016] In one implementation, Alzheimer's disease is early, intermediate, or late-stage Alzheimer's disease.

[0017] In one embodiment, the composition comprises a first reagent or first device for detecting metabolites of a deuterated molecular probe in the brain, preferably a magnetic resonance imaging system.

[0018] In one embodiment, the deuterium-labeled glucose is 1, 2, 3, 4, 5, or 6-deuterated glucose. In one embodiment, the deuterium-labeled glucose is [2, 3, 4, 6, 6'- 2 H5]-glucose. In one embodiment, the deuterated acetate is a 1, 2, 3, or 4-deuterated acetate, preferably [ 2 [H3]-acetate. In one embodiment, the deuterium-labeled amino acid is [ 2 [H]-amino acid. In one embodiment, the acetate is selected from sodium acetate and potassium acetate.

[0019] In one embodiment, the metabolite is selected from water, glutamate or glutamate, glutamine, and lactic acid. In another embodiment, the metabolite is glutamate or glutamate and / or glutamine.

[0020] In one embodiment, the composition includes a second reagent or a second device for detecting a deuterated molecular probe, preferably a magnetic resonance imaging system.

[0021] In one embodiment, the composition includes magnetic resonance. 2 H-signal acquisition component. In one implementation, 2 In the H signal acquisition component 1 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 2; 2 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 4.

[0022] In one embodiment, the composition includes a memory storing instructions. These instructions can be executed by a processor during magnetic resonance. 2 Short repetition time (TR) imaging sequences for H-wave spectroscopy, such as steady-state free precession inductively decayed signal sequences or equilibrium steady-state free precession sequences. Instructions can be executed by the processor during magnetic resonance imaging. 2 Quantitative estimation method for H signal.

[0023] In one implementation, an increase in the intensity, level, amount, or rate of production of metabolites in a subject's brain compared to a reference level indicates that the subject has Alzheimer's disease or is at risk of having Alzheimer's disease; preferably, the reference level is the level in a normal subject or the normal level in the brain.

[0024] On one hand, a method is provided for the accurate diagnosis of Alzheimer's disease or dynamic monitoring of Alzheimer's disease progression or determination of the risk of developing Alzheimer's disease in subjects, including administering a deuterium-labeled molecular probe to the subject, wherein the deuterium-labeled molecular probe is selected from deuterium-labeled glucose, deuterium-labeled acetate or deuterium-labeled amino acid.

[0025] In one implementation, the deuterium-labeled molecular probe is administered orally or by injection.

[0026] In one implementation, the subject is a human or a mammal.

[0027] In one implementation, Alzheimer's disease is early, intermediate, or late-stage Alzheimer's disease.

[0028] In one embodiment, the method further includes detecting in the subject the metabolites of the deuterated molecular probe in the brain using a first reagent or a first device, preferably a magnetic resonance imaging system.

[0029] In one embodiment, the deuterium-labeled glucose is 1, 2, 3, 4, 5, or 6-deuterated glucose. In one embodiment, the deuterium-labeled glucose is [2, 3, 4, 6, 6'- 2H5]-glucose. In one embodiment, the deuterated acetate is a 1, 2, 3, or 4-deuterated acetate, preferably [ 2 [H3]-acetate. In one embodiment, the deuterium-labeled amino acid is [ 2 [H]-amino acid. In one embodiment, the acetate is selected from sodium acetate and potassium acetate.

[0030] In one embodiment, the metabolite is selected from water, glutamic acid or glutamate, glutamine, and lactic acid.

[0031] In one implementation, the metabolite is glutamate or glutamate and / or glutamine.

[0032] In one embodiment, the method further includes detecting the deuterated molecular probe with a second reagent or a second device, preferably a magnetic resonance imaging system.

[0033] In one embodiment, the first device and / or the second device includes a magnetic resonance imaging (MRI) sensor. 2 H-signal acquisition component. In one implementation, 2 In the H signal acquisition component 1 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 2; 2 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 4.

[0034] In one embodiment, the first and / or second means includes a memory storing instructions. These instructions can be executed by the processor during magnetic resonance imaging. 2 Short repetition time (TR) imaging sequences for H-wave spectroscopy, such as steady-state free precession inductively decayed signal sequences or equilibrium steady-state free precession sequences. Instructions can be executed by the processor during magnetic resonance imaging. 2 Quantitative estimation method for H signal.

[0035] In one embodiment, an increase in the intensity, level, amount, or rate of production of metabolites in the subject's brain (hypothalamic region) compared to a reference level indicates that the subject has Alzheimer's disease or is at risk of developing Alzheimer's disease. Preferably, the reference level is the level in a normal subject or a normal level in the brain. In one embodiment, an increase in the rate of production of glutamate or glutamate and / or glutamine compared to a reference level indicates that the subject has Alzheimer's disease or is at risk of developing Alzheimer's disease.

[0036] In another aspect, a device is provided for the accurate diagnosis of Alzheimer's disease or for determining the risk of a subject developing Alzheimer's disease, comprising:

[0037] The detection unit is configured to perform magnetic resonance imaging (MRI) of the subject's brain at one or more time points before and / or after substrate administration. 2 H-imaging;

[0038] Collection unit, configured to collect magnetic resonance images of the subject's brain. 2 H-imaging information;

[0039] A computing unit configured to calculate the substrate consumption rate and the metabolite generation rate, and compare them with those of a normal subject, wherein if the detected substrate consumption rate is reduced and / or the metabolite generation rate is increased compared with a normal subject, the subject being tested has or is at risk of having Alzheimer's disease; wherein the substrate is a deuterium-labeled molecular probe.

[0040] The device may also have a storage unit for storing instructions that, when executed by the processor, will produce magnetic resonance. 2 Short repetition time (TR) imaging sequences for H-wave spectral imaging, such as those based on steady-state free precession inductively decaying signal sequences or equilibrium steady-state free precession sequences, and / or the instructions described herein are executed by the processor to produce magnetic resonance imaging. 2 Quantitative calculation method for H signal.

[0041] The detection unit may include magnetic resonance 2 H-signal acquisition component. In one implementation, 2 In the H signal acquisition component 1 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 2; 2 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 4.

[0042] In one embodiment, the deuterium-labeled molecular probe is selected from deuterium-labeled glucose, deuterium-labeled acetate, or deuterium-labeled amino acids. In one embodiment, the subject is a human or mammal. In one embodiment, the Alzheimer's disease is early, intermediate, or late-stage Alzheimer's disease.

