Compositions and methods for detection and imaging of amyloid fibrils, amyloid plaques, RNA, and nucleoli

Abstract

The compounds are used for the detection and imaging of amyloid plaques or both of proteins or peptides, for screening or testing the efficacy of inhibitors against amyloidosis and / or fibrillary growth of proteins or peptides, and / or for the detection of RNA and nucleolar imaging. The compounds are d 8 or d 10 Metal complexes or salts thereof. The metal complexes of said compounds can bind to amyloid proteins or peptides, plaques or both and / or RNA, nucleoli or both. This binding induces the accumulation and supramolecular self-assembly of the metal complexes, thereby causing changes in the photophysical properties of the metal complexes.

Description

Invention Field

[0001] This invention generally relates to the detection of analytes and / or imaging of analytes, and more specifically to the detection and imaging of amyloid proteins or peptides, plaques, or both. This invention also belongs to the field of screening or testing the efficacy of inhibitors of amyloid fibrillation and / or amyloid plaque formation, as well as the field of detecting and / or imaging amyloid fibrillation and plaque formation associated with neurodegenerative diseases, dementia, and other related diseases or conditions. This invention also relates to the detection of RNA and imaging of the nucleolus. Background of the Invention

[0003] Amyloids are linear aggregates of proteins or peptides that are typically arranged in a β-sheet conformation, allowing abnormal proteins or peptides to accumulate in tissues. They are associated with many conditions with a wide range of symptoms, including Alzheimer's disease, type 2 diabetes, Huntington's disease, Parkinson's disease, dementia, and other related diseases or conditions. These diseases or conditions are collectively referred to as amyloidosis or protein disorders, and have been identified as arising from the abnormal aggregation of various proteins or peptides that thereby impair tissue and / or organ function.

[0004] Alzheimer's disease is one of the most common neurodegenerative diseases caused by protein disorders and a leading cause of dementia. The formation and deposition of extracellular amyloid-β peptide are considered major events in the disease process, followed by hyperphosphorylation of τ protein and the formation of neurofibrillary tangles (Hamley, Chem. Rev., 112:5147-5192 (2012)). These factors may initially lead to impaired biochemical communication in neurons, subsequently resulting in neuronal death.

[0005] Parkinson's disease is another representative example of a neurodegenerative disease caused by protein disorders. The social impact of Parkinson's disease increases with the growing elderly population. Typically, Parkinson's disease is associated with the abnormal accumulation of α-synuclein aggregates to form amyloid fibrils and the premature death of dopamine-producing neurons in the midbrain (Singleton et al., Science, 302:841-841 (2003)). These lead to a rapid depletion of dopamine in the striatum. Because Parkinson's disease primarily affects the motor system, the most prominent symptoms are balance disorders, bradykinesia, spasticity, and tremor.

[0006] Although amyloidosis and protein disorders constitute important topics in biomedical research, especially in neuroscience, early diagnosis of these conditions remains an unresolved challenge. Various methods exist for detecting amyloid fibrillation, including colorimetric staining, fluorescent staining, and enzyme-linked immunosorbent assay (ELISA). However, these existing methods have limitations. For example, colorimetric staining using dyes such as Congo red typically requires polarized light microscopy, and the birefringence of the dye is difficult to interpret (Biancalana et al., Biochim. Biophys. Acta, 1804:1405-1412 (2010)). Fluorescent staining using thioflavone T is widely used to identify and stain misfolded protein aggregates. However, the emission signal of thioflavone T is not in the red or near-infrared (NIR) region. Therefore, the evaluation of thioflavone T is complicated by the autofluorescence of various biomolecules due to the overlap of fluorescence emission spectra (Anderson et al., J. Clin. Pathol., 27:656-663 (1974)). Furthermore, thioflavin T has an unfavorable small Stokes shift, further limiting the detection of amyloid proteins and plaques. ELISA also has inherent drawbacks because it requires the use of expensive enzyme-linked antibodies and carcinogens in the chemiluminescent detection process (Yu et al., Angew. Chem., Int. Ed., 53:12832-12835 (2014)). It also carries the risk of underestimating or falsely identifying amyloid levels (Stenh et al., Ann. Neurol., 58:147-150 (2005); Sehlin et al., J. Alzheimers Dis., 21:1295-1301 (2010)).

[0007] As a key component and the largest structure in the nucleus of eukaryotic cells, the nucleolus is the most well-known site of ribosomal biogenesis (Olson et al., Trends Cell Biol., 10:189-196 (2000); Németh et al., Trends Genet., 27:149-156 (2011); O'Sullivan et al., Biomol. Concepts, 4:277-286 (2013)). The nucleolus participates in the transcription and processing of ribosomal RNA (rRNA) and plays a role in the assembly of ribosomal proteins. Abnormal morphological changes or alterations in a relevant number of nucleoli may be the cause of certain types of cancer and other human diseases (Busch et al., Cancer Res., 23:313-339 (1963); Kelemen et al., Cancer, 65:1017-1020 (1990); Krystosek, Exp. Cell Res., 241:202-209 (1998); Derenzini et al., J. Pathol., 191:181-186 (2000); Lammerding et al., J. Cell Biol., 170:781-791 (2005); Quin et al., Biochim. Biophys. Acta, 1842:802-816 (2014); Woods et al., Biochim. Biophys. Acta, 1849:821-829 (2015)). As a result, the nucleolus has been identified as a diagnostic biomarker for pathological detection of malignant lesions and is being investigated as a target for cancer chemotherapy. Ribonucleases (RNases) are a class of nucleases that catalyze the degradation of RNA into smaller components (Raines, Chem. Rev., 98:1045-1066 (1998)). For example, pancreatic RNase, commonly abbreviated as RNase A, is a major endonuclease in human organs and tissues (Huang et al., PLoS One, 9:e96490 (2014)). It has been found to play a role in autoimmune diseases, renal failure, and pancreatic diseases. Antitumor activity has also been reported, as some members of the RNase A family exhibit both cytotoxic and cellular inhibitory effects. They show different cytotoxicity against tumor cells rather than normal cells, as normal cells are protected due to their high affinity for RNase inhibitors (Gaur et al., J. Biol. Chem., 276:24978-24984 (2001)).

[0008] Although the nucleolus plays a crucial role in the diagnostics of disease treatment, to date, only one commercially available probe for nucleolus imaging, namely SYTO, is available. TM RNASelectTM A green fluorescent cell staining agent. It is practically non-emissive in the absence of nucleic acids, but exhibits bright green fluorescence when bound to RNA (Yu et al., J. Mater. Chem. B, 4:2614-2619 (2016)). Although SYTO... TM RNASelect TM Green fluorescent cell staining agents are the only commercially available probes for nucleolar imaging, but they have many drawbacks, including high cost, poor photostability, small Stokes shift, and stringent storage requirements. The photostability of biological probes is crucial for the accuracy of organelle staining and the quality of confocal images. However, a major problem associated with the use of most organic dyes or fluorophores is photobleaching, which permanently renders them incapable of fluorescence (O'Mara et al., Talanta, 176:130-139 (2018)).

[0009] There is an urgent need to develop a method for the rapid detection and / or imaging of amyloid proteins or peptides, plaques, or both, for the early diagnosis of diseases and symptoms caused by amyloidosis and protein disorders. There is also an urgent need to develop a method for the rapid screening or evaluation of the efficacy of inhibitors against amyloidosis and / or fibrillary growth of proteins or peptides. Furthermore, there is an urgent need to develop a method for the rapid detection and nucleolar imaging of RNA for the early diagnosis of specific types of cancer and other human diseases.

[0010] The object of the present invention is to provide compounds for detecting analytes and / or imaging or screening them and / or testing inhibitors, particularly (1) detecting amyloid, plaques or both of proteins or peptides and / or imaging them, (2) screening or testing the efficacy of inhibitors against amyloidosis and / or fibrillary growth of proteins or peptides, and / or (3) detecting RNA and imaging the nucleolus.

[0011] Another object of the present invention is to provide methods for detecting analytes and / or imaging or screening them and / or testing inhibitors, particularly for (1) detecting amyloid, plaques or both of proteins or peptides and / or imaging them, (2) screening or testing the efficacy of inhibitors against amyloidosis and / or fibrillary growth of proteins or peptides, and / or (3) detecting RNA and imaging the nucleolus.

[0012] Another object of the present invention is to provide kits for detecting analytes and / or imaging or screening them and / or testing inhibitors, particularly for (1) detecting amyloid, plaques or both of proteins or peptides and / or imaging them, (2) screening or testing the efficacy of inhibitors against amyloidosis and / or fibrillation of proteins or peptides, and / or (3) detecting RNA and imaging nucleoli. Invention Overview

[0014] Compounds, mixtures, compositions, kits, and methods for detecting analytes and / or imaging or screening them and / or testing inhibitors are disclosed.

[0015] For example, in some forms, the compound is d 8 or d 10 Metal complexes or salts thereof, comprising:

[0016] (a) Metal atoms with coordination numbers of 2, 3, or 4, selected from Pt(II), Pd(II), Ni(II), Ir(I), Rh(I), Au(III), Ag(III), Cu(III), Ni(0), Pd(0), Pt(0), Cu(I), Ag(I), Au(I), Zn(II), Cd(II), and Hg(II); and

[0017] (b) One or more ligands having donor atoms, the donor atoms being independently selected from carbon (C), nitrogen (N), oxygen (O), phosphorus (P), sulfur (S), arsenic (As) and selenium (Se).

[0018] Metal complexes can possess planar or partially planar structures. They can bind to analytes, with the binding induced by non-covalent metal-metal interactions leading to aggregation and supramolecular self-assembly. Non-covalent interactions, such as π-π stacking interactions, electrostatic interactions, hydrogen bonding, hydrophobic interactions, and combinations thereof, can facilitate the binding of metal complexes to analytes, resulting in aggregation and supramolecular self-assembly.

[0019] In some forms, the analyte is an amyloid protein or peptide, a plaque, or both. In other forms, the analyte is RNA, a nucleolus, or both.

[0020] Aggregation and supramolecular self-assembly of metal complexes can produce changes in their photophysical properties. In some forms, these changes may include variations in absorbance, luminescence, resonant light scattering (RLS), or combinations thereof. In some forms, changes in luminescence may include increases in luminescence quantum yield and / or emission intensity. In some forms, changes in luminescence may be or include shifts in emission energy or wavelength, preferably redshifts.

[0021] In some forms, metal complexes bind to analytes through non-covalent interactions, such as, but not limited to, π-π stacking interactions, electrostatic interactions, hydrogen bonding interactions, hydrophobic interactions, and combinations thereof. This metal complex-analyte ensemble then brings the metal complexes together very tightly to form aggregates, thereby enhancing the non-covalent metal-metal interactions between the molecules of the metal complex and causing changes in the photophysical properties of the metal complex, such as luminescence.

[0022] The specificity of metal complexes for a given analyte is based on a combination of non-covalent interactions between them. As demonstrated by the following description and examples, non-covalent interactions between metal complexes and analytes can be engineered through molecular engineering. 8 or d 10 The planar or partially planar structure of metal complexes gives them a tendency to form highly ordered oligomeric structures. This characteristic can be used to detect and / or image a wide variety of analytes. Based on the structural properties of both the analyte and the metal complex, potential non-covalent interactions between them can be predicted. Therefore, the supramolecular self-assembly behavior of the metal complex with respect to the analyte can be estimated.

[0023] By selecting the functional groups on the metal center and / or ligands of the metal center, especially the ligands of metal complexes, d can be designed and / or modified. 8 or d 10 Metal complexes bind to target analytes. In some forms, the presence of specific functional groups on one or more ligands can induce or promote specific interactions between the metal complex and the target analyte.

[0024] Preferably, the analyte has a repeating structure to enable aggregation and supramolecular self-assembly of the metal complex thereon. In some forms, the analyte is electrostatically attracted to the metal complex, and the electrostatic interaction between the analyte and the metal complex can be one of the driving forces for binding. In some forms, the analyte is electrically neutral or electrostatically repulsive to the metal complex, and the metal complex can bind to such an analyte through other types of non-covalent interactions, such as, but not limited to, π-π stacking interactions, hydrogen bonding interactions, hydrophobic interactions, and combinations thereof.

[0025] The compound may have the structure of Formula I:

[0026]

[0027] in

[0028] (a) M represents a metal atom selected from Pt(II), Pd(II), Ni(II), Ir(I), Rh(I), Au(III), Ag(III) and Cu(III).

[0029] (b) L1, L2, L3, and L4 represent ligands, where each ligand provides a donor atom to coordinate to a metal atom.

[0030] (c) n+ / - indicates the number of positive or negative charges carried by the metallic complex in the formula, where n is zero or a positive integer, such as 1, 2, 3, 4, and 5.

[0031] (d)X m- / + This represents a counterion that maintains a neutral charge, where X m- / + It has a charge opposite to that of the metallic complex, and where m is zero or a positive integer, such as 1, 2, 3, 4, and 5, and where m = n or m ≠ n.

[0032] (e) The expression represents the stoichiometry of the counter ions.

[0033] (f) Dashed lines represent covalent bonds between any two ligands, fusion of any ring portions from the two ligands, or combinations thereof.

[0034] In some forms, L1, L2, and L3 are optionally substituted and / or optionally deprotonated C6-C. 50 Aromatics or C3-C 50 Heteroaromatic hydrocarbons, such as benzene, pyridine, thiophene, furan, pyrazole, imidazole, oxazole, isoxazole, thiazole, isothiazine, isoquinoline, pyrrole, pyrazine, pyridazine, pyrimidine, benzimidazole, benzofuran, benzothiazole, indole, naphthalene, anthracene, pyrene, triazole, tetrazolium, pyran, thiamphenicol, oxadiazole, triazine, tetrazine, carbazole, dibenzothiophene, dibenzofuran, fluorene and their derivatives.

[0035] The compound may have the structure of Formula II:

[0036]

[0037] Where M′ represents a metal atom selected from Ni(0), Pd(0), Pt(0), Cu(I), Ag(I), Au(I), Zn(II), Cd(II), and Hg(II).

[0038] L5 and L6 represent ligands, with each ligand providing a donor atom to coordinate to the metal atom.

[0039] The compound may also have the structure of Formula III:

[0040]

[0041] L7, L8, and L9 represent ligands, with each ligand providing a donor atom to coordinate to a metal atom.

[0042] Methods for preparing exemplary compounds are disclosed. These methods are compatible with a wide variety of functional groups, ligands, metal complexes, and compounds, and therefore a variety of derivatives can be obtained by the disclosed methods.

[0043] A method for detecting amyloid protein, plaques, or both of proteins or peptides in a sample is disclosed. The method comprises (a) combining one or more of the disclosed compounds with the sample, and (b) detecting changes in the photophysical properties of metal complexes of the compounds. Detection of changes in the photophysical properties of the metal complexes indicates the presence of aggregation and supramolecular self-assembly of the metal complexes, wherein the presence of aggregation and supramolecular self-assembly of the metal complexes indicates the presence of amyloid protein, plaques, or both of proteins or peptides in the sample.

[0044] A method for imaging amyloid proteins, plaques, or both of proteins or peptides in a sample containing proteins or peptides is disclosed. The method comprises (a) combining one or more disclosed compounds with a sample under conditions that allow a metal complex of the compound to bind to the amyloid proteins, plaques, or both of the proteins or peptides and subsequently undergo aggregation and supramolecular self-assembly of the metal complex, wherein the aggregation and supramolecular self-assembly of the metal complex produces a change in the photophysical properties of the metal complex of the compound; and (b) imaging the amyloid proteins, plaques, or both of the proteins or peptides based on one or more photophysical properties specific to the aggregated and supramolecularly self-assembled metal complex.

[0045] In some forms, the sample comprises human or non-human animal body fluids, human or non-human animal tissues, or combinations thereof. The body fluids may be cerebrospinal fluid; the tissues may be brain tissue. In some forms, the sample contains amyloid proteins or peptides, plaques, or both, as well as linear aggregates of proteins or peptides arranged in a β-sheet conformation.

[0046] A method for testing the efficacy of inhibitors against amyloidosis and / or fibrillary growth of proteins or peptides is disclosed. The method includes (a) combining one or more disclosed compounds with an inhibitor-treated sample containing a protein or peptide, and separately combining the compound with an untreated sample containing the protein or peptide, and (b) comparing the photophysical properties of the metal complexes of the compounds between the two samples. The magnitude of the difference in the photophysical properties of the metal complexes between the two samples indicates the degree of change in the aggregation and supramolecular self-assembly state of the metal complexes; the degree of change in the aggregation and supramolecular self-assembly state of the metal complexes indicates the efficacy of the inhibitor.

[0047] A method for detecting RNA, nucleolus, or both in a sample is disclosed. The method comprises (a) combining one or more disclosed compounds with a sample, and (b) detecting changes in the photophysical properties of metal complexes of the compounds. Detection of changes in the photophysical properties of the metal complexes indicates the presence of aggregation and supramolecular self-assembly of the metal complexes, wherein the presence of aggregation and supramolecular self-assembly of the metal complexes indicates the presence of RNA, nucleolus, or both in the sample.

[0048] A method for imaging nucleoli in a sample is disclosed. The method includes (a) combining one or more disclosed compounds with a sample under conditions that allow metal complexes of the compounds to bind to the nucleoli and subsequently undergo aggregation and supramolecular self-assembly of the metal complexes, wherein the aggregation and supramolecular self-assembly of the metal complexes produce changes in the photophysical properties of the metal complexes of the compounds, and (b) imaging the nucleoli based on one or more photophysical properties specific to the aggregated and supramolecularly self-assembled metal complexes.

[0049] In some forms, the sample contains eukaryotic cells. The cells may be, but are not limited to, 3T3 cells, A549 cells, Chinese hamster ovary (CHO) cells, HEK 293 cells, HeLa cells, Hep G2 cells, and HT1080 cells.

[0050] Also disclosed are kits for detecting amyloid proteins, plaques, or both, and / or imaging them; for screening or testing the efficacy of inhibitors against amyloidosis and / or fibrillary growth of proteins or peptides; and / or for detecting RNA and imaging the nucleolus. The kits may contain one or more of the disclosed compounds and optional instructions for use in one or more containers. The kits may also contain a vector.

[0051] Other advantages of the disclosed compounds, mixtures, compositions, kits, and methods will be set forth in part in the description which follows, and in part will be understood from the description, or may be realized by practice of the disclosed compounds, mixtures, compositions, kits, and methods. The advantages of the disclosed compounds, mixtures, compositions, kits, and methods will be realized and obtained through the elements and combinations particularly pointed out in the appended claims. It should be understood that both the foregoing general description and the following detailed description are exemplary and illustrative only and do not limit the claimed invention. It should also be understood that the disclosed compounds, mixtures, compositions, kits, and methods are not limited to the specific methodologies, schemes, and / or reagents described, as they can vary. Brief description of the attached diagram

[0053] Several embodiments of the disclosed compounds, mixtures, compositions, kits, and methods are illustrated in conjunction with the accompanying drawings, which are included in and form part of this specification, and together with the description, serve to explain the principles of the disclosed compounds, mixtures, compositions, kits, and methods.

[0054] Figure 1 The UV-Vis absorption spectra of complex 1-Pt in dimethylformamide (DMF) (line a) and water (line b) solutions at 298 K are shown.

[0055] Figure 2 The normalized emission spectra of the complex 1-Pt in DMF (line a) and water (line b) solutions at 298 K are shown.

[0056] Figure 3A The UV-Vis absorption spectra of complex 1-Pt (50 μM) are shown after adding different amounts of insulin amyloid protein (0-10 μM) to PBS buffer. Arrows indicate trends in spectral changes. Figure 3B The graph shows the relationship between absorbance at 550 nm and the concentration of insulin amyloid protein.

[0057] Figure 4A The corrected emission spectra of complex 1-Pt (50 μM) are shown after adding different amounts of insulin amyloid protein (0-10 μM) to PBS buffer. Arrows indicate trends in spectral changes. Figure 4B The graph shows the relationship between the relative emission intensity at 650 nm and the concentration of insulin amyloid protein.

[0058] Figure 5A The RLS spectra of complex 1-Pt (50 μM) are shown after adding different amounts of insulin amyloid protein (0-10 μM) to PBS buffer. Arrows indicate trends in spectral changes. Figure 5BThe graph shows the relationship between relative RLS intensity at 550 nm and insulin amyloid concentration.

[0059] Figure 6A The UV-Vis absorption spectra of complex 1-Pt (50 μM) are shown after adding different amounts of natural insulin (0-10 μM) to PBS buffer. Figure 6B The graph shows the relationship between absorbance at 550 nm and the concentration of natural insulin.

[0060] Figure 7A The corrected emission spectra of complex 1-Pt (50 μM) are shown after adding different amounts of natural insulin (0-10 μM) to PBS buffer. Figure 7B The graph shows the relationship between the relative emission intensity at 650 nm and the concentration of natural insulin.

[0061] Figure 8A The RLS spectra of complex 1-Pt (50 μM) are shown after adding different amounts of natural insulin (0-10 μM) to PBS buffer. Figure 8B The graph shows the relationship between relative RLS intensity at 550 nm and the concentration of natural insulin.

[0062] Figure 9A The corrected emission spectra of thioflavone T (10 μM) after the addition of an insulin sample (10 μM) are shown at different incubation times. Arrows indicate trends in spectral variation. Figure 9B The graph shows the relationship between the relative emission intensity at 490 nm and the incubation time.

