WO2016123841A1 - 老年痴呆和心脑血管疾病检测的多模态成像的制剂及应用 - Google Patents

老年痴呆和心脑血管疾病检测的多模态成像的制剂及应用 Download PDF

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WO2016123841A1
WO2016123841A1 PCT/CN2015/074688 CN2015074688W WO2016123841A1 WO 2016123841 A1 WO2016123841 A1 WO 2016123841A1 CN 2015074688 W CN2015074688 W CN 2015074688W WO 2016123841 A1 WO2016123841 A1 WO 2016123841A1
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imaging
cardiovascular
preparation
senile dementia
disease
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French (fr)
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王雪梅
来兰梅
赵春秋
苏美娜
李晓琦
姜晖
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东南大学
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Priority claimed from CN201510056250.1A external-priority patent/CN104689344B/zh
Priority claimed from CN201510060160.XA external-priority patent/CN104587493B/zh
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/04—X-ray contrast preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/22—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations

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  • the invention relates to a preparation and application of multimodal imaging for detecting dementia and cardiovascular and cerebrovascular diseases, which consists of a transition metal ion, a noble metal ion, a nanocluster thereof and a porphyrin derivative excellent in biocompatibility, through in situ Biological action fixed-point synthesis
  • the imaging detection reagent for precise targeted rapid labeling of relevant lesions can achieve non-invasive real-time dynamic high-sensitivity rapid tracing and monitoring, and has no biological side effects, and the detection method is simple and easy.
  • Alzheimer's disease is a central nervous system degenerative disease characterized by cognitive and visual spatial disorders, and has become one of the major diseases that threaten the health of the elderly.
  • AD Alzheimer's disease
  • the incidence of AD is increasing year by year, which has brought a series of medical and social problems, which has attracted the attention of the medical community and the community.
  • Alzheimer's disease is the most common type of Alzheimer's disease.
  • AD Alzheimer's disease
  • preclinical AD ie, high-risk AD
  • MCI mild cognitive impairment
  • Cardiovascular and cerebrovascular diseases are diseases involving the circulatory system.
  • the circulatory system refers to organs and tissues that transport blood in the human body, including the heart and blood vessels (arteries, veins, microvessels), etc., which can be subdivided into acute and chronic, generally with arteries. Hardening related. These diseases have similar causes, disease processes and treatments. However, to date, there has been no particularly effective method for the treatment of cardiovascular and cerebrovascular diseases. Therefore, early diagnosis has become an important way to improve the survival rate of patients with cardiovascular and cerebrovascular diseases.
  • cardiovascular and cerebrovascular diseases mainly include stroke, atherosclerosis, myocardial infarction, cerebral ischemia and cardiac ischemia and reperfusion injury, diabetes, brain tumors and the like.
  • Stroke is also called stroke, which is a general term for Chinese medicine for acute cerebrovascular diseases. It is a kind of cerebral blood circulation disorder disease that is fainted, unconscious, accompanied by skewed mouth and poor language and half-length is the main symptom. Due to the high incidence of stroke, high mortality, high disability rate, high recurrence rate and many complications, the medical profession ranks it as one of the three major diseases that threaten human health.
  • Atherosclerosis is the most common and important group of vascular diseases called arteriosclerosis. It is characterized by thickening and hardening of the arterial wall, loss of elasticity and shrinkage of the lumen. The appearance of lipids accumulated on the intima of the artery is yellow atheroma, which is called atherosclerosis. Mainly involving large and medium-sized arteries, its clinical manifestations are mainly based on the disease of the affected organs. According to the location of the affected arteries, the disease includes: aorta and its main branches, coronary artery, carotid artery, cerebral artery, renal artery, mesenteric artery and atherosclerosis of extremities. Generally manifested as the possibility of mental and physical decline.
  • Myocardial infarction is also called myocardial infarction.
  • Myocardial infarction is coronary occlusion and blood flow is interrupted, causing partial necrosis of some myocardium due to severe persistent ischemia.
  • AD Alzheimer's disease and cardiovascular and cerebrovascular diseases bring enormous social and economic burdens to individuals and society as a whole.
  • the existing imaging methods commonly used to diagnose AD mainly include MRI (magnetic resonance imaging), SPE-CT (single photon emission tomography), and PET (positron emission tomography).
  • MRI magnetic resonance imaging
  • SPE-CT single photon emission tomography
  • PET positron emission tomography
  • Commonly used in the diagnosis of cardiovascular and cerebrovascular diseases and related diseases are X-ray, electrocardiogram, conventional CT (computed tomography), magnetic resonance imaging, etc.
  • CT computed tomography
  • magnetic resonance imaging etc.
  • these commonly used diagnosis of senile dementia and cardiovascular and cerebrovascular diseases There are certain limitations in the method.
  • MRI provides a basis for morphology and certain functional studies for the diagnosis of AD, it lacks specificity; the sensitivity and specificity of SPE-CT are not strong, and early diagnosis is not achieved.