[0043] In one embodiment, the deuterium-labeled glucose is a 1, 2, 3, 4, 5, or 6-deuterated glucose, preferably [2, 3, 4, 6, 6'- 2 H5]-glucose. In one embodiment, the deuterated acetate is a 1, 2, 3, or 4-deuterated acetate, preferably [ 2 [H3]-acetate. In one embodiment, the deuterium-labeled amino acid is [ 2 [H]-amino acid. In one embodiment, the acetate is selected from sodium acetate and potassium acetate.

[0044] In one embodiment, the metabolite is selected from water, glutamate or glutamate, glutamine, and lactic acid. In another embodiment, the metabolite is glutamate or glutamate and / or glutamine.

[0045] In one implementation, the detection unit is a magnetic resonance imaging system, preferably a 3-10T magnetic resonance imaging system, such as a 4, 5, 6, 7, 8 or 9T magnetic resonance imaging system.

[0046] In one implementation, the substrate consumption rate and metabolite generation rate of normal subjects.

[0047] In one embodiment, an increase in the intensity, level, amount, or rate of production of metabolites in the subject's brain (hypothalamic region) compared to a reference level indicates that the subject has Alzheimer's disease or is at risk of developing Alzheimer's disease. Preferably, the reference level is the level in a normal subject or a normal level in the brain. In one embodiment, an increase in the rate of production of glutamate or glutamate and / or glutamine compared to a reference level indicates that the subject has Alzheimer's disease or is at risk of developing Alzheimer's disease.

[0048] In another aspect, a method for constructing a model for determining the progression of Alzheimer's disease is provided. This method includes using magnetic resonance deuterium spectroscopy imaging to track the dynamic processes of metabolites from a deuterated probe in different brain regions of a subject at one or more time points, wherein the subject has been administered the deuterated molecular probe, which is selected from deuterated glucose, deuterated acetate, or deuterated amino acids. In one embodiment, the subject is a human or mammal, preferably a rhesus monkey. In one embodiment, the subject is a subject with early, middle, or late Alzheimer's disease, or a normal subject at different ages, such as a naturally aging subject. In one embodiment, the metabolites are selected from water, glutamate or glutamate, glutamine, and lactate. In one embodiment, the dynamic processes include the dynamic processes of the deuterated probe consumption rate and the metabolite production rate.

[0049] In one implementation, the method further includes classifying the subject's disease progression into different stages using gait measurement, cerebrospinal fluid biomarkers, and PET / MRI imaging; and combining magnetic resonance imaging... 2 H metabolic flow imaging was used to compare metabolic flow characteristics at different stages of Alzheimer's disease and establish a metabolic flow signature map. In one implementation, cerebrospinal fluid biomarkers were selected from Aβ and tau proteins.

[0050] In one embodiment, an increase in the intensity, level, amount, or rate of production of metabolites in the subject's brain (hypothalamic region) compared to a reference level indicates that the subject has Alzheimer's disease or is at risk of developing Alzheimer's disease. Preferably, the reference level is the level in a normal subject or a normal level in the brain. In one embodiment, an increase in the rate of production of glutamate or glutamate and / or glutamine compared to a reference level indicates that the subject has Alzheimer's disease or is at risk of developing Alzheimer's disease.

[0051] In another aspect, a device for dynamically monitoring the progression of Alzheimer's disease in subjects is provided, comprising:

[0052] The detection unit is configured to perform magnetic resonance imaging (MRI) of the subject's brain at one or more time points before and / or after substrate administration. 2 H-imaging;

[0053] Collection unit, configured to collect magnetic resonance images of the subject's brain. 2 H-imaging information;

[0054] The computing unit is configured to calculate the consumption rate of the substrate and the generation rate of the metabolites, and compare them with the model described herein. If the detected consumption rate of the substrate and the generation rate of the metabolites are consistent with the consumption rate of the substrate and the generation rate of the metabolites in a specific disease stage in the model, the subject is considered to be in that specific disease stage.

[0055] The subject was given the deuterium-labeled molecular probe, which was selected from deuterium-labeled glucose, deuterium-labeled acetate, or deuterium-labeled amino acids.

[0056] The detection unit may include magnetic resonance 2 H-signal acquisition component. In one implementation, 2 In the H signal acquisition component 1 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 2; 2 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 4.

[0057] The device may also have a storage unit for storing instructions that, when executed by the processor, will produce magnetic resonance. 2 Short repetition time (TR) imaging sequences for H-wave spectral imaging, such as those based on steady-state free precession inductively decaying signal sequences or equilibrium steady-state free precession sequences, and / or the instructions described herein are executed by the processor to produce magnetic resonance imaging. 2 Quantitative calculation method for H signal.

[0058] In one embodiment, the subject is a human or mammal, preferably a rhesus monkey. In one embodiment, the subject is a subject with early, intermediate, or late-stage Alzheimer's disease. In one embodiment, the metabolite is selected from water, glutamate or glutamate, glutamine, and lactate. In one embodiment, the metabolite is glutamate or glutamate and / or glutamine.

[0059] In one embodiment, an increase in the intensity, level, amount, or rate of production of metabolites in the subject's brain (hypothalamic region) compared to a reference level indicates that the subject has Alzheimer's disease or is at risk of developing Alzheimer's disease. Preferably, the reference level is the level in a normal subject or a normal level in the brain. In one embodiment, an increase in the rate of production of glutamate or glutamate and / or glutamine compared to a reference level indicates that the subject has Alzheimer's disease or is at risk of developing Alzheimer's disease.

[0060] In this document, deuterium-labeled glucose can be 1, 2, 3, 4, 5, or 6-deuterated glucose, preferably [2, 3, 4, 6, 6'- 2 H5]-glucose such as [2,3,4,6,6'- 2 [H5]-D-glucose. In this document, the deuterated acetate can be a 1, 2, 3, or 4-deuterated acetate, preferably [ 2 [H3]-acetate. Deuterium-labeled amino acids can [ 2 [H]-amino acid. Acetate can be selected from sodium acetate and potassium acetate.

[0061] The advantages of this invention include at least the following:

[0062] (1). Develop a method for visualizing and quantifying the glucose metabolism pathway in the human brain based on magnetic resonance molecular imaging technology, and use this technology to guide the early diagnosis of AD and tumor-related diseases;

[0063] (2) For individuals, data on glucose metabolism distribution and metabolic flux can be established to guide reasonable diet and exercise, improve physical fitness, and reduce the incidence of AD.

[0064] (3) Traditional cognitive scales and structural imaging can only be used to detect patients in the middle and late stages of AD. Detection methods based on molecular markers such as Aβ and tau proteins can detect early abnormalities, but they do not have the ability to predict disease progression and have limited clinical value. In view of the shortcomings of existing methods, this invention develops a magnetic resonance deuterium (2H) imaging method to explore the pathogenesis of AD from the perspective of human brain energy substance metabolism pathways and metabolic flow characteristics, realize early diagnosis and reflect disease progression. It is expected to fill the gap in the current diagnosis of AD disease by lacking information on abnormal brain energy substance metabolism activities, and provide a new approach to elucidating the pathogenesis of AD from the perspective of molecular metabolic abnormalities, realizing early diagnosis of AD and discovering new targets for AD treatment.