[0063] Figure 10A The UV-Vis absorption spectra of complex 1-Pt (50 μM) after the addition of an insulin sample (10 μM) are shown at different incubation times. Arrows indicate trends in spectral variation. Figure 10B The graph shows the relationship between absorbance at 550 nm and incubation time.

[0064] Figure 11A The corrected emission spectra of complex 1-Pt (50 μM) after the addition of an insulin sample (10 μM) are shown at different incubation times. Arrows indicate trends in spectral variation. Figure 11B The graph shows the relationship between relative emission intensity at 650 nm and incubation time.

[0065] Figure 12A The RLS spectra of complex 1-Pt (50 μM) after the addition of an insulin sample (10 μM) are shown at different incubation times. Arrows indicate trends in spectral variation. Figure 12B The graph shows the relationship between the relative RLS intensity at 550 nm and the incubation time.

[0066] Figure 13 Showing the use of d 8 or d 10 A schematic diagram of the design principle of a luminescence on-off assay for detecting and / or imaging amyloid fibrillation and plaque formation using metal complexes.

[0067] Figure 14A The corrected emission spectra of the complex 1-Pt (0–50 μM) with different amounts of insulin amyloid protein (10 μM) added to PBS buffer are shown. Arrows indicate trends in spectral changes. Figure 14B The graph shows the relationship between the relative emission intensity at 650 nm and the concentration of the complex 1-Pt.

[0068] Figure 15A This shows a luminescent confocal image prepared from insulin amyloid protein (10 μM) stained with complex 1-Pt (50 μM) in PBS buffer. Figure 15B This shows a bright-field confocal image prepared from insulin amyloid protein (10 μM) stained with complex 1-Pt (50 μM) in PBS buffer. Figure 15C This shows a merged confocal image prepared from insulin amyloid protein (10 μM) stained with complex 1-Pt (50 μM) in PBS buffer.

[0069] Figure 16 The graph shows the relative emission intensity at 490 nm as a function of incubation time for aggregates of thioflavone T (10 μM) and mixtures of different insulin samples (10 μM). Insulin samples were incubated in denaturing buffer at different concentrations of L-ascorbic acid (0 (■), 10 (●), 20 (▲), 50 (▼), 70). 100 Incubate in the presence of mM).

[0070] Figure 17 The graph shows the relative emission intensity at 650 nm as a function of incubation time for aggregates of complex 1-Pt (50 μM) and mixtures of different insulin samples (10 μM). Insulin samples were incubated in denaturing buffer at different concentrations of L-ascorbic acid (0 (■), 10 (●), 20 (▲), 50 (▼), 70...). 100 Incubate in the presence of mM).

[0071] Figure 18This is a bar graph showing the relative emission intensities at 650 nm of an aggregate containing a mixture of complex 1-Pt (50 μM), insulin amyloid (10 μM), and various metal ions (100 μM) in PBS buffer. (A) No metal ions, (B) Mg 2+ (C)Ca 2+ (D)Mn 2+ (E)Fe 2+ (F)Fe 3+ (G)Cu 2+ and (H)Zn 2+ The “Pt” group represents the negative control, which contains the complex 1-Pt (50 μM) but does not contain insulin amyloid.

[0072] Figure 19A This is a bar graph showing the relative emission intensities at 650 nm for solutions containing complex 1-Pt (50 μM) and different biomolecules in PBS buffer. (A) α-Amylase (10 μM), (B) Albumin from bovine serum (10 μM), (C) Albumin from human serum (10 μM), (D) Alkaline phosphatase (10 μM), (E) Trypsin (10 μM), (F) DNA (10 μg / mL) -1 (G)RNA (10 μg mL) and (G)RNA -1 The “Pt” group represents the negative control, which contains complex 1-Pt (50 μM) but does not contain insulin amyloid. The “Amyloid” group represents the positive control, which contains complex 1-Pt (50 μM) and insulin amyloid (10 μM). Figure 19B This is a bar graph showing the relative emission intensities at 650 nm of an aggregate containing complex 1-Pt (50 μM), insulin amyloid (10 μM), and a mixture of various biomolecules in PBS buffer. (A) α-Amylase (10 μM), (B) Albumin from bovine serum (10 μM), (C) Albumin from human serum (10 μM), (D) Alkaline phosphatase (10 μM), (E) Trypsin (10 μM), (F) DNA (10 μg / mL) -1 (G)RNA (10 μg mL) and (G)RNA -1 The “Pt” group represents the negative control, which contains complex 1-Pt (50 μM) but does not contain insulin amyloid. The “Amyloid” group represents the positive control, which contains complex 1-Pt (50 μM) and insulin amyloid (10 μM).

[0073] Figure 20AThe bar graph shows the cell viability of HeLa cells after incubation at 37°C for 24 hours with different concentrations of the complex 1-Pt (0, 6.25, 12.5, 25, 50, 100 μM). Figure 20B The bar graph shows the cell viability of CHO cells after incubation at 37°C for 24 hours with different concentrations of the complex 1-Pt (0, 6.25, 12.5, 25, 50, 100 μM).

[0074] Figure 21 The UV-Vis absorption spectrum of the complex 2-Pt in aqueous solution at 298 K is shown.

[0075] Figure 22 The normalized emission spectrum of the complex 2-Pt in aqueous solution at 298 K is shown.

[0076] Figure 23A This shows the addition of different amounts of RNA (0–10 μg / mL) to PBS buffer. -1 The UV-Vis absorption spectrum of the complex 2-Pt (20 μM) is shown. The arrows indicate the trend of spectral changes. Figure 23B The graph shows the relationship between absorbance at 550 nm and RNA concentration.

[0077] Figure 24A This shows the addition of different amounts of RNA (0–10 μg / mL) to PBS buffer. -1 Corrected emission spectrum of the complex 2-Pt (20 μM). Arrows indicate the trend of spectral changes. Figure 24B The graph shows the relationship between the relative emission intensity at 670 nm and the concentration of RNA.

[0078] Figure 25A This shows the addition of different amounts of RNA (0–10 μg / mL) to PBS buffer. -1 RLS spectrum of the complex 2-Pt (20 μM). Arrows indicate the trend of spectral changes. Figure 25B The graph shows the relationship between relative RLS intensity at 550 nm and RNA concentration.

[0079] Figure 26 This is a bar graph showing the addition of different amounts of RNA (0-10 μg / mL) to PBS buffer. -1 The zeta potential of the complex 2-Pt (20 μM) was determined.

[0080] Figure 27A This shows the addition of RNA (10 μg / mL) to PBS buffer. -1 Corrected emission spectra of different amounts of the complex 2-Pt (0-20 μM) were obtained. Arrows indicate the trend of spectral changes. Figure 27BThe graph shows the relationship between the relative emission intensity at 670 nm and the concentration of the complex 2-Pt.

[0081] Figure 28A The image shows a luminescent confocal image of fixed HeLa cells stained with complex 2-Pt (20 μM) at 37 °C for 1 hour. Figure 28B Bright-field confocal images of fixed HeLa cells stained with complex 2-Pt (20 μM) for 1 hour at 37 °C are shown. Figure 28C This shows a merged confocal image of fixed HeLa cells stained with complex 2-Pt (20 μM) for 1 hour at 37 °C.

[0082] Figure 29A The image shows a luminescent confocal image of fixed CHO cells stained with complex 2-Pt (20 μM) at 37 °C for 1 hour. Figure 29B Bright-field confocal images of fixed CHO cells stained with complex 2-Pt (20 μM) for 1 hour at 37 °C are shown. Figure 29C This shows a merged confocal image of fixed CHO cells stained with complex 2-Pt (20 μM) at 37°C for 1 hour.

[0083] Figure 30 Showing the use of d 8 or d 10 A schematic diagram illustrating the design principle of luminescence on-off assay for RNA detection and nucleolar imaging using metal complexes.

[0084] Figure 31A The image shows a luminescent confocal image of fixed HeLa cells stained with complex 2-Pt (20 μM) at 37 °C for 1 hour. Figure 31B Shown from Figure 31A The overall relative emission intensity distribution of fixed HeLa cells. The x-axis represents the scanning distance.

[0085] Figure 32A The image shows a luminescent confocal image of fixed CHO cells stained with complex 2-Pt (20 μM) at 37 °C for 1 hour. Figure 32B Shown from Figure 32A The overall relative emission intensity distribution of fixed CHO cells. The x-axis represents the scanning distance.

[0086] Figure 33A The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with RNase (30 μg / mL) at 37 °C. －1 A confocal image of fixed HeLa cells incubated for 2 hours. Figure 33BThe procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with RNase (30 μg / mL) at 37 °C. －1 Bright-field confocal images of fixed HeLa cells incubated for 2 hours. Figure 33C The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with RNase (30 μg / mL) at 37 °C. -1 Merged confocal images of fixed HeLa cells incubated for 2 hours. Figure 33D The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with DNase (30 μg mL) at 37 °C. －1 A confocal image of fixed HeLa cells incubated for 2 hours. Figure 33E The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with DNase (30 μg mL) at 37 °C. －1 Bright-field confocal images of fixed HeLa cells incubated for 2 hours. Figure 33F The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with DNase (30 μg mL) at 37 °C. －1 Merged confocal images of fixed HeLa cells incubated for 2 hours. Figure 33G The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with RNase and DNase (30 μg mL each) at 37 °C. －1 A confocal image of fixed HeLa cells after incubation for 2 hours. Figure 33H The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37°C for 1 hour, followed by staining with RNase and DNase (30 μg / mL each) at 37°C. -1 Bright-field confocal images of fixed HeLa cells after incubation for 2 hours. Figure 33I The procedure was shown to involve staining with 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with both RNase and DNase (30 μg / mL each) at 37 °C. -1 Merged confocal images of fixed HeLa cells incubated for 2 hours.

[0087] Figure 34A The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with RNase (30 μg / mL) at 37 °C. -1 ) A confocal image of luminescent CHO cells that have been incubated for 2 hours. Figure 34B The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with RNase (30 μg / mL) at 37 °C. -1Bright-field confocal images of fixed CHO cells incubated for 2 hours. Figure 34C The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with RNase (30 μg / mL) at 37 °C. -1 A confocal image of fixed CHO cells incubated for 2 hours. Figure 34D The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with DNase (30 μg mL) at 37 °C. -1 ) A confocal image of luminescent CHO cells that have been incubated for 2 hours. Figure 34E The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with DNase (30 μg / mL) at 37 °C. -1 Bright-field confocal images of fixed CHO cells incubated for 2 hours. Figure 34F The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with DNase (30 μg mL) at 37 °C. -1 Merged confocal images of fixed CHO cells incubated for 2 hours. Figure 34G The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with RNase and DNase (30 μg mL each) at 37 °C. －1 A confocal image of luminescent CHO cells fixed after incubation for 2 hours. Figure 34H The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with RNase and DNase (30 μg mL each) at 37 °C. -1 Bright-field confocal images of fixed CHO cells after incubation for 2 hours. Figure 34I The procedure was shown to involve staining with 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with both RNase and DNase (30 μg / mL each) at 37 °C. -1 A confocal image of fixed CHO cells incubated for 2 hours.

[0088] Figure 35A The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with SYTO at 37 °C. TM RNASelect TM Immobilized HeLa cells incubated with green fluorescent cell staining agent (500 nM) for 20 minutes. The emission was collected at 620–720 nm. Figure 35B The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with SYTO at 37 °C. TM RNASelect TMImmobilized HeLa cells were incubated with green fluorescent cell staining agent (500 nM) for 20 minutes. The emission was collected at 505–555 nm. Figure 35C The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with SYTO at 37 °C. TM RNASelect TM Immobilized HeLa cells were incubated with green fluorescent cell staining agent (500 nM) for 20 minutes. The emission was collected at 620–720 nm and 505–555 nm.

[0089] Figure 36A The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with SYTO at 37 °C. TM RNASelect TM Implanted CHO cells incubated with green fluorescent cell staining agent (500 nM) for 20 minutes. The emission was collected at 620–720 nm. Figure 36B The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with SYTO at 37 °C. TM RNASelect TM Implanted CHO cells incubated with green fluorescent cell staining agent (500 nM) for 20 minutes. The emission was collected at 505-555 nm. Figure 36C The procedure was shown to involve staining with complex 2-Pt (20 μM) at 37 °C for 1 hour, followed by staining with SYTO at 37 °C. TM RNASelect TM Implanted CHO cells incubated with green fluorescent cell staining agent (500 nM) for 20 minutes. The emission was collected at 620–720 nm and 505–555 nm.

[0090] Figure 37A The bar graph shows the cell viability of HeLa cells after incubation at 37°C for 24 hours with different concentrations of the complex 2-Pt (0, 1.25, 2.5, 5, 10, 20 μM). Figure 37B The bar graph shows the cell viability of CHO cells after incubation at 37°C for 24 hours with different concentrations of the complex 2-Pt (0, 1.25, 2.5, 5, 10, 20 μM). Invention Details

[0092] Compounds, mixtures, compositions, and kits for detecting analytes and / or imaging or screening them and / or testing inhibitors are disclosed, particularly for (1) detecting amyloid, plaques, or both of proteins or peptides and / or imaging them, (2) screening or testing the efficacy of inhibitors against amyloid degeneration and / or fibrillation of proteins or peptides, and / or (3) detecting RNA and imaging nucleoli.

[0093] In some forms, the compound contains d that can bind to the analyte. 8 or d 10 Metal complexes. The analyte can be an amyloid protein, plaque, or both of the protein or peptide. The analyte can also be RNA, nucleolus, or both. The binding occurs via non-covalent metal-metal interactions, resulting in a luminescent signal in the red to near-infrared (NIR) region through the aggregation and supramolecular self-assembly of the metal complex. Non-covalent interactions, such as π-π stacking interactions, electrostatic interactions, hydrogen bonding, hydrophobic interactions, and combinations thereof, can facilitate the binding of the metal complex to the analyte, leading to aggregation and supramolecular self-assembly of the metal complex. The accompanying visible light excitation and large Stokes shift reduce interference associated with autofluorescence commonly encountered in the presence of various biological substrates, making the compound suitable for bioassays.

[0094] The disclosed compounds, mixtures, compositions, kits, and methods can be more readily understood by referring to the following detailed description of specific embodiments, the examples included, the accompanying drawings, and the preceding and following descriptions. Unless otherwise indicated or clearly contradicted by the context, all methods described herein can be performed in any suitable order.

[0095] Unless otherwise required, the use of any and all instances or exemplary language (e.g., “such”) provided herein is intended only to better illustrate the invention and does not constitute a limitation on the scope of the invention. No language in the specification should be construed as indicating that any unclaimed element is essential for carrying out the invention.

[0096] The disclosed compounds, mixtures, compositions, and kits can be used with, in combination with, the disclosed methods, and can be used to prepare products of, or products of, the disclosed methods. It should be understood that when combinations, subsets, interactions, groups, etc., of these compounds, mixtures, compositions, and kits are disclosed, although every different individual and common combination of these materials may not be explicitly mentioned, each is specifically considered and described herein. For example, if a compound is disclosed and discussed, and many modifications that can be made to a number of molecules containing that compound are discussed, then every combination and arrangement of the compounds and possible modifications will be specifically considered unless otherwise specifically indicated. Thus, if a class of molecules A, B, and C and a class of molecules D, E, and F are disclosed, and an example of the combination molecule AD is disclosed, then each is considered individually and collectively, even if each molecule is not described individually. Therefore, in this example, each of the combinations AE, AF, BD, BE, BF, CD, CE, and CF is specifically considered and should be considered as disclosed by the disclosures of A, B, and C; D, E, and F; and the example combination AD. Similarly, any subset or combination of these is also specifically considered and disclosed. Thus, for example, subgroups of AE, BF, and CE are specifically considered and should be considered as disclosed by the disclosures of A, B, and C; D, E, and F; and the example combination AD. Furthermore, each of the compounds, mixtures, compositions, kits, components, etc., considered and disclosed above may also be specifically and independently included or excluded from any group, subgroup, list, set, etc., of such material. These concepts apply to all aspects of this application, including but not limited to steps in methods for preparing and using the disclosed compounds, compositions, mixtures, and kits. Therefore, if multiple additional steps are possible, it should be understood that each of these additional steps can be performed using any specific embodiment or combination of embodiments of the disclosed method, and each such combination is specifically considered and should be considered as disclosed.

[0097] Throughout the specification and claims of this application, the word “comprise” and variations thereof, such as “comprising” and “comprises”, mean “including but not limited to” and are not intended to exclude, for example, other additives, components, integers or steps.

[0098] Any discussion of documents, actions, materials, devices, articles, etc., already included in this application shall not be construed as an admission that any or all of these matters constitute part of the prior art or are common general knowledge in the field relating to this disclosure, simply because they existed prior to the priority date of each claim of this application.

[0099] I. Definition

[0100] Unless the context clearly indicates otherwise, the singular forms “a,” “an,” and “the” all include plural references. For example, a reference to “a compound” includes multiple compounds, and a reference to “the compound” is a reference to one or more compounds and their equivalents known to those skilled in the art.

[0101] Unless otherwise expressly stated in the context, the terms “may,” “may be,” “can,” and “may be,” as well as related terms, are intended to indicate that the subject matter in question is optional (that is, the subject matter exists in some embodiments but not in others) and does not imply the capability or probability of the subject matter.

[0102] The terms “optional” and “optionally” mean that the event, situation or substance described below may or may not occur or not exist, and the description includes both the possibility that the event, situation or substance may or may not occur or not exist.

[0103] The term "about" is used to describe values ​​that are higher or lower than the stated value within a range of about + / - 10%; in other embodiments, the value may be higher or lower than the stated value within a range of about + / - 5%; in other embodiments, the value may be higher or lower than the stated value within a range of about + / - 2%; in other embodiments, the value may be higher or lower than the stated value within a range of about + / - 1%. The foregoing ranges are intended to be clear from the context and do not imply further limitations.

[0104] A range may be expressed herein as from “about” one specific value and / or to “about” another specific value. When expressing such a range, unless the context explicitly indicates otherwise, what is specifically considered and regarded as disclosed is a range from one specific value and / or to another specific value. Similarly, when a value is expressed as an approximate value using the antecedent “about,” it will be understood that, unless the context explicitly indicates otherwise, that specific value constitutes another specific consideration of an implementation that should be regarded as disclosed. It will also be understood that, unless the context explicitly indicates otherwise, the endpoints of each range are obvious relative to the other endpoints and are independent of the other endpoints. It should be understood that, unless the context explicitly indicates otherwise, all individual values ​​and subranges of values ​​contained within a explicitly disclosed range are also specifically considered and should be regarded as disclosed. Finally, it should be understood that all ranges refer to the range as a range, and as a set of individual numbers from a first endpoint (inclusive) to a second endpoint (inclusive). In the latter case, it should be understood that any individual number may be chosen as a form of quantity, value, or characteristic referred to by the range. In this way, a scope describes a set of numbers or values ​​from a first endpoint (inclusive) to a second endpoint (inclusive), from which a single member (i.e., a single number) can be selected as the quantity, value, or characteristic referred to by the scope. The foregoing applies regardless of whether some or all of these embodiments are explicitly disclosed in a particular context.

[0105] Carbon range (e.g., C1-C) 10 The aim is to disclose each possible carbon value and / or subrange covered therein individually. For example, C1–C 10 The carbon length range disclosed includes C1, C2, C3, C4, C5, C6, C7, C8, C9 and C6. 10 And it disclosed the sub-ranges it covers, such as C2-C9, C3-C8, C1-C5, etc.

[0106] The terms "derivative" and "multiple derivatives" refer to a chemical compound / part that has a structure similar to that of a parent compound / part, but differs from it in one or more components, functional groups, atoms, etc. Derivatives can be formed from a parent compound / part through (multiple) chemical reactions. Differences between derivatives and parent compounds / parts may include, but are not limited to, the substitution of one or more functional groups with one or more different functional groups, or the introduction or removal of one or more substituents containing hydrogen atoms. Derivatives may also differ from parent compounds / parts in their protonated state.

[0107] As used in this article, "halogen" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), iodine (I), or astatine (At).

[0108] The term "alkyl" refers to a monovalent group derived from an alkane by removing a hydrogen atom from any carbon atom. Alkanes represent saturated hydrocarbons, including those that are cyclic (monocyclic or polycyclic). Alkyl groups can be straight-chain, branched, or cyclic. Preferred alkyl groups have 1 to 30 carbon atoms, i.e., C1-C2. 30 Alkyl group. In some forms, C1-C 30 Alkyl groups can be straight-chain C1-C 30 Alkyl, branched C1-C 30 Alkyl, cyclic C1-C 30 Alkyl, straight-chain or branched C1-C 30 Alkyl, straight-chain or cyclic C1-C 30 Alkyl, branched or cyclic C1-C 30 Alkyl groups, or straight-chain, branched, or cyclic C1-C 30 alkyl.

[0109] The term "heteroalkyl" refers to an alkyl group in which one or more carbon atoms are replaced by heteroatoms, such as O, N, or S. Heteroalkyl groups can be straight-chain, branched, or cyclic (monocyclic or polycyclic). Preferred heteroalkyl groups have 1 to 30 carbon atoms, i.e., C1-C2. 30 Heteroalkyl. In some forms, C1-C 30 Heteroalkyl groups can be straight-chain C1-C 30 Heteroalkyl, branched C1-C 30 Heteroalkyl, cyclic C1-C 30 Heteroalkyl, straight-chain or branched C1-C 30 Heteroalkyl, straight-chain or cyclic C1-C 30 Heteroalkyl, branched or cyclic C1-C 30 Heteroalkyl, or straight-chain, branched, or cyclic C1-C 30 Heteroalkyl groups.