  • the present invention discloses senile dementia and cardiovascular and cerebrovascular diseases. Formulation and application of rapid detection and multimodal imaging. These preparations are composed of transition metal ions, noble metal ions, nano-clusters and porphyrin derivatives with excellent biocompatibility, and are integrated into the relevant lesions by in situ biological action. Accurately targeting fast-labeled imaging detection reagents enables non-invasive real-time dynamic, highly sensitive and fast tracing and monitoring without biotoxic side effects, and the detection method is simple and easy.
  • the present invention provides a preparation for early rapid detection and multimodal imaging of senile dementia and cardiovascular and cerebrovascular diseases, the imaging preparation being composed of transition metal ions, noble metal ions and their excellent biocompatibility
  • the nanocluster is composed of a porphyrin derivative, and the imaging detection reagent for accurately targeting the relevant lesions by precise in situ biosynthesis is synthesized.
  • the transition metal ion is one of cerium and zinc ions or a mixture of any two.
  • the noble metal ion is one of a gold ion, a silver ion, and a platinum ion, or a mixture of any two.
  • cardiovascular and cerebrovascular diseases refer to diseases such as stroke, atherosclerosis, myocardial infarction, cerebral ischemia and cardiac ischemia and reperfusion injury, diabetes, brain tumor and the like.
  • the nanoclusters are one or a mixture of any two of gold nanoclusters, silver nanoclusters, platinum nanoclusters, and the like.
  • porphyrin derivative is tetra-p-sulfonic acid-phenylporphyrin.
  • the above detection reagents are used in the early detection of senile dementia and cardiovascular and cerebrovascular diseases.
  • a transition metal ion, a noble metal ion, a nanocluster thereof and a porphyrin reagent excellent in biocompatibility are prepared, and the specific steps are as follows:
  • the above imaging preparations are used in the early detection of senile dementia and cardiovascular and cerebrovascular diseases.
  • the above application comprises the following steps: when the preparation is used for imaging senile dementia and cardiovascular and cerebrovascular diseases, metal ions and their nanocluster reagents and porphyrins are injected into the normal control model rats and the elderly by tail vein injection or local injection.
  • metal ions, their nanoclusters and porphyrin reagents can be rapidly enriched in the lesions, using small animal in vivo fluorescence imager, CT (computed tomography), Ultrasound and MRI (magnetic resonance imaging) were used to image the model mice.
  • Rats or stroke model mice after 1 to 72 hours of non-invasive real-time dynamic high-sensitivity rapid tracing and monitoring, observe the circulation of the body in the Alzheimer's disease model mouse or stroke model mice to achieve drug in the lesion Targeted enrichment and precise targeted rapid marker imaging of relevant lesion sites;
  • Imaging the lesions with a living fluorescence imager, CT (computed tomography) and MRI (magnetic resonance imaging) and qualitatively and quantitatively.
  • the above injection method is tail vein injection or local injection.
  • the reagent of the invention has good biocompatibility, no toxic and side effects, and can realize early, rapid, Alzheimer's disease (especially Alzheimer's disease) and cardiovascular and cerebrovascular diseases through specific enrichment in the lesion site. Multimodal imaging detection has broad medical application prospects.
  • the preparations used in this study consisted of transition metal ions with excellent biocompatibility, noble metal ions and their in situ biosynthesis of nanoclusters and porphyrin derivatives, targeting and targeting in vivo ischemia by targeting biologically active molecules.
  • Oxygen and other specific micro-environment can achieve accurate targeted labeling and rapid imaging detection of relevant lesions, and can perform non-invasive real-time dynamic high-sensitivity rapid tracing and monitoring, without biological side effects, and the detection method is simple and easy.
  • imaging detection such as CT (computed tomography), ultrasound, fluorescence and MRI (magnetic resonance imaging) can quickly identify the pathogenesis of the model mice and enrich in the region to achieve the target lesions. Quasi-marking, and the degree of enrichment in Alzheimer's model mice increased with increasing age.
  • Alzheimer's disease and cardiovascular and cerebrovascular diseases stroke, atherosclerosis, myocardial infarction, cerebral ischemia, cardiac ischemia and reperfusion injury, diabetes, brain tumors, etc.
  • rapid, multimodal such as CT (computed tomography), ultrasound, fluorescence and MRI (magnetic resonance imaging), etc.
  • CT computed tomography
  • MRI magnetic resonance imaging
  • the invention can realize the imaging of Alzheimer's disease and cardio-cerebral vascular diseases without damage, in situ and real-time dynamic targeting, and perform multi-modal and multi-modal simultaneous diagnosis and accurate targeted positioning and treatment, and has broad medical application. prospect.
  • Figure 1 is a graph showing changes in fluorescence intensity in the head lesions of different strokes at different time points.
  • a transition metal ion, a noble metal ion, a nanocluster thereof and a porphyrin reagent excellent in biocompatibility are prepared, and the specific steps are as follows:
  • the metal ion is one of a gold ion, a silver ion, a platinum ion, a zinc ion, or the like, or a mixture of any two;
  • the nanocluster is one of a gold nanocluster, a silver nanocluster, and a platinum nanocluster Or a mixture of any two;
  • the porphyrin is tetra-p-sulfonic acid-phenyl porphyrin (TSPP).
