A compound based on crbn ligand-induced ubiquitination of hsp90a, preparation method and use for treating cancer

Abstract

A compound based on CRBN ligand-induced ubiquitination degradation of Hsp90α and its application in cancer treatment. The compound has the structural formula shown as X-Y-Z, where X represents the ligand of HSP90α protein, Z represents the ligand of E3 ligase, and Y represents the chain connecting X and Z. This compound can be used to treat or prevent tumors.

Description

Technical Field

[0001] This invention relates to the field of antitumor drug preparation, specifically to a compound that targets and degrades HSP90α protein, its preparation method, and its application. Background Technology

[0002] Molecular chaperones are a class of proteins that regulate protein homeostasis by assisting in the conformational maturation of nascent polypeptides (called client proteins) and the refolding of denatured proteins. Heat shock proteins (HSPs) are a highly conserved and widely expressed group of molecular chaperone proteins in eukaryotes. The main function of HSPs in the body is to participate in maintaining the correct folding of their client proteins, enabling them to form the conformations required for physiological functions. They play a crucial role in regulating the balance between protein synthesis and degradation, as well as protein localization. Based on homology and molecular weight, heat shock proteins are mainly divided into five families: the HSP90 family (83-90 kDa); the HSP70 family (66-78 kDa); the HSP60 family (approximately 60 kDa); the large molecule HSP family (100-110 kDa); and the small molecule HSP family (15-30 kDa), among which HSP90 is currently a hot research topic.

[0003] HSP90 has four main isoforms: HSP90α and HSP90β are primarily found in the cytoplasm; glucose-regulated protein 94 (Grp94) is located in the endoplasmic reticulum; and tumor necrosis factor receptor-associated protein 1 (Trap1) is located in the mitochondrial matrix. The four isoforms have almost identical functions. The most widely distributed isoforms in the human body are HSP90α and HSP90β, which exist primarily as homodimers within cells and consist of three main domains: an N-terminal domain, a middle domain, and a C-terminal domain. In tumor cells, HSP90 is overexpressed. The N-terminal domain binds to and hydrolyzes ATP to provide energy. The three domains work synergistically to act as molecular chaperones for HSP90, folding, transporting, and assembling its client proteins. Current research indicates that HSP90 has more than 280 client proteins, many of which are expression products or signal transduction factors in tumor-related signaling pathways, such as Her2, EGFR, Raf-1, and Akt.

[0004] Furthermore, studies have shown that HSP90α expression is more specific in tumor cells compared to HSP90β, and plasma HSP90α levels in clinical cancer patients are positively correlated with tumor malignancy, making it a potential diagnostic marker for malignant tumors. HSP90α not only plays a role in tissue repair in normal cells but also promotes tumor cell migration and metastasis; blocking HSP90α secretion can significantly inhibit tumor metastasis. Daniel G's research found that HSP90α can interact with plasminogen in vitro to form plasmin, increasing plasmin-dependent cell motility and enhancing cancer cell invasiveness. Tze S's research group discovered that extracellular HSP90α participates in the CD91 / 1κK / NF-κB cascade reaction, inducing overexpression of TCF12 protein, leading to cadherin downregulation and enhanced invasiveness of colon cancer cells.

[0005] The above studies indicate that HSP90 protein promotes tumor cell growth and metastasis through different mechanisms, and inhibits or reduces the levels of HSP90 inside and outside tumor cells, especially the extracellular HSP90α level, which helps to inhibit tumor cell growth and metastasis. Therefore, using PROTAC technology to reduce the level of HSP90α protein can play a key role in killing tumor cells, but there are currently no related reports. Summary of the Invention

[0006] To overcome the toxic side effects caused by the pan-subtype inhibition of existing HSP90 inhibitors, this invention provides compounds based on CRBN ligand-induced ubiquitination and degradation of Hsp90α, and their application in cancer treatment. These compounds utilize the PROTAC (proteolysis-targeting chimeras) technology to induce HSP90α protein degradation through the ubiquitin-proteasome system, thereby killing tumor cells. One end of these molecules targets and binds to the E3 ligase CRBN, while the other end targets and binds to the HSP90α protein, with the two ends linked by a linker to form a bifunctional molecule. This compound ubiquitinates the target protein via the E3 ligase and guides the target protein into the proteasome degradation system for specific degradation.

[0007] The compound provided by this invention is a selective degrader of HSP90α protein, with no significant effect on other subtypes (HSP90β), and exhibits high safety. Therefore, