Phosphorylated tau–gold nanoparticle complex, method for preparing same, and drug screening method using same

Abstract

The present invention relates to a phosphorylated tau-gold nanoparticle complex, a method for manufacturing the same, and a drug screening method using same. The phosphorylated tau-gold nanoparticle complex manufactured according to the method for manufacturing a phosphorylated tau-gold nanoparticle complex of the present invention enables evaluation of how effectively a specific drug degrades phosphorylated tau aggregates and thus can be advantageously used for high-throughput screening of drugs for preventing or treating primary tauopathies.

Description

Phosphorylated tau-gold nanoparticle complex, method for preparing the same, and drug screening method using the same

[0001] The present application claims priority based on Korean Patent Application No. 10-2024-0166328 filed on November 20, 2024, and all contents disclosed in the specification and drawings of said application are incorporated into the present application.

[0002] The present invention relates to a phosphorylated tau-gold nanoparticle complex, a method for preparing the same, and a drug screening method using the same.

[0003] As average life expectancy increases, the aging of society in Korea is accelerating. By 2050, the number of people aged 65 or older is expected to reach approximately 19 million out of a total projected population of about 47.74 million, accounting for 39.8% of the total population—more than double the current figure. In this accelerating aging society, the population with degenerative brain diseases is also increasing in proportion. As of 2018, approximately 750,000 people, representing 10% of the national population of about 7.5 million people aged 65 or older, were diagnosed with degenerative brain diseases.

[0004] In line with this trend, the domestic and international markets for treatments for degenerative brain diseases are also growing rapidly. The global market for dementia treatments grew at an average annual rate of 8.5% from $12.8598 billion in 2014, and is projected to reach $21.1548 billion in 2020 and form a market of approximately $12.6 billion in 2024, representing more than a fourfold increase. The domestic market for dementia treatments is expected to grow from 400 billion won in 2012 to approximately 1.4 trillion won in 2020, with a projected average annual growth rate of 20% in the future.

[0005] Tauopathy is a degenerative brain disease caused by abnormal changes in tau protein in the brain. While normal tau protein plays a role in stabilizing microtubules in brain cells, in tauopathy, tau protein becomes hyperphosphorylated or abnormally modified. As a result, tau protein clumps together to form toxic structures called 'neurofibrillary tangles' inside nerve cells, which can impair nerve cell function and eventually lead to cell death.

[0006] Existing tau protein aggregate screening systems designed to identify drug candidates effective for tauopathy face several limitations. AI-based screening systems struggle to accurately assess the degradation effects on tau aggregates when drug information is insufficient. Furthermore, methods utilizing Fluorescence Resonance Energy Transfer (FRET) technology carry the risk of altering the intrinsic structure of proteins and are susceptible to fluorescence noise affecting results; additionally, they are unsuitable for high-throughput, high-cost experiments due to their high cost and time requirements. Given that cell-based experiments are time-consuming and require specialized personnel, improvements in tau protein aggregate screening methods are currently necessary in terms of accuracy and efficiency.

[0007] The inventors have completed the present invention by confirming that a drug screening method for the prevention or treatment of tauopathy using a phosphorylated tau-gold nanoparticle complex enables high-speed, high-volume drug screening by compensating for the shortcomings of existing technologies in terms of accuracy, efficiency, and time.

[0008] The object of the present invention is to provide a phosphorylated tau-gold nanoparticle complex comprising gold nanoparticles coated with phosphorylated tau oligomers.

[0009] In addition, another objective of the present invention is to provide a method for manufacturing a phosphorylated tau-gold nanoparticle complex.

[0010] In addition, another objective of the present invention is to provide a drug screening method for the prevention or treatment of tauopathy.

[0011] To achieve the above objectives, the present invention provides a phosphorylated tau-gold nanoparticle composite comprising gold nanoparticles coated with a phosphorylated tau oligomer.

[0012] Furthermore, the present invention provides a method for preparing a phosphorylated tau-gold nanoparticle complex comprising: (A) a step of dialyzing phosphorylated tau; (B) a step of preparing a phosphorylated tau solution by adding an ATP solution to the solution of step (A); and (C) a step of incubating a gold nanoparticle solution after adding a gold nanoparticle solution to the phosphorylated tau solution of step (B).