  • Alzheimer's disease model mice metal ions and their nanoclusters and porphyrin reagents for early, rapid and multimodal detection of Alzheimer's disease.
  • This reagent is used for imaging Alzheimer's disease with metal ions and their nanocluster reagents.
  • porphyrins are injected into the normal control model rats and Alzheimer's disease model mice by tail vein injection or local injection.
  • Metal ions, their nanoclusters and porphyrin reagents can be rapidly enriched in the brain lesions, using a small animal living fluorescence imager.
  • the model mouse is imaged; the specific steps are:
  • the transition metal ion is a mixture of zinc ions and noble metal ions as gold ions
  • the nanoclusters are gold nanoclusters
  • the porphyrin is tetra-p-sulfonic acid-phenyl porphyrin (TSPP);
  • the brain is imaged and qualitatively and quantitatively analyzed using a living fluorescence imager.
  • the images were imaged by a living fluorescence imager at different time points such as 1 hour, 4 hours, 8 hours, 12 hours, 24 hours, and 48 hours, and then the experimental model rats were gas anesthetized with 5% isoflurane. It is placed on the small animal living imager operating platform, and images of brain lesions can be acquired by selecting wavelength excitations such as 420 nm and 480 nm.
  • Example 2 The same as in Example 2, except that 0.1 mL of a sterile transition metal ion of 100 mmol/L and 100 mmol/L of noble metal ions were used.
  • the transition metal ion is a cerium ion
  • the noble metal ion is a platinum ion
  • the nanocluster is a silver nanocluster
  • the porphyrin is a tetra-p-sulfonic acid-phenyl porphyrin (TSPP).
  • TSPP tetra-p-sulfonic acid-phenyl porphyrin
  • Example 2 Same as Example 2, except that 0.5 mL of a sterile transition metal ion having a concentration of 0.1 mmol/L was used. And 100 mmol/L precious metal ions.
  • the transition metal ion is a zinc ion; the noble metal ion is a silver ion; the nanocluster is a platinum nanocluster; and the porphyrin is a tetra-p-sulfonic acid-phenyl porphyrin (TSPP).
  • TSPP tetra-p-sulfonic acid-phenyl porphyrin
  • a metal ion and a nanocluster and a porphyrin derivative reagent are prepared.
  • the specific steps are: dissolving the metal ion, its nanocluster and the porphyrin derivative reagent in ultrapure water, respectively, and preparing the concentration to be 0.1 to 100 mmol/L.
  • the metal ion is one of a gold ion, a silver ion, a platinum ion, a zinc ion, or the like, or a mixture of any two.
  • the nanoclusters are one of a gold nanocluster, a silver nanocluster, a platinum nanocluster or a mixture of any two; the porphyrin is a sulfonic acid porphyrin derivative.
  • the metal ion and its nanocluster reagent and porphyrin derivative reagent are injected into the animal body such as the normal control model mouse and the vascular injury-induced stroke model mouse by the tail vein injection or local injection, metal ion Its nanocluster and porphyrin derivative reagents can be rapidly enriched in the brain, such as ischemia and hypoxia, combined with small animal in vivo fluorescence imaging, CT (computed tomography), ultrasound and MRI (magnetic resonance imaging). ) Imagery such as model rats are imaged. Related specific steps include:
  • the experimental model rats are gas anesthetized with 5% isoflurane. It was placed on a small animal living imager operating platform, and wavelengths such as 420 nm and 480 nm were selected to excite images collected from the brain lesion area of living animals.
  • a reagent solution such as a metal ion, a nanocluster reagent and a porphyrin derivative is injected into a normal control model mouse and an atherosclerotic model mouse through a tail vein injection or a local injection.
  • Metal ions, their nanoclusters and porphyrin derivative reagents can be rapidly enriched in the lesions of blood vessels, using small animal in vivo fluorescence imager, CT (computed tomography), ultrasound, fluorescence and MRI (magnetic resonance imaging) The imager is imaged by an imager.
  • CT computed tomography
  • ultrasound fluorescence
  • MRI magnetic resonance imaging
  • the experimental model rats are gas anesthetized with 5% isoflurane. It was placed on a small animal living imager operating platform, and wavelengths such as 420 nm and 480 nm were selected to excite images of lesions collected on living animals.
  • Step 2) The metal ion is a mixture of gold ions and zinc ions; the nanoclusters are gold nanoclusters; and the porphyrin is a sulfonic acid porphyrin derivative.
  • Example 6 Same as Example 6, except that 0.1 mL of a sterile metal ion platinum ion having a concentration of 100 mmol/L is used; the nanocluster is a silver nanocluster; and the porphyrin derivative is a sulfonic acid porphyrin derivative. Things. The results were the same as in Example 6.
  • Example 6 The same as in Example 6, except that 0.5 mL of a sterile metal ion having a concentration of 0.1 mmol/L was used.
  • the metal ion is a mixture of silver ions and zinc ions; the nanoclusters are platinum nanoclusters; and the porphyrin derivative is a sulfonic acid porphyrin derivative.
