HSP90 alpha selective inhibitor as well as synthesis method and application thereof

Abstract

The invention discloses an HSP90 alpha selective inhibitor as well as a synthesis method and application thereof, and belongs to the technical field of medicines. The synthesis method comprises the following steps: by taking UP302 and (4-bromobutyl) triphenylphosphonium bromide as raw materials, anhydrous acetonitrile as a solvent and sodium iodide and cesium carbonate as catalysts, stirring and heating for 5 hours under the protection of nitrogen at 50 DEG C, and separating to obtain the HSP90 alpha selective inhibitor. The compound UP302 with HSP90 inhibition potential is separated from dianella ensifolia, but the antitumor activity of the compound UP302 needs to be improved. Based on a mitochondrial targeting strategy, triphenylphosphine (TPP) is used for modifying UP302, and two triphenylphosphine derivatives Mito-UP302-1 and Mito-UP302-2 which are novel in structure are successfully synthesized. According to the present invention, the Mito-UP302-1 can selectively inhibit the HSP90 alpha, can effectively inhibit the mitochondrial functional activity, and can provide the significant anti-tumor effect; the discovery not only solves the problem of insufficient HSP90 inhibitor subtype selectivity, but also provides a new strategy for research and development of a novel and efficient HSP90 alpha selective inhibitor and cancer treatment.

Description

Technical Field

[0001] The present invention belongs to the field of medicines, and in particular relates to a HSP90α selective inhibitor and a synthesis method and application thereof. Background Art

[0002] Heat shock protein 90 (HSP90) is an important molecular chaperone that can interact with other proteins and help other proteins (called substrates or clients) gain functional activity. HSP90 controls protein function and activity by promoting protein folding, binding of ligands to their receptors or targets, or assembly of multiprotein complexes, and becomes a key factor in regulating protein homeostasis in eukaryotic cells under physiological and stress conditions. HSP90 chaperones interact with a large number of diverse client proteins, many of which are important regulatory molecules for tumorigenesis, immunosuppression, invasion and metastasis, such as EGFR, P-53, Raf-1 and IKK. HSP90 is highly expressed in many types of cancer, including gastric cancer, breast cancer, nasopharyngeal carcinoma, etc. Overexpression of HSP90 can participate in the occurrence and development of cancer by promoting cell growth and proliferation and affecting the glycolysis process.

[0003] In mammalian cells, HSP90 has four subtypes: HSP90α / β, glucose-regulated protein 94 (GRP94) and tumor necrosis factor receptor-associated protein 1 (TRAP1). Among them, HSP90α is an inducible protein, and its functional activity is closely related to the development of tumors. As the malignancy of tumors increases, the secretion of cellular HSP90α also increases, making it a new tumor marker for early detection of tumors and evaluation of therapeutic efficacy. HSP90β is a constitutive protein used to maintain normal cell growth and function. Compared with HSP90β, HSP90α can be secreted into the cell membrane and outside the cell, and plays a key role in cell signaling and protein homeostasis, especially in tumor growth and cell differentiation. These characteristics make HSP90α an important drug target in cancer treatment. Therefore, the synthesis of HSP90α selective inhibitors has potential application prospects in the field of tumor treatment.