[0065] (4) Accurate detection of 2H-labeled probes and their downstream metabolites is the prerequisite and foundation for visualizing the metabolic pathways of energy substances in the human brain and quantifying metabolic flow. However, the concentration of 2H-labeled metabolites in the human body is extremely low (3-4 orders of magnitude lower than that of 1H), and it changes dynamically with metabolic activity. Furthermore, the metabolic characteristics of different brain regions differ, thus placing extremely high demands on the sensitivity, temporal resolution, and spatial resolution of 2H detection. In terms of imaging principles, detection sensitivity and high temporal and spatial resolution are inherently mutually restrictive, and traditional methods struggle to achieve all three simultaneously. This study will collaboratively innovate in multiple aspects, including signal acquisition components, signal acquisition methods, and signal reconstruction methods. Starting from the physical principles of magnetic resonance imaging and cutting-edge technologies such as artificial intelligence, it aims to break the mutual constraints between detection sensitivity, temporal resolution, and spatial resolution, and achieve innovative methods for high spatiotemporal resolution detection of low-concentration 2H-labeled metabolites and accurate quantification of metabolic flow.

[0066] (5) This invention can diagnose, predict or determine the likelihood or risk of a subject developing AD at an early stage. Attached Figure Description

[0067] Figure 1: Typical deuterium spectra of the hypothalamus in 5xFAD and C57 mice at high-resolution 1H images 80 minutes after infusion. Glx (glutamate and / or glutamine) levels in this region were higher in 5xFAD mice than in C57 mice. Figure 1(A) is an anatomical image of the mouse brain. (B) 2H spectra of pixel 1 marked in 5xFAD and C57 mice (A) at 80 minutes. Signal intensities of HDO, glucose (Glc), and Glx are shown in Figure 2. The average signal intensity was taken across all pixels in the slice containing the largest brain volume. Higher Glx production was also observed in 5xFAD mice, particularly after 80 minutes. Throughout the experiment, 5xFAD mice had a higher average Glc intensity, but the consumption rates were similar in both groups.

[0068] Figure 2: Mean signal intensity of HDO, Glc, and Glx over time. * indicates statistically significant difference between the two groups, p < 0.05.

[0069] Figure 3: Statistical plot of signal intensity of water, glucose, and Glx at 80 min in the 6 pixels annotated in Figure 1, A. The comparison shows that 5xFAD mice have higher levels of Glx production, especially in two regions of the hypothalamus. Detailed Implementation

[0070] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions in the embodiments of this invention will be clearly and completely described below in conjunction with the embodiments of this invention. Obviously, the described embodiments are only some embodiments of this invention, not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0071] As used herein, molecular probes are a class of tool molecules that can specifically recognize or bind to specific molecules (such as proteins, nucleic acids, metabolites, etc.) in a living organism, and achieve the detection, localization, or functional analysis of the target molecule through signal output (fluorescence, radioactivity, magnetic signals, etc.). In the embodiments described herein, the molecular probe is selected from deuterium-labeled glucose, deuterium-labeled acetate, or deuterium-labeled amino acids, preferably in oral or injectable form. The deuterium-labeled glucose can be 1, 2, 3, 4, 5, or 6-deuterated glucose, preferably [2, 3, 4, 6, 6'- 2 [H5]-glucose, such as [2,3,4,6,6'- 2 [H5]-D-glucose. The deuterated acetate can be a 1, 2, 3, or 4-deuterated acetate, preferably [ 2 H3]-acetate. The labeled amino acid can be [ 2 [H]-amino acids, preferably selected from sodium acetate and potassium acetate.

[0072] As used herein, metabolites refer to products formed by the molecular probe in a subject (e.g., in the subject's brain, such as the hypothalamus region). A subject's disease state or condition can be determined by comparing the levels of metabolites in the corresponding region of a normal subject or the levels of normal metabolites. In this study, Alzheimer's disease or the risk of developing Alzheimer's disease can be diagnosed at a very early or early stage based on the levels (e.g., elevated levels or rates of production) and / or reduced rates of substrate consumption of metabolites (e.g., water, glutamate or glutamate salts, glutamine, and lactate, especially glutamate or glutamate salts and / or glutamine).

[0073] As used herein, the production rate or generation rate in the context of metabolites refers to the amount of metabolites produced per unit time (e.g., within 120 minutes).

[0074] As used in this article, Alzheimer's disease (AD) is a common neurodegenerative disease of the elderly, characterized by progressive impairment of memory and other cognitive functions, and is the most common type of dementia in the elderly. Clinical manifestations include progressive recent and distant memory impairment, decline in analytical and judgment abilities, changes in personality and behavior, and even confusion. The natural course of Alzheimer's disease is usually 5–10 years. The concept of Alzheimer's disease can encompass a continuous process including preclinical AD, mild cognitive impairment (MCI) stage, and dementia stage. There is no strict division between stages, but all stages present with AD-related pathological manifestations. Alzheimer's disease can be divided into early, middle, or late Alzheimer's disease. Early Alzheimer's disease clinically manifests as recent memory impairment, decreased verbal communication ability, and difficulty with advanced daily living activities. Middle Alzheimer's disease clinically manifests as complete inability to learn and recall new information; impaired but not completely lost distant memory, poor concentration, and decline in daily living abilities. Late-stage Alzheimer's disease is clinically characterized by complete dependence on caregivers for daily living and a tendency towards cognitive decline. Advantageously, this invention can be used to diagnose or identify early-stage Alzheimer's disease.

[0075] As used in this article, magnetic resonance imaging (MRI) systems are medical imaging devices based on the principle of nuclear magnetic resonance. They utilize magnetic fields and radio frequency pulses to excite hydrogen protons in human tissues to generate resonance signals, which are then spatially encoded and reconstructed by computer to form non-invasive, high-resolution tomographic images. These images are characterized by no ionizing radiation, high soft tissue contrast, and multiplanar imaging, and are widely used for disease diagnosis. In MRI, the 2H (deuterium) signal acquisition component is a radio frequency system component specifically adapted to the characteristics of the deuterium nucleus. Its core is a radio frequency coil tuned to the 2H resonant frequency, along with dedicated power amplifiers, preamplifiers, and other components to address the low natural abundance and low gyromagnetic ratio of 2H. The 2H (deuterium) signal acquisition component is readily available to those skilled in the art.