  • the results were the same as in Example 6.

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Abstract

一种检测老年痴呆和心血管疾病的多模态成像的制剂，所述制剂由生物相容性优良的过渡金属离子、贵金属离子及其纳米簇与卟啉衍生物组成，通过原位生物作用定点合成对相关病灶部位精准靶向快速标记的成像检测试剂。

Description

老年痴呆和心脑血管疾病检测的多模态成像的制剂及应用 技术领域
本发明涉及老年痴呆和心脑血管疾病检测的多模态成像的制剂及应用，这些试剂由生物相容性优良的过渡金属离子、贵金属离子及其纳米簇与卟啉衍生物组成，通过原位生物作用定点合成对相关病灶部位精准靶向快速标记的成像检测试剂，可以实现非侵入式的实时动态的高灵敏快速示踪和监测，且无生物毒副作用，检测方法简便易行。
背景技术
老年期痴呆是以认知和视觉空间障碍为主要特征的中枢神经系统退行性疾病,已经成为威胁老年人健康的主要疾病之一。目前，随着全球人口老龄化的加快，AD(老年性痴呆症)发病率呈逐年升高的趋势，已经带来了一系列的医学和社会问题，引起了医学界和社会各界的重视。
阿尔茨海默病是老年痴呆症最常见的类型。2011年，美国国立老龄研究所和阿尔茨海默病协会根据疾病进程将AD(老年性痴呆症)分为临床前AD(即AD高危人群)、轻度认知功能障碍(mild cognitive impairment，MCI)及AD痴呆，并分别发布了3个阶段的诊断指南，重点关注了AD的早期诊断和治疗。然而，到目前为止，对于AD的治疗，仍无特别有效方法。所以，早期诊断成为提高AD患者生存率的重要途径。
心脑血管疾病是涉及循环系统的疾病，循环系统指人体内运送血液的器官和组织，主要包括心脏、血管(动脉、静脉、微血管)等，可以细分为急性和慢性，一般都是与动脉硬化有关。这些疾病都有着相似的病因、病发过程及治疗方法。然而，到目前为止，对于心脑血管疾病的治疗，仍无特别有效方法。所以，早期诊断成为提高心脑血管疾病患者生存率的重要途径。
常见的心脑血管疾病主要包括中风、动脉粥样硬化、心肌梗死、脑缺血和心脏缺血及其再灌注损伤、糖尿病、脑瘤等。
中风也叫脑卒中，是中医学对急性脑血管疾病的统称。它是以猝然昏倒，不省人事，伴发口角歪斜、语言不利而出现半身不遂为主要症状的一类脑血液循环障碍性疾病。由于中风发病率高、死亡率高、致残率高、复发率高以及并发症多的特点，所以医学界把它同冠心病、癌症并列为威胁人类健康的三大疾病之一。
因发病急骤，症见多端，病情变化迅速，与风之善行、数变特点相似，故名中风、卒中。 本病常留有后遗症，发病年龄也趋向年轻化，因此，是威胁人类生命和生活质量的重大疾患。
动脉粥样硬化是一组称为动脉硬化的血管病中最常见、最重要的一种。其特点是动脉管壁增厚变硬、失去弹性和管腔缩小，由于在动脉内膜上积聚的脂质外观呈黄色粥样，因此称为动脉粥样硬化。主要累及大中型动脉，其临床表现主要以受累器官的病象为主。按受累动脉部位的不同，本病有：主动脉及其主要分支、冠状动脉、颈动脉、脑动脉、肾动脉、肠系膜动脉和四肢动脉粥样硬化等类别。一般表现为可能出现脑力与体力衰退。
心肌梗死又叫心肌梗塞，心肌梗塞是冠状动脉闭塞，血流中断，使部分心肌因严重的持久性缺血而发生局部坏死。临床上有剧烈而较持久的胸骨后疼痛，发热、白细胞增多、红细胞沉降率加快，血清心肌酶活力增高及进行性心电图变化，可发生心律失常、休克或心力衰竭。
脑缺血和心脏缺血及其再灌注损伤，近年来.随着休克治疗的进步以及动脉搭桥术、溶栓疗法、经皮腔内冠脉血管成形术、心脏外科体外循环、心肺脑复苏，断肢再植和器官移植等方法的建立和推广应用，使许多组织器官缺血后重新得到血液再灌注。多数情况下，缺血后再灌注可使组织器官功能得到恢复，损伤的结构得到修复，患者病情好转康复；但有时缺血后再灌注.不仅不能使组织、器官功能恢复，反而加重组织、器官的功能障碍和结构损伤。这种在缺血基础上恢复血流后组织损伤反而加重，甚至发生不可逆性损伤的现象称为缺血再灌注损伤。