[0013] In addition, the present invention provides a drug screening method for the prevention or treatment of tauopathy, comprising: 1) a step of treating a phosphorylated tau-gold nanoparticle complex of claim 1 with a candidate drug for the treatment of tauopathy; and 2) a step of comparing the color change of a phosphorylated tau-gold nanoparticle complex treated with the candidate drug of step 1) with that of a phosphorylated tau-gold nanoparticle complex not treated.

[0014] The phosphorylated tau-gold nanoparticle complex prepared according to the method for preparing a phosphorylated tau-gold nanoparticle complex of the present invention can be used to verify how effectively a specific drug degrades phosphorylated tau aggregates, and thus can be usefully used for high-speed high-throughput screening of drugs for the prevention or treatment of primary tauopathy.

[0015] Figure 1 is a figure showing the results of analyzing the molecular weight of tau phosphorylated by GSK-3β in one embodiment of the present invention.

[0016] Figure 2 is a figure showing the result of reacting phosphorylated tau without dialysis with gold nanoparticles in one embodiment of the present invention.

[0017] Figure 3 is a figure showing the result of reacting dialysis-treated phosphorylated tau with gold nanoparticles in one embodiment of the present invention.

[0018] Figure 4 is a figure showing the results of comparing the degree of aggregation of gold nanoparticles in dialysis-treated phosphorylated tau (left) and general DW (right) in one embodiment of the present invention.

[0019] Figure 5 is a figure showing the results of reacting phosphorylated tau, which has been made electrically neutral by adding ATP, with gold nanoparticles at different concentrations in one embodiment of the present invention.

[0020] Figure 6 is a figure showing the results of confirming the stability under salt conditions of a gold nanoparticle complex (p-TACON) prepared by adding a high concentration of phosphorylated tau in one embodiment of the present invention.

[0021] Figure 7 is a figure showing the results of confirming the stability under salt conditions of a gold nanoparticle complex (p-TACON) prepared by adding a low concentration of phosphorylated tau in one embodiment of the present invention.

[0022] FIG. 8 is a figure showing the results of measuring the hydrodynamic diameter of general gold nanoparticles and gold nanoparticle complexes (p-TACON) using dynamic light scattering in one embodiment of the present invention.

[0023] FIG. 9 is a figure showing the results of measuring the size of general gold nanoparticles and gold nanoparticle complexes (p-TACON) through transmission electron microscopy analysis in one embodiment of the present invention.

[0024] FIG. 10 is a figure showing the results of measuring the size of ordinary gold nanoparticles and gold nanoparticle complexes (p-TACON) using UV-Vis spectroscopy in one embodiment of the present invention.

[0025] FIG. 11 is a figure showing the results of measuring the zeta potential of general gold nanoparticles and a gold nanoparticle complex (p-TACON) in one embodiment of the present invention.

[0026] FIG. 12 is a figure showing the reaction results of a gold nanoparticle complex (p-TACON) and protease type XIV (P14) at different concentrations in one embodiment of the present invention.

[0027] Hereinafter, the present invention will be described in detail with reference to the attached drawings for embodiments of the present invention. However, the following embodiments are presented as examples of the present invention, and if it is determined that a detailed description of a technology or configuration well known to those skilled in the art may unnecessarily obscure the essence of the present invention, such detailed description may be omitted, and the present invention is not limited by this. The present invention is capable of various modifications and applications within the scope of the claims set forth below and the equivalent scope interpreted therefrom.

[0028] Furthermore, the terminology used in this specification is used to appropriately describe preferred embodiments of the present invention, and may vary depending on the intent of the user or operator, or the conventions of the field to which the present invention belongs. Accordingly, the definitions of these terms should be based on the content throughout this specification. Throughout the specification, when a part is described as "comprising" a certain component, unless specifically stated otherwise, this means that it does not exclude other components but may include additional components.

[0029] Throughout this specification, '%' used to indicate the concentration of a particular substance is (w / w) % for solid / solid, (w / v) % for solid / liquid, and (v / v) % for liquid / liquid, unless otherwise noted.

[0030]

[0031] In one aspect, the present invention provides a phosphorylated tau-gold nanoparticle composite comprising gold nanoparticles coated with a phosphorylated tau oligomer.

[0032] In one embodiment of the present invention, the phosphorylated tau may be phosphorylated by glycogen synthase kinase-3 beta (GSK-3β), but is not limited thereto.