[0110] The term "alkenyl" refers to a monovalent group derived from an alkene by removing a hydrogen atom from any carbon atom. An alkene is an unsaturated hydrocarbon containing at least one carbon-carbon double bond. The alkenyl group can be straight-chain, branched, or cyclic (monocyclic or polycyclic). Preferred alkenyl groups have 2 to 30 carbon atoms, i.e., C2-C. 30 Alkenyl. In some forms, C2-C 30 Alkenes can be straight-chain C2-C 30 Alkenyl, branched C2-C 30 Alkenyl, cyclic C2-C 30 Alkenyl, straight-chain or branched C2-C 30 Alkenyl, straight-chain or cyclic C2-C 30 Alkenyl, branched or cyclic C2-C 30 Alkenyl, or straight-chain, branched, or cyclic C2-C 30 Alkenyl group.

[0111] The term "heterenyl" refers to an alkenyl group in which one or more double-bonded carbon atoms are replaced by heteroatoms. Heterenyl groups can be straight-chain, branched, or cyclic (monocyclic or polycyclic). Preferred heteroenyl groups have 1 to 30 carbon atoms, i.e., C1-C2. 30 Heteroalkenyl groups. In some forms, C1-C 30 Heterene groups can be straight-chain C1-C 30 Heteroalkenyl, branched C1-C 30 Heteroalkenyl, cyclic C1-C 30 Heteroalkenyl, straight-chain or branched C1-C 30 Heteroalkenyl, straight-chain or cyclic C1-C 30 Heteroalkenyl, branched or cyclic C1-C 30 Heteroalkenyl, or straight-chain, branched, or cyclic C1-C 30 Heterene group.

[0112] The term "alkynyl" refers to a monovalent group derived from alkynes by removing a hydrogen atom from any carbon atom. Alkynes are unsaturated hydrocarbons containing at least one carbon-carbon triple bond. The alkynyl group can be straight-chain, branched, or cyclic (monocyclic or polycyclic). Preferred alkynyl groups have 2 to 30 carbon atoms, i.e., C2-C. 30 Alkyne group. In some forms, C2-C 30 The alkynyl group can be a straight-chain C2-C 30 Alkyne group, branched C2-C 30 Alkyne group, cyclic C2-C 30 Alkyne group, straight-chain or branched C2-C 30 Alkyne group, straight-chain or cyclic C2-C 30 Alkyne group, branched or cyclic C2-C 30 Alkyne group, or straight-chain, branched, or cyclic C2-C 30 Alkyne group.

[0113] The term "heterynyl" refers to an ynyl group in which one or more trivalent carbon atoms are replaced by heteroatoms. Heterynyl groups can be straight-chain, branched, or cyclic (monocyclic or polycyclic). Preferred heterynyl groups have 1 to 30 carbon atoms, i.e., C1-C2. 30 Neyne group. In some forms, C1-C 30 The heteroyne group can be a straight-chain C1-C 30 Heterynyl group, branched C1-C 30 Heterynyl group, cyclic C1-C 30 Heterynyl group, straight-chain or branched C1-C 30 Heterynyl group, straight-chain or cyclic C1-C 30 Heterynyl, branched or cyclic C1-C 30 Heterynyl group, or straight-chain, branched, or cyclic C1-C30 Zeyne group.

[0114] The term "aryl" refers to a monovalent group derived from an aromatic hydrocarbon by removing hydrogen atoms from a ring atom. Aromatic hydrocarbons are monocyclic or polycyclic aromatic hydrocarbons. In polycyclic aromatic hydrocarbons, the rings may be connected sideways or fused together. Preferred aromatic hydrocarbons have 6 to 50 carbon atoms, i.e., C6-C6. 50 Aromatic hydrocarbons. In some forms, C6-C 50 Aromatic hydrocarbons can be branched C6-C 50 Aromatic hydrocarbons, monocyclic C6-C 50 Aromatic hydrocarbons, polycyclic C6-C 50 Aromatic hydrocarbons, branched polycyclic C6-C 50 Aromatic hydrocarbons, fused polycyclic C6-C 50 Aromatic or branched fused polycyclic C6-C 50 Aromatic hydrocarbons. Therefore, in polycyclic aryl groups, the rings can be connected together laterally or can be fused. Preferred aryl groups have 6 to 50 carbon atoms, i.e., C6-C6. 50 Aryl. In some forms, C6-C 50 The aryl group can be a branched C6-C group. 50 Aryl, monocyclic C6-C 50 Aryl, polycyclic C6-C 50 Aryl, branched polycyclic C6-C 50 Aryl, fused polycyclic C6-C 50 Aryl or branched fused polycyclic C6-C 50 Aryl.

[0115] The term "heteroaryl" refers to a monovalent group derived from heteroaromatic hydrocarbons by removing hydrogen atoms from the ring atoms. Heteroaromatic hydrocarbons are heterocyclic compounds derived from aromatic hydrocarbons by substituting one or more methine (-C=) and / or vinylidene (-CH=CH-) groups with trivalent or divalent heteroatoms, maintaining the continuous π-electron system characteristics of aromatic systems and a number of out-of-plane π-electrons corresponding to Hückel's rule (4n+2). Heteroaromatic hydrocarbons can be monocyclic or polycyclic. In polycyclic heteroaromatic hydrocarbons, the rings can be connected sideways or fused together. Preferred heteroaromatic hydrocarbons have 3 to 50 carbon atoms, i.e., C3-C4. 50 heteroaromatic hydrocarbons. In some forms, C3-C 50 heteroaromatics can be branched C3-C 50 heteroaromatics, monocyclic C3-C 50 heteroaromatics, polycyclic C3-C 50 heteroaromatics, branched polycyclic C3-C 50 heteroaromatics, fused polycyclic C3-C 50 heteroaromatics or branched fused polycyclic C3-C 50Heteroaromatic hydrocarbons. Therefore, in a polycyclic heteroaryl group, the rings can be connected together laterally or can be fused. Preferred heteroaryl groups have 3 to 50 carbon atoms, i.e., C3-C4. 50 Mixed aryl groups. In some forms, C3-C 50 Heteroaryl groups can be branched C3-C 50 Heteroaryl, monocyclic C3-C 50 heteroaryl, polycyclic C3-C 50 heteroaryl, branched polycyclic C3-C 50 Heteroaryl, fused polycyclic C3-C 50 Heteroaryl or branched fused polycyclic C3-C 50 Mixed aromatic compounds.

[0116] The term "arylene" refers to a divalent group derived from an aromatic hydrocarbon by removing hydrogen atoms from two ring carbon atoms. In polycyclic arylene groups, the rings can be connected sideways or fused together. Preferred arylene groups have 6 to 50 carbon atoms, i.e., C6-C6. 50 Aromatic. In some forms, C6-C 50 Arylidene groups can be branched C6-C. 50 aryl, monocyclic C6-C 50 aryl, polycyclic C6-C 50 aryl, branched polycyclic C6-C 50 aryl, fused polycyclic C6-C 50 aryl or branched fused polycyclic C6-C 50 Alpha-aryl.

[0117] The term "heteroaryl" refers to a divalent group derived from heteroaryl hydrocarbons by removing hydrogen atoms from two ring atoms. In polycyclic heteroaryl groups, the rings can be connected sideways or fused together. Preferred heteroaryl groups have 3 to 50 carbon atoms, i.e., C3-C4. 50 Hybrid aryl. In some forms, C3-C 50 Heteroaryl groups can be branched C3-C 50 Heteroarylene, monocyclic C3-C 50 Heteroarylene, polycyclic C3-C 50 Heteroarylene, branched polycyclic C3-C 50 Heteroaryl, fused polycyclic C3-C 50 Heteroarylene or branched fused polycyclic C3-C 50 Hybrid aryl.

[0118] The term "aminooxy" refers to -O-NH2, in which the hydrogen atom can be replaced by a substituent.

[0119] The term "hydroxyamino" refers to -NH-OH, in which hydrogen atoms can be replaced by substituents.

[0120] The term "hydroxamic acid" refers to -C(=O)NH-OH, in which the hydrogen atom can be substituted by a substituent.

[0121] The term "conjugated system" refers to a molecular entity whose structure can be represented as a system of alternating single and multiple bonds, such as CH2=CH-CH=CH2 or CH2=CH-C≡N. In such a system, conjugation is the interaction between one p orbital and another p- orbital across an intermediate σ bond. Conjugated systems can be or contain aromatic and / or heteroaromatic moieties.

[0122] As used herein, the term "substituted" refers to a substituent that substituted a chemical group or portion for one or more hydrogen atoms in a chemical group or portion. Such substituents include, but are not limited to:

[0123] Halogen atoms, alkyl groups, heteroalkyl groups, alkenyl groups, heteroalkenyl groups, alkynyl groups, heteroalkynyl groups, aryl groups, heteroaryl groups, -OH, -SH, -NH2, -N3, -OCN, -NCO, -ONO2, -CN, -NC, -ONO, -CONH2, -NO, -NO2, -ONH2, -SCN, -SNCS, -CF3, -CH2CF3, -CH2Cl, -CHCl2, -CH2NH2, -NHCOH, -CHO, -COCl, -COF, -COBr, -COOH, -SO3H, -CH2SO2CH3, -PO3H2, -OPO3H2, -P(=O)(OR G1′ (OR) G2′ -OP(=O)(OR) G1′ (OR) G2′ ), -BR G1′ (OR G2′ -B(OR) G1′ (OR) G2′ ) or -G′R G1′ Where -G′ represents -O-, -S-, or -NR. G2′ -, -C(=O)-, -S(=O)-, -SO2-, -C(=O)O-, -C(=O)NR G2′ -、-OC(=O-、-NR G2′ C(=O)-, -OC(=O)O-, -OC(=O)NR G2′ -、-NR G2′ C(=O)O-、-NR G2′ C(=O)NR G3′ -, -C(=S)-, -C(=S)S-, -SC(=S)-, -SC(=S)S-, -C(=NR G2′)-、-C(=NR G2′ )O-、-C(=NR G2′ )NR G3′ -、-OC(=NR G2′ )-、-NR G2′ C(=NR G3′ )-、-NR G2′ SO2-, -C(=NR) G2′ )NR G3′ -、-OC(=NR G2′ )-、-NR G2′ C(=NR G3′ )-、-NR G2′ SO2-, -NR G2′ SO2NR G3′ -、-NR G2′ C(=S)-、-SC(=S)NR G2′ -、-NR G2′ C(=S)S-、-NR G2′ C(=S)NR G3′ -、-SC(=NR G2′ )-、-C(=S)NR G2′ -、-OC(=S)NR G2′ -、-NR G2′ C(=S)O-、-SC(=O)NR G2′ -、-NR G2′ C(=O)S-, -C(=O)S-, -SC(=O)-, -SC(=O)S-, -C(=S)O-, -OC(=S)-, -OC(=S)O-, -SO2NR G2′ -、-BR G2′ - or -PR G2′ -,

[0124] Among them, R G1′ R G2′ R G3′ Each time it appears, it is independently a hydrogen atom, halogen atom, alkyl group, heteroalkyl group, alkenyl group, heteroalkenyl group, alkynyl group, heteroalkynyl group, aryl group, or heteroaryl group.

[0125] In some cases, "substituted" also refers to one or more carbon atoms in a carbon chain (e.g., alkyl, alkenyl, alkynyl, and aryl groups) being substituted once or multiple times by heteroatoms such as, but not limited to, nitrogen, oxygen, and sulfur.

[0126] It should be understood that “substitution” or “substituted” includes the implicit condition that such substitution is based on the permissible valence states of the substituted atoms and substituents, and that the substitution produces a stable compound, i.e., a compound that does not spontaneously undergo transformations such as rearrangement, cyclization, elimination, etc.

[0127] The term "d" 8 or d 10 "metal complexes" and "multiple d" 8 or d 10 "Metal complex" refers to a compound containing at least one metal complex with d 8 or d 10 Any metal complex with a metal atom of a specific electronic configuration. The term "d" refers to... 8 or d 10 "Metal complex aggregates" refer to the aggregates near the analyte. 8 or d 10 Localized concentration enrichment of metal complexes occurs. The analyte can be amyloid protein, plaque, or both of the protein or peptide. The analyte can also be RNA, nucleolus, or both. 8 or d 10 Non-covalent metal-metal interactions between molecules in metal complexes can lead to localized concentration enrichment. Non-covalent interactions, such as π-π stacking interactions, electrostatic interactions, hydrogen bonding interactions, and hydrophobic interactions, and their combinations, can facilitate the interaction between analytes and d... 8 or d 10 Between metal complexes and d 8 or d 10 The bonding between different molecules in a metal complex. In some forms, d 8 or d 10 Metal complex aggregates can be bound to the analyte via d 8 or d 10 The aggregation of metal complexes and supramolecular self-assembly are formed.

[0128] The terms "ligand" and "multiple ligands" refer to ions or molecules that form metal complexes by bonding to a central metal atom through one or more donor atoms. The nature of metal ligand bonding ranges from covalent to ionic. The bond order of metal ligands can range from one to three. Bonding to a metal atom generally involves the formal donation of one or more electron pairs from a donor atom. Donor atoms can be carbon (C), nitrogen (N), oxygen (O), phosphorus (P), sulfur (S), arsenic (As), and selenium (Se).

[0129] The term "coordination number" refers to the total number of donor atoms that coordinate with the central metal atom in a metal complex.

[0130] As used herein, the terms “amyloid” and “plaque” can refer to linear aggregates of any protein or peptide. Aggregates may be arranged in a β-sheet conformation; they may be distributed in solution or immobilized on the surface of a solid support. The term “plaque” also refers to fibrous deposits of protein, peptide, or amyloid.

[0131] The term "RNA" as used in this article refers to ribonucleic acid, a polymer molecule essential for various biological functions in the coding, decoding, regulation, and expression of genes. It can be distributed in solution, located in organelles (such as the nucleolus), or immobilized on the surface of a solid carrier.

[0132] As used in this article, the term "nucleolus" refers to the largest structure in the nucleus of eukaryotic cells. The nucleolus is composed of proteins, DNA, and RNA. It is well known that they serve as the site for the synthesis and processing of ribosomal RNA (rRNA).

[0133] The term "luminescence" refers to the emission of light from a substance that is not caused by heat. It can be caused by chemical reactions, electrical energy, subatomic motion, or stress on a crystal, ultimately resulting in spontaneous emission. It can refer to chemiluminescence, which is light emitted due to a chemical reaction. It can also refer to photoluminescence, which is light emitted due to the absorption of photons. Photoluminescence includes fluorescence and phosphorescence.

[0134] The terms "carrier" and "multiple carriers" refer to all components present in a formulation or composition other than the active ingredient or multiple active ingredients. These may include, but are not limited to, diluents, binders, lubricants, disintegrants, fillers, plasticizers, pigments, colorants, stabilizers, and flow aids.

[0135] As used herein, “subject” includes, but is not limited to, human or non-human mammals. The term does not indicate a specific age or sex. Therefore, it is intended to cover adult and newborn subjects, as well as fetuses, whether male or female. A patient is a subject suffering from a disease or condition. The term “patient” includes both human and non-human mammal subjects.

[0136] It should be understood that, unless otherwise stated, the disclosed methods and compositions are not limited to specific synthetic methods, specific analytical techniques, or specific reagents, and therefore can be varied. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0137] II.Compounds

[0138] This article discloses compounds for detecting analytes and / or imaging or screening them and / or testing inhibitors.

[0139] In some forms, the analyte is amyloid protein or peptide, plaque, or both. In other forms, the analyte is RNA, nucleolus, or both. The compounds can be used to screen for or test the efficacy of inhibitors against amyloidosis and / or fibrillary growth of proteins or peptides. The compounds can also be used for nucleolar imaging.

[0140] For example, in some forms, the compound is d 8 or d 10 Metal complexes or salts thereof, comprising:

[0141] (a) Metal atoms with coordination numbers of 2, 3, or 4, selected from Pt(II), Pd(II), Ni(II), Ir(I), Rh(I), Au(III), Ag(III), Cu(III), Ni(0), Pd(0), Pt(0), Cu(I), Ag(I), Au(I), Zn(II), Cd(II), and Hg(II); and

[0142] (b) One or more ligands having donor atoms, the donor atoms being independently selected from carbon (C), nitrogen (N), oxygen (O), phosphorus (P), sulfur (S), arsenic (As) and selenium (Se).

[0143] In some forms, the metal atom is not Au(III).

[0144] In some forms, the ligand does not have the following structure:

[0145]

[0146] In some forms, the metal atom is Pt(II), and the ligands do not have the following structure:

[0147]

[0148] 1. Metal complexes

[0149] Metal complexes of compounds can have planar or partially planar structures. It is noteworthy that square planar d... 8 or d 10 Metal complexes tend to form solid, highly ordered, extended linear or oligomeric structures.

[0150] Metal complexes can bind to analytes, with the binding occurring through non-covalent metal-metal interactions that induce aggregation and supramolecular self-assembly. Non-covalent interactions, such as π-π stacking interactions, electrostatic interactions, hydrogen bonding, hydrophobic interactions, and combinations thereof, can facilitate the binding of metal complexes to analytes, leading to aggregation and supramolecular self-assembly. As a result, aggregates of metal complexes can form.

[0151] In some forms, metal complexes can bind to one or more proteins or peptides, including but not limited to amyloid-β peptide, α-synuclein, insulin, huntingtin, tau protein, hyperphosphorylated tau protein (pτ), prion protein, IAPP (insulin amyloid peptide), calcitonin, and PrP. Sc Atrial natriuretic factor, apolipoprotein A1, serum amyloid A, medin, prolactin, transthyretin, lysozyme, β-2 microglobulin, coagulant, karatoepithelialin, lens protein, desmin, selenoprotein, actin, cystatin, immunoglobulin light chain AL, S-IBM, and myosin. In some forms, metal complexes can bind to amyloid-β peptides, plaques, or both.

[0152] In some forms, metal complexes cannot bind to or form self-assembled aggregates on native proteins or peptides. Preferably, metal complexes cannot form self-assembled aggregates through non-covalent metal-metal interactions on native proteins or peptides.

[0153] In some forms, metal complexes can bind to one or more types of RNA, including but not limited to messenger RNA (mRNA), transfer RNA (tRNA), ribosomal RNA (rRNA), transfer messenger RNA (tmRNA), antisense RNA (asRNA), enhancer RNA (eRNA), guide RNA (gRNA), ribosomes, short hairpin RNA (shRNA), hourly sequence RNA (stRNA), small interfering RNA (siRNA), and trans-acting siRNA (ta-siRNA).

[0154] In some forms, metal complexes cannot bind to or form self-assembled aggregates on double-stranded DNA. Preferably, metal complexes cannot form self-assembled aggregates via non-covalent metal-metal interactions on double-stranded DNA. In some forms, metal complexes can bind to double-stranded DNA via non-covalent interactions, such as π-π stacking interactions, electrostatic interactions, hydrogen bonding interactions, or combinations thereof; however, due to the double-stranded structure of DNA, aggregation or self-assembly of the metal complex cannot be induced. Metal complexes may undergo insertion, such that they are inserted between base pairs of DNA, preventing aggregation or self-assembly via non-covalent metal-metal interactions.

[0155] The aggregation and supramolecular self-assembly of metal complexes can lead to changes in their photophysical properties. In some forms, these changes can include variations in absorbance, luminescence, resonant light scattering (RLS), or combinations thereof.

[0156] In some forms, changes in luminescence can be or include increases in the luminescence quantum yield and / or emission intensity, such as Figure 13 and 30 As shown. In some forms, the change in luminescence can be or includes a shift in emission energy or wavelength compared to non-aggregated or non-supramolecular self-assembled forms, preferably a redshift. In some forms, the increase in luminescence quantum yield and / or emission intensity and / or the shift in emission energy or wavelength may be caused by aggregation and supramolecular self-assembly of the metal complex via non-covalent metal-metal interactions, similar to exciton coupling. Non-covalent interactions, such as π-π stacking interactions, electrostatic interactions, hydrogen bonding interactions, hydrophobic interactions, and combinations thereof, can facilitate the binding of the metal complex to the analyte, leading to aggregation and supramolecular self-assembly of the metal complex. The increase in luminescence quantum yield and / or emission intensity can be correlated with the luminescence signal in the red to near-infrared (NIR) region, for example, between about 600 nm and about 1000 nm. Preferably, the luminescence signal is correlated with a large Stokes shift. In some forms, the Stokes shift is greater than 100 nm, greater than 150 nm, greater than 200 nm, greater than 250 nm, greater than 300 nm, greater than 350 nm, or greater than 400 nm. More preferably, the Stokes shift is greater than 400 nm. Compared to non-aggregated or non-supramolecular self-assembled forms, the change in luminescence can be or includes a shift in emission energy or wavelength, preferably a redshift. The luminescence signal can originate from a transition between a singlet excited state and a singlet ground state, or from a triplet excited state and a singlet ground state.

[0157] In some forms, changes to RLS can be or include an increase in the RLS signal strength.

[0158] In some forms, metal complexes bind to analytes through non-covalent interactions, such as, but not limited to, π-π stacking interactions, electrostatic interactions, hydrogen bonding interactions, hydrophobic interactions, and combinations thereof. This metal complex-analyte aggregate then causes the metal complexes to assemble tightly into aggregates, thereby enhancing the non-covalent metal-metal interactions between the molecules of the metal complex and causing changes in the photophysical properties of the metal complex, such as luminescence.