总而言之，老年痴呆和心脑血管疾病无论对于个人和整个社会都带来了巨大的社会和经济负担，随着科学技术的进步，已经有多种影像学方法及生物标志物应用于临床检测。现有的常用于诊断AD的影像学方法主要包括MRI(核磁共振成像)，SPE-CT(单光子发射断层成像)，PET(正电子发射断层成像)。常用于诊断心脑血管及其相关疾病的方法有X线、心电图、常规CT(计算机X线断层扫描)、磁共振成像等多种，然而，上述这些常用的诊断老年痴呆和心脑血管疾病的方法均有一定的局限性，例如：MRI虽然对AD的诊断提供了形态学和一定功能学的依据，但缺乏特异性；SPE-CT的敏感性和特异性也不强，达不到早期诊断AD的目的，而PET则因为价格昂贵，限制了它的应用。因此，寻找一种敏感又特异的微创检测技术对老年痴呆和心脑血管疾病的早期诊断尤为重要。但到目前为止，仍未见能对老年痴呆和心脑血管疾病进行早期、快速、实时、多模态成像的试剂相关的报道。
发明内容
技术问题：针对目前现有检测技术的缺陷，本发明公开了老年痴呆和心脑血管疾病早 期快速检测及多模态成像的制剂及应用，这些制剂由生物相容性优良的过渡金属离子、贵金属离子及其纳米簇与卟啉衍生物组成，通过原位生物作用定点合成对相关病灶部位精准靶向快速标记的成像检测试剂，可以实现非侵入式的实时动态的高灵敏快速示踪和监测，且无生物毒副作用，检测方法简便易行。
技术方案：为了解决上述技术问题，本发明提供了老年痴呆和心脑血管疾病早期快速检测及多模态成像的制剂，所述影像制剂由生物相容性优良的过渡金属离子、贵金属离子及其纳米簇与卟啉衍生物组成，并通过原位生物作用定点合成对相关病灶部位精准靶向快速标记的成像检测试剂。
其中，上述过渡金属离子是铈、锌离子中的一种或任意两种的混合物。
其中，上述贵金属离子是金离子、银离子、铂离子中的一种或任意两种的混合物。
其中，上述心脑血管疾病是指脑卒中、动脉粥样硬化、心肌梗死、脑缺血和心脏缺血及其再灌注损伤、糖尿病、脑瘤等疾病。
其中，上述纳米簇是金纳米簇、银纳米簇、铂纳米簇等中的一种或任意两种的混合物。
其中，上述卟啉衍生物为四-对-磺酸基-苯基卟啉。
其中，上述检测试剂在老年痴呆和心脑血管疾病的早期快速检测方面的应用。
具体制备措施如下：
首先制备生物相容性优良的过渡金属离子、贵金属离子及其纳米簇和卟啉试剂，具体步骤是：
1)将相关的过渡金属离子、贵金属离子及其纳米簇和卟啉试剂分别溶于超纯水，均配制成一定的浓度。
2)用荧光光谱仪、紫外分光光度仪等对相关的过渡金属离子、贵金属离子及其纳米簇和卟啉试剂的荧光发射、紫外吸收光谱等进行表征。对生物相容性优良的过渡金属离子、贵金属离子及其纳米簇和卟啉衍生物等试剂与靶向分子如Aβ蛋白等蛋白分子及相关生物活性物质之间的相互作用进行电化学、荧光和紫外吸收光谱等检测，获得相关制剂作用的标准曲线。
3)利用已配制的过渡金属离子、贵金属离子及其纳米簇和卟啉衍生物等检测试剂对实验组的体液(如血液或血清等)或者尿液等进行测试，相比较于标准溶液，老年痴呆症和心脑血管相关疾病的实验组的光学以及电化学特性发生了显著的变化，而正常对照组无明显变化。
上述的影像制剂在老年痴呆和心脑血管疾病的早期快速检测方面的应用。
其中，上述的应用，包括以下步骤：将该制剂用于老年痴呆和心脑血管疾病成像时，将金属离子及其纳米簇试剂和卟啉通过尾静脉注射或局部注射到正常对照模型鼠和老年痴呆症模型鼠或血管损伤诱发的脑卒中模型鼠体内，金属离子及其纳米簇和卟啉试剂可快速在病灶部位富集，使用小动物活体荧光成像仪、CT(计算机X线断层扫描)、超声以及MRI(核磁共振成像)对模型鼠进行成像。
上述的应用，其具体步骤是：
1)构建老年痴呆症模型鼠；
2)利用血管损伤诱发构建脑卒中模型鼠等动物模型；
3)将0.1～0.5mL无菌的浓度为0.1～100mmol/L的过渡金属离子、0.1～100mmol/L贵金属离子及其纳米簇和卟啉试剂溶液，注射到正常对照模型鼠和老年痴呆症模型鼠或脑卒中模型鼠上，经过1～72小时的非侵入式的实时动态的高灵敏快速示踪和监测，观测体内循环在老年痴呆症模型鼠或脑卒中模型鼠上实现药物在病变部位的特异性富集与相关病灶部位的精准靶向快速标记成像；
4)用活体荧光成像仪、CT(计算机X线断层扫描)及MRI(核磁共振成像)对病灶部位进行成像并对其进行定性及定量分析。
其中，上述的注射方法为尾静脉注射或局部注射。