[0004] HSP90α is a highly conserved ATP-dependent molecular chaperone protein composed of an N-terminal domain (NTD), a middle domain (MD), and a C-terminal domain (CTD). It participates in the metabolic transformation process of mitochondria in cancer cells and is closely related to mitochondrial functional activity. The mitochondrial targeting strategy is a specific treatment method for mitochondria, which can deliver drugs specifically to mitochondria and improve the therapeutic effect by inhibiting mitochondrial functional activity. Therefore, the synthesis of mitochondrial targeted drugs provides a potential new approach for the development of new HSP90α inhibitors. There are two methods for designing mitochondrial delivery drugs: directly conjugating targeting ligands / groups to drugs and directly conjugating targeting ligands to nanocarriers. It has been reported that many direct conjugates can be used as targeting groups for mitochondrial delivery of anticancer drugs, including lipophilic cations (triphenylphosphine, rhodamine 123, and divalent ions) and peptides (mitochondrial penetrating peptide (MPP), mitochondrial targeting sequence peptide (MTS), and Szeto-Schiller peptide (SS)). The principle of this method is based on the fact that the mitochondrial membrane potential of tumor cells is higher than that of their cytosol and non-cancer cells. The most widely used targeting group is triphenylphosphine (TPP), which is a lipophilic cation that relies on a high mitochondrial membrane potential difference to selectively accumulate drugs in mitochondria. At the same time, the driving force of lipophilic cations in tumor cells is more than 10 times that of normal cells, which provides an effective means for the development of anti-tumor drugs. According to literature analysis, there is a class of HSP90 inhibitors that belong to the resorcinol class of compounds, and the resorcinol core in its structure is the key part of inhibiting HSP90 activity. Summary of the invention

[0005] The present invention provides a HSP90α selective inhibitor and a synthesis method and application thereof, so as to solve the problem that the existing HSP90 inhibitors often lack selectivity for different subtypes of HSP90 (such as HSP90α and HSP90β); and solve the problem that although the original UP302 compound has the potential to inhibit HSP90, its anti-tumor activity needs to be improved.

[0006] The present invention provides a HSP90α selective inhibitor represented by the following structural formula: Among them, when R1 is , R2 is H, Mito-UP302-1; When R1 is H, R2 is , Mito-UP302-2.

[0007] The present invention also provides a use of a HSP90α selective inhibitor of the structural formula shown in the above formula in the treatment of cancer.

[0008] The present invention also provides a method for synthesizing a HSP90α selective inhibitor of the structural formula shown in the above formula, comprising: Using UP302 and (4-bromobutyl)triphenylphosphonium bromide as raw materials, anhydrous acetonitrile as solvent, sodium iodide and cesium carbonate as catalysts, and stirring and heating at 50°C for 5 hours under nitrogen protection, a selective inhibitor of HSP90α was isolated. The reaction formula of the synthesis method is: Among them, when R1 is , R2 is H, Mito-UP302-1; When R1 is H, R2 is , Mito-UP302-2.

[0009] The compound dimethoxytolyl-4-propylresorcinol (UP302) isolated from the mountain sedge orchid contains a resorcinol skeleton in its structure and has the potential to inhibit HSP90, but its anti-tumor activity needs to be improved. We use mitochondrial targeting technology to modify it in a targeted manner to improve its activity. When the two phenolic hydroxyl groups of the resorcinol compound are replaced separately, the compound will selectively inhibit the subtypes of HSP90 due to the different sites where the phenolic hydroxyl groups are retained. The structures of existing HSP90α selective inhibitors were analyzed, and it was found that retaining the para-hydroxyl group may be the key to achieving selective inhibition of HSP90α. Therefore, the present invention uses triphenylphosphine (TPP) to replace the ortho- and para-phenolic hydroxyl groups to synthesize two triphenylphosphine derivatives: (4-(2-(3-(2,4-dimethoxy-3-methylphenyl)propyl)-5-hydroxyphenoxy)butyl)triphenylphosphonium bromide formate (Mito-UP302-1) and (4-(4-(3-(2,4-dimethoxy-3-methylphenyl)propyl)-3-hydroxyphenoxy)butyl)triphenylphosphonium bromide formate (Mito-UP302-2), and their activities are verified to explore new and effective HSP90α selective inhibitors.