[0033] In one embodiment of the present invention, the gold nanoparticle composite may have a diameter of 1 to 100 nm, preferably 5 to 40 nm, more preferably 15 to 30 nm, and even more preferably 22 to 23 nm, but is not limited thereto.

[0034] In one embodiment of the present invention, the phosphorylated tau oligomer may be coated on the surface of gold nanoparticles with a thickness of 1 to 5 nm, preferably 2 to 4 nm, more preferably 2.5 to 3.5 nm, but is not limited thereto.

[0035] In one embodiment of the present invention, a phosphorylated tau oligomer may be coated in a corona form on the surface of the gold nanoparticle composite, but is not limited thereto.

[0036] In one embodiment of the present invention, the gold nanoparticle composite may not undergo an aggregation reaction under salt conditions, but is not limited thereto.

[0037]

[0038] In one aspect, the present invention provides a method for preparing a phosphorylated tau-gold nanoparticle complex comprising: (A) a step of dialyzing phosphorylated tau; (B) a step of preparing a phosphorylated tau solution by adding an ATP solution to the solution of step (A); and (C) a step of incubating a gold nanoparticle solution after adding a gold nanoparticle solution to the phosphorylated tau solution of step (B).

[0039] In one embodiment of the present invention, the buffer of phosphorylated tau may be exchanged through the dialysis of step (A), but is not limited thereto.

[0040] In one embodiment of the present invention, the concentration of the ATP solution in step (B) may be 0.01 to 10 mg / mL, preferably 0.1 to 5 mg / mL, preferably 1 mg / mL, but is not limited thereto.

[0041] In one embodiment of the present invention, the addition of the ATP solution in step (B) may prevent the aggregation reaction under salt conditions, but is not limited thereto.

[0042] In one embodiment of the present invention, the concentration of tau protein contained in the phosphorylated tau solution of step (B) may be 73 nM or more, preferably 102 nM or more, more preferably 117.648 nM, but is not limited thereto.

[0043] In one embodiment of the present invention, the phosphorylated tau solution and the gold nanoparticle solution of step (C) may be mixed in a volume ratio of 0.1 to 5:1, preferably 1 to 3:1, more preferably 2:1, but are not limited thereto.

[0044] In one embodiment of the present invention, the incubation of step (C) may be performed at 30 to 45°C, preferably 35 to 40°C, more preferably 37°C, at 50 to 200 rpm, preferably 100 to 150 rpm, more preferably 120 rpm, for 12 to 36 hours, preferably 18 to 30 hours, more preferably 24 hours, but is not limited thereto.

[0045]

[0046] In one aspect, the present invention provides a drug screening method for the prevention or treatment of tauopathy, comprising: 1) treating a phosphorylated tau-gold nanoparticle complex of claim 1 with a candidate drug for the treatment of tauopathy; and 2) comparing the color change of a phosphorylated tau-gold nanoparticle complex treated with the candidate drug of step 1) with that of a phosphorylated tau-gold nanoparticle complex not treated.

[0047] The term "tauopathy" used in this invention refers to a neurodegenerative disease caused by abnormal changes in tau protein in the brain. Normal tau protein plays a role in stabilizing microtubules in brain cells, but in tauopathy, tau protein is hyperphosphorylated or abnormally modified. As a result, tau protein clumps together to form toxic structures called "neurofibrillary tangles" inside nerve cells, which can lead to a decline in nerve cell function and eventually cell death.

[0048] Tauopathy can be classified into primary and secondary tauopathy. In primary tauopathy, the abnormal accumulation of tau protein is the primary cause of the disease and the core pathological phenomenon; abnormal structural changes in the hyperphosphorylated ring of tau protein act as the main pathological mechanism, leading to neuronal damage and degenerative symptoms. In secondary tauopathy, the accumulation of tau protein occurs secondarily due to other pathological phenomena. In other words, rather than tau protein accumulation being the primary cause, it is a pathological phenomenon induced through other degenerative processes.

[0049] In one embodiment of the present invention, the tauopathy may be primary tauopathy, but is not limited thereto.

[0050] In one embodiment of the present invention, the color change in step 2) may be measured by the naked eye, a spectrometer, or a colorimeter (colorimeter), but is not limited thereto.

[0051] In one embodiment of the present invention, the method may be capable of high-speed mass screening, but is not limited thereto.