本发明所述试剂生物相容性好，无毒副作用，可以通过在病变部位的特异性富集，实现对老年痴呆症(尤其是阿尔兹海默病)和心脑血管疾病进行早期、快速、多模态成像检测，具有广阔的医学应用前景。
有益效果：本发明与现有技术方法相比，具有以下优点和效果：
本研究采用的制剂由生物相容性优良的过渡金属离子、贵金属离子及其原位生物合成的纳米簇与卟啉衍生物组成，通过与靶向生物活性分子定点作用和靶向体内缺血缺氧等特异性微环境实现相关病灶部位的精准靶向标记与快速成像检测，可进行活体非侵入式的实时动态的高灵敏快速示踪和监测，且无生物毒副作用，检测方法简便易行，有效避免了传统纳米材料合成过程中引入的化学试剂以及纳米材料稳定剂对有机体造成的生物毒性，同时避免了传统纳米材料易被网状内皮系统捕获后被清除从而不能到达病灶组织的缺点，以便实现活体靶向成像分析。结果表明：通过CT(计算机X线断层扫描)、超声、荧光以及MRI(核磁共振成像)等成像检测能快速识别模型鼠的发病部位并富集在该区域，实现靶向病灶部位的精 准标记，而且在老年痴呆模型鼠上富集的程度随着月龄的增大而增加。以上结果说明该类药物可以实现对老年痴呆和心脑血管疾病(脑卒中、动脉粥样硬化、心肌梗死、脑缺血、心脏缺血及其再灌注损伤、、糖尿病、脑瘤等)的早期、快速、多模态(如CT(计算机X线断层扫描)、超声、荧光以及MRI(核磁共振成像)等)成像检测。该发明可实现无损伤、原位、实时动态靶向的老年痴呆症和心脑血管相关疾病的成像，进行多形态与多模态的同步诊断及准确靶向定位与治疗，具有广阔的医学应用前景。
附图说明
图1为脑卒中模型鼠尾静脉注射，不同时间点头部病灶区荧光强度变化图。
具体实施方式
以下的说明本质上仅仅是示例性的而并不是为了限制本公开、应用或用途。
实施例1
首先制备生物相容性优良的过渡金属离子、贵金属离子及其纳米簇和卟啉试剂，具体步骤是：
1)将相关的过渡金属离子、贵金属离子及其纳米簇和卟啉试剂分别溶于超纯水，均配制成一定的浓度。
2)用荧光光谱仪、紫外分光光度仪等对相关的过渡金属离子、贵金属离子及其纳米簇和卟啉试剂的荧光发射、紫外吸收光谱等进行表征。对生物相容性优良的过渡金属离子、贵金属离子及其纳米簇和卟啉衍生物等试剂与靶向分子如Aβ蛋白等蛋白分子及相关生物活性物质之间的相互作用进行电化学、荧光和紫外吸收光谱等检测，获得相关制剂特异性作用的依据。
3)利用已配制的过渡金属离子、贵金属离子及其纳米簇和卟啉衍生物等检测试剂对实验组的体液(如血液或血清等)或者尿液等进行测试，相比较于标准溶液，老年痴呆症实验组的光学以及电化学特性发生了显著的变化，而正常对照组无明显变化。
步骤1)所述金属离子是金离子、银离子、铂离子、锌离子等中的一种或任意两种的混合物；所述纳米簇是金纳米簇、银纳米簇、铂纳米簇中的一种或任意两种的混合物；所述卟啉为四-对-磺酸基-苯基卟啉(TSPP)。
实施例2
老年痴呆症模型鼠活体成像分析：应用于老年痴呆早期、快速、多模态检测的金属离子及其纳米簇和卟啉试剂，该试剂用于老年痴呆成像时，将金属离子及其纳米簇试剂和卟啉通过尾静脉注射或局部注射到正常对照模型鼠和老年痴呆症模型鼠体内，金属离子及其纳米簇和卟啉试剂可快速在脑的病灶部位富集，使用小动物活体荧光成像仪对模型鼠进行成像；其具体步骤是：
(1)构建老年痴呆症模型鼠；
(2)将0.3mL无菌的浓度为10mmol/L的过渡金属离子、10mmol/L的贵金属离子及其纳米簇和卟啉试剂溶液，注射到正常对照模型鼠和老年痴呆症模型鼠上，经过1～72小时的非侵入式的实时动态的高灵敏快速示踪和监测，观测体内循环在老年痴呆症模型鼠上实现药物在病变部位的特异性富集与相关病灶部位的精准靶向快速标记成像；所述过渡金属离子是锌离子、贵金属离子为金离子的混合物；所述纳米簇是金纳米簇；所述卟啉为四-对-磺酸基-苯基卟啉(TSPP)；
3)利用活体荧光成像仪对脑部进行成像并对其进行定性及定量分析。
具体为，利用活体荧光成像仪成像，分别选取1小时、4小时、8小时、12小时、24小时、48小时等不同时间点，然后将该实验模型鼠用5％异氟烷进行气体麻醉，将其置于小动物活体成像仪操作平台上，选择420nm和480nm等波长激发即可采集到脑部病变区域的图像。