[0159] The specificity of metal complexes for a given analyte is based on a combination of non-covalent interactions between them. As demonstrated by the following description and examples, non-covalent interactions between metal complexes and analytes can be engineered through molecular engineering. 8 or d 10 The planar or partially planar structure of metal complexes gives them a tendency to form highly ordered oligomeric structures. This characteristic can be used to detect and / or image various analytes. Based on the structural properties of both the analyte and the metal complex, those skilled in the art can predict possible non-covalent interactions between them. Consequently, the supramolecular self-assembly behavior of the metal complex with respect to the analyte can be estimated.

[0160] By selecting the functional groups on the metal center and / or ligands of the metal center, especially the ligands of metal complexes, d can be designed and / or modified. 8 or d 10 Metal complexes bind to target analytes. In some forms, the presence of specific functional groups on one or more ligands can induce or promote specific interactions between the metal complex and the target analyte.

[0161] Preferably, the analyte has a repeating structure to enable aggregation and supramolecular self-assembly of the metal complex thereon. In some forms, the analyte is electrostatically attracted to the metal complex, and the electrostatic interaction between the analyte and the metal complex can be one of the driving forces for binding. In some forms, the analyte carries a neutral charge or has an electrostatic repulsive force on the metal complex, and the metal complex can bind to such an analyte through other types of non-covalent interactions, such as, but not limited to, π-π stacking interactions, hydrogen bonding interactions, hydrophobic interactions, and combinations thereof.

[0162] 2. Ligands of metal complexes

[0163] The bonding between a ligand and a metal atom in a metal complex generally involves the formal donation of one or more electron pairs from a donor atom of the ligand. Donor atoms can be carbon (C), nitrogen (N), oxygen (O), phosphorus (P), sulfur (S), arsenic (As), and selenium (Se).

[0164] Exemplary ligands include optionally substituted C6-C 50Aromatics or C3-C 50 Heteroaromatic hydrocarbons, such as benzene, pyridine, thiophene, furan, pyrazole, imidazole, oxazole, isoxazole, thiazole, isothiazine, isoquinoline, pyrrole, pyrazine, pyridazine, pyrimidine, benzimidazole, benzofuran, benzothiazole, indole, naphthalene, anthracene, pyrene, triazole, tetrazolium, pyran, thiamphenicol, oxadiazole, triazine, tetrazine, carbazole, dibenzothiophene, dibenzofuran, fluorene and their derivatives.

[0165] Exemplary ligands also include halide ions and SCN. - (Donor atom: S), O-NO2 - (Donor atom: O), N3 - O 2- S 2- H2O, O-NO - (Donor atom: O), NCS - (Donor atom: N), NH3, NO2 - (Donor atom: N), N≡C - (Donor atom: N), C≡N - (Donor atom: C), CO (Donor atom: C), C≡CR - OR - SR - 、SeR - 、SeR 1 R 2 N3R, N≡CR (donor atom: N), NR 1 R 2 R 3 PR 1 R 2 R 3 and AsR 1 R 2 R 3 In some forms, one or more ligands of the metal complex are C≡CR. - .

[0166] In some forms of these ligands, R, R 1 R 2 and R 3 Independently:

[0167] Hydrogen atom, halogen atom, sulfonic acid, azide group, cyanate group, isocyanate group, nitrate ester group, nitrile group, isonitrile group, nitrosoxy group, nitroso group, nitro group, aldehyde group, acyl halide group, carboxylic acid group, carboxylic acid ester group, optionally substituted alkyl group, optionally substituted heteroalkyl group, optionally substituted alkenyl group, optionally substituted heteroalkenyl group, optionally substituted alkynyl group, optionally substituted heteroalkynyl group, optionally substituted aryl group, optionally substituted heteroaryl group;

[0168] A hydroxyl group that optionally contains a substituent on a hydroxyl oxygen, wherein the substituent is an optionally substituted alkyl group, an optionally substituted heteroalkyl group, an optionally substituted alkenyl group, an optionally substituted heteroalkenyl group, an optionally substituted alkynyl group, an optionally substituted heteroalkynyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group.

[0169] A thiol group optionally containing one substituent on a thiol thiosulfate, wherein the substituent is an optionally substituted alkyl group, an optionally substituted heteroalkyl group, an optionally substituted alkenyl group, an optionally substituted heteroalkenyl group, an optionally substituted alkynyl group, an optionally substituted heteroalkynyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group.

[0170] Contains an alkyl group, a heteroalkyl group, an alkenyl group, a heteroalkenyl group, an alkynyl group, a heteroalkynyl group, an aryl group, or a sulfonyl group that has been optionally substituted;

[0171] An amino group that optionally contains one or two substituents on an amino nitrogen, wherein the substituents are optionally substituted alkyl groups, optionally substituted heteroalkyl groups, optionally substituted alkenyl groups, optionally substituted heteroalkenyl groups, optionally substituted alkynyl groups, optionally substituted heteroalkynyl groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, or combinations thereof.

[0172] An amide group optionally comprising one or two substituents on the amide nitrogen, wherein the substituents are optionally substituted alkyl groups, optionally substituted heteroalkyl groups, optionally substituted alkenyl groups, optionally substituted heteroalkenyl groups, optionally substituted alkynyl groups, optionally substituted heteroalkynyl groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, or combinations thereof.

[0173] An azo group containing an optionally substituted alkyl group, an optionally substituted heteroalkyl group, an optionally substituted alkenyl group, an optionally substituted heteroalkenyl group, an optionally substituted alkynyl group, an optionally substituted heteroalkynyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group.

[0174] An acyl group containing an optionally substituted alkyl group, an optionally substituted heteroalkyl group, an optionally substituted alkenyl group, an optionally substituted heteroalkenyl group, an optionally substituted alkynyl group, an optionally substituted heteroalkynyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group.

[0175] An ester group containing an optionally substituted alkyl group, an optionally substituted heteroalkyl group, an optionally substituted alkenyl group, an optionally substituted heteroalkenyl group, an optionally substituted alkynyl group, an optionally substituted heteroalkynyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group.

[0176] Carbonate groups containing optionally substituted alkyl groups, optionally substituted heteroalkyl groups, optionally substituted alkenyl groups, optionally substituted heteroalkenyl groups, optionally substituted alkynyl groups, optionally substituted heteroalkynyl groups, optionally substituted aryl groups, or optionally substituted heteroaryl groups.

[0177] An ether group containing an optionally substituted alkyl group, an optionally substituted heteroalkyl group, an optionally substituted alkenyl group, an optionally substituted heteroalkenyl group, an optionally substituted alkynyl group, an optionally substituted heteroalkynyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group.

[0178] An aminooxy group optionally comprising one or two substituents on an amino nitrogen atom, wherein the substituents are optionally substituted alkyl groups, optionally substituted heteroalkyl groups, optionally substituted alkenyl groups, optionally substituted heteroalkenyl groups, optionally substituted alkynyl groups, optionally substituted heteroalkynyl groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, or combinations thereof; or

[0179] A hydroxylamino group optionally comprising one or two substituents, wherein the substituents are optionally substituted alkyl groups, optionally substituted heteroalkyl groups, optionally substituted alkenyl groups, optionally substituted heteroalkenyl groups, optionally substituted alkynyl groups, optionally substituted heteroalkynyl groups, optionally substituted aryl groups, substituted heteroaryl groups, or combinations thereof.

[0180] In some forms, R, R 1 R 2 R 3 Its organic substituents are optionally and independently replaced by one or more groups, wherein each such group is independently:

[0181] Halogen atoms, alkyl groups, heteroalkyl groups, alkenyl groups, heteroalkenyl groups, alkynyl groups, heteroalkynyl groups, aryl groups, heteroaryl groups, -OH, -SH, -NH2, -N3, -OCN, -NCO, -ONO2, -CN, -NC, -ONO, -CONH2, -NO, -NO2, -ONH2, -SCN, -SNCS, -CF3, -CH2CF3, -CH2Cl, -CHCl2, -CH2NH2, -NHCOH, -CHO, -COCl, -COF, -COBr, -COOH, -SO3H, -CH2SO2CH3, -PO3H2, -OPO3H2, -P(=O)(ORG1′ )(OR G2′ )、 -OP(=O)(OR G1′ )(OR G2′ )、 -BR G1′ (OR G2′ )、 -B(OR G1′ )(OR G2′ ) or -G′R G1′ , where -G′ is -O-, -S-, -NR G2′ -, -C(=O)-, -S(=O)-, -SO2-, -C(=O)O-, -C(=O)NR G2′ -, -OC(=O)-, -NR G2′ C(=O)-, -OC(=O)O-, -OC(=O)NR G2′ -, -NR G2′ C(=O)O-, -NR G2′ C(=O)NR G3′ -, -C(=S)-, -C(=S)S-, -SC(=S)-, -SC(=S)S-, -C(=NR G2′ )-, -C(=NR G2′ )O-, -C(=NR G2′ )NR G3′ -, -OC(=NR G2′ )-, -NR G2′ C(=NR G3′ )-, -NR G2′ SO2-, -C(=NR<​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​C(=O)S-, -C(=O)S-, -SC(=O)-, -SC(=O)S-, -C(=S)O-, -OC(=S)-, -OC(=S)O-, -SO2NR G2′ -、-BR G2′ -or-PR G2′ -,

[0182] Among them, R G1′ R G2′ and R G3′ Each time it appears, it is independently a hydrogen atom, halogen atom, alkyl group, heteroalkyl group, alkenyl group, heteroalkenyl group, alkynyl group, heteroalkynyl group, aryl group, or heteroaryl group.

[0183] In some forms, the ligand does not have the following structure:

[0184]

[0185] In some forms, the metal atom is Pt(II), and the ligands do not have the following structure:

[0186]

[0187] Each ligand can exist independently in its natural or deprotonated form.

[0188] 3. Exemplary formulas and structures of compounds

[0189] In some forms, metal complexes have a molecular geometry with a square planar shape containing monodentate, dipentate, tripentate, or tetradentate ligands. In some forms, the compound can have the structure of Formula I:

[0190]

[0191] in

[0192] (a) M represents a metal atom selected from Pt(II), Pd(II), Ni(II), Ir(I), Rh(I), Au(III), Ag(III) and Cu(III).

[0193] (b) L1, L2, L3, and L4 represent ligands, where each ligand provides a donor atom to coordinate to a metal atom.

[0194] (c) n+ / - indicates the number of positive or negative charges carried by the metallic complex in the formula, where n is zero or a positive integer, such as 1, 2, 3, 4, and 5.

[0195] (d)X m- / + This represents a counterion that maintains a neutral charge, where X m- / +It has a charge opposite to that of the metallic complex, and where m is zero or a positive integer, such as 1, 2, and 3, m = n or m ≠ n.

[0196] (e) The expression represents the stoichiometry of the counter ions.

[0197] (f) Dashed lines represent covalent bonds between any two ligands, fusion of any ring portions from two ligands, or combinations thereof.

[0198] When X m- / + It is an anion, i.e., X m- At that time, it can be selected from chloride ions (Cl... - ), hexafluorophosphate (PF6) - ), nitrate (NO3) - ), perchlorate (ClO4) - ), tetrafluoroborate (BF4) - ), tetraphenylborate (B(C6H5)4 - ), trifluoromethanesulfonate (CF3SO3) - ), dihydrogen phosphate (H2PO4) - ), sulfate (SO4 2- ), hydrogen phosphate (HPO4) 2- ), phosphate (PO4) 3- ) and its derivatives. When X m- / + It is a cation, namely X m+ At that time, it can be selected from K + Na + Ca 2+ Mg 2+ bis(triphenylphosphine)imine ion ([(C6H5)3P)2N] + ), phosphonium, pyridinium ([C5H5NH) + ), quaternary ammonium cations and their derivatives.

[0199] An exemplary structure of Formula I includes the following:

[0200]

[0201] The curves represent covalent bonds between any two ligands, fusion of any ring portions from the two ligands, or combinations thereof.

[0202] In some forms, the compound does not have the following structure:

[0203]

[0204] In some forms, L1, L2, and L3 are independently selected from optionally substituted and / or optionally deprotonated C6-C. 50Aromatics or C3-C 50 Heteroaromatic hydrocarbons, such as 5-membered aromatic hydrocarbons, 6-membered aromatic hydrocarbons, 5-membered heteroaromatic hydrocarbons, and 6-membered heteroaromatic hydrocarbons. Examples of L1, L2, and L3 include benzene, pyridine, thiophene, furan, pyrazole, imidazole, oxazole, isoxazole, thiazole, isothiazine, isoquinoline, pyrrole, pyrazine, pyridazine, pyrimidine, benzimidazole, benzofuran, benzothiazole, indole, naphthalene, anthracene, pyrene, triazole, tetraazole, pyran, thiamphenicol, oxadiazole, triazine, tetraazine, carbazole, dibenzothiophene, dibenzofuran, fluorene, and their derivatives.

[0205] In some forms, L4 is selected from benzene, pyridine, thiophene, furan, pyrazole, imidazole, oxazole, isoxazole, thiazole, isothiazole, isoquinoline, pyrrole, pyrazine, pyridazine, pyrimidine, benzimidazole, benzofuran, benzothiazole, indole, naphthalene, anthracene, pyrene, triazole, tetrazolium, pyran, thiazole, oxadiazole, triazine, tetrazine, carbazole, dibenzothiophene, dibenzofuran, fluorene, halides, alkylamines, arylamines, alkylphosphines, arylphosphines, alkylarsine, arylarsine, C≡CR - SR - OR - 、SeR - and its derivatives, wherein R is as defined above. For example, R is selected from H or substituted or unsubstituted C1-C. 30 Alkyl, C2-C 30 alkenyl, C2-C 30 alkynyl group, C3-C 30 Aryl, C3-C 30 heteroaryl, C1-C 30 Alkoxy, C3-C 30 Aryloxy group, C3-C 30 Arylthio, C1-C 30 Alkylthio, C2-C 30 carbonyl, C1-C 30 Carboxyl, amino, amide, or polyaryl (containing fused or non-fused ring moieties). In some forms, L4 is C≡CR. - .

[0206] In some forms, L1 and L2 are connected by covalent bonds, fusion of the ring portions of the two ligands, or a combination thereof. In some forms, L2 and L3 are further connected by covalent bonds, fusion of the ring portions of the two ligands, or a combination thereof. In some forms, L1 and L4 are further connected by covalent bonds, fusion of the ring portions of the two ligands, or a combination thereof. In some forms, L3 and L4 are further connected by covalent bonds, fusion of the ring portions of the two ligands, or a combination thereof.

[0207] In some forms, the metal atom is not Au(III). In some forms, the metal atom is Au(III), and the compound does not have the following structure:

[0208]

[0209] In some forms, the ligand does not have the following structure:

[0210]

[0211] In some forms, the metal atom is Pt(II), and the ligands do not have the following structure:

[0212]

[0213] In some forms, metal complexes have a linear, planar molecular geometry. In some forms, the compound may have the structure of Formula II:

[0214]

[0215] Where M′ represents a metal atom selected from Ni(0), Pd(0), Pt(0), Cu(I), Ag(I), Au(I), Zn(II), Cd(II), and Hg(II).

[0216] L5 and L6 represent ligands, with each ligand providing a donor atom to coordinate to the metal atom.

[0217] In some forms, metal complexes possess trigonal planar molecular geometries with monodentate, bidentate, or tripentate ligands. In some forms, the compounds can also have the structure of Formula III:

[0218]

[0219] L7, L8, and L9 represent ligands, with each ligand providing a donor atom to coordinate to a metal atom.

[0220] An exemplary structure of Formula III includes the following:

[0221]

[0222]

[0223] The curves represent covalent bonds between any two ligands, fusion of any ring portions from the two ligands, or combinations thereof.

[0224] Exemplary structures of metal complexes in compounds of formula I, II, or III are shown below.

[0225]

[0226] Where M is Pt(II) (complex 1-Pt), Pd(II) (complex 1-Pd), Ni(II) (complex 1-Ni), Ir(I) (complex 1-Ir), Rh(I) (complex 1-Rh), Au(III) (complex 1-Au), Ag(III) (complex 1-Ag), or Cu(III) (complex 1-Cu).

[0227] Where n is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0228] Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0229] in It is the stoichiometry of the counter ions in the formula.

[0230]

[0231] Where M is Pt(II) (complex 2-Pt), Pd(II) (complex 2-Pd), Ni(II) (complex 2-Ni), Ir(I) (complex 2-Ir), Rh(I) (complex 2-Rh), Au(III) (complex 2-Au), Ag(III) (complex 2-Ag), or Cu(III) (complex 2-Cu).

[0232] Where n+ is the number of positive charges carried by the metallic complex in the formula, and n is a positive integer.

[0233] Where X m－ It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n.

[0234] in It is the stoichiometry of the counter ions in the formula.

[0235]

[0236] Where M is Pt(II) (complex 3-Pt), Pd(II) (complex 3-Pd), Ni(II) (complex 3-Ni), Ir(I) (complex 3-Ir), Rh(I) (complex 3-Rh), Au(III) (complex 3-Au), Ag(III) (complex 3-Ag), or Cu(III) (complex 3-Cu).

[0237] Where n+ / - is the number of positive or negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0238] Where X m- / +It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0239] in It is the stoichiometry of the counter ions in the formula.

[0240]

[0241] Where M is Pt(II) (4-Pt complex), Pd(II) (4-Pd complex), Ni(II) (4-Ni complex), Ir(I) (4-Ir complex), Rh(I) (4-Rh complex), Au(III) (4-Au complex), Ag(III) (4-Ag complex), or Cu(III) (4-Cu complex).

[0242] Where n- is the number of negative charges carried by the metal complex in the formula, and n is a positive integer.

[0243] Where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n.

[0244] in It is the stoichiometry of the counter ions in the formula.

[0245]

[0246] Where M is Pt(II) (5-Pt complex), Pd(II) (5-Pd complex), Ni(II) (5-Ni complex), Ir(I) (5-Ir complex), Rh(I) (5-Rh complex), Au(III) (5-Au complex), Ag(III) (5-Ag complex), or Cu(III) (5-Cu complex).

[0247] Where n is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0248] Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0249] in It is the stoichiometry of the counter ions in the formula.

[0250]

[0251] Where M is Pt(II) (complex 6-Pt), Pd(II) (complex 6-Pd), Ni(II) (complex 6-Ni), Ir(I) (complex 6-Ir), Rh(I) (complex 6-Rh), Au(III) (complex 6-Au), Ag(III) (complex 6-Ag), or Cu(III) (complex 6-Cu).

[0252] Where n is the number of negative charges carried by the metal complex in the formula, and n is a positive integer.

[0253] Where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n.

[0254] in It is the stoichiometry of the counter ions in the formula.

[0255]

[0256] Where M is Pt(II) (complex 7-Pt), Pd(II) (complex 7-Pd), Ni(II) (complex 7-Ni), Ir(I) (complex 7-Ir), Rh(I) (complex 7-Rh), Au(III) (complex 7-Au), Ag(III) (complex 7-Ag), or Cu(III) (complex 7-Cu).

[0257] Where n is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0258] Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0259] in It is the stoichiometry of the counter ions in the formula.

[0260]

[0261] Where M is Pt(II) (complex 8-Pt), Pd(II) (complex 8-Pd), Ni(II) (complex 8-Ni), Ir(I) (complex 8-Ir), Rh(I) (complex 8-Rh), Au(III) (complex 8-Au), Ag(III) (complex 8-Ag), or Cu(III) (complex 8-Cu).

[0262] Where n is the number of negative charges carried by the metal complex in the formula, and n is a positive integer.

[0263] Where X m+It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n.

[0264] in It is the stoichiometry of the counter ions in the formula.

[0265]

[0266] Where M is Pt(II) (complex 9-Pt), Pd(II) (complex 9-Pd), Ni(II) (complex 9-Ni), Ir(I) (complex 9-Ir), Rh(I) (complex 9-Rh), Au(III) (complex 9-Au), Ag(III) (complex 9-Ag), or Cu(III) (complex 9-Cu).

[0267] Where n is the number of negative charges carried by the metal complex in the formula, and n is a positive integer.

[0268] Where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n.

[0269] in It is the stoichiometry of the counter ions in the formula.

[0270]

[0271] Where M is Pt(II) (10-Pt complex), Pd(II) (10-Pd complex), Ni(II) (10-Ni complex), Ir(I) (10-Ir complex), Rh(I) (10-Rh complex), Au(III) (10-Au complex), Ag(III) (10-Ag complex), or Cu(III) (10-Cu complex).

[0272] Where n is the number of negative charges carried by the metallic complex in the formula, and n is a positive integer, where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n.

[0273] in It is the stoichiometry of the counter ions in the formula.

[0274]

[0275] Where M is Pt(II) (complex 11-Pt), Pd(II) (complex 11-Pd), Ni(II) (complex 11-Ni), Ir(I) (complex 11-Ir), Rh(I) (complex 11-Rh), Au(III) (complex 11-Au), Ag(III) (complex 11-Ag), or Cu(III) (complex 11-Cu).

[0276] Where n is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0277] Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0278] in It is the stoichiometry of the counter ions in the formula.