[0076] As used in this article, precision diagnosis refers to a method of disease diagnosis that comprehensively considers the clinical characteristics of a disease and its underlying biological changes, as well as individual differences in genetic background, environment, and lifestyle. Compared to traditional clinical diagnosis, precision diagnosis is an evaluation system guided by disease-related biomarkers established based on large-scale bioinformatics data. Biomarkers can come from high-throughput, multimodal bioinformatics big data from large population cohorts, which can be used to perform fine-grained population stratification and characteristic classification of diseases.

[0077] As used in this article, dynamic monitoring in the context of disease refers to the continuous and regular observation and data collection of disease-related physiological indicators, pathological characteristics, or treatment responses, and the analysis of their changing trends to assess disease progression and guide treatment adjustments.

[0078] As used in this article, early diagnosis refers to the timely identification of a disease in its preclinical or early symptom stages through specific testing methods, screening approaches, or clinical assessments. Prospective diagnostic approaches occur before the full manifestation of typical disease symptoms, or when only mild, nonspecific symptoms are present.

[0079] In-depth research into the pathogenesis of Alzheimer's disease (AD) from the perspective of brain energy metabolism pathways and metabolic flux characteristics, to achieve early diagnosis and reflect disease progression, has become an inevitable trend. However, currently available clinical methods for detecting human molecular metabolism, such as PET and MRI, cannot reflect abnormal brain metabolic activity at the level of metabolic pathways and metabolic flux. To address this challenge, this paper proposes an innovative approach using deuterium (2H) magnetic resonance imaging to visualize brain energy metabolism pathways and quantify metabolic flux. This aims to overcome the major deficiency in current AD diagnosis—the lack of information on abnormal brain molecular metabolic activity—and achieve accurate early diagnosis of AD. Furthermore, it provides a novel approach to elucidating the pathogenesis of AD from the perspective of molecular metabolic abnormalities and discovering new therapeutic targets for AD.

[0080] Energy metabolism is fundamental to the normal functioning of the brain and its vital functions such as antioxidant defense. Neurons are highly sensitive to energy fluctuations; impaired glucose metabolism disrupts energy homeostasis, leading to neuronal dysfunction and cognitive impairment. Simultaneously, Aβ-induced neurotoxicity is also related to insufficient neuronal energy. However, the specific processes underlying these correlations—including the pathological characteristics of metabolic pathways such as glycolysis and the TCA cycle, the catalytic and balancing roles of pyruvate and lactate, and their changing trends with the progression of Alzheimer's disease (AD)—remain unclear. This application utilizes high spatiotemporal resolution magnetic resonance deuterium imaging to explore the changes in metabolic concentrations and kinetics of metabolites such as lactate and glutamate in the early stages of AD, constructing specific metabolic pathological models of glucose, acetate, and amino acids, and elucidating the progression pathway of AD, revealing the unique role of energy metabolism in the pathogenesis, diagnosis, and intervention of AD.

[0081] in human body 2The concentration of H-labeled metabolites is extremely low (3-4 orders of magnitude lower than that of 1H) and dynamically changes with metabolic activity. Furthermore, metabolic characteristics differ across different brain regions, thus placing extremely high demands on the sensitivity, temporal resolution, and spatial resolution of 2H detection. However, in imaging principles, detection sensitivity and high temporal and spatial resolution are inherently mutually restrictive, and traditional methods struggle to achieve all three simultaneously. Collaborative innovation is needed in signal acquisition components, methods, and reconstruction techniques to overcome these constraints and achieve high spatiotemporal resolution detection of low-concentration 2H-labeled metabolites, as well as precise quantification of metabolic flux.

[0082] Method Introduction

[0083] (1) High spatiotemporal resolution magnetic resonance driven by artificial intelligence and physical models 2 H imaging method

[0084] This application addresses the challenge of high spatiotemporal resolution detection of low-concentration 2H-labeled metabolites from three aspects: magnetic resonance signal acquisition components, spectral acquisition methods, and signal reconstruction. Based on electromagnetic field simulation, a dual-core, multi-channel signal acquisition component (2H (metabolic information) / 1H (structural information)) is developed to achieve uniform and highly sensitive acquisition of dual-core signals. According to the characteristics of magnetic resonance spectroscopy and its temporal variation, a high spatiotemporal resolution spectral imaging sequence is developed, and specific sampling trajectories are designed to improve sampling efficiency. Based on physical models such as 2H magnetic resonance spectroscopy and brain metabolic dynamics, an artificial intelligence-based physical model-driven metabolic flow parameter estimation framework is constructed, ultimately achieving accurate estimation of brain metabolic flow characteristics.

[0085] (2) Research on the correlation between brain energy metabolism characteristics in Alzheimer's disease (AD) and the progression of AD, and a diagnostic model.

[0086] This application utilizes a developed high spatiotemporal resolution magnetic resonance deuterium (2H) imaging method and novel multi-target 2H-labeled probes to visualize the metabolic pathways of substances such as glucose, amino acids, and acetate in the brain of large animal AD models, and to quantitatively analyze metabolic fluxes such as reaction rates and clearance rates. It explores the metabolic flux characteristics of the brain under normal and AD pathological states, constructing a metabolic flux feature atlas. Through longitudinal studies, it characterizes the pathological changes and pathways of metabolic flux at different stages of AD development, revealing the relationship between the dynamic characteristics of brain metabolic flux and AD disease progression. Based on a convolutional neural network model with a connection attention mechanism, it learns multi-level metabolic features for early AD diagnosis and establishes an AD diagnostic model. Through metabolic imaging and the establishment of a metabolic flux pathological model in non-human primates, it explores the transfer from animal to human applications, and ultimately, based on a 5.0T ultra-high field human magnetic resonance imaging system, it explores the clinical application of imaging technology and deuterium 2H probes.

[0087] Research Methods and Technical Approach

[0088] High spatiotemporal resolution magnetic resonance 2H imaging method driven by artificial intelligence and physical models

[0089] 1) Magnetic Resonance 2H Signal Acquisition Component: The performance of the magnetic resonance signal acquisition component determines the signal-to-noise ratio and detection sensitivity of the magnetic resonance signal. A component capable of simultaneously acquiring 1H (structural information) and 2H (metabolic information) signals needs to be designed. However, the interactions between different nuclear channels within the signal acquisition component, the coupling between channels, and the complex interactions between electromagnetic waves and biological tissue in high-intensity electromagnetic fields can severely reduce data transmission efficiency and receiving sensitivity.