[0052] As described above, specific embodiments of the present invention have been described in detail; however, those skilled in the art who understand the spirit of the present invention will be able to easily propose other inventions that are inferior or other embodiments included within the scope of the spirit of the present invention by adding, changing, or deleting other components within the same spirit. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive. The scope of the present invention is defined by the claims set forth below rather than by the detailed description above, and all modifications or variations derived from the meaning and scope of the claims and equivalent concepts should be interpreted as being included within the scope of the present invention.

[0053] <Example 1> Preparation of Phosphorylated Tau Oligomer Coated Gold Nanoparticle Composite

[0054] 1-1. Preparation of Phosphorylated Tau Oligomer Coated Gold Nanoparticle Composites

[0055] To prepare a phosphorylated-tau amyloid corona-shelled nanoparticle (p-TACON) complex, phosphorylated tau protein and gold nanoparticles were used.

[0056] Phosphorylated tau protein, phosphorylated by GSK-3β (Glycogen synthase kinase-3 beta) and having a molecular weight of 64–68 kDa (Fig. 1), was purchased from Sino Biological and used. Additionally, gold nanoparticles were synthesized using the Turkevich method and had a hydrodynamic diameter of 20 nm; they were concentrated by centrifugation at 10,500 rpm for 16 minutes and 30 seconds before use.

[0057] Since phosphorylated tau is dissolved in a buffer containing 50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM DTT, and 10% glycerol, to prevent the aggregation of gold nanoparticles by salt (Fig. 2), a dialysis kit (Pur-A-Lyzer TM Buffer change of phosphorylated tau was performed using a Mini Dialysis Kit (Cat# PURN25005, sigma aldrich). The dialysis kit has a Molecular Weight Cut-Off (MWCO) of 25,000 Da. 100 μL of phosphorylated tau was added to the kit, immersed in 500 mL of distilled water, and dialysis was performed at 100 rpm for 3 hours. The phosphorylated tau solution after dialysis had a volume of 130–150 μL, and since proteins in the solution could not pass through the membrane, the amount of protein before and after dialysis remained the same. After adding distilled water to the phosphorylated tau solution to bring the total volume to 1 mL, it was confirmed that no aggregation caused by salt occurred when reacted with gold nanoparticles (Fig. 3).

[0058] However, gold nanoparticles to which phosphorylated tau solution was added exhibited aggregation compared to ordinary gold nanoparticles, resulting in a relatively darker color (Fig. 4). This aggregation occurred because phosphorylated tau, although phosphorylated, carries a positive charge and reacts with negatively charged gold nanoparticles. To resolve this, a 1 mg / mL ATP solution was added to the dialyzed phosphorylated tau solution instead of DW to make the total volume 1 mL. Since ATP carries a negative charge, it was mixed with the phosphorylated tau solution to create an electrically neutral solution. It was confirmed that the phosphorylated tau solution with added ATP did not cause a color change even when reacting with gold nanoparticles (Fig. 5). The table in Fig. 5 shows the final concentration of phosphorylated tau. A gold nanoparticle complex (p-TACON) was prepared by incubating the solution in which the phosphorylated tau solution with added ATP was reacted with gold nanoparticles at 37°C and 120 rpm for one day.

[0059] 1-2. Confirmation of the Stability of the Gold Nanoparticle Composite

[0060] To primarily check whether the surface of the gold nanoparticles was coated with protein, 800 μL of 1X PBS was added, and a stability test of the gold nanoparticle complex (p-TACON) was conducted under 0.8X PBS conditions.

[0061] Specifically, the volume of phosphorylated tau protein was varied to 100, 110, 120, 130, 140, 150, and 160 μL (tau protein concentration: 147.06, 161.76, 176.47, 191.18, 205.88, 220.59, 235.29 nM), and after adding PBS, the absorbance of the solution was measured at wavelengths of 400–800 nm using UV-Vis spectroscopy after 0, 1, 4, 8, and 24 hours.

[0062] As shown in Figure 6, the stability of the gold nanoparticle complex was confirmed to be maintained, as the peak shape was maintained even though the concentration of tau protein was varied by adding 100 to 160 μL of phosphorylated tau protein in 10 μL increments to a total volume of 200 μL.

[0063] In order to confirm whether stability is maintained even when the concentration of tau protein is lowered, a stability test was conducted using the method described above with the concentration of tau protein lowered.

[0064] Specifically, the volume of phosphorylated tau protein was 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, and 120 μL out of a total of 200 μL (tau protein concentration: 14.706–176.472 nM) (see table in Figure 7), and stability tests were performed under 0.8X PBS conditions.