[0279]

[0280] Where M is Pt(II) (complex 12-Pt), Pd(II) (complex 12-Pd), Ni(II) (complex 12-Ni), Ir(I) (complex 12-Ir), Rh(I) (complex 12-Rh), Au(III) (complex 12-Au), Ag(III) (complex 12-Ag), or Cu(III) (complex 12-Cu).

[0281] Where n is the number of negative charges carried by the metallic complex in the formula, and n is a positive integer, where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n.

[0282] in It is the stoichiometry of the counter ions in the formula.

[0283]

[0284] Where M′ is Ni(0) (complex 13-Ni), Pd(0) (complex 13-Pd), Pt(0) (complex 13-Pt), Cu(I) (complex 13-Cu), Ag(I) (complex 13-Ag), Au(I) (complex 13-Au), Zn(II) (complex 13-Zn), Cd(II) (complex 13-Cd), or Hg(II) (complex 13-Hg).

[0285] Where n is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0286] Where Xm+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0287] in It is the stoichiometry of the counter ions in the formula.

[0288]

[0289] Where M′ is Ni(0) (complex 14-Ni), Pd(0) (complex 14-Pd), Pt(0) (complex 14-Pt), Cu(I) (complex 14-Cu), Ag(I) (complex 14-Ag), Au(I) (complex 14-Au), Zn(II) (complex 14-Zn), Cd(II) (complex 14-Cd), or Hg(II) (complex 14-Hg).

[0290] Where n is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0291] Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0292] in It is the stoichiometry of the counter ions in the formula.

[0293] Compounds of Formula I, II, or III can be readily synthesized using techniques commonly known to synthetic organic and inorganic chemists. Exemplary methods for synthesizing specific compounds of Formula I, namely complex 1-Pt and complex 2-Pt, are described in the disclosed embodiments.

[0294] III. Mixtures and Compositions

[0295] Mixtures, compositions, and kits formed by implementing or preparing to implement the disclosed methods are disclosed.

[0296] 1. Mixtures and compositions

[0297] For example, mixtures comprising multiple compounds for detecting analytes and / or imaging or screening them and / or testing inhibitors are disclosed. In some forms, the analyte is amyloid protein or peptide, plaque, or both. In some forms, the analyte is RNA, nucleolus, or both. The mixture can be used to screen or test the efficacy of inhibitors against amyloidosis and / or fibrillary growth of proteins or peptides. The mixture can also be used for nucleolar imaging.

[0298] In some forms, the mixture contains multiple compounds having the structure of formula I, II or III.

[0299] In some forms, the compounds in the mixture may have different specificities for different types of amyloid proteins or plaques. Alternatively, the compounds in the mixture may have different specificities for different proteins or peptides of amyloid, plaques, or both. The compounds in the mixture may exhibit different photophysical properties after aggregation and supramolecular self-assembly, thereby enabling the simultaneous detection and / or imaging of different types of amyloid proteins or plaques and / or the simultaneous detection and / or imaging of different proteins or peptides of amyloid, plaques, or both.

[0300] In some forms, compounds in a mixture can exhibit different specificities for different types of RNA. After aggregation and supramolecular self-assembly, compounds in a mixture can display different photophysical properties, thus enabling the simultaneous detection of different types of RNA.

[0301] In another example, a composition comprising one or more disclosed compounds, such as compounds having the structure of formula I, II, or III, and one or more other compounds, solvents, or materials is disclosed. In some forms, one or more other compounds, solvents, or materials may improve the properties of the disclosed compound and / or increase its stability. The composition may be in the form of a solution, suspension, emulsion, powder, or solid agglomerate.

[0302] 2. Reagent kit

[0303] The above-described compounds, mixtures, and compositions can be packaged together with other components in any suitable combination as a kit for carrying out or assisting in carrying out the disclosed methods. A given kit is useful if the components in it are designed and suitable for use in the disclosed methods.

[0304] The kit contains one or more disclosed compounds, mixtures, and compositions in one or more containers. The kit may also contain one or more other components, such as compounds, solvents, and materials, as a carrier. The carrier does not interfere with the effectiveness of the disclosed compounds in performing its function. The kit may include instructions for use.

[0305] The kit can be used to detect and / or image analytes. In some forms, the analyte is amyloid protein, plaque, or both. In other forms, the analyte is RNA, nucleolus, or both. The kit can be used to screen or test the efficacy of inhibitors against amyloidosis and / or fibrillary growth of proteins or peptides. The kit can also be used for nucleolar imaging.

[0306] The kit may also contain one or more positive controls. In some forms, the positive control is a solution, suspension, or dry powder of one or more proteins or peptides, amyloid plaques, or both. In some forms, the positive control is a solution, suspension, or dry powder of RNA.

[0307] IV. Instructions for Use

[0308] Methods for detecting analytes and / or imaging or screening them and / or testing inhibitors are disclosed.

[0309] In some forms, the analyte is amyloid protein or peptide, plaque, or both. In other forms, the analyte is RNA, nucleolus, or both. The compounds can be used to screen or test the efficacy of inhibitors against amyloidosis and / or fibrillary growth of proteins or peptides. The compounds can also be used for nucleolar imaging.

[0310] For example, methods for detecting analytes in a sample may include (a) combining one or more disclosed compounds with the sample, and (b) detecting changes in the photophysical properties of metal complexes of the compounds. The detection of changes in the photophysical properties of the metal complexes indicates the presence of aggregation and supramolecular self-assembly of the metal complexes, which in turn indicates the presence of the analyte in the sample.

[0311] For example, a method for imaging an analyte in a sample may include: (a) combining one or more disclosed compounds with the sample under conditions that allow the metal complexes of the compounds to bind to the analyte and subsequently aggregate and supramolecularly self-assemble, wherein the aggregation and supramolecularly self-assembly of the metal complexes results in a change in the photophysical properties of the metal complexes; and (b) imaging the analyte based on one or more photophysical properties specific to the aggregated and supramolecularly self-assembled metal complexes.

[0312] 1. Detection of amyloid proteins or peptides, plaques, or both, and / or imaging of them.

[0313] A method for detecting and / or imaging one or more amyloid proteins, plaques, or both in a sample containing proteins or peptides is disclosed. The method comprises (a) combining one or more of the disclosed compounds with the sample, and (b) detecting changes in the photophysical properties of metal complexes of the compounds. Detection of changes in the photophysical properties of the metal complexes indicates the presence of aggregation and supramolecular self-assembly of the metal complexes, wherein the presence of aggregation and supramolecular self-assembly of the metal complexes indicates the presence of amyloid proteins, plaques, or both in the sample.

[0314] Depending on the type of change in the photophysical properties of the metal complex, different techniques, such as colorimetric assays, luminescence assays, RLS analysis, or combinations thereof, can be used to detect and / or image amyloids, plaques, or both of proteins or peptides.

[0315] In some forms, it is possible to use, such as Figure 13 The illustrated luminescence-on assay is used to detect and / or image amyloid plaques or both of proteins or peptides. Increased luminescence intensity can be induced by the aggregation and supramolecular self-assembly of metal complexes induced by binding to amyloid plaques or both of proteins or peptides. This increase in luminescence intensity can be caused by aggregation and supramolecular self-assembly of metal complexes through non-covalent metal-metal interactions, similar to the effects of exciton coupling. Non-covalent interactions, such as π-π stacking interactions, electrostatic interactions, hydrogen bonding interactions, hydrophobic interactions, and combinations thereof, can facilitate the binding of metal complexes to amyloid plaques or both of proteins or peptides, leading to aggregation and supramolecular self-assembly of the metal complexes.

[0316] In some forms, samples are measured using non-imaging spectrometers, such as those from cuvettes, small sample holders, or multi-well plates. In other forms, samples are measured using imaging spectrometers, such as confocal microscopes.

[0317] In some forms, the method further includes parallel measurements on one or more samples of amyloid, plaques, or both of other proteins or peptides, or on one or more samples of the same protein or peptide, wherein the structural properties of the amyloid, plaques, or both in these samples are previously known and / or characterized. By performing such measurements using amyloid or plaques with known structural properties, the combined set of information can provide a pathway to inferring the structural properties of the amyloid, plaques, or both in the sample under study.

[0318] A method for studying the formation of amyloid fibrils and plaques of one or more proteins or peptides under different conditions is also disclosed. The method includes (a) combining one or more disclosed compounds with samples of proteins or peptides collected or prepared at different times and / or under different conditions, and (b) comparing the photophysical properties of the metal complexes between the samples. Differences in the photophysical properties of the metal complexes between the samples indicate differences in the degree or stage of amyloid fibril and plaque formation. The kinetics of amyloid fibril and plaque formation can be derived from a time-dependent comparison of the photophysical properties of the metal complexes between samples collected or prepared at different times.

[0319] 2. Evaluate the efficacy of inhibitors against amyloidosis and / or fibrillary growth of proteins or peptides.

[0320] A method for testing the efficacy of inhibitors against amyloidosis and / or fibrillary growth of one or more proteins or peptides is disclosed. The method includes (a) combining one or more disclosed compounds with an inhibitor-treated sample containing a protein or peptide, and separately combining them with an untreated sample containing a protein or peptide, and (b) comparing the photophysical properties of the metal complexes of the compounds between the two samples. The magnitude of the difference in the photophysical properties of the metal complexes between the two samples indicates the degree of change in the aggregation and supramolecular self-assembly state of the metal complexes; the degree of change in the aggregation and supramolecular self-assembly state of the metal complexes indicates the efficacy of the inhibitor.

[0321] In some forms, inhibitor-treated samples can be prepared by treating samples containing proteins and peptides with one or more inhibitors for a sufficient period of time for the inhibitors to take effect. The inhibitors can be added before, during, or after the amyloidosis and / or fibrillation of the proteins or peptides.

[0322] A method for screening inhibitors against amyloidosis and / or fibrillary growth of one or more proteins or peptides is also disclosed. The method includes evaluating and then comparing the efficacy of the inhibitors against amyloidosis and / or fibrillary growth of the proteins or peptides.

[0323] 3. RNA detection and nucleolar imaging

[0324] A method for detecting RNA, nucleolus, or both in a sample is disclosed. The method comprises (a) combining one or more disclosed compounds with a sample, and (b) detecting changes in the photophysical properties of metal complexes of the compounds. Detection of changes in the photophysical properties of the metal complexes indicates the presence of aggregation and supramolecular self-assembly of the metal complexes, wherein the presence of aggregation and supramolecular self-assembly of the metal complexes indicates the presence of RNA, nucleolus, or both in the sample.

[0325] Depending on the type of change in the photophysical properties of the metal complex, different techniques, such as colorimetry, luminescence assay, RLS analysis, or combinations thereof, can be used for RNA detection and nucleolar imaging.

[0326] In some forms, it is possible to use, such as Figure 30The luminescence-on assay shown is used for RNA detection and nucleolar imaging. The aggregation and supramolecular self-assembly of metal complexes induced by their binding to RNA, nucleolar, or both can induce an increase in luminescence intensity. This increase in luminescence intensity can be caused by aggregation and supramolecular self-assembly of metal complexes through non-covalent metal-metal interactions, similar to exciton coupling. Non-covalent interactions, such as π-π stacking interactions, electrostatic interactions, hydrogen bonding, hydrophobic interactions, and combinations thereof, can facilitate the binding of metal complexes to RNA, nucleolar, or both, leading to aggregation and supramolecular self-assembly of the metal complexes.

[0327] In some forms, non-imaging spectrometers such as cuvettes, small sample holders, or multi-well plates are used to measure samples. In other forms, imaging spectrometers such as confocal microscopes are used to measure samples.

[0328] A method for imaging nucleoli in a sample is disclosed. The method includes (a) combining one or more disclosed compounds with a sample under conditions that allow metal complexes of the compound to bind to the nucleolus and subsequently undergo aggregation and supramolecular self-assembly of the metal complexes, wherein the aggregation and supramolecular self-assembly of the metal complexes produce changes in the photophysical properties of the metal complexes of the compound, and (b) imaging the nucleolus based on one or more photophysical properties specific to the aggregated and supramolecularly self-assembled metal complexes.

[0329] In some forms, the sample contains eukaryotic cells. The cells may be, but are not limited to, 3T3 cells, A549 cells, Chinese hamster ovary (CHO) cells, HEK 293 cells, HeLa cells, Hep G2 cells, and HT1080 cells.

[0330] In some forms, imaging spectrometers, such as confocal microscopes, are used to image the sample.

[0331] 4. Use in combination

[0332] The disclosed methods also include the combined use of more than one of the disclosed compounds. The compounds can be combined to form mixtures or compositions as described above.

[0333] In some forms, the compounds in the mixture may have different specificities for different types of amyloid proteins or plaques. Alternatively, the compounds in the mixture may have different specificities for different proteins or peptides of amyloid, plaques, or both. The compounds in the mixture may exhibit different photophysical properties after aggregation and supramolecular self-assembly, thereby enabling the simultaneous detection and / or imaging of different types of amyloid proteins or plaques, and / or the simultaneous detection and / or imaging of different proteins or peptides of amyloid, plaques, or both.

[0334] In some forms, compounds in a mixture can exhibit different specificities for different types of RNA. After aggregation and supramolecular self-assembly, compounds in a mixture can display different photophysical properties, thus enabling the simultaneous detection of different types of RNA.

[0335] 5. Sample

[0336] In some forms, the sample contains one or more proteins or peptides. The proteins or peptides may be isolated proteins or peptides. In some forms, the sample containing proteins or peptides may contain human or non-human animal body fluids, human or non-human animal tissues, or combinations thereof. Exemplary body fluids include saliva, sputum, serum, blood, urine, mucus, vaginal lubricating secretions, pus, cerebrospinal fluid, and wound exudate. In some forms, the body fluid is cerebrospinal fluid. Exemplary tissues include organ tissues and non-organ tissues, such as brain tissue, heart tissue, kidney tissue, liver tissue, eye tissue, tongue tissue, and pancreatic tissue. In some forms, the tissue is brain tissue. Human or non-human animal tissues may be lysed to prepare the sample.

[0337] The sample may contain amyloid proteins or peptides, plaques, or both, and may include linear aggregates of proteins or peptides arranged in a β-sheet conformation.

[0338] Amyloids, plaques, or both of proteins or peptides in a sample can be analyzed directly, or they can be amplified prior to analysis. In some forms, one or more additional steps can be performed to induce the formation of amyloids, plaques, or both of proteins or peptides in the sample. In some forms, the formation of amyloids, plaques, or both of proteins or peptides may involve denaturation of the protein or peptide. In some forms, the formation of amyloids, plaques, or both of proteins or peptides can be induced by chemical methods, physical methods, or both. Exemplary chemical methods include adding one or more compounds, such as transition metal ions, adding a specific solvent, such as ethanol and methanol, preparing the sample by dissolving the protein or peptide in a specific solvent, altering the pH of the sample, and preparing the sample by dissolving the protein or peptide under specific pH conditions, such as acidic or alkaline conditions. Exemplary physical methods include altering the temperature of the sample, such as increasing the temperature, and introducing physical disturbances into the sample, such as stirring, shaking, or vortexing the sample.

[0339] In some forms, the sample contains one or more proteins or peptides selected from, but not limited to, amyloid-β peptide, α-synuclein, insulin, huntingtin, tau protein, hyperphosphorylated tau protein (pτ), prion protein, IAPP (insulin amyloid peptide), calcitonin, and PrP. ScThe sample contains: atrial natriuretic factor, apolipoprotein A1, serum amyloid A, medin, prolactin, transthyretin, lysozyme, β-2 microglobulin, coagulin, corneal epithelin, lens protein, desmin, selenoprotein, actin, cystatin, immunoglobulin light chain AL, S-IBM, and myosin. In some forms, the sample contains amyloid, plaques, or both of these proteins or peptides. In some forms, the sample contains amyloid, plaques, or both of amyloid-β peptide.

[0340] In some forms, the sample is obtained from the patient. In some forms, the patient has one or more diseases or conditions associated with amyloidosis and proteinosis. In some forms, the disease or condition is selected from, but is not limited to, Alzheimer's disease, Parkinson's disease, injection-specific amyloidosis, Huntington's disease, mild cognitive impairment, cerebral amyloid angiopathy, myopathy, neuropathy, traumatic brain injury, frontotemporal dementia, Pick's disease, multiple sclerosis, prion disease, type 2 diabetes, fatal familial insomnia, arrhythmia, isolated atrial amyloidosis, atherosclerosis, rheumatoid arthritis, familial amyloid polyneuropathy, hereditary nonneurogenic systemic amyloidosis, Finnish amyloidosis, lattice corneal dystrophy, systemic AL amyloidosis, and Down syndrome. In some forms, the disease or condition is Alzheimer's disease.

[0341] In some forms, the sample contains RNA, nucleolus, or both. In some forms, the RNA is RNA isolated from a biological source. In some forms, the sample contains or is derived from eukaryotic cells. Exemplary eukaryotic cells include, but are not limited to, 3T3 cells, A549 cells, Chinese hamster ovary (CHO) cells, HEK 293 cells, HeLa cells, Hep G2 cells, and HT1080 cells.

[0342] The RNA in the sample can be analyzed directly, or it can be amplified before analysis.

[0343] In some forms, the sample contains one or more types of RNA selected from, but not limited to, messenger RNA (mRNA), transfer RNA (tRNA), ribosomal RNA (rRNA), transfer messenger RNA (tmRNA), antisense RNA (asRNA), enhancer RNA (eRNA), guide RNA (gRNA), ribosomes, short hairpin RNA (shRNA), hourly sequence RNA (stRNA), small interfering RNA (siRNA), and trans-acting siRNA (ta-siRNA).

[0344] 6. Diagnosis of amyloidosis and proteinosis

[0345] Methods for diagnosing one or more diseases or conditions associated with amyloidosis and proteinosis in patients in need are disclosed.

[0346] In some forms, the disease or condition is selected from, but is not limited to, Alzheimer's disease, Parkinson's disease, injection-induced amyloidosis, Huntington's disease, mild cognitive impairment, cerebral amyloid angiopathy, myopathy, neuropathy, traumatic brain injury, frontotemporal dementia, Pick's disease, multiple sclerosis, prion diseases, type 2 diabetes, fatal familial insomnia, arrhythmia, isolated atrial amyloidosis, atherosclerosis, rheumatoid arthritis, familial amyloid polyneuropathy, hereditary nonneurogenic systemic amyloidosis, Finnish amyloidosis, lattice corneal dystrophy, systemic AL amyloidosis, and Down syndrome. In some forms, the disease or condition is Alzheimer's disease.

[0347] In some forms, the method includes: (a) extracting a sample from a patient; and (b) detecting and / or imaging the presence of amyloid, plaques, or both of one or more disclosed compounds using one or more disclosed compounds. In some forms, step (b) further includes quantifying the amount of amyloid, plaques, or both of the protein or peptide. In some forms, the method includes detecting and / or imaging the presence of amyloid, plaques, or both of one or more disclosed compounds using one or more disclosed compounds. In some forms, the method further includes quantifying the amount of amyloid, plaques, or both of the protein or peptide.

[0348] In some forms, samples from a patient include bodily fluids, tissues, or combinations thereof. Bodily fluids may be cerebrospinal fluid; tissues may be brain tissue.

[0349] In some forms, the sample from the patient contains one or more proteins or peptides selected from, but not limited to, amyloid-β peptide, α-synuclein, insulin, huntingtin, tau protein, hyperphosphorylated tau protein (pτ), prion protein, IAPP (insulin amyloid peptide), calcitonin, and PrP. Sc The sample contains: atrial natriuretic factor, apolipoprotein A1, serum amyloid A, medin, prolactin, transthyretin, lysozyme, β-2 microglobulin, coagulin, corneal epithelin, lens protein, desmin, selenoprotein, actin, cystatin, immunoglobulin light chain AL, S-IBM, and myosin. In some forms, the sample contains one or more of these proteins or peptides, amyloid plaques, or both. In some forms, the sample contains amyloid-β peptide, amyloid plaques, or both.

[0350] In some forms, the method includes in vivo imaging. In some forms, in vivo imaging involves (a) administering one or more disclosed compounds to the patient whole body or to a specific body region; and (b) using fluorescence imaging to detect the presence of amyloid, plaques, or both of one or more proteins or peptides whole body or in a specific body region. In some forms, the specific body region is in the brain.

[0351] The disclosed compositions and methods can be further understood through the following numbered paragraphs.

[0352] 1. A compound for detecting and / or imaging an analyte, wherein the compound is d 8 or d 10 Metal complexes or salts thereof, said compounds comprising:

[0353] (a) Metal atoms with coordination numbers of 2, 3, or 4, selected from Pt(II), Pd(II), Ni(II), Ir(I), Rh(I), Au(III), Ag(III), Cu(III), Ni(0), Pd(0), Pt(0), Cu(I), Ag(I), Au(I), Zn(II), Cd(II), and Hg(II); and

[0354] (b) One or more ligands having donor atoms, said donor atoms being independently selected from carbon (C), nitrogen (N), oxygen (O), phosphorus (P), sulfur (S), arsenic (As), and selenium (Se).

[0355] The metal complex is bound to the analyte, and the binding of the metal complex to the analyte induces the aggregation and supramolecular self-assembly of the metal complex through non-covalent metal-metal interactions.

[0356] 2. The compound described in paragraph 1, wherein the compound has the structure of formula I:

[0357]

[0358]

[0359] in

[0360] (a) M represents a metal atom selected from Pt(II), Pd(II), Ni(II), Ir(I), Rh(I), Au(III), Ag(III) and Cu(III).

[0361] (b) L1, L2, L3, and L4 represent ligands, where each ligand provides a donor atom to coordinate to a metal atom.