[0010] Based on the mitochondrial targeting strategy, the present invention modified the resorcinol compound UP302 with triphenylphosphine and synthesized two novel compounds, namely (4-(2-(3-(2,4-dimethoxy-3-methylphenyl)propyl)-5-hydroxyphenoxy)butyl)triphenylphosphonium formate (Mito-UP302-1) and (4-(4-(3-(2,4-dimethoxy-3-methylphenyl)propyl)-3-hydroxyphenoxy)butyl)triphenylphosphonium bromide formate (Mito-UP302-2). Among them, Mito-UP302-1 can inhibit the functional activity of mitochondria, selectively inhibit HSP90α, exert a significant anti-tumor effect, solve the important problem of insufficient subtype selectivity of HSP90 inhibitors, provide a new method for actively developing new and efficient HSP90α selective inhibitors, and provide an effective new strategy for cancer treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 is a reaction formula for a method for synthesizing a HSP90α selective inhibitor provided in an embodiment of the present invention; Figure 2 The compound Mito-UP302-1 provided in the embodiment of the present invention 1 H-NMR spectrum; Figure 3 The compound Mito-UP302-1 provided in the embodiment of the present invention 13 C-NMR spectrum; Figure 4 is the HMQC diagram of the compound Mito-UP302-1 provided in the examples of the present invention; Figure 5 is the HMBC diagram of the compound Mito-UP302-1 provided in the examples of the present invention; Figure 6 is a HR-ESI-MS graph of the compound Mito-UP302-1 provided in an embodiment of the present invention; Figure 7 The compound Mito-UP302-2 provided in the embodiment of the present invention 1 H-NMR spectrum; Figure 8 The compound Mito-UP302-2 provided in the embodiment of the present invention 13 C-NMR spectrum; Fig. 9 is the HMQC diagram of the compound Mito-UP302-2 provided in the examples of the present invention; Fig.10 is the HMBC diagram of the compound Mito-UP302-2 provided in the examples of the present invention; Fig.11is a HR-ESI-MS graph of the compound Mito-UP302-2 provided in an embodiment of the present invention; Fig.12 The left figure is a Western Blot measurement diagram of the changes in the levels of HSP90α and HSP90β proteins in HT29 cells after treatment with the compound Mito-UP302-1 provided in the examples of the present invention for 24 hours; Fig.12 The right figure is a bar graph showing the changes in the protein contents of HSP90α and HSP90β and their relative ratio (HSP90α / HSP90β) in HT29 cells after treatment with the compound Mito-UP302-1 provided in the examples of the present invention for 24 hours; Fig.13 The left figure is a Western Blot diagram of the changes in the levels of HSP90α and HSP90β proteins in HT29 cells after treatment with the compound Mito-UP302-2 provided in the examples of the present invention for 24 hours; Fig.13 The right figure is a bar graph showing the changes in the protein contents of HSP90α and HSP90β and their relative ratio (HSP90α / HSP90β) in HT29 cells after treatment with the compound Mito-UP302-2 provided in the examples of the present invention for 24 hours; Fig.14 This is a graph showing the mitochondrial membrane potential detection of the compound Mito-UP302-1 provided in an embodiment of the present invention; Fig.15 This is a graph showing changes in ATP content in HT29 cells treated with Mito-UP302-1 provided in an embodiment of the present invention. DETAILED DESCRIPTION

[0012] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with specific embodiments and with reference to the accompanying drawings. It should be understood that these descriptions are only exemplary and are not intended to limit the scope of the present invention. In addition, in the following description, the description of well-known structures and technologies is omitted to avoid unnecessary confusion of the concept of the present invention.

[0013] Example 1 Synthesis of (4-(2-(3-(2,4-dimethoxy-3-methylphenyl)propyl)-5-hydroxyphenoxy)butyl)triphenylphosphonium formate (Mito-UP302-1): The synthetic route is as follows: 1. Take a 100 mL eggplant-shaped bottle, add UP302 (240 mg, 0.79 mM, 1 equivalent, Nanjing Myermi Biotechnology Co., Ltd.), (4-bromobutyl) triphenylphosphonium bromide (455.84 mg, 0.95 mM, 1.2 equivalent, Shanghai Titan Technology Co., Ltd.), cesium carbonate (513.5 mg, 1.58 mM, 2.0 equivalent, Bid Pharmaceuticals), sodium iodide (59.25 mg, 0.40 mM, 0.5 equivalent, Komi Reagent), add 5 mL of anhydrous acetonitrile (Annaiji Chemical), heat to 50 degrees under nitrogen protection and stir for 5 hours.