[0090] To address the aforementioned challenges, a multi-channel frequency coil structure supporting 2H / 1H dual-core imaging was designed and fabricated based on radio frequency electromagnetic field simulation methods. This structure comprehensively utilizes low-input-impedance, low-noise preamplifiers and induced current decoupling to reduce mutual interference between channels, achieving uniform excitation and high-sensitivity acquisition of 2H / 1H dual-core signals. This application will be based on a 5T human magnetic resonance imaging system: the 1H excitation coil will have at least 2 channels, and the receiving coil will have at least 2 channels; the 2H excitation coil will have at least 2 channels, and the receiving coil will have at least 4 channels.

[0091] 2) Magnetic Resonance 2H Spectroscopy Acquisition Method: The magnetic resonance spectroscopy acquisition method determines the spatiotemporal resolution of metabolite detection. The ultimate goal is to quantify the metabolic flow characteristics of different brain regions, thus placing extremely high demands on the spatiotemporal resolution of 2H signal detection. To achieve high spatial resolution detection, the data acquisition speed inevitably slows down, resulting in insufficient temporal resolution and difficulty in accurately observing the dynamic changes of metabolites. To address this challenge, a high spatiotemporal resolution spectral imaging sequence was developed. Based on the characteristics of magnetic resonance spectroscopy and its temporal variation, a specific sampling trajectory was designed to reduce unnecessary sampling points and improve sampling efficiency, providing a methodological basis for achieving high spatiotemporal resolution deuterium (2H) imaging.

[0092] High spatiotemporal resolution 2H magnetic resonance imaging (MRI) requires optimization in three aspects: sequence design, sampling trajectory, and undersampling scheme. Compared with traditional 1H MRI, 2H has a shorter longitudinal relaxation time. Based on this characteristic, imaging sequences with shorter repetition times (TRs) can be selected for 2H MRI, such as steady-state free precession free induction attenuation signal (SSFP-FID) sequences or balanced steady-state free precession (balanced SSFP, bSSFP) sequences. In addition, 2H MRI sampling requires high spectral resolution but not high requirements for the spectral frequency range. Based on this characteristic, corresponding sampling trajectories such as Cartesian, concentric ring, or rosette can be selected. Furthermore, given the relatively small number of 2H RF coil channels, undersampling schemes will mainly focus on the time dimension. The project will select the optimal acquisition sequence, corresponding sampling trajectory, and undersampling scheme based on acquisition time, signal-to-noise ratio, spectral resolution, and other indicators for different application scenarios.

[0093] 3) Quantitative estimation method of 2H signal in magnetic resonance imaging: The quantitative estimation method of magnetic resonance signal must ensure the accuracy of metabolite detection and metabolic flux quantification. By using exogenous 2H-labeled probes, the downstream metabolites of the probes are dynamically detected, thereby reflecting the molecular metabolic activity characteristics of various brain regions. However, the concentration of 2H probes within the safe range for the human body is extremely low, making the detection of 2H-labeled metabolites easily affected by background noise, thus posing a significant challenge to accurate detection.

[0094] To address this issue, a framework for estimating metabolic flux parameters based on artificial intelligence will be constructed, starting from physical models such as the 2H magnetic resonance spectroscopy of the probe and brain metabolic dynamics, to achieve accurate estimation of brain metabolic flux characteristics. After the 2H-labeled probe is ingested, the probe and its downstream metabolites can be detected on the deuterium (2H) magnetic resonance spectrum. The signal model can be expressed as:

[0095] Where S represents the overall spectral signal intensity obtained from the reconstruction, Amm(xx,TT) is the content of metabolite mm at spatial location xx and time TT, M is the number of metabolites, and LLmm represents the distribution characteristics of metabolites in the frequency domain. Directly solving the above equations will result in a large fitting error due to the low signal-to-noise ratio, leading to inaccurate estimation of the content of each metabolite.

[0096] In the above model, since the probe and its metabolites are known, the frequency domain distribution characteristics Lm of each metabolite can be learned in advance using the acquired spectral data:

[0097] Among them, a n,m These are weighting coefficients. The number of spectral sampling points is 2Ns, the echo time is TE, and the sampling interval is Δf. The chemical shift of each metabolite is fm, and the main magnetic field offset in the actual sampling is Δfm. The chemical shift fm of each metabolite can be obtained based on prior knowledge. Based on the actual acquired spectral data and the above physical model, the frequency domain distribution characteristics Lm of each metabolite are learned in advance.

[0098] In addition to the frequency domain characteristics of metabolites, glucose and its downstream metabolites also satisfy pharmacokinetic constraints, and this prior information will also be used in the construction of the network model. A simplified kinetic model of deuterated glucose, as described in the literature (Neuroimage Clin. 2022; 33:102932), includes downstream products such as lactic acid, glutamate + glutamine, and water. The concentration variations of glucose and its downstream products are influenced by factors such as reaction rate and clearance rate.

[0099] Where [Glc], [Lac], and [Glx] are the concentrations (mM) of glucose, lactate, and glutamate + glutamine, respectively; kg is the rate of glucose transfer between blood and brain tissue (min). -1 ); denoted as ρ, where ρ is the concentration of glucose in plasma (mM); f is the enrichment of deuterium; v is the extracellular volume fraction outside the blood vessels; Vmax = Vlac + Vglx, meaning the rate of glucose consumption (Vmax) is the sum of the rates of lactate and glutamate + glutamine synthesis (Vlac and Vglx); km is the constant for glucose absorption; klac and kglx are the clearance rates of lactate and glutamate + glutamine, respectively.

[0100] Association between brain metabolic flow characteristics and AD disease progression and diagnostic model research

[0101] 1) Construction of normal and AD pathological models in non-human primates

[0102] Currently, transgenic mice are the primary animal models used in Alzheimer's disease (AD) research. However, mouse brain tissue differs significantly from the human brain in terms of structure, function, and genetic background, making it difficult to directly apply research findings from mice to humans. Non-human primates, on the other hand, share a high degree of similarity with humans in terms of genetic background, brain structure, pathological characteristics, and aging process, making them ideal experimental animals for AD research. Naturally aging monkeys exhibiting AD pathological characteristics and behavioral symptoms were selected as experimental subjects.

[0103] Deuterium (2H)-labeled probes (including [2,3,4,6,6'-2H5]-glucose, [2H3]-acetate, and [2H]-amino acids, etc.) were used, and magnetic resonance deuterium (2H) spectroscopy was employed to track the dynamic metabolic processes of deuterium (2H)-labeled probes in different brain regions of an animal model. Taking the glucose and acetate metabolic pathways as an example, magnetic resonance 2H spectroscopy could quantify the concentrations of downstream metabolites such as lactate, glutamate, and water in different brain regions at various time points. This enabled visualization of brain energy metabolism pathways and quantitative analysis of metabolic flux, thereby constructing and comparing the characteristics and differences in energy uptake and metabolic activities in the brain under normal and Alzheimer's disease conditions.