[0362] (c) n+ / - represents the number of positive or negative charges carried by the metallic complex in the formula, where n is zero or a positive integer.

[0363] (d)X m- / + This represents a counterion that maintains a neutral charge, where X m- / + It has a charge opposite to that of the metallic complex, and where m is zero or a positive integer, m = n or m ≠ n.

[0364] (e) The expression represents the stoichiometry of the counter ions.

[0365] (f) Dashed lines represent covalent bonds between any two ligands, fusion of any ring portions from the two ligands, or combinations thereof.

[0366] 3. The compounds described in paragraph 2, wherein L1, L2, and L3 are optionally substituted and / or optionally deprotonated C6-C. 50 Aromatics or C3-C 50 Heteroaromatic hydrocarbons include benzene, pyridine, thiophene, furan, pyrazole, imidazole, oxazole, isoxazole, thiazole, isothiazole, isoquinoline, pyrrole, pyrazine, pyridazine, pyrimidine, benzimidazole, benzofuran, benzothiazole, indole, naphthalene, anthracene, pyrene, triazole, tetrazolium, pyran, thiamphenicol, oxadiazole, triazine, tetrazine, carbazole, dibenzothiophene, dibenzofuran, fluorene and their derivatives.

[0367] 4. The compound described in paragraph 2 or paragraph 3, wherein L1 and L2 are connected by covalent bonds, fusion of the cyclic portions of the two ligands, or a combination thereof.

[0368] 5. The compound described in paragraph 1, wherein the compound has the structure of formula II:

[0369]

[0370] Where M′ represents a metal atom selected from Ni(0), Pd(0), Pt(0), Cu(I), Ag(I), Au(I), Zn(II), Cd(II), and Hg(II).

[0371] L5 and L6 represent ligands, with each ligand providing a donor atom to coordinate to the metal atom.

[0372] 6. The compound described in paragraph 1, wherein the compound has the structure of formula III:

[0373]

[0374] L7, L8, and L9 represent ligands, with each ligand providing a donor atom to coordinate to a metal atom.

[0375] 7. The compound described in any one of paragraphs 1-6, wherein the metal complex is bound to the analyte via a non-covalent interaction, wherein the non-covalent interaction includes electrostatic interaction, hydrogen bonding interaction, hydrophobic interaction, or a combination thereof.

[0376] 8. The compound described in any one of paragraphs 1-7, wherein the metal complex has a planar or partially planar structure.

[0377] 9. The compound described in any one of paragraphs 1-8, wherein the aggregation and supramolecular self-assembly of the metal complexes result in one or more changes in the photophysical properties of the metal complexes.

[0378] 10. The compounds described in paragraph 9, wherein the changes in the photophysical properties include changes in absorbance, luminescence, resonant light scattering (RLS), or combinations thereof.

[0379] 11. The compound described in paragraph 10, wherein the change in luminescence includes an increase in luminescence quantum yield and / or emission intensity, and / or a shift in emission energy or wavelength.

[0380] 12. Any one of the compounds described in paragraphs 1-11, wherein the compound is selected from:

[0381]

[0382] Where M is Pt(II) (complex 1-Pt), Pd(II) (complex 1-Pd), Ni(II) (complex 1-Ni), Ir(I) (complex 1-Ir), Rh(I) (complex 1-Rh), Au(III) (complex 1-Au), Ag(III) (complex 1-Ag), or Cu(III) (complex 1-Cu).

[0383] Where n is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0384] Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0385] in This is the stoichiometry of the counter ions in the formula;

[0386]

[0387] Where M is Pt(II) (complex 2-Pt), Pd(II) (complex 2-Pd), Ni(II) (complex 2-Ni), Ir(I) (complex 2-Ir), Rh(I) (complex 2-Rh), Au(III) (complex 2-Au), Ag(III) (complex 2-Ag), or Cu(III) (complex 2-Cu).

[0388] Where n+ is the number of positive charges carried by the metallic complex in the formula, and n is a positive integer.

[0389] Where X m－ It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n, where This is the stoichiometry of the counter ions in the formula;

[0390]

[0391] Where M is Pt(II) (complex 3-Pt), Pd(II) (complex 3-Pd), Ni(II) (complex 3-Ni), Ir(I) (complex 3-Ir), Rh(I) (complex 3-Rh), Au(III) (complex 3-Au), Ag(III) (complex 3-Ag), or Cu(III) (complex 3-Cu).

[0392] Where n+ / - is the number of positive or negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0393] Where X m- / + It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0394] in This is the stoichiometry of the counter ions in the formula;

[0395]

[0396] Where M is Pt(II) (4-Pt complex), Pd(II) (4-Pd complex), Ni(II) (4-Ni complex), Ir(I) (4-Ir complex), Rh(I) (4-Rh complex), Au(III) (4-Au complex), Ag(III) (4-Ag complex), or Cu(III) (4-Cu complex).

[0397] Where n is the number of negative charges carried by the metal complex in the formula, and n is a positive integer.

[0398] Where X m+It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n, where This is the stoichiometry of the counter ions in the formula;

[0399]

[0400] Where M is Pt(II) (5-Pt complex), Pd(II) (5-Pd complex), Ni(II) (5-Ni complex), Ir(I) (5-Ir complex), Rh(I) (5-Rh complex), Au(III) (5-Au complex), Ag(III) (5-Ag complex), or Cu(III) (5-Cu complex).

[0401] Where n is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0402] Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0403] in It is the stoichiometry of the counter ions in the formula.

[0404]

[0405] Where M is Pt(II) (complex 6-Pt), Pd(II) (complex 6-Pd), Ni(II) (complex 6-Ni), Ir(I) (complex 6-Ir), Rh(I) (complex 6-Rh), Au(III) (complex 6-Au), Ag(III) (complex 6-Ag), or Cu(III) (complex 6-Cu).

[0406] Where n is the number of negative charges carried by the metal complex in the formula, and n is a positive integer.

[0407] Where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n.

[0408] in It is the stoichiometry of the counter ions in the formula.

[0409]

[0410] Where M is Pt(II) (complex 7-Pt), Pd(II) (complex 7-Pd), Ni(II) (complex 7-Ni), Ir(I) (complex 7-Ir), Rh(I) (complex 7-Rh), Au(III) (complex 7-Au), Ag(III) (complex 7-Ag), or Cu(III) (complex 7-Cu).

[0411] Where n is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0412] Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0413] in It is the stoichiometry of the counter ions in the formula.

[0414]

[0415] Where M is Pt(II) (complex 8-Pt), Pd(II) (complex 8-Pd), Ni(II) (complex 8-Ni), Ir(I) (complex 8-Ir), Rh(I) (complex 8-Rh), Au(III) (complex 8-Au), Ag(III) (complex 8-Ag), or Cu(III) (complex 8-Cu).

[0416] Where n is the number of negative charges carried by the metal complex in the formula, and n is a positive integer.

[0417] Where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n.

[0418] in It is the stoichiometry of the counter ions in the formula.

[0419]

[0420] Where M is Pt(II) (complex 9-Pt), Pd(II) (complex 9-Pd), Ni(II) (complex 9-Ni), Ir(I) (complex 9-Ir), Rh(I) (complex 9-Rh), Au(III) (complex 9-Au), Ag(III) (complex 9-Ag), or Cu(III) (complex 9-Cu).

[0421] Where n is the number of negative charges carried by the metal complex in the formula, and n is a positive integer.

[0422] Where X m+It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n, where It is the stoichiometry of the counter ions in the formula.

[0423]

[0424] Where M is Pt(II) (10-Pt complex), Pd(II) (10-Pd complex), Ni(II) (10-Ni complex), Ir(I) (10-Ir complex), Rh(I) (10-Rh complex), Au(III) (10-Au complex), Ag(III) (10-Ag complex), or Cu(III) (10-Cu complex).

[0425] Where n is the number of negative charges carried by the metallic complex in the formula, and n is a positive integer, where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n.

[0426] in It is the stoichiometry of the counter ions in the formula.

[0427]

[0428] Where M is Pt(II) (complex 11-Pt), Pd(II) (complex 11-Pd), Ni(II) (complex 11-Ni), Ir(I) (complex 11-Ir), Rh(I) (complex 11-Rh), Au(III) (complex 11-Au), Ag(III) (complex 11-Ag), or Cu(III) (complex 11-Cu).

[0429] Where n is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0430] Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0431] in It is the stoichiometry of the counter ions in the formula.

[0432]

[0433] Where M is Pt(II) (complex 12-Pt), Pd(II) (complex 12-Pd), Ni(II) (complex 12-Ni), Ir(I) (complex 12-Ir), Rh(I) (complex 12-Rh), Au(III) (complex 12-Au), Ag(III) (complex 12-Ag), or Cu(III) (complex 12-Cu).

[0434] Where n is the number of negative charges carried by the metallic complex in the formula, and n is a positive integer, where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m = n or m ≠ n.

[0435] in It is the stoichiometry of the counter ions in the formula.

[0436]

[0437] Where M′ is Ni(0) (complex 13-Ni), Pd(0) (complex 13-Pd), Pt(0) (complex 13-Pt), Cu(I) (complex 13-Cu), Ag(I) (complex 13-Ag), Au(I) (complex 13-Au), Zn(II) (complex 13-Zn), Cd(II) (complex 13-Cd), or Hg(II) (complex 13-Hg).

[0438] Where n is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0439] Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0440] in It is the stoichiometry of the counter ions in the formula.

[0441]

[0442] Where M′ is Ni(0) (complex 14-Ni), Pd(0) (complex 14-Pd), Pt(0) (complex 14-Pt), Cu(I) (complex 14-Cu), Ag(I) (complex 14-Ag), Au(I) (complex 14-Au), Zn(II) (complex 14-Zn), Cd(II) (complex 14-Cd), or Hg(II) (complex 14-Hg).

[0443] Where n is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer.

[0444] Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m = n or m ≠ n.

[0445] in It is the stoichiometry of the counter ions in the formula.

[0446] 13. The compound described in any one of paragraphs 1-12, wherein the analyte is selected from (1) amyloid protein, plaque or both of a protein or peptide, and (2) RNA, nucleolus or both.

[0447] 14. A method for detecting an analyte in a sample, comprising:

[0448] (a) Combine any one of the compounds described in paragraphs 1-13 with the sample.

[0449] (b) Detecting changes in the photophysical properties of metal complexes.

[0450] The detection of changes in the photophysical properties of the metal complex indicates the presence of aggregation and supramolecular self-assembly of the metal complex, which in turn indicates the presence of the analyte in the sample.

[0451] 15. The method described in paragraph 14, wherein the analyte is selected from (1) amyloid protein, plaque, or both of a protein or peptide, and (2) RNA, nucleolus, or both.

[0452] 16. The method described in paragraph 14 or paragraph 15, wherein the sample comprises human or non-human animal bodily fluids, human or non-human animal tissues, or combinations thereof.

[0453] 17. The method described in paragraph 16, wherein the fluid is cerebrospinal fluid.

[0454] 18. The method described in paragraph 16, wherein the tissue is brain tissue.

[0455] 19. The method of any one of paragraphs 14 to 18, wherein the analyte is an amyloid protein, plaque, or both of a protein or peptide, wherein the amyloid protein, plaque, or both of the protein or peptide in the sample comprises linear aggregates of the protein or peptide arranged in a β-sheet conformation.

[0456] 20. A method for testing the efficacy of an inhibitor against amyloidosis and / or fibrillation of proteins or peptides, comprising:

[0457] (a) Combining any one of the compounds described in paragraphs 1-13 with an inhibitor-treated sample containing a protein or peptide, and combining it separately with an untreated sample containing a protein or peptide.

[0458] (b) Compare the photophysical properties of the metal complexes between the two samples.

[0459] The magnitude of the difference in photophysical properties between the two samples indicates the degree of change in the aggregation and supramolecular self-assembly states of the metal complexes, and the degree of change in the aggregation and supramolecular self-assembly states of the metal complexes indicates the efficacy of the inhibitor.

[0460] 21. A method for imaging an analyte in a sample, comprising:

[0461] (a) Combining any one of the compounds described in paragraphs 1-13 with a sample under conditions that allow the metal complexes of the compound to bind to the analyte and subsequently undergo aggregation and supramolecular self-assembly of the metal complexes, wherein the aggregation and supramolecular self-assembly of the metal complexes result in changes in the photophysical properties of the metal complexes.

[0462] (b) Imaging of analytes based on one or more photophysical properties specific to aggregated and supramolecular self-assembled metal complexes.

[0463] 22. The method described in paragraph 21, wherein the analyte is selected from (1) amyloid protein, plaque, or both of a protein or peptide, and (2) RNA, nucleolus, or both.

[0464] 23. The method described in paragraph 21 or paragraph 22, wherein the sample comprises eukaryotic cells, the eukaryotic cells optionally selected from 3T3 cells, A549 cells, Chinese hamster ovary (CHO) cells, HEK 293 cells, HeLa cells, Hep G2 cells, and HT1080 cells.

[0465] 24. A kit for detecting and / or imaging an analyte, comprising in one or more containers one or more of the compounds described in any one of paragraphs 1-13 and optional instructions for use.

[0466] 25. The kit described in paragraph 24, wherein the analyte is selected from (1) amyloid protein, plaque or both of a protein or peptide, and (2) RNA, nucleolus or both.

[0467] 26. The kit described in paragraph 24 or paragraph 25, further comprising a carrier.

[0468] 27. The kit of any one of paragraphs 24 to 26, wherein the presence of the analyte can induce aggregation and supramolecular self-assembly of the metal complex thereon after binding, wherein the aggregation and supramolecular self-assembly of the metal complex can be detected by changes in the photophysical properties of the metal complex.

[0469] V. Example

[0470] Example 1. Synthesis and characterization of complex 1-Pt.

[0471] Materials and methods

[0472] Complex 1-Pt was prepared by stirring a mixture of [Pt{bzimpy(PrSO3)2}Cl]PPN (100 mg, 0.076 mmol), HC≡C-C6H3-(CH2OH)2-3,5 (40 mg, 0.247 mmol), copper iodide (I) (catalytic amount), and triethylamine (1 mL) in degassed methanol (100 mL) for one day under nitrogen atmosphere at 100 °C. After removing the solvent by rotary evaporation, the solid residue was dissolved in methanol. After filtration, the complex was recrystallized from the diethyl ether-methanol solution. The final water-soluble complex was then prepared by a salt metathesis reaction with potassium hexafluorophosphate. The precipitate was separated by centrifugation and washed successively with acetonitrile and dichloromethane. The final product was obtained as an orange solid.

[0473] Using tetramethylsilane as an internal standard, proton nuclear magnetic resonance (NMR) was recorded on a Bruker AVANCE 400 Fourier transform NMR spectrometer (400 MHz). 1 ¹H NMR spectroscopy. Infrared (IR) spectroscopy was performed using a Shimadzu IRAffinity-1 Fourier transform infrared spectrophotometer (7800–350 cm⁻¹). -1 The results were obtained on a KBr disk. Fast atom bombardment (FAB) mass spectra were recorded on a Thermo Fisher Scientific DFS high-resolution magnetic sector mass spectrometer. Elemental analysis was performed on a Thermo Fisher Scientific Flash EA 1112 elemental analyzer at the Institute of Chemistry, Chinese Academy of Sciences.

[0474] result

[0475] The chemical characterization data of the complex 1-Pt are as follows.

[0476] Yield: 40 mg (56%). 1¹H NMR (400MHz, [D6]DMSO, 298K, δ / ppm): δ 2.17 (m, 4H, -CH2-), 2.70 (t, J = 6.3Hz, 4H, -CH2SO3), 4.60 (d, J = 5.7Hz, 4H, -CH2O), 4.88 (m, 4H, -CH2N-), 5.42 (t, J = 6.0Hz, 2H, -OH), 7.18 (s, 1H, phenyl), 7.23 (s, 2H, phenyl), 7.51 (m, 4H, benzimidazolyl), 7.83 (m, 2H, benzimidazolyl), 8.15 (t, J = 8.2Hz, 1H, pyridyl), 8.39 (m, 2H, benzimidazolyl), 8.69ppm (d, J = 8.2Hz, 2H, pyridyl). IR(KBr):ν=2120cm -1 (w;ν(C≡C)). Negative FAB-MS:m / z 910[MK] - C 35 H 32 Elemental analysis of KN5O8PtS2·2CH2Cl2: Calculated values ​​(%): C, 39.72; H, 3.24; N, 6.26; Measured values: C, 39.52; H, 3.16; N, 6.17.

[0477] The analysis results confirmed the high purity of the complex 1-Pt.

[0478] Example 2. Photophysical properties of complex 1-Pt.

[0479] Materials and methods

[0480] The photophysical properties of the complex 1-Pt were measured at a concentration of 30 μM. As described in (1) Van Houten et al., J. Am. Chem. Soc., 98:4853-4858 (1976), (2) Caspar et al., J. Am. Chem. Soc., 105:5583-5590 (1983), and (3) Wallace et al., Inorg. Chem., 32:3836-3843 (1993), the luminescence quantum yield in degassed DMF and aqueous solution was measured using the optical dilution method reported in Crosby et al., J. Phys. Chem., 75:991-1024 (1971), with degassed acetonitrile and aqueous solution of [Ru(bpy)3]Cl2 as references. The photoexcitation wavelength was 436 nm.

[0481] result

[0482] The UV-Vis absorption spectra of the complex 1-Pt in both DMF and aqueous solution at 298 K showed an absorption tail at 470-480 nm, which is attributed to metal-to-ligand charge transfer (MLCT) [dπ(Pt)→π*(bzimpy)] transitions, with some ligand-to-ligand charge transfer (LLCT) [π(C≡C)→π*(bzimpy)] characteristics. Figure 1 The 1-Pt complex in degassed DMF solution at 298 K exhibits an emission band at 566 nm, representing the electronic vibrational structure. Figure 2 The emission band originates from the triplet ligand ( 3 The IL)[π→π*(bzimpy)] excited state. The complex 1-Pt in degassed aqueous solution at 298 K exhibits a Gaussian emission band at 673 nm, attributed to charge transfer originating from the triplet metal-metal to ligand state. 3 MMLCT) excited state.

[0483] Example 3. Insulin amyloid protein can induce the aggregation and supramolecular self-assembly of complex 1-Pt in aqueous solution.

[0484] Materials and methods

[0485] To induce amyloid fibrillation, insulin was administered at a dose of 1.0 mg / mL. -1 The concentration of [1-Pt] was dissolved in acidic buffer ([NaCl] = 137 mM, [KCl] = 2.7 mM, pH = 2.0). The solution was incubated at 65 °C and stirred at 300 rpm for 120 min to form insulin amyloid. Different amounts of insulin amyloid or native insulin (0-10 μM) were added to a solution of complex 1-Pt (50 μM) in PBS buffer (10.0 mM, pH = 7.4, 10% DMSO). The UV-Vis absorption, emission, and RLS spectra of different amounts of insulin amyloid or native insulin were recorded at 25 °C. The emission spectra were recorded at an excitation wavelength of 400 nm.

[0486] result

[0487] Figure 3A The UV-Vis absorption spectrum of complex 1-Pt (50 μM) is shown, along with the corresponding absorbance changes when mixed with increased amounts of insulin-like amyloid (0–10 μM). Adding insulin-like amyloid to complex 1-Pt results in an increase in absorbance at a low-energy absorption tail at approximately 550 nm. Figure 3B This is due to the aggregation of metal complexes and supramolecular self-assembly.

[0488] Figure 4AThe corrected emission spectrum of complex 1-Pt (50 μM) is shown, along with the corresponding changes in emission intensity when mixed with increased amounts of insulin-like amyloid (0–10 μM). Addition of insulin-like amyloid to complex 1-Pt resulted in luminescence activation at 650 nm. Figure 4B This is due to the aggregation of metal complexes and supramolecular self-assembly.

[0489] Figure 5A The RLS spectrum of complex 1-Pt (50 μM) is shown, along with the corresponding changes in RLS intensity when mixed with increased amounts of insulin-like amyloid (0–10 μM). Addition of insulin-like amyloid to complex 1-Pt resulted in a significant enhancement of RLS intensity at approximately 550 nm. Figure 5B This is due to the aggregation of metal complexes and supramolecular self-assembly.

[0490] When mixed with natural insulin, the UV-Vis absorption spectrum of the complex 1-Pt is ( Figure 6A and 6B ), emission spectrum ( Figure 7A and 7B ) and RLS spectrum ( Figure 8A and 8B No significant spectral changes were observed. These results indicate that insulin amyloid can induce the aggregation and supramolecular self-assembly of its complex 1-Pt in buffered aqueous solution, while native insulin cannot.

[0491] Example 4. Complex 1-Pt can be used to study the kinetics of insulin amyloid fibrillation.

[0492] Materials and methods

[0493] To induce amyloid fibrillation, insulin was administered at a dose of 1.0 mg / mL. -1 The insulin was dissolved in acidic buffer ([NaCl] = 137 mM, [KCl] = 2.7 mM, pH = 2.0). The solution was incubated at 65 °C with stirring at 300 rpm. Insulin samples with different incubation times were added to a solution of thioflavone T in PBS buffer (10.0 mM, pH = 7.4). The final concentrations of insulin and thioflavone T were both 10 μM. Emission spectra were recorded at an excitation wavelength of 440 nm at 25 °C. Similarly, insulin samples with different incubation times were added to a solution of complex 1-Pt in PBS buffer (10.0 mM, pH = 7.4, 10% DMSO). The final concentrations of insulin and complex 1-Pt were 10 μM and 50 μM, respectively. UV-Vis absorption, emission, and RLS spectra were recorded at 25 °C; emission spectra were recorded at an excitation wavelength of 400 nm.