[0065] As shown in Figure 7, gold nanoparticle complexes prepared by adding low concentrations of phosphorylated tau protein (14.706–58.824 nM) (10–40 μL out of a total of 200 μL) could not withstand the salt and aggregated, losing the characteristic peak shape of 20 nm gold nanoparticles. However, starting from a phosphorylated tau protein concentration of 73.53 nM (50 μL), the complexes were able to withstand salt conditions, and it was confirmed that gold nanoparticle complexes prepared by adding a phosphorylated tau protein concentration of 102.942 nM (70 μL) or higher withstood 0.8X PBS conditions for more than 24 hours.

[0066] Through this, it was confirmed that p-TACON synthesis is possible by dialyzing phosphorylated tau to change the buffer, adding 80 μL of phosphorylated tau solution with ATP solution to 40 μL of concentrated gold nanoparticle solution to achieve a final protein concentration of 117.648 nM, and incubating for one day at 37 ℃ and 120 rpm.

[0067] <Experimental Example 1> Measurement of Size Change of Gold Nanoparticle Complex

[0068] 1-1. Hydrodynamic Diameter Measurement Using Dynamic Light Scattering

[0069] The hydrodynamic diameter of the gold nanoparticle composite (p-TACON) prepared in Example 1 above was measured using Dynamic Light Scattering (DLS).

[0070] As shown in Figure 8, the gold nanoparticles (Bare AuNP) used in the synthesis of p-TACON have a diameter of approximately 19.765 nm, while p-TACON has a diameter of approximately 22.725 nm.

[0071] Through this, it was confirmed that phosphorylated tau aggregates were coated on the surface of gold nanoparticles with a size of 2.96 nm.

[0072] 1-2. Size Measurement via Transmission Electron Microscopy Analysis

[0073] The size of the gold nanoparticle composite (p-TACON) prepared in Example 1 above was measured visually through transmission electron microscopy (TEM) analysis.

[0074] As shown in Figure 9, compared to the gold nanoparticles (Bare AuNP) used in the synthesis of p-TACON, it was visually confirmed that phosphorylated tau protein aggregates were coated on the surface of p-TACON in a corona form with a thickness of about 3 nm.

[0075] 1-3. Size Analysis Using UV-Vis Spectroscopy

[0076] The change in size of the gold nanoparticle composite (p-TACON) prepared in Example 1 above was analyzed using UV-Vis spectroscopy.

[0077] As shown in Figure 10, the gold nanoparticles (Bare AuNP) used in the synthesis of p-TACON showed maximum absorbance at a wavelength of 518 nm, whereas p-TACON showed maximum absorbance at a wavelength of 521 nm.

[0078] Through this, it was confirmed that the maximum absorbance changed as the surface plasmon resonance frequency shifted to a longer wavelength range due to the increase in particle size caused by the protein coating.

[0079] <Experimental Example 2> Measurement of Dispersion Stability of Gold Nanoparticle Composite

[0080] To confirm the dispersion stability of the gold nanoparticle composite (p-TACON) prepared in Example 1 above in solution, the zeta potential was measured.

[0081] Zeta potential is a measure used to determine the stability of dispersed particles, such as colloids or nanoparticles, within a solvent, and is related to the surface inversion of the particles. Particle stability is considered good when the zeta potential is between ±40 and 60 mV, while it is considered extremely stable when it is ±60 mV or higher.

[0082] As shown in Figure 11, the gold nanoparticles (Bare AuNP) used in the synthesis of p-TACON exhibited a zeta potential of -53.322 mV, while p-TACON had a zeta potential of -57.792 mV, which is lower than that of ordinary gold nanoparticles.

[0083] Through this, it was confirmed that phosphorylated tau was coated on the surface of gold nanoparticles, enhancing the stability of the particles.

[0084] <Experimental Example 3> Confirmation of Applicability of Gold Nanoparticle Complex to Drug Screening Platform

[0085] Additional experiments were conducted to confirm the potential of the gold nanoparticle complex (p-TACON) prepared in Example 1 above as a drug screening platform.

[0086] Since the gold nanoparticles in p-TACON are protected by phosphorylated tau, they do not aggregate and are well dispersed even under salt conditions; however, if the protein layer on the surface of the gold nanoparticles is degraded, they will aggregate due to the salt. To confirm this, protease type XIV (P14), a cocktail of three types of proteases, was dissolved in PBS and reacted with p-TACON at different concentrations.