[0014] 2. LCMS detection showed that most of the raw materials disappeared.

[0015] 3. The reaction solution was filtered through a triangular funnel and washed three times with acetonitrile. The filtrate was concentrated under reduced pressure and separated by preparative HPLC (machine model: Waters 3767, column: Gemini 5u C18 150 x 21.2 mm, flow rate: 20 mL / min, mobile phase: methanol (62%-95%) / water (0.1% formic acid) (38%-5%)).

[0016] 4. Concentrate and separate to obtain the target product: (4-(2-(3-(2,4-dimethoxy-3-methylphenyl)propyl)-5-hydroxyphenoxy)butyl)triphenylphosphonium formate (Mito-UP302-1) (50 mg, yield 9.48%) and (4-(4-(3-(2,4-dimethoxy-3-methylphenyl)propyl)-3-hydroxyphenoxy)butyl)triphenylphosphonium bromide formate (Mito-UP302-2) (50 mg, yield 9.48%) Signals assigned to Mito-UP302-1: white solid. 1H-NMR (400M, DMSO- d 6) MHz, H-1′), 3.74 (s, 2H, 4-OMe), 3.53 (s, 3H, 2-OMe), 3.29 (m, 1H, 5-triphenylphosphine), 3.85 (d, 1H, J = 8.4 Hz, H-6), 3.83 (d, 1H, J = 8.0 Hz, H-6′), 3.70 (m, 1H, 5-triphenylphosphine), 3.19 (s, 3H, 4-OMe), 3.28 (m, 1H, 5-triphenylphosphine), 3.50 (s, 3H, 2-OMe), 3.28 (m, 1H, 5-triphenylphosphine), 3.53 (m, 1H, 5-triphenylphosphine), 3.85 (d, 1H, J = 8.4 Hz, H-6), 3.83 (d, 1H, J = 8.0 Hz, H-6′), 3.83 (m, 1H, 4H, H-7, H-9), 2.00 (s, 3H, 3-Me), 1.90 (m, 2H, H-2′′), 1.71 (m, 2H, H-8), 1.58 (m, 2H, H-4′′). 13C-NMR (100M, DMSO- d6 ) C 158.2 (C-4′), 157.8 (C-2′), 157.7 (C-2), 157.3 (C-4), 131.1~135.8 (13C, C-1′′′~6′′′), 130.6 (C-6′), 127.7 (C-1), 127.6(C-6), 120.4 (C-1′), 119.9 (C-1′′′~6′′′), 119.2 (C-3), 119.0 (C-1′′′~6′′′), 107.6 (C-5′), 107.1 (C-5), 100.8 (C-3′), 66.4 (C-1′′), 61.0 (C-2-OMe), 56.4(C-4-OMe), 31.7 (C-8), 30.5 (C-4′′), 30.3 (C-2′′), 30.0 (C-7), 29.4 (C-9), 20.8 (C-3′′), 10.0 (C-10). (+)-HR-ESI- MS m / z 619.2953[M-Br] + . Mito-UP302-2: white solid. 1H-NMR (400M, DMSO- d6 ) 8.44 (s, 1H, 2′-OH),7.73~7.92 (m, 15H, H-triphenylphosphine), 6.97 (d, 1H, J = 8.4 Hz, H-6), 6.89 (d, 1H, J = 8.4 Hz, H-6′), 6.66 (d, 1H, J = 8.8 Hz, H-5), 6.45 (d, 1H, J = 2.0 Hz, H-3′), 6.21 (dd, 1H, J = 2.0, 8.4 Hz, H-5′), 3.93 (t, 2H, J = 6.0 Hz, H-1′′),3.71 (s, 3H, 4-OMe), 3.58 (s, 3H, 2-OMe), 2.48 (m, 4H, H-7, H-9), 2.04 (s,3H, 3-Me), 1.88 (m, 2H, H-2′′), 1.66~1.76 (m, 4H, H-8, H-4′′). 13C-NMR (100M, DMSO-d6) C 158.3 (C-4′), 157.7 (C-2′), 157.4 (C-2), 157.3 (C-4), 131.1~135.9 (13C, C-1′′′~6′′′), 130.6 (C-6′), 127.8 (C-1), 127.7 (C-6), 121.6 (C-1′), 119.9 (C-1′′′~6′′′), 119.2 (C-3), 119.0 (C-1′′′~6′′′), 107.6 (C-5), 104.9 (C-5′), 102.9 (C-3′), 66.4 (C-1′′), 61.1 (C-2-OMe), 56.4 (C- 4-OMe), 31.8 (C-8), 30.3 (C-4′′), 30.1 (C-2′′, C-9), 29.7 (C-7), 20.8 (C-3′′), 10.1(C-10). (+)-ESI-MS m / z 619.2954 [M-Br] + . like Figure 1 As shown, the embodiment of the present invention provides a reaction formula for the synthesis method.