[0494] Kinetically, the formation of insulin-like amyloid fibrils can be modeled using a sigmoid function, which has three distinct phases: a hysteresis phase, a logarithmic phase, and a stationary phase. The sigmoid function is used to determine the k-axis of insulin-like amyloid fibril formation. app and t 迟滞 The value is based on the equation shown below (see similar examples of data fitting in Nielsen et al., Biochemistry, 40:6036-6046 (2001); Hwang et al., J. Biol. Chem., 285:41701-41711 (2010); and Donabedian et al., ACS Chem. Neurosci., 6:1526-1535 (2015)).

[0495]

[0496] Where y is absorbance, emission intensity, or RLS intensity; A1 is absorbance, emission intensity, or RLS intensity before amyloid fibrils form; A2 is absorbance, emission intensity, or RLS intensity after amyloid fibrils form; x is incubation time; x o It is the incubation time when absorbance, emission intensity, or RLS intensity is at half its maximum value; d x It is a time constant. Therefore, the apparent rate constant k app equals 1 / d x hysteresis time t 迟滞 By x o -2d x Provided.

[0497] result

[0498] Figure 9A The corrected emission spectrum of thioflavone T (10 μM) is shown, along with the corresponding changes in emission intensity when mixed with insulin samples (10 μM) incubated for different times. Amyloid fibril formation occurs via a nucleation-dependent mechanism, as evidenced by the sigmoid curve. Figure 9B ).

[0499] Figure 10A , 11A Figures 1 and 12A show the UV-Vis absorption, emission, and RLS spectra of the complex 1-Pt (50 μM), respectively, and the corresponding spectral changes when mixed with insulin samples (10 μM) incubated for different times. All obtained data can be fitted using a sigmoid function. Figure 10B , 11B And 12B). For example Figure 13As shown, the kinetics of insulin amyloid fibrillation can be divided into three phases: a hysteresis phase, a logarithmic phase, and a stationary phase. During the formation of insulin amyloid fibrillation, aggregation induced by the complex 1-Pt and supramolecular self-assembly occur, leading to significant changes in its photophysical properties.

[0500] Table 1 summarizes the calculated apparent rate constants and hysteresis times for insulin-induced amyloid fibrillation. As shown in the figure, regardless of the spectroscopic method used for detection, the kinetic parameters reported by complex 1-Pt are comparable to those reported by thioflavin T.

[0501] Table 1. Apparent rate constants (kJ / kb) of insulin-amyloid fibrillation reported by thioflavin T and its complex 1-Pt app ) and lag time (t) 迟滞 ).

[0502]

[0503] Example 5. Complex 1-Pt exhibits high binding affinity for amyloid protein and plaque.

[0504] Materials and methods

[0505] Different amounts of the complex 1-Pt (0-50 μM) were added to insulin amyloid protein (10 μM) in a solution of PBS buffer (10.0 mM, pH 7.4, 10% DMSO). Emission spectra were recorded at an excitation wavelength of 400 nm at 25 °C. The experimental data were fitted using the Hill equation shown below (see similar examples of data fitting in Donabedian et al., ACS Chem. Neurosci., 6:1526-1535 (2015); Goutelle et al., Fundam. Clin. Pharmacol., 22:633-648 (2008); and Gesztelyi et al., Arch. Hist. Exact Sci., 66:427-438 (2012)).

[0506]

[0507] Where y is the relative emission intensity; x is the concentration of complex 1-Pt; n is the Hill coefficient, which describes the synergistic binding with insulin-like amyloid protein; K d It is the apparent dissociation constant. The apparent binding constant K a It is K d The reciprocal of.

[0508] result

[0509] Figure 14AThe corrected emission spectra of the complex 1-Pt at different concentrations are shown after adding the same amount of insulin amyloid. The obtained binding curves were fitted to the Hill equation ( Figure 14B The apparent binding constant between complex 1-Pt and insulin-like amyloid protein was found to be 5.46 × 10⁻⁶. 4 M -1 This is on the same order of magnitude as the apparent binding constant between thioflavone T and insulin amyloid protein measured under similar assay conditions.

[0510] Example 6. Complex 1-Pt can become strongly luminescent after binding to insulin amyloid protein.

[0511] Materials and methods

[0512] Insulin amyloid (10 μM) was added to a solution of complex 1-Pt (50 μM) in PBS buffer (10.0 mM, pH 7.4, 10% DMSO). Samples for confocal microscopy were prepared by placing aliquots of the mixture onto a microscope slide and then placing a coverslip on top. Confocal microscopy experiments were performed on a Carl Zeiss LSM700 confocal scanning microscope. Confocal images were captured using a solid-state laser with an excitation wavelength of 555 nm under a 20x objective lens, and the emission was collected in the 600–700 nm range.

[0513] result

[0514] A comparison of images obtained from laser excitation and bright-field luminescence showed that luminescence of complex 1-Pt was observed only in regions where amyloid fibrils were present (Fig. 15). Therefore, it can be concluded that complex 1-Pt aggregates upon binding to insulin amyloid, resulting in strong luminescence of the metal complex.

[0515] Example 7. Complex 1-Pt can be used to screen inhibitors targeting protein aggregation.

[0516] Materials and methods

[0517] In the presence of different amounts (0, 10, 20, 50, 70, 100 mM) of L-ascorbic acid, insulin was administered at a dose of 1.0 mg / mL. -1Dissolved in acidic buffer ([NaCl] = 137 mM, [KCl] = 2.7 mM, pH = 2.0). The solution was incubated at 65 °C with stirring at 300 rpm. Aliquots were removed at desired time intervals. Insulin samples (10 μM) incubated for different times were added to a solution of thioflavone T (10 μM) in PBS buffer (10.0 mM, pH = 7.4). Emission spectra were recorded at an excitation wavelength of 440 nm at 25 °C. Similarly, insulin samples (10 μM) incubated for different times were added to a solution of complex 1-Pt (50 μM) in PBS buffer (10.0 mM, pH = 7.4, 10% DMSO). Emission spectra were recorded at an excitation wavelength of 400 nm at 25 °C. The k-value for insulin amyloid fibrillation was determined by fitting using the sigmoid equation listed above. app and t 迟滞 value.

[0518] result

[0519] The effect of L-ascorbic acid on the formation of insulin-like amyloid fibrils was examined by thioflavone T fluorescence assay and complex 1-Pt luminescence assay. Figure 16 The results show the variation in the relative emission intensity of thioflavone T at 490 nm under different concentrations of L-ascorbic acid and different incubation times. Figure 17 The relative emission intensity of complex 1-Pt at 650 nm is shown to vary with different concentrations of L-ascorbic acid and different incubation times. Table 2 summarizes the calculated apparent rate constants and hysteresis times for insulin-like amyloid fibrillation in the presence of different concentrations of L-ascorbic acid. Clearly, the two assays yielded very similar results. (See Table 2 and...) Figure 16 and 17 As shown in the curve, the inhibitory effect of L-ascorbic acid on the formation of amyloid fibrils is concentration-dependent.

[0520] Table 2. Apparent rate constants (kJ / kJ) of insulin-like amyloid fibrillation in the presence of different concentrations of L-ascorbic acid, as determined by thioflavin T and complex 1-Pt. app ) and lag time (t) 迟滞 ).

[0521]

[0522] Example 8. The addition of metal ions does not interfere with the properties of the complex 1-Pt.

[0523] Materials and methods

[0524] Different metal ions (100 μM), including Mg 2+Ca 2+ Mn 2+ Fe 2+ Fe 3+ Cu 2+ and Zn 2+ The complex 1-Pt (50 μM) was mixed separately with PBS buffer (10.0 mM, pH 7.4, 10% DMSO). Insulin amyloid protein (10 μM) was added to each mixture. Emission spectra were recorded at an excitation wavelength of 400 nm at 25 °C.

[0525] result

[0526] The relative emission intensities of the complex 1-Pt were measured in the presence of insulin-like amyloid protein and different metal ions. It was found that the relative emission intensities remained almost constant. Figure 18 This result indicates that the amyloid detection performance of complex 1-Pt is not affected by the presence of these metal ions, which is based on the d 8 / d 10 One of the most important advantages of metal complex probes over conventionally used probes such as thioflavin T is that they are metal complex probes.

[0527] Example 9. The addition of biomolecules does not interfere with the performance of the complex 1-Pt.

[0528] Materials and methods

[0529] Insulin amyloid protein (10 μM) and / or other biomolecules, including α-amylase (10 μM), albumin from bovine serum (10 μM), albumin from human serum (10 μM), alkaline phosphatase (10 μM), trypsin (10 μM), and DNA (10 μg / mL). -1 ) and RNA (10 μg / mL) -1 The complex 1-Pt (50 μM) was added to a solution of PBS buffer (10.0 mM, pH 7.4, 10% DMSO). The emission spectrum was recorded at an excitation wavelength of 400 nm at 25 °C.

[0530] result

[0531] It was found that emission intensity increased only after the addition of insulin amyloid protein, while emission was not activated after the introduction of other biomolecules. Figure 19A The change in emission intensity produced by simultaneously adding insulin amyloid and a mixture of various interfering biomolecules is similar to the change in emission intensity produced by adding insulin amyloid alone. Figure 19B These results demonstrate that this assay is a highly selective and specific sensing platform.

[0532] Example 10. The complex 1-Pt exhibits low cytotoxicity.

[0533] Materials and methods

[0534] HeLa cells were attached to 96-well plates at approximately 10,000 cells per well and cultured at 37°C in a humidified incubator in Dulbecco modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS) (100 μL) for 24 hours, with the carbon dioxide level maintained at a constant 5%. Different amounts of the complex 1-Pt (0, 6.25, 12.5, 25, 50, 100 μM) were added to the DMEM, and the cells were incubated at 37°C for 24 hours. Similarly, CHO cells were attached to 96-well plates at approximately 10,000 cells per well and cultured at 37°C in a humidified incubator in Ham's F-12 nutrient mixture supplemented with 10% FBS (100 μL) for 24 hours, with the carbon dioxide level maintained at a constant 5%. Different amounts of complex 1-Pt (0, 6.25, 12.5, 25, 50, 100 μM) were added to Ham's F-12 nutrient mixture, and cells were incubated at 37°C for 24 hours. Wells containing cells without complex 1-Pt served as controls. Subsequently, 10 μL of 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazole bromide (MTT) in PBS buffer (5 mg / mL) was added to each well. -1 The solution was prepared and the plate was incubated at 37°C for 3 hours. The solution was removed, and the precipitated formazan was dissolved in DMSO (200 μL). After dissolution, the absorbance of the formazan at 570 nm was measured using a microplate absorbance reader. Cell viability was expressed as the percentage of absorbance of cells treated with complex 1-Pt to that of the control.

[0535] result

[0536] The results showed that after incubation with the complex 1-Pt at a concentration as high as 50 μM, HeLa cells maintained more than 98% cell viability. Figure 20A When the concentration of the complex 1-Pt was increased to 100 μM, cell viability was found to decrease slightly, but remained above 94%.

[0537] On the other hand, the results showed that after incubation with the complex 1-Pt at a concentration as high as 100 μM, CHO cells maintained more than 94% cell viability. Figure 20B These results demonstrate the low cytotoxicity of the complex 1-Pt.

[0538] Example 11. Synthesis and characterization of complex 2-Pt.

[0539] Materials and methods

[0540] Complex 2-Pt was prepared by stirring a mixture of [Pt{bzimpy(C4H9)2}Cl]Cl (100 mg, 0.145 mmol), [HC≡C-C6H4-{NHC(NH2)(=NH2)}-4]Cl (100 mg, 0.435 mmol), copper iodide (I) (catalytic amount), and triethylamine (1 mL) in degassed methanol (100 mL) for one day under nitrogen atmosphere at 100 °C. After removing the solvent by rotary evaporation, the solid residue was dissolved in methanol. After filtration, the complex was recrystallized from the diethyl ether-methanol solution. The precipitate was washed successively with acetonitrile and dichloromethane. The final product was obtained as an orange solid.

[0541] Using tetramethylsilane as an internal standard, proton nuclear magnetic resonance (NMR) was recorded on a Bruker AVANCE 400 Fourier transform NMR spectrometer (400 MHz). 1 ¹H NMR spectroscopy. Infrared (IR) spectroscopy was performed using a Shimadzu IRAffinity-1 Fourier transform infrared spectrophotometer (7800–350 cm⁻¹). -1 The samples were obtained on a KBr disk. Fast atom bombardment (FAB) mass spectra were recorded on a Thermo Fisher Scientific DFS high-resolution magnetic sector mass spectrometer. Elemental analysis was performed on a Thermo Fisher Scientific Flash EA 1112 elemental analyzer at the Institute of Chemistry, Chinese Academy of Sciences.

[0542] result

[0543] The chemical characterization data of the complex 2-Pt are as follows.

[0544] Yield: 70 mg (57%). 1 ¹H NMR (400MHz, [D6]DMSO, 298K, δ / ppm): δ 0.92 (t, J = 7.3Hz, 6H, -CH3), 1.44 (m, 4H, -CH2-), 1.91 (m, 4H, -CH2-), 4.92 (t, J = 7.0Hz, 4H, -CH2N-), 7.31 (d, J = 8.5Hz, 2H, phenyl), 7.44 (s, 4H, -NH2), 7.58 (d, J = 8.5Hz, 2H, phenyl), 7.65 (m, 4H, benzimidazolyl), 8.07 (d, J = 8.1Hz, 2H, benzimidazolyl), 8.53 (d, J = 8.1Hz, 2H, benzimidazolyl), 8.62 (m, 3H, pyridyl), 9.76ppm (s, 1H, -NH-). IR(KBr):ν=2110cm -1(w;ν(C≡C)). Positive FAB-MS: m / z 389[M-2Cl] 2+ C 36 H 38 Elemental analysis of Cl2N8Pt·CH2Cl2: Calculated values ​​(%): C, 47.60; H, 4.32; N, 12.00; Measured values: C, 47.61; H, 4.63; N, 12.06.

[0545] The analysis results confirmed the high purity of the complex 2-Pt.

[0546] Example 12. Photophysical properties of complex 2-Pt.

[0547] Materials and methods

[0548] The photophysical properties of the complex 2-Pt were measured at a concentration of 30 μM. As described in (1) Van Houten et al., J. Am. Chem. Soc., 98:4853-4858 (1976), (2) Caspar et al., J. Am. Chem. Soc., 105:5583-5590 (1983), and (3) Wallace et al., Inorg. Chem., 32:3836-3843 (1993), the luminescence quantum yield in degassed methanol and aqueous solution at 298 K was measured using the optical dilution method reported in Crosby et al., J. Phys. Chem., 75:991-1024 (1971), with degassed acetonitrile and aqueous solution of [Ru(bpy)3]Cl2 as references. The photoexcitation wavelength was 436 nm.

[0549] result

[0550] The UV-Vis absorption spectra of the 2-Pt complex at 298 K in both methanol and aqueous solution show an absorption tail at approximately 450–470 nm, attributed to metal-to-ligand charge transfer (MLCT) transitions [dπ(Pt)→π*(bzimpy)], with some ligand-to-ligand charge transfer (LLCT) characteristics [π(C≡C)→π*(bzimpy)]. Figure 21 The 2-Pt complex in degassed methanol solution at 298 K exhibits an emission band at 564 nm, representing the electronic vibrational structure. This emission band originates from the triplet ligand ( 3 The IL)[π→π*(bzimpy)] excited state. The complex 2-Pt in degassed aqueous solution at 298 K exhibits a Gaussian emission band at 683 nm, attributed to charge transfer originating from the triplet metal-metal to ligand state. 3 MMLCT) excited state ( Figure 22 ).

[0551] Example 13. RNA can induce the aggregation and supramolecular self-assembly of complex 2-Pt in aqueous solution.

[0552] Materials and methods

[0553] Different amounts of RNA (0-10 μg / mL) were used. -1 The complex 2-Pt (20 μM) was added to a solution of PBS buffer (10 mM, pH 7.4). UV-Vis absorption, emission, RLS, and zeta potential data of the samples were recorded at 37 °C. The emission spectrum was recorded at an excitation wavelength of 360 nm.

[0554] result

[0555] Figure 23A The UV-Vis absorption spectrum of the complex 2-Pt (20 μM) is shown, as well as the absorption spectrum when combined with an increased amount of RNA (0-10 μg / mL). -1 The corresponding absorbance changes upon mixing. Adding RNA to the 2-Pt complex resulted in an increase in absorbance at the low-energy absorption tail at approximately 550 nm. Figure 23B This is due to the aggregation of metal complexes and supramolecular self-assembly.

[0556] Figure 24A The corrected emission spectrum of the complex 2-Pt (20 μM) is shown, as well as that when combined with an increased amount of RNA (0-10 μg / mL). -1 The corresponding emission intensity changes upon mixing. Adding RNA to the 2-Pt complex resulted in luminescence activation at 670 nm. Figure 24B This is due to the aggregation of metal complexes and supramolecular self-assembly.

[0557] Figure 25A The RLS spectrum of the complex 2-Pt (20 μM) is shown, as well as the spectrum when combined with increased amounts of RNA (0-10 μg / mL). -1 The corresponding changes in RLS intensity upon mixing. Adding RNA to the 2-Pt complex resulted in a significant increase in RLS intensity at approximately 550 nm. Figure 25B This is due to the aggregation of metal complexes and supramolecular self-assembly.

[0558] Figure 26 This shows the addition of different amounts of RNA (0-10 μg / mL) to PBS buffer. -1 The zeta potential data for the complex 2-Pt (20 μM) are shown. Adding RNA to the complex 2-Pt results in a more negative zeta potential, which is due to the binding of the metal complex to the RNA.

[0559] Example 14. Complex 2-Pt has a high binding affinity for RNA.

[0560] Materials and methods

[0561] Different amounts of the complex 2-Pt (0-20 μM) were added to RNA (10 μg / mL). -1 The sample was prepared in PBS buffer (10.0 mM, pH 7.4). Emission spectra were recorded at an excitation wavelength of 360 nm at 37 °C. The experimental data were fitted using the Hill equation shown below.

[0562]

[0563] Where y is the relative emission intensity; x is the concentration of the complex 2-Pt; n is the Hill coefficient, which describes the cooperativity in binding with RNA; K d It is the apparent dissociation constant. The apparent binding constant K a It is K d The reciprocal of.

[0564] result

[0565] Figure 27A The corrected emission spectra of the complex 2-Pt at different concentrations after adding the same amount of RNA are shown. The obtained binding curves were fitted to the Hill equation ( Figure 27B The apparent binding constant between the complex 2-Pt and insulin-like amyloid protein was found to be 6.01 × 10⁻⁶. 4 M -1 .

[0566] Example 15. The complex 2-Pt can become strongly luminescent after binding to RNA and nucleolus.

[0567] Materials and methods

[0568] HeLa cells were cultured in a humidified incubator at 37°C with DMEM supplemented with 10% FBS, while maintaining a constant CO2 level of 5%. Similarly, CHO cells were cultured in a humidified incubator at 37°C with Ham's F-12 nutrient mixture supplemented with 10% FBS, while maintaining a constant CO2 level of 5%. Cells were then attached to sterile coverslips in 35 mm cell culture dishes and cultured for 48 hours. The culture medium was removed, and the cells were fixed in pre-chilled methanol at -20°C for 10 minutes. After washing the cells three times in PBS buffer (1 mL), a solution of complex 2-Pt (20 μM) in PBS buffer was applied, and the cells were incubated at 37°C for 1 hour. After staining, the labeling solution was removed, and the cells were washed three times in PBS buffer (1 mL). Coverslips were placed on sterile microscope slides. Confocal microscopy experiments were performed on a Leica TCSSPE confocal scanning microscope. Confocal images were captured using a solid-state laser with an excitation wavelength of 488 nm under a 63x objective lens, and the emission was collected in the 620-720 nm range.

[0569] result

[0570] Imaging results showed that bright red luminescent spots were clearly visible in the nuclei of HeLa cells stained with complex 2-Pt. Figure 28A -C).

[0571] On the other hand, bright red luminescent spots were also clearly visible in the nuclei of CHO cells stained with complex 2-Pt. Figure 29A -C).

[0572] Figure 30 Showing the use of d 8 or d 10 A schematic diagram illustrating the design principle of luminescence activation assays using metal complexes for RNA detection and nucleolar imaging. The aggregation and supramolecular self-assembly of metal complexes on RNA induce luminescence activation.

[0573] Figure 31A The image shows a luminescent confocal image of fixed HeLa cells stained with complex 2-Pt (20 μM) at 37 °C for 1 hour. Figure 31B Shown from Figure 31A The overall relative emission intensity distribution of fixed HeLa cells was observed. The luminescence of the 2-Pt complex was found to be primarily located in the nucleolus (corresponding to...). Figure 31B The emission peaks between approximately 12 and 25 μm indicate that selective nucleolar imaging in HeLa cells is achieved through the complex 2-Pt.

[0574] Figure 32AThe image shows a luminescent confocal image of fixed CHO cells stained with complex 2-Pt (20 μM) at 37°C for 1 hour. Figure 32B Shown from Figure 32A The overall relative emission intensity distribution of fixed CHO cells was observed. The luminescence of the 2-Pt complex was found to be primarily located in the nucleolus (corresponding to...). Figure 32B The emission peaks between approximately 7 and 12 μm indicate that selective nucleolar imaging in CHO cells is achieved through the complex 2-Pt.