[0087] As shown in Figure 12, low concentrations of P14 did not sufficiently decompose the phosphorylated tau aggregates on the surface of p-TACON and thus did not induce aggregation of p-TACON, but high concentrations of P14 decomposed the phosphorylated tau aggregates and induced aggregation of p-TACON.

[0088] Through this, it was confirmed that the gold nanoparticle complex (p-TACON) can be used as a platform to evaluate the efficacy of drugs targeting phosphorylated tau, and to measure how effectively the drug degrades phosphorylated tau aggregates. Furthermore, based on this, it was confirmed that drug candidates effective for primary tauopathy-related diseases can be screened.

[0089] The present invention has been described above with reference to preferred embodiments and experimental examples. Those skilled in the art will understand that the present invention may be embodied in modified forms without departing from the essential characteristics of the invention. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the invention is defined by the claims, not by the foregoing description, and all variations within the scope of the claims should be interpreted as being included in the invention.

Claims

1. Phosphorylated tau-gold nanoparticle complex comprising gold nanoparticles coated with phosphorylated tau oligomers.

2. In Paragraph 1, Phosphorylated tau-gold nanoparticle complex characterized in that the above phosphorylated tau is phosphorylated by glycogen synthase kinase-3 beta (GSK-3β).

3. In Paragraph 1, Phosphorylated tau-gold nanoparticle composite, characterized in that the gold nanoparticle composite has a diameter of 1 to 100 nm.

4. In Paragraph 1, A phosphorylated tau-gold nanoparticle composite characterized by the above phosphorylated tau oligomer being coated on the surface of gold nanoparticles to a thickness of 1 to 5 nm.

5. In Paragraph 1, Phosphorylated tau-gold nanoparticle composite characterized by having a phosphorylated tau oligomer coated in a corona form on the surface of the gold nanoparticle composite.

6. In Paragraph 1, The above-mentioned gold nanoparticle complex is a phosphorylated tau-gold nanoparticle complex characterized by not undergoing an aggregation reaction under salt conditions. 7.(A) Step of dialysis of phosphorylated tau; (B) a step of preparing a phosphorylated tau solution by adding an ATP solution to the solution of step (A) above; and (C) a step of adding a gold nanoparticle solution to the phosphorylated tau solution of step (B) above, followed by incubation; a method for preparing a phosphorylated tau-gold nanoparticle complex.

8. In Paragraph 7, A method for preparing a phosphorylated tau-gold nanoparticle complex, characterized by exchanging the buffer of phosphorylated tau through the dialysis of step (A) above.

9. In Paragraph 7, A method for preparing a phosphorylated tau-gold nanoparticle complex, characterized in that the concentration of the ATP solution in step (B) above is 0.01 to 10 mg / mL.

10. In Paragraph 7, A method for preparing a phosphorylated tau-gold nanoparticle complex, characterized by preventing aggregation reactions under salt conditions through the addition of an ATP solution in step (B) above.

11. In Paragraph 7, A method for preparing a phosphorylated tau-gold nanoparticle complex, characterized in that the concentration of tau protein contained in the phosphorylated tau solution of step (B) above is 73 nM or higher.

12. In Paragraph 7, A method for preparing a phosphorylated tau-gold nanoparticle composite, characterized in that the incubation of step (C) above is performed at 30 to 45 ℃ at 50 to 200 rpm for 12 to 36 hours. 13.1) A step of treating the phosphorylated tau-gold nanoparticle complex of claim 1 with a candidate drug for tauopathy treatment; and 2) a step of comparing the color change of a phosphorylated tau-gold nanoparticle complex treated with the candidate drug of step 1) and a phosphorylated tau-gold nanoparticle complex not treated; comprising a drug screening method for the prevention or treatment of tauopathy.

14. In Paragraph 13, A drug screening method for the prevention or treatment of tauopathy, characterized in that the above tauopathy is primary tauopathy.

15. In Paragraph 13, A drug screening method for the prevention or treatment of tauopathy, characterized by measuring the color change in step 2) above with the naked eye, a spectrometer, or a colorimeter (colorimeter).

16. In Paragraph 13, The above method is a drug screening method for the prevention or treatment of tauopathy, characterized by being capable of high-speed, high-volume screening.

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