结果显示，正常对照模型鼠头部无荧光产生，而老年痴呆症模型鼠上病变区域的荧光强度随着时间呈现先增加后减弱的变化，说明试剂中的离子或者纳米簇等制剂可以特意性的原位合成或者富集在病变区域，而后又逐渐代谢。同时荧光强度显示相同条件下，老年痴呆症月龄大的模型鼠富集的更多，荧光强度更大，说明病变的程度更深。小动物活体成像仪对正常组和实验组在420nm激发下，获取的荧光信号强度对比结果参见图1。
实施例3
与实施例2相同，所不同的是采用的0.1mL无菌的浓度为100mmol/L的过渡金属离子和100mmol/L贵金属离子。所述过渡金属离子是铈离子、贵金属离子是铂离子；所述纳米簇是银纳米簇；所述卟啉为四-对-磺酸基-苯基卟啉(TSPP)。结果与实施例2相同。
实施例4
与实施例2相同，所不同的是采用的0.5mL无菌的浓度为0.1mmol/L的过渡金属离子 和100mmol/L贵金属离子。所述过渡金属离子是锌离子；贵金属离子是银离子；所述纳米簇是铂纳米簇；所述卟啉为四-对-磺酸基-苯基卟啉(TSPP)。结果与实施例2相同。
实施例5
应用于脑卒中早期、快速、实时、多模态检测的金属离子及其纳米簇和卟啉衍生物的检测试剂
首先制备生物相容性优良的金属离子及其纳米簇和卟啉衍生物试剂，具体步骤是：将金属离子及其纳米簇和卟啉衍生物试剂分别溶于超纯水，均配制成浓度为0.1～100mmol/L。所述金属离子是金离子、银离子、铂离子、锌离子等中的一种或任意两种的混合物。所述纳米簇是金纳米簇、银纳米簇、铂纳米簇中的一种或任意两种的混合物；所述卟啉为磺酸基卟啉衍生物。
该试剂用于脑卒中成像时，将金属离子及其纳米簇试剂和卟啉衍生物试剂通过尾静脉注射或局部注射到正常对照模型鼠和血管损伤诱发的脑卒中模型鼠等动物体内，金属离子及其纳米簇和卟啉衍生物试剂可快速在脑的缺血缺氧等病灶部位富集，结合使用小动物活体荧光成像仪、CT(计算机X线断层扫描)、超声以及MRI(核磁共振成像)等影像仪对模型鼠等动物进行成像。相关具体步骤包括：
1)利用血管损伤诱发构建脑卒中模型鼠；
2)将0.1～0.5mL无菌的浓度为0.1～100mmol/L的金属离子及其纳米簇和卟啉衍生物试剂溶液，注射到正常对照模型鼠和脑卒中模型鼠上，经过1～72小时的非侵入式的实时动态的高灵敏快速示踪和监测，观测体内循环在脑卒中模型鼠上实现药物在病变部位的特异性富集与相关病灶部位的精准靶向快速标记成像；
3)用活体荧光成像仪、CT(计算机X线断层扫描)、超声、荧光以及MRI(核磁共振成像)等影像仪对活体动物及其脑部进行成像并对其进行定性及定量分析。
例如，利用活体荧光成像仪成像时，分别选取1小时、4小时、8小时、12小时、24小时、48小时等不同时间点，然后将该实验模型鼠用5％异氟烷进行气体麻醉，将其置于小动物活体成像仪操作平台上，选择420nm和480nm等波长激发采集到活体动物脑部病变区域的图像。
结果显示，正常对照模型鼠头部无荧光产生，而脑卒中模型鼠上病变区域的荧光强度随着时间呈现先增加后减弱的变化，说明试剂中的离子或者纳米簇等制剂可以特异性地定点作 用或者富集在缺血缺氧等病变区域，而后又逐渐代谢。脑卒中模型鼠尾静脉注射，不同时间点头部病灶区荧光强度变化参见图1。
实施例6
应用于动脉粥样硬化早期、快速、实时、多模态检测的金属离子及其纳米簇和卟啉衍生物试剂。
该试剂用于动脉粥样硬化成像时，将金属离子及其纳米簇试剂和卟啉衍生物等试剂溶液通过尾静脉注射或局部注射到正常对照模型鼠和动脉粥样硬化模型鼠等动物体内，金属离子及其纳米簇和卟啉衍生物试剂可快速在血管内的病灶部位富集，使用小动物活体荧光成像仪、CT(计算机X线断层扫描)、超声、荧光以及MRI(核磁共振成像)等影像仪对模型鼠进行成像。相关具体步骤包括：
1)构建动脉粥样硬化模型鼠等动物模型；
2)将0.3mL无菌的浓度为50mmol/L的金属离子及其纳米簇和卟啉衍生物等试剂溶液，注射到正常对照模型鼠和动脉粥样硬化模型鼠等动物模型上，经过1～72小时的非侵入式的实时动态的高灵敏快速示踪和监测，观测体内循环在动脉粥样硬化模型动物上实现药物在病变部位的特异性富集与相关病灶部位的精准靶向快速成像标记；
3)用活体荧光成像仪、CT(计算机X线断层扫描)、超声、荧光以及MRI(核磁共振成像)等影像仪对活体动物身上病变区域进行成像并对其进行定性及定量分析。