[0017] like Figure 2-Figure 6 As shown, the synthetic product Mito-UP302-1 is provided. 1 H-NMR spectrum, 13C-NMR spectrum, HMQC spectrum, HMBC spectrum, and HR-ESI-MS spectrum.

[0018] like Figure 7-Figure 11 As shown, the synthetic product Mito-UP302-2 is provided. 1 H-NMR spectrum, 13 C-NMR spectrum, HMQC spectrum, HMBC spectrum, and HR-ESI-MS spectrum.

[0019] Example 2 CCK8 assay to determine the inhibitory effect of Mito-UP302-1 / 2 on different tumors 1. Cell lines Human colon cancer cell line HT29, human chronic myeloid leukemia cell line K562, human lung adenocarcinoma cell line A549, human esophageal cancer cell line K30, human colon adenocarcinoma cell line LOVO, human colorectal cancer cell line HCT116, mouse colon cancer cell line CT26 2. Implementation steps 1. In a clean bench, HT29 cells, K562 cells, A549 cells, K30 cells, LOVO cells, HCT116 cells, and CT26 cells in the logarithmic growth phase were cultured at 6×10 3 The cells were inoculated at a density of 100 / well in a 96-well plate and cultured overnight in a CO2 incubator at 37°C.

[0020] 2. Mito-UP302-1 / 2 was prepared into a 20mM stock solution with DMSO, and different concentrations of drug-containing solutions were prepared with complete culture medium. The culture medium in the 96-well plate was discarded, and 100μL of the drug solution prepared with complete culture medium was added to each well, and the plates were placed in a 37°C, 5% CO2 incubator for 48h. A control group without drug culture medium was set up, and a blank group with only complete culture medium and no cells or drug culture medium was set up.

[0021] 3. After the culture is completed, observe the survival of each group of cells under an inverted microscope.

[0022] 4. In the clean bench, discard the drug culture solution in each well, add 100 μL PBS to wash once, prepare CCK-8 reagent and serum-free culture medium at a volume ratio of 1:10, add 100 μL reagent to each well of the experimental group, control group, and blank group, place in a 37°C incubator for 1.5-2.5 hours, measure the absorbance (OD value) at 450 nm with an enzyme reader, and calculate the IC value of Mito-UP302-1 / 2 for different tumors with GraphPadPrism. 50 value.

[0023] 3. Experimental Results IC of Mito-UP302-1 / 2 against different tumors 50 value As shown in the figure, Mito-UP302-1 / 2 has significant anti-tumor activity as determined by CCK-8 assay.