[0104] 2) Metabolic flux profiles of AD disease progression

[0105] Researching and establishing the metabolic characteristics of Alzheimer's disease (AD) at different stages of progression is of great significance for the diagnosis and evaluation of treatment efficacy. This study explores the metabolic flow characteristics corresponding to different stages of AD progression. Rhesus monkeys of different ages (middle-aged: around 10 years old, elderly: around 20 years old) were selected, and AD disease progression was divided into different stages using traditional gait measurements, cerebrospinal fluid biomarkers (Aβ, tau protein), and PET / MRI imaging (Alzheimers Dement, 2011; 7:270–279). Using these animal models, combined with 2H magnetic resonance imaging (MRI), the metabolic flow characteristics of different stages of AD were compared to establish a metabolic flow feature atlas. 3) Research on an AD diagnostic model based on CoT feature pyramid deep learning: Based on 2H MRI metabolic signals and dynamic feature information, a feature pyramid model based on Contextual Transformer (CoT) Block was constructed to learn multi-level features in AD 2H MRI metabolic flow feature images and 1H structural images.

[0106] The model mainly consists of four parts: an encoder, a decoder, skip connections, and feature combination. In the encoder, a series of feature maps at different resolutions are extracted, followed by normalization, activation functions, and max pooling layers. In the decoder, each step involves upsampling the feature maps, halving the number of feature channels, concatenating them with the corresponding feature maps from the encoder, and finally merging the resulting feature maps to generate classification probability values.

[0107] The encoder and decoder sections introduce a novel attention module, CoT, which fully leverages contextual information to guide the learning of a dynamic attention matrix, thereby enhancing the ability to represent image features. CoT integrates contextual information mining and self-attention learning into a unified architecture. It first performs k×k×k sets of convolutions on all adjacent key values ​​to spatially associate each key value with its context. The learned key values ​​with contextual information are denoted by K1. This module also uses two consecutive 1×1×1 convolutions to obtain the relation matrix A. The two convolutions are denoted by Wθ and Wδ, respectively. Before this, K1 is concatenated with Q to obtain the relation matrix A = [K1, Q]W. θ W δ This process no longer creates isolated QK pairs, but rather relational mappings rich in contextual feature information. Next, this relational matrix A is multiplied by V to obtain K2. K2 is a dynamic contextual representation of the input, capturing the dynamic feature interactions between inputs. Finally, an attention mechanism is used to fuse K1 and K2 to obtain the output.

[0108] Example

[0109] The following embodiments are provided to illustrate the present invention. Those skilled in the art should understand that the embodiments are merely illustrative and not restrictive. The invention is limited only by the scope of the appended claims.

[0110] Example 1:

[0111] method

[0112] A mouse AD model of 5xFAD (n=3, male, 6 months old) was used and compared with normal mice C57 (n=3, male, 6 months old). T2-weighted high-resolution anatomical images were obtained on a 9.4T animal system (uMR 9.4T, United Imaging Life Science Instruments, Wuhan, China) using a dedicated mouse brain coil with parameters of TR / TE=3000ms / 35.56ms, matrix=370*370, FOV=17mm*17mm, and slice thickness=0.2mm.

[0113] Deuterium magnetic resonance imaging was performed using a Bruker 11.7T MR system and a self-made 2H / 1H coil (Du F, Yuan J, Li N, et al. Deuterium metabolism imaging of rat brain at 9.4T using a double-nuclear transceiver [C] / / Proc.Intl.Soc.Mag.Reson.Med.2022,3243). All mice were anesthetized with isoflurane during imaging acquisition. The dose was dynamically adjusted within the range of 0.5-1.8% to maintain a respiratory rate of 30-40 breaths per minute throughout the experiment. All mice were injected via tail vein [2,3,4,6,6'- 2 [H5]-D-glucose (Dingbang Biotechnology Co., Ltd., Shenzhen, China), dissolved in physiological saline at a dose of 3 g / kg. T2-weighted anatomical images were also obtained at 11.7T, but due to coil […]. 1 Impaired H-imaging performance and degraded image quality. Chemical shift imaging (CSI) sequences were used to detect deuterated labeled substrates in the brain. Imaging parameters were: TR = 150 ms, matrix = 8x8x8, field of view = 16 mm x 16 mm x 16 mm, bandwidth = 2 kHz, acquisition points = 256, average value = 150, acquisition time = 16.5 min. The imaging sequence was repeated 7 times: once before glucose infusion as baseline, and 6 times after infusion.

[0114] Whole-brain spectral data were exported, and MATLAB code was used to perform inverse Fourier transform, line broadening, and phase correction. Then, the spectra were fitted to a Lorentzian model using a least-squares fitting algorithm (Zou C, Ruan Y, Li H, et al. A new deuterium-labeled compound [2,3,4,6,6'-). 2 [H5]-D-glucose for deuterium magnetic resonance metabolic imaging[J].NMR in Biomedicine,2023,36(7):e4890). Signal intensity was normalized relative to HDO obtained before glucose infusion. The signal intensity was normalized by using HDO obtained at 11.7T. 1 The H image was registered with the image at 9.4T to determine... 2 Anatomical localization using H-spectral data.

[0115] result

[0116] Figure 1 shows typical deuterium spectra in the hypothalamus region of 5xFAD and C57 mice at high resolution 1H images 80 minutes after infusion. Glx levels in this region are higher in 5xFAD mice than in C57 mice. Figure 1(A) is an anatomical image of the mouse brain. (B) 2H spectra of pixel 1 (marked) in 5xFAD and C57 mice at 80 minutes.

[0117] The signal intensities of HDO, glucose (Glc), and Glx are shown in Figure 2. The average signal intensity was taken across all pixels in the slice containing the largest brain volume. Higher Glx production was also observed in the 5xFAD mice, particularly after 80 minutes. Throughout the experiment, the 5xFAD mice had a higher average Glc intensity, but the consumption rates were similar in both groups.

[0118] Figure 2 shows the average signal intensities of HDO, Glc, and Glx over time. * indicates statistically significant differences between the two groups, p < 0.05.

[0119] Figure 3 shows a statistical plot of the signal intensities of water, glucose, and Glx at 80 min in the six pixels annotated in Figure A of Figure 1. The comparison shows that 5xFAD mice have higher levels of Glx production, particularly in two regions of the hypothalamus.