例如，利用活体荧光成像仪成像时，分别选取1小时、4小时、8小时、12小时、24小时、48小时等不同时间点，然后将该实验模型鼠用5％异氟烷进行气体麻醉，将其置于小动物活体成像仪操作平台上，选择420nm和480nm等波长激发采集到活体动物身上病变区域的图像。
结果显示，正常对照模型鼠身上无荧光产生，而动脉粥样硬化模型鼠身上可见斑块状的荧光，说明试剂中的离子或者纳米簇等制剂可以特异性地定点作用或者富集在缺血缺氧等病变区域。
步骤2)所述金属离子是金离子、锌离子的混合物；所述纳米簇是金纳米簇；所述卟啉为磺酸基卟啉衍生物。
实施例7
与实施例6相同，所不同的是采用的0.1mL无菌的浓度为100mmol/L的金属离子铂离子；所述纳米簇是银纳米簇；所述卟啉衍生物为磺酸基卟啉衍生物。结果与实施例6相同。
实施例8
与实施例6相同，所不同的是采用的0.5mL无菌的浓度为0.1mmol/L的金属离子。所述金属离子是银离子、锌离子的混合物；所述纳米簇是铂纳米簇；卟啉衍生物为磺酸基卟啉衍生物。结果与实施例6相同。
以上显示和描述了本发明的基本原理和主要特征和本发明的优点。本行业的技术人员应该了解，本发明不受上述实施例的限制，上述实施例和说明书中描述的只是说明本发明的原理，在不脱离本发明精神和范围的前提下，本发明还会有各种变化和改进，这些变化和改进都落入要求保护的本发明范围内。本发明要求保护范围由所附的权利要求书及其等效物界定。

Claims (8)

  1. 一种老年痴呆和心脑血管疾病检测的多模态成像的制剂，其特征在于，所述影像制剂由生物相容性优良的过渡金属离子、贵金属离子及其纳米簇与卟啉衍生物组成，其中的任意一种或任意两种物质的量比为1:1的混合物，并通过原位生物作用定点合成对相关病灶部位精准靶向快速标记的成像检测试剂。
  2. 根据权利要求1所述的老年痴呆和心脑血管疾病检测的多模态成像的制剂，其特征在于，所述心脑血管疾病是指脑卒中、动脉粥样硬化、心肌梗死、脑缺血、心脏缺血及其再灌注损伤、糖尿病或脑瘤疾病。
  3. 根据权利要求1所述的老年痴呆和心脑血管疾病检测的多模态成像的制剂，其特征在于，所述过渡金属离子是铈或锌离子中的一种或任意两种的混合物。
  4. 根据权利要求1所述的老年痴呆和心脑血管疾病检测的多模态成像的制剂，其特征在于，所述贵金属离子是金离子、银离子或铂离子中的一种或任意两种的混合物。
  5. 根据权利要求1所述的老年痴呆和心脑血管疾病检测的多模态成像的制剂，其特征在于，所述纳米簇是金纳米簇、银纳米簇或铂纳米簇中的一种或任意两种的混合物。
  6. 根据权利要求1所述的老年痴呆和心脑血管疾病检测的多模态成像的制剂，其特征在于，所述卟啉衍生物为四-对-磺酸基-苯基卟啉。
  7. 一种如权利要求1所述的老年痴呆和心脑血管疾病检测的多模态成像的制剂的应用，其特征在于该应用包括：将该制剂用于老年痴呆症和心脑血管相关疾病成像时，将金属离子及其纳米簇试剂和卟啉通过尾静脉注射或局部注射到正常对照模型鼠和老年痴呆症或血管损伤诱发的脑卒中模型鼠体内，金属离子及其纳米簇和卟啉试剂快速在病灶部位富集，使用小动物活体荧光成像仪、CT(计算机X线断层扫描)、超声以及MRI(核磁共振成像)对模型鼠进行成像。
  8. 根据权利要求7所述的老年痴呆和心脑血管疾病检测的多模态成像的制剂的应用，其特征在于该方法的具体步骤是：
    1)构建老年痴呆症模型鼠；
    2)构建心脑血管疾病的模型鼠：利用血管损伤诱发构建脑卒中模型鼠；
    3)将0.1～0.5mL无菌的浓度为0.1～100mmol/L的过渡金属离子、0.1～100mmol/L贵金属离子及其纳米簇和卟啉试剂溶液，注射到正常对照模型鼠和老年痴呆症模型鼠或脑卒中模型鼠上，经过1～72小时的非侵入式的实时动态的高灵敏快速示踪和监测，观测体内循环在老年痴呆症模型鼠或脑卒中模型鼠上实现药物在病变部位的特异性富集与相关病灶部位的精 准靶向快速标记成像；
    4)用活体荧光成像仪、计算机X线断层扫描CT及核磁共振成像MRI对病灶部位进行成像并对其进行定性及定量分析。
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