[0575] In addition to the glowing spots originating from the cell nucleus, smaller glowing spots were also observed in HeLa and CHO cells. These smaller glowing spots represent RNA in the cytoplasm.

[0576] Example 16. Complex 2-Pt can be used to detect RNA degradation catalyzed by RNase.

[0577] Materials and methods

[0578] After staining fixed HeLa cells and / or fixed CHO cells with 2-Pt complex (20 μM) at 37°C for 1 hour, RNase and / or DNase (30 μg / mL) were applied. -1 Cells were incubated in a PBS buffer solution at 37°C for 2 hours. After RNase and / or DNase digestion, the enzyme solution was removed, and the cells were washed three times in PBS buffer (1 mL). A coverslip was placed on a sterile microscope slide. Confocal images were captured using a solid-state laser with an excitation wavelength of 488 nm under a 63x objective lens, and the emission was collected in the 620–720 nm range.

[0579] result

[0580] Confocal images of HeLa cells show that the red luminescence signal in the nucleolus was almost completely lost after RNase treatment. Figures 33A-33C Conversely, treatment with DNase did not induce a significant loss of the red luminescence signal from the nucleolus. Figure 33D-33F Simultaneously using RNase and DNase (both 30 μg / mL) -1 The processing also resulted in the almost complete loss of the red luminescence signal from the nucleolus. Figure 33G-33I ).

[0581] On the other hand, confocal images of CHO cells showed that after treatment with RNase, the red luminescence signal in the nucleolus was almost completely lost. Figures 34A-34C Conversely, treatment with DNase did not induce a significant loss of the red luminescence signal from the nucleolus. Figure 34D-34F Simultaneously using RNase and DNase (both 30 μg / mL) -1The processing also resulted in the almost complete loss of the red luminescence signal from the nucleolus. Figure 34G-34I These results indicate that the luminescence activation assay using complex 2-Pt is specific for RNA detection but not for DNA detection.

[0582] Example 17. Complex 2-Pt can be used to selectively stain RNA and nucleoli.

[0583] Materials and methods

[0584] After staining fixed HeLa cells and / or fixed CHO cells with complex 2-Pt (20 μM) at 37°C for 1 hour, SYTO was applied. TM RNASelect TM A solution of green fluorescent cell staining agent (500 nM) in PBS buffer was prepared, and the cells were incubated at 37°C for 20 minutes. After staining, the labeling solution was removed, and the cells were washed three times in PBS buffer (1 mL). A coverslip was placed on a sterile microscope slide. Confocal images were captured under a 63x objective lens using a solid-state laser with an excitation wavelength of 488 nm. For the complex 2-Pt, emission was collected in the 620-720 nm range, while for SYTO... TM RNASelect TM Green fluorescent cell staining agent is collected and emitted at 505-555 nm.

[0585] result

[0586] Fixed HeLa cells and / or fixed CHO cells were treated with complex 2-Pt and SYTO. TM RNASelect TM Co-staining with a green fluorescent cell staining agent, which is a commercially available probe targeting the nucleolar.

[0587] Confocal images of HeLa cells (Fig. 35) and CHO cells (Fig. 36) show red fluorescence from the complex 2-Pt and SYTO from the nucleolus. TM RNASelect TM The green emission of the green fluorescent cell staining agent showed good colocalization.

[0588] Example 18. The complex 2-Pt exhibits low cytotoxicity.

[0589] Materials and methods

[0590] HeLa cells were attached to 96-well plates at approximately 10,000 cells per well and cultured at 37°C in a humidified incubator with DMEM supplemented with 10% FBS (100 μL) for 24 hours, while maintaining a constant CO2 level of 5%. Different amounts of the complex 2-Pt (0, 1.25, 2.5, 5, 10, 20 μM) were added to the DMEM, and the cells were incubated at 37°C for 24 hours. Similarly, CHO cells were attached to 96-well plates at approximately 10,000 cells per well and cultured at 37°C in a humidified incubator with Ham's F-12 nutrient mixture supplemented with 10% FBS (100 μL) for 24 hours, while maintaining a constant CO2 level of 5%. Different amounts of complex 2-Pt (0, 1.25, 2.5, 5, 10, 20 μM) were added to Ham's F-12 nutrient mixture, and cells were incubated at 37°C for 24 hours. Wells containing cells without complex 2-Pt served as controls. Subsequently, 10 μL of MTT in PBS buffer (5 mg / mL) was added to each well. -1 The solution was prepared and the plate was incubated at 37°C for 3 hours. The solution was removed, and the precipitated formazan was dissolved in DMSO (200 μL). After dissolution, the absorbance of formazan at 570 nm was measured using a microplate absorbance reader. Cell viability is expressed as the percentage of absorbance of cells treated with complex 2-Pt to that of the control.

[0591] result

[0592] The results showed that after incubation with the complex 2-Pt at a concentration as high as 10 μM, HeLa cells maintained more than 96% cell viability. Figure 37A When the concentration of the 2-Pt complex was increased to 20 μM, cell viability decreased slightly, but remained above 81%.

[0593] On the other hand, the results showed that after incubation with the complex 2-Pt at a concentration as high as 20 μM, CHO cells maintained more than 94% cell viability. Figure 37B These results demonstrate the low cytotoxicity of the complex 2-Pt.

[0594] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed invention pertains. All publicly available documents cited herein and materials that cite them are expressly incorporated by reference.

[0595] Those skilled in the art will recognize that many equivalents of the specific embodiments of the invention described herein can be determined using only conventional experiments. Such equivalents are intended to be covered by the following claims.

Claims

1. A compound for detecting and / or imaging an analyte, said analyte being selected from (1) amyloid protein, plaque, or both proteins or peptides, and (2) RNA, wherein said compound is d 8 or d 10 Metal complexes or salts thereof, comprising: (a) Metal atoms with coordination numbers of 2, 3, or 4, selected from Pt(II), Pd(II), Ni(II), Ir(I), Rh(I), Au(III), Ag(III), Cu(III), Ni(0), Pd(0), Pt(0), Cu(I), Ag(I), Au(I), Zn(II), Cd(II), and Hg(II); and (b) One or more ligands having donor atoms, the donor atoms being independently selected from carbon (C), nitrogen (N), oxygen (O), phosphorus (P), sulfur (S), arsenic (As) and selenium (Se). The metal complex is bound to the analyte, and the binding of the metal complex to the analyte induces the aggregation and supramolecular self-assembly of the metal complex through non-covalent metal-metal interactions. in, The compound has the structure of Formula I: in (a) M represents a metal atom selected from Pt(II), Pd(II), Ni(II), Ir(I), Rh(I), Au(III), Ag(III), and Cu(III). (b) L1, L2, L3, and L4 represent ligands, where each ligand provides a donor atom to coordinate to a metal atom. (c) The expression represents the number of positive or negative charges carried by the metallic complex, where n is zero or a positive integer. (d) This refers to counterions that maintain a neutral charge, among which It has a charge opposite to that of the metal complex, and where m is zero or a positive integer, m = n or m ≠ n. (e) The expression represents the stoichiometry of the counter ions. (f) Dashed lines represent covalent bonds between any two ligands, fusion of any ring portions from the two ligands, or combinations thereof; L4 includes optionally substituted C6-C 50 Aromatics or C3-C 50 The L4 group further includes groups selected from the following: halogen atoms, alkyl groups, heteroalkyl groups, alkenyl groups, heteroalkenyl groups, alkynyl groups, heteroalkynyl groups, aryl groups, heteroaryl groups, etc. Wherein, the optional substituted C6-C 50 Aromatics or C3-C 50 Substituents in heteroaromatic hydrocarbons include: halogen atoms, alkyl groups, heteroalkyl groups, alkenyl groups, heteroalkenyl groups, alkynyl groups, heteroalkynyl groups, aryl groups, and heteroaryl groups. Among them, R G1’ R G2’ R G3’ Independently constitutes a hydrogen atom, halogen atom, alkyl group, heteroalkyl group, alkenyl group, heteroalkenyl group, ynyl group, heteroynyl group, aryl group, or heteroaryl group. Wherein, the alkyl group is C1-C 30 Alkyl groups, wherein the heteroalkyl groups are C1-C 30 Heteroalkyl groups, wherein the alkenyl groups are C2-C 30 alkenyl, wherein the heteroalkenyl group is C2-C 30 Heteroalkenyl, wherein the alkynyl group is C2-C 30 The alkynyl group, wherein the heteroyynyl group is C2-C 30 The aryl group is C6-C. 50 Aryl, wherein the heteroaryl group is C3-C 50 Mixed aromatics; Alternatively, the compound is selected from one of the following complexes: Complex 3 in is the number of positive or negative charges carried by the metallic complex in the formula, where n is zero or a positive integer. in It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m=n or m≠n. in This is the stoichiometry of the counter ions in the formula; Complex 5 Where n- is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer. Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m=n or m≠n. in This is the stoichiometry of the counter ions in the formula; Complex 6 Where n- is the number of negative charges carried by the metal complex in the formula, and n is a positive integer. Where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m=n or m≠n. in This is the stoichiometry of the counter ions in the formula; Complex 7 Where n- is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer. Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m=n or m≠n. in This is the stoichiometry of the counter ions in the formula; Complex 8 Where n- is the number of negative charges carried by the metal complex in the formula, and n is a positive integer. Where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m=n or m≠n. in This is the stoichiometry of the counter ions in the formula; Complex 9 Where n- is the number of negative charges carried by the metal complex in the formula, and n is a positive integer. Where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m=n or m≠n. in This is the stoichiometry of the counter ions in the formula; Complex 10 Where n- is the number of negative charges carried by the metal complex in the formula, and n is a positive integer. Where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m=n or m≠n. in This is the stoichiometry of the counter ions in the formula; Complex 11 Where n- is the number of negative charges carried by the metallic complex in the formula, and n is zero or a positive integer. Where X m+ It is a counterion that maintains charge neutrality, where m is zero or a positive integer, m=n or m≠n. in This is the stoichiometry of the counter ions in the formula; Complex 12 Where n- is the number of negative charges carried by the metal complex in the formula, and n is a positive integer. Where X m+ It is a counterion that maintains charge neutrality, where m is a positive integer, m=n or m≠n. in It is the stoichiometry of the counter ions in the formula.

2. The compound of claim 1, wherein L1, L2, and L3 are optionally substituted and / or optionally deprotonated C6-C. 50 Aromatics or C3-C 50 Heteroaromatic hydrocarbons, including benzene, pyridine, thiophene, furan, pyrazole, imidazole, oxazole, isoxazole, thiazole, isothiazine, isoquinoline, pyrrole, pyrazine, pyridazine, pyrimidine, benzimidazole, benzofuran, benzothiazole, indole, naphthalene, anthracene, pyrene, triazole, tetrazolium, pyran, thiamphenicol, oxadiazole, triazine, tetrazine, carbazole, dibenzothiophene, dibenzofuran, fluorene and their derivatives.

3. The compound of claim 1 or claim 2, wherein L1 and L2 are connected by covalent bonds, fusion of the cyclic portions of the two ligands, or a combination thereof.

4. The compound of any one of claims 1-3, wherein the metal complex is bound to the analyte via a non-covalent interaction, wherein the non-covalent interaction includes electrostatic interaction, hydrogen bonding interaction, hydrophobic interaction, or a combination thereof.

5. The compound of any one of claims 1-4, wherein the metal complex has a planar structure or a partially planar structure.

6. The compound of any one of claims 1-5, wherein the aggregation and supramolecular self-assembly of the metal complex produce one or more changes in the photophysical properties of the metal complex.

7. The compound of claim 6, wherein the change in the photophysical property includes changes in absorbance, luminescence, resonant light scattering (RLS), or a combination thereof.

8. The compound of claim 7, wherein the change in luminescence includes an increase in luminescence quantum yield and / or emission intensity, and / or a shift in emission energy or wavelength.

9. The compound according to any one of claims 1-8, wherein the compound is: Complex 2 Where n+ is the number of positive charges carried by the metallic complex in the formula, and n is a positive integer. Where X m－ It is a counterion that maintains charge neutrality, where m is a positive integer, m=n or m≠n. in It is the stoichiometry of the counter ions in the formula.

10. A method for detecting an analyte in a sample, comprising: (a) Combining the metal complex with the sample, (b) Detecting changes in the photophysical properties of metal complexes, The detection of changes in the photophysical properties of the metal complex indicates the presence of aggregation and supramolecular self-assembly of the metal complex, which in turn indicates the presence of the analyte in the sample. The analytes are selected from (1) amyloid protein, plaque, or both proteins or peptides, and (2) RNA; The metal complex described herein has the structure of Formula I: in (a) M represents a metal atom selected from Pt(II), Pd(II), Ni(II), Ir(I), Rh(I), Au(III), Ag(III), and Cu(III). (b) L1, L2, L3, and L4 represent ligands, where each ligand provides a donor atom to coordinate to a metal atom. (c) The expression represents the number of positive or negative charges carried by the metallic complex, where n is zero or a positive integer. (d) This refers to counterions that maintain a neutral charge, among which It has a charge opposite to that of the metal complex, and where m is zero or a positive integer, m = n or m ≠ n. (e) The expression represents the stoichiometry of the counter ions. (f) Dashed lines represent covalent bonds between any two ligands, fusion of any ring portions from the two ligands, or combinations thereof. L4 includes optionally substituted C6-C 50 Aromatics or C3-C 50 The L4 group further includes groups selected from the following: halogen atoms, alkyl groups, heteroalkyl groups, alkenyl groups, heteroalkenyl groups, alkynyl groups, heteroalkynyl groups, aryl groups, heteroaryl groups, etc. Wherein, the optional substituted C6-C 50 Aromatics or C3-C 50 Substituents in heteroaromatic hydrocarbons include: halogen atoms, alkyl groups, heteroalkyl groups, alkenyl groups, heteroalkenyl groups, alkynyl groups, heteroalkynyl groups, aryl groups, and heteroaryl groups. Among them, R G1’ R G2’ R G3’ Independently constitutes a hydrogen atom, halogen atom, alkyl group, heteroalkyl group, alkenyl group, heteroalkenyl group, ynyl group, heteroynyl group, aryl group, or heteroaryl group. Wherein, the alkyl group is C1-C 30 Alkyl groups, wherein the heteroalkyl groups are C1-C 30 Heteroalkyl groups, wherein the alkenyl groups are C2-C 30 alkenyl, wherein the heteroalkenyl group is C2-C 30 Heteroalkenyl, wherein the alkynyl group is C2-C 30 The alkynyl group, wherein the heteroyynyl group is C2-C 30 The aryl group is C6-C. 50 Aryl, wherein the heteroaryl group is C3-C 50 Mixed aromatic compounds.

11. The method of claim 10, wherein the sample comprises human or non-human animal bodily fluids, human or non-human animal tissues, or combinations thereof.

12. The method of claim 11, wherein the bodily fluid is cerebrospinal fluid.

13. The method of claim 11, wherein the tissue is brain tissue.

14. The method of any one of claims 10 to 13, wherein the analyte is a protein or peptide of amyloid, plaque, or both, wherein the protein or peptide comprises linear aggregates of proteins or peptides arranged in a β-sheet conformation.

15. A method for testing the efficacy of an inhibitor against amyloidosis and / or fibrillation of proteins or peptides, comprising: (a) Combining a metal complex with an inhibitor-treated sample containing a protein or peptide, and combining it separately with an untreated sample containing a protein or peptide. (b) Compare the photophysical properties of the metal complexes between the two samples. The magnitude of the difference in photophysical properties between the two samples of metal complexes indicates the degree of change in the aggregation and supramolecular self-assembly state of the metal complexes, and the degree of change in the aggregation and supramolecular self-assembly state of the metal complexes indicates the efficacy of the inhibitor. The metal complex described herein has the structure of Formula I: in (a) M represents a metal atom selected from Pt(II), Pd(II), Ni(II), Ir(I), Rh(I), Au(III), Ag(III), and Cu(III). (b) L1, L2, L3, and L4 represent ligands, where each ligand provides a donor atom to coordinate to a metal atom. (c) The expression represents the number of positive or negative charges carried by the metallic complex, where n is zero or a positive integer. (d) This refers to counterions that maintain a neutral charge, among which It has a charge opposite to that of the metal complex, and where m is zero or a positive integer, m = n or m ≠ n. (e) The expression represents the stoichiometry of the counter ions. (f) Dashed lines represent covalent bonds between two optional ligands, fusion of optional ring portions from two ligands, or combinations thereof, wherein L4 includes optionally substituted C6-C. 50 Aromatics or C3-C 50 The L4 group further includes groups selected from the following: halogen atoms, alkyl groups, heteroalkyl groups, alkenyl groups, heteroalkenyl groups, alkynyl groups, heteroalkynyl groups, aryl groups, heteroaryl groups, etc. Wherein, the optional substituted C6-C 50 Aromatics or C3-C 50 Substituents in heteroaromatic hydrocarbons include: halogen atoms, alkyl groups, heteroalkyl groups, alkenyl groups, heteroalkenyl groups, alkynyl groups, heteroalkynyl groups, aryl groups, and heteroaryl groups. Among them, R G1’ R G2’ R G3’ Independently constitutes a hydrogen atom, halogen atom, alkyl group, heteroalkyl group, alkenyl group, heteroalkenyl group, ynyl group, heteroynyl group, aryl group, or heteroaryl group. Wherein, the alkyl group is C1-C 30 Alkyl groups, wherein the heteroalkyl groups are C1-C 30 Heteroalkyl groups, wherein the alkenyl groups are C2-C 30 alkenyl, wherein the heteroalkenyl group is C2-C 30 Heteroalkenyl, wherein the alkynyl group is C2-C 30 The alkynyl group, wherein the heteroyynyl group is C2-C 30 The aryl group is C6-C. 50 Aryl, wherein the heteroaryl group is C3-C 50 Mixed aromatic compounds.

16. A method for imaging an analyte in a sample, comprising: (a) Combining the metal complex with the sample under conditions that allow the metal complex of the compound to bind to the analyte and subsequently undergo aggregation and supramolecular self-assembly of the metal complex, wherein the aggregation and supramolecular self-assembly of the metal complex results in a change in the photophysical properties of the metal complex. (b) Imaging of the analyte based on one or more photophysical properties specific to aggregated and supramolecular self-assembled metal complexes. The analytes are selected from (1) amyloid protein, plaque, or both proteins or peptides, and (2) RNA; The metal complex described herein has the structure of Formula I: in (a) M represents a metal atom selected from Pt(II), Pd(II), Ni(II), Ir(I), Rh(I), Au(III), Ag(III), and Cu(III). (b) L1, L2, L3, and L4 represent ligands, where each ligand provides a donor atom to coordinate to a metal atom. (c) The expression represents the number of positive or negative charges carried by the metallic complex, where n is zero or a positive integer. (d) This refers to counterions that maintain a neutral charge, among which It has a charge opposite to that of the metal complex, and where m is zero or a positive integer, m = n or m ≠ n. (e) The expression represents the stoichiometry of the counter ions. (f) Dashed lines represent covalent bonds between any two ligands, fusion of any ring portions from the two ligands, or combinations thereof. L4 includes optionally substituted C6-C 50 Aromatics or C3-C 50 The L4 group further includes groups selected from the following: halogen atoms, alkyl groups, heteroalkyl groups, alkenyl groups, heteroalkenyl groups, alkynyl groups, heteroalkynyl groups, aryl groups, heteroaryl groups, etc. Wherein, the optional substituted C6-C 50 Aromatics or C3-C 50 Substituents in heteroaromatic hydrocarbons include: halogen atoms, alkyl groups, heteroalkyl groups, alkenyl groups, heteroalkenyl groups, alkynyl groups, heteroalkynyl groups, aryl groups, and heteroaryl groups. Among them, R G1’ R G2’ R G3’ Independently constitutes a hydrogen atom, halogen atom, alkyl group, heteroalkyl group, alkenyl group, heteroalkenyl group, ynyl group, heteroynyl group, aryl group, or heteroaryl group. Wherein, the alkyl group is C1-C 30 Alkyl groups, wherein the heteroalkyl groups are C1-C 30 Heteroalkyl groups, wherein the alkenyl groups are C2-C 30 alkenyl, wherein the heteroalkenyl group is C2-C 30 Heteroalkenyl, wherein the alkynyl group is C2-C 30 The alkynyl group, wherein the heteroyynyl group is C2-C 30 The aryl group is C6-C. 50 Aryl, wherein the heteroaryl group is C3-C 50 Mixed aromatic compounds.

17. The method of claim 16, wherein the sample comprises eukaryotic cells selected from 3T3 cells, A549 cells, Chinese hamster ovary (CHO) cells, HEK 293 cells, HeLa cells, Hep G2 cells, and HT1080 cells.

18. A kit for detecting and / or imaging an analyte, comprising in one or more containers one or more compounds according to any one of claims 1-9 and optional instructions for use, wherein the analyte is selected from (1) amyloid, plaque, or both proteins or peptides, and (2) RNA.

19. The kit of claim 18, further comprising a carrier.

20. The kit of claim 18 or 19, wherein the presence of the analyte can induce aggregation and supramolecular self-assembly of the metal complex thereon after binding, wherein the aggregation and supramolecular self-assembly of the metal complex can be detected by changes in the photophysical properties of the metal complex.

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