[0120] In AD models, whole-brain Glx production rates are higher. However, in this study, this significant difference was observed as early as 6 months of age (instead of 14 months). Furthermore, 3D CSI revealed consistently higher Glx production rates in the hypothalamus of 5xFAD mice. Additionally, there was no difference in glucose consumption between the two groups. This observation may differ from the 18F-FDG-PET study (Bouter C, Henniges P, Franke TN, et al.). 18F-FDG-PET detects drastic changes in brain metabolism in the Tg4-42 model of Alzheimer's disease[J]. Frontiers in aging neuroscience, 2019, 10: 425). However, in another study using a 7-month-old 3xTgAD mouse model with 13C-labeled glucose, the flux of 13C-labeled glucose was found to be even higher than that in normal mice (Sancheti H, Patil I, Kanamori K, et al. Hypermetabolic state in the 7-month-old triple transgenic mouse model of Alzheimer's disease and the effect of lipoic acid: a 13C-NMR study[J]. Journal of Cerebral Blood Flow & Metabolism, 2014, 34(11): 1749-1760). Further investigation into the inconsistent findings will be conducted. Comparisons will be extended comprehensively to PET and even behavioral characteristics of animal models. In addition, larger groups will continue to complete this preliminary study.

[0121] Early impairment of brain energy metabolism is a driving factor in the progression of Alzheimer's disease (AD). This embodiment explores the potential of using deuterium magnetic resonance imaging (dMRI) to characterize glucose metabolism in an AD mouse model. The method of this invention uses [2,3,4,6,6'- 2[H5]-D-glucose was used for deuterium magnetic resonance imaging in 5xFAD and C57 mice. Preliminary results showed that 5xFAD mice had higher glutamine / glutamate (Glx) production rates compared to normal mice, particularly in the hypothalamus region. This work may inspire the development of new methods for early detection of Alzheimer's disease (AD) based on abnormal glucose metabolism. Alzheimer's disease (AD) is a neurodegenerative disease with high mortality and disability rates. Early diagnosis is the most effective way to control disease progression, but it remains a huge challenge (Anand R, Gill KD, Mahdi A A. Therapeutics of Alzheimer's disease: Past, present and future[J]. Neuroopharmacology,2014,76:27-50). Recent studies have found that brain energy metabolism disorders occur many years before the onset of clinical symptoms and play an important role in disease progression (Jagust W. Imaging the evolution and pathophysiology of Alzheimer disease[J]. Nature Reviews Neuroscience, 2018, 19(11):687-700). In this study, we explored the potential of using deuterium metabolism imaging to reveal the characteristics of glucose metabolism in Alzheimer disease using an animal model (De Feyter HM, Behar KL, Corbin ZA, et al. Deuterium metabolic imaging (DMI) for MRI-based 3D mapping of metabolism in vivo[J]. Science advances, 2018, 4(8):eaat7314).

[0122] While the invention has been described with respect to several embodiments at considerable length and in some detail, it is not intended to limit it to any such details or embodiments or any specific implementation. Rather, it should be interpreted with reference to the appended claims, which are intended to provide the broadest possible interpretation based on the prior art and thus effectively cover the intended scope of the invention. It should be understood that although the invention has been described in conjunction with its detailed description, the foregoing description is intended to clarify, rather than limit, the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the appended claims.

Claims

1. Use of a deuterium-labeled molecular probe or a composition containing a deuterium-labeled molecular probe in the preparation of a kit or system for the precise diagnosis of Alzheimer's disease in a subject or for the dynamic monitoring of Alzheimer's disease progression or for determining the risk of developing Alzheimer's disease in a subject, wherein the deuterium-labeled molecular probe is selected from deuterium-labeled glucose, deuterium-labeled acetate, or deuterium-labeled amino acids, preferably in oral or injectable form; preferably, the subject is a human or mammal; preferably, the Alzheimer's disease is early, intermediate, or late-stage Alzheimer's disease.

2. The use according to claim 1, wherein the composition comprises a first reagent or a first device for detecting metabolites of deuterated molecular probes in the brain, preferably a magnetic resonance imaging system.

3. The use according to claim 1 or 2, wherein the deuterium-labeled glucose is 1, 2, 3, 4, 5, or 6-deuterated glucose, preferably [2, 3, 4, 6, 6'- 2 [H5]-glucose, such as [2,3,4,6,6'- 2 H5]-D-glucose; the deuterated acetate is a 1, 2, 3, or 4-deuterated acetate, preferably [ 2 H3]-acetate; the deuterium-labeled amino acid is [ 2 [H]-amino acid; preferably, the acetate is selected from sodium acetate and potassium acetate.

4. The use according to claim 2 or 3, wherein the metabolite is selected from water, glutamate or glutamate salt, glutamine and lactic acid; preferably wherein an increase in the intensity, level, amount or rate of production of the metabolite in the subject's brain compared to a reference level indicates that the subject has Alzheimer's disease or is at risk of having Alzheimer's disease; preferably, the reference level is the level in a normal subject or the normal level in the brain.

5. The use according to any one of claims 1-4, wherein the composition comprises a second reagent or a second device for detecting deuterium-labeled molecular probes, preferably a magnetic resonance imaging system; Preferably, the composition comprises magnetic resonance. 2 H signal acquisition component, preferably 2 In the H signal acquisition component 1 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 2; 2 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 4; The composition includes a memory storing instructions that, when executed by a processor, perform high spatiotemporal resolution magnetic resonance. 2 H-wave spectral acquisition, including sequence design, sampling trajectory, and undersampling process; Preferably, the sequence design includes magnetic resonance. 2 Short repetition time (TR) imaging sequences for H-wave spectral imaging, such as steady-state free precession free induction attenuation signal sequences or equilibrium steady-state free precession sequences. Preferably, the sampling trajectory is a sampling trajectory selected from Cartesian, concentric ring, or rosette and / or The undersampling process is an undersampling scheme focused on the time dimension; and / or The instructions are executed by the processor to produce magnetic resonance. 2 Quantitative estimation method for H signal.

6. A device for accurately diagnosing Alzheimer's disease or determining the risk of a subject developing Alzheimer's disease, comprising: The detection unit is configured to perform magnetic resonance imaging (MRI) of the subject's brain at one or more time points before and / or after substrate administration. 2 H-imaging; Collection unit, configured to collect magnetic resonance images of the subject's brain. 2 H-imaging information; preferably, the collection unit includes magnetic resonance imaging. 2 H signal acquisition component, preferably 2 In the H signal acquisition component 1 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 2; 2 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 4; The computing unit is configured to calculate the substrate consumption rate and the metabolite generation rate and compare them with the substrate consumption rate and metabolite generation rate of a normal subject, wherein if the detected substrate consumption rate is reduced and / or the metabolite generation rate is increased compared with a normal subject, then the subject being tested has or is at risk of having Alzheimer's disease. The substrate is a deuterium-labeled molecular probe; Optionally, a storage unit stores instructions that, when executed by the processor, perform high spatiotemporal resolution magnetic resonance. 2 H-wave spectral acquisition, including sequence design, sampling trajectory, and undersampling process; Preferably, the sequence design includes magnetic resonance. 2 Short repetition time (TR) imaging sequences for H-wave spectral imaging, such as steady-state free precession free induction attenuation signal sequences or equilibrium steady-state free precession sequences. Preferably, the sampling trajectory is a sampling trajectory selected from Cartesian, concentric ring, or rosette and / or The undersampling process is an undersampling scheme focused on the time dimension; and / or The instructions are executed by the processor to produce magnetic resonance. 2 Quantitative estimation method for H signal.