[0024] Example 3 Western blot detection of Mito-UP302-1 / 2 selectively inhibiting HSP90α Experimental steps: 1. Discard the culture medium of the drug-treated cells, rinse once with pre-cooled PBS, add 500 μL PBS, scrape off the cells with a cell scraper, place them in an EP tube, and centrifuge at 1000 rpm for 3 minutes.

[0025] 2. Discard the supernatant, add an appropriate amount of RIPA lysis buffer containing PMSF, and gently blow to suspend the cell pellet.

[0026] 3. Disrupt the cells with an ultrasonic disruptor and place on ice for 50 minutes.

[0027] 4. Centrifuge each group of cells at 4°C (12500rpm, 20 minutes). After centrifugation, carefully aspirate the supernatant and transfer it to a new EP tube.

[0028] 5. Protein concentration detection: Take 2μL or 5μL of supernatant, add it to a 96-well plate that has been pre-added with 18μL or 15μL PBS, add it to the prepared BCA working solution, and incubate at 37℃ in the dark for 30 minutes. Use an enzyme reader to measure the absorbance value of each group of proteins at a wavelength of 562nm, and then use the standard curve to calculate the protein concentration of each sample.

[0029] 6. Preparation of protein samples for electrophoresis: Add the corresponding amount of 5× loading buffer to the protein solution, mix gently and heat in a 100℃ metal bath for ten minutes. Cool to room temperature and store at -20℃.

[0030] 7. Prepare electrophoresis fluid, transfer solution (pre-cooled), and 10% SDS-PAGE gel.

[0031] 8. Load 25 μg protein per well, and use the following voltages for electrophoresis: 80 V for concentrated gel and 120 V for separation gel. Use sandwich wet transfer, 250 mA, for 90 minutes.

[0032] Block with 9.5% skim milk for 1-1.5 h, wash with TBST, then incubate with primary antibody and shake on a tilting shaker at 4°C overnight.

[0033] 10. After the primary antibody incubation, place the PVDF membrane in TBST and wash it three times, 15 minutes each time, incubate the secondary antibody at room temperature for 1 hour, and immediately wash it three times with TBST at room temperature after the incubation, 10-15 minutes each time.

[0034] Prepare the luminescent solution at a ratio of 11.1:1, then develop with ECL and analyze the results with ImageJ.

[0035] The experimental results are as follows Fig.12 , Fig.13 As shown, Western Blot was used to determine the levels of HSP90α and Hsp90β proteins in human colorectal cancer cells HT29 after treatment with Mito-UP302-1 / 2 for 24 hours, where HSP90α / HSP90β represented the relative ratio of HSP90α to HSP90β protein expression levels after 24 hours of drug action. The HSP90α / HSP90β ratio was significantly reduced after Mito-UP302-1 treatment, and the results showed that only Mito-UP302-1 had selective inhibition of HSP90α.

[0036] Example 4 To evaluate the inhibitory effect of Mito-UP302-1 on mitochondrial activity in HT29 cells.

[0037] 1. Flow cytometry was used to determine the effect of Mito-UP302-1 on mitochondrial membrane potential in HT29 cells 1. Implementation steps ⑴ The experiment used the Mitochondrial Membrane Potential Detection Kit (TMRE), which is a kit that uses TMRE as a fluorescent probe to detect changes in mitochondrial membrane potential in cells, tissues or purified mitochondria. HT29 cells were cultured at 3.5×10 5 The cells were seeded in a 6-well plate at a density of 100 cells / mL and placed in a CO2 incubator for overnight culture. The Mito-UP302-1 drug solution was prepared in complete culture medium containing 10% fetal bovine serum (FBS) in the experimental group and in DMSO in the control group, and then added to each group for 24 h.

[0038] ⑵ Digest with trypsin and collect each group of cells in a 15ml centrifuge tube. Centrifuge at 600rpm for 5min and discard the supernatant. Wash with pre-cooled PBS and transfer the cells to a 1.5ml centrifuge tube. Try to avoid mechanical damage to the cells during the operation.

[0039] ⑶ Prepare TMRE staining working solution: dilute TMRE according to the ratio of adding 1 ml detection buffer to every 1 µL TMRE (1000X), and mix by pipetting.

[0040] (4) Add 0.5 ml TMRE staining solution to each tube of sample, incubate in a 37°C incubator for 20 min, centrifuge at 600 rpm for 5 min at room temperature, discard the supernatant, and wash the cells twice with 37°C preheated culture medium.

[0041] ⑸Take 0.5 ml of culture medium and gently resuspend the cells. Detect using flow cytometer. Keep on ice before loading. Analyze the data using FlowJo.

[0042] 2. Experimental results Fig.14 As shown, flow cytometry was used to detect the mitochondrial membrane potential of HT29 cells after treatment with Mito-UP302-1, and the results showed that the mitochondrial membrane potential was significantly decreased.

[0043] 2. ELISA was used to detect the ATP concentration in HT29 cells treated with Mito-UP302-1.

[0044] 1. Experimental Procedure ⑴ The experiment uses an ATP content test kit, the principle of which is that creatine kinase catalyzes adenosine triphosphate and creatine to generate phosphocreatine, and the amount of phosphorus generated is detected by phosphomolybdic acid colorimetry to calculate the ATP content. Sample preparation: HT29 cells are treated with Mito-UP302-1 and then collected by centrifugation. ATP in the cells is extracted by lysis on ice using a special lysis buffer. After lysis, the cell debris is removed by centrifugation.

[0045] (2) Preparation of standard curve: According to the reagent manufacturer's instructions, prepare a series of ATP standard solutions with known concentrations for drawing the standard curve.

[0046] ⑶ Preparation of ATP detection working solution: dilute the ATP detection reagent with an appropriate amount of detection buffer.

[0047] (4) ATP concentration determination: Add 100 μL of ATP detection working solution into the detection well and place it at room temperature for 3-5 minutes to ensure that the background ATP is consumed.

[0048] ⑸ Quickly add 10-100 μL of sample or standard, mix gently for 2 seconds using a micropipette, wait for 2 seconds and immediately measure using an ELISA reader.

[0049] ⑹Calculate the ATP concentration in the sample based on the standard curve and analyze the data.

[0050] 2. Experimental results are as follows Fig.15 As shown, the ATP content in HT29 cells treated with Mito-UP302-1, the results showed that Mito-UP302-1 significantly inhibited the ATP content in the cells.

[0051] It can be seen from the above test examples that the compounds of the present invention have significant anti-tumor activity, can inhibit mitochondrial activity, and can selectively inhibit the activity of HSP90α, unlike HSP90β. They have ideal HSP90 subtype selection ability, which expands and supplements the current research status and can be further studied as potential anti-tumor drugs.

[0052] The embodiments described above are merely descriptions of preferred implementation modes of the present invention and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A HSP90α selective inhibitor represented by the following structural formula, characterized in that: Among them, when R1 is , R2 is H, Mito-UP302-1; When R1 is H, R2 is , Mito-UP302-2.

2. Use of a HSP90α selective inhibitor of the structural formula as shown in claim 1 in the preparation of cancer drugs.

3. A method for synthesizing a HSP90α selective inhibitor of the structural formula as shown in claim 1, characterized in that: include: Using UP302 and (4-bromobutyl)triphenylphosphine bromide as raw materials, anhydrous acetonitrile as solvent, sodium iodide and cesium carbonate as catalysts, and stirring and heating at 50°C for 5 hours under nitrogen protection, a selective inhibitor of HSP90α was isolated, wherein the molar ratio of UP302: (4-bromobutyl)triphenylphosphine bromide: cesium carbonate: sodium iodide was 1:1.2:2:0.5; The reaction formula of the synthesis method is: Among them, when R1 is , R2 is H, Mito-UP302-1; When R1 is H, R2 is , Mito-UP302-2.

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