7. The apparatus of claim 6, wherein the deuterium-labeled molecular probe is selected from deuterium-labeled glucose, deuterium-labeled acetate, or deuterium-labeled amino acids; preferably, the subject is a human or mammal; preferably, the Alzheimer's disease is early, middle, or late Alzheimer's disease.

8. The apparatus of claim 7, wherein the deuterium-labeled glucose is 1, 2, 3, 4, 5, or 6-deuterated glucose, preferably [2, 3, 4, 6, 6'- 2 H5]-glucose such as [2,3,4,6,6'- 2 H5]-D-glucose; the deuterated acetate is a 1, 2, 3, or 4-deuterated acetate, preferably [ 2 H3]-acetate; the deuterium-labeled amino acid is [ 2 [H]-amino acid; preferably, the acetate is selected from sodium acetate and potassium acetate; Preferably, the metabolites are selected from water, glutamic acid or glutamate, glutamine, and lactic acid; Preferably, an increase in the intensity, level, amount, or rate of production of metabolites in the subject's brain compared to a reference level indicates that the subject has Alzheimer's disease or is at risk of having Alzheimer's disease; preferably, the reference level is the level in a normal subject or the normal level in the brain.

9. The apparatus according to any one of claims 6-8, wherein the detection unit is a magnetic resonance imaging system, preferably a 3-10T magnetic resonance imaging system, such as a 4, 5, 6, 7, 8 or 9T magnetic resonance imaging system.

10. The apparatus according to any one of claims 6-9, wherein the substrate consumption rate and metabolite generation rate of a normal subject.

11. A method for constructing a model for determining the progression of Alzheimer's disease, the method comprising using magnetic resonance deuterium spectroscopy imaging to track the dynamic processes of metabolites of a deuterated probe in different brain regions of a subject at one or more time points, wherein the subject has been administered the deuterated molecular probe, the deuterated molecular probe being selected from deuterated glucose, deuterated acetate, or deuterated amino acids; preferably, the subject is a human or mammal, preferably a rhesus monkey; preferably, the subject is a patient with early, intermediate, or late-stage Alzheimer's disease. Subjects, normal subjects of different ages, such as naturally aging subjects; preferably, wherein the metabolites are selected from water, glutamate or glutamate, glutamine and lactate; preferably, the dynamic process includes a dynamic process of the consumption rate of the deuterated probe and the generation rate of the metabolites; preferably, wherein an increase in the intensity, level, amount or generation rate of the metabolites in the subject's brain compared to a reference level indicates that the subject has Alzheimer's disease or is at risk of having Alzheimer's disease; preferably, the reference level is the level in normal subjects or the normal level in the brain; Preferably, the method includes magnetic resonance imaging. 2 H-wave spectroscopy employs imaging sequences with short repetition times (TR), such as those based on steady-state free precession inductively decaying signal sequences or equilibrium steady-state free precession sequences, and / or performs magnetic resonance imaging. 2 Quantitative estimation method for H signal.

12. The method of claim 11, further comprising classifying the subject's disease progression into different stages using gait measurement, cerebrospinal fluid biomarkers, and PET / MRI imaging; and combining magnetic resonance imaging... 2 H metabolic flow imaging was used to compare the metabolic flow characteristics at different stages of Alzheimer's disease and establish a metabolic flow characteristic map; preferably, the cerebrospinal fluid biomarkers were selected from Aβ and tau proteins.

13. A device for dynamically monitoring the progression of Alzheimer's disease in a subject, comprising: The detection unit is configured to perform magnetic resonance imaging (MRI) of the subject's brain at one or more time points before and / or after substrate administration. 2 H-imaging; Collection unit, configured to collect magnetic resonance images of the subject's brain. 2 H-imaging information; preferably, the collection unit includes magnetic resonance imaging. 2 H signal acquisition component, preferably 2 In the H signal acquisition component 1 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 2; 2 The number of channels in the H-excitation coil shall not be less than 2, and the number of channels in the receiving coil shall not be less than 4; A computing unit configured to calculate the consumption rate of the substrate and the generation rate of the metabolites, and to compare them with the model described in claim 11 or 12, wherein if the detected consumption rate of the substrate and the generation rate of the metabolites are consistent with the consumption rate of the substrate and the generation rate of the metabolites in a specific disease stage in the model, the subject is considered to be in that specific disease stage. Optionally, a storage unit stores instructions that, when executed by the processor, perform high spatiotemporal resolution magnetic resonance. 2 H-wave spectral acquisition, including sequence design, sampling trajectory, and undersampling process; The subject is given the deuterium-labeled molecular probe, which is selected from deuterium-labeled glucose, deuterium-labeled acetate, or deuterium-labeled amino acids. Preferably, the subject is a human or mammal, preferably a macaque. Preferably, the subject is a subject with early, middle, or late Alzheimer's disease. Preferably, the metabolite is selected from water, glutamate or glutamate, glutamine, and lactate. Preferably, an increase in the intensity, level, amount, or rate of production of the metabolite in the subject's brain compared to a reference level indicates that the subject has Alzheimer's disease or is at risk of having Alzheimer's disease. Preferably, the reference level is the level in a normal subject or the normal level in the brain. Preferably, the sequence design includes magnetic resonance. 2 Short repetition time (TR) imaging sequences for H-wave spectral imaging, such as steady-state free precession free induction attenuation signal sequences or equilibrium steady-state free precession sequences. Preferably, the sampling trajectory is a sampling trajectory selected from Cartesian, concentric ring, or rosette and / or The undersampling process is an undersampling scheme focused on the time dimension; and / or The instructions are executed by the processor to produce magnetic resonance. 2 Quantitative estimation method for H signal; Preferably, the deuterium-labeled glucose is 1, 2, 3, 4, 5, or 6-deuterated glucose, more preferably [2, 3, 4, 6, 6'- 2 H5]-glucose such as [2,3,4,6,6'- 2 H5]-D-glucose; the deuterated acetate is a 1, 2, 3, or 4-deuterated acetate, preferably [ 2 H3]-acetate; the deuterium-labeled amino acid is [ 2 [H]-amino acid; preferably, the acetate is selected from sodium acetate and potassium acetate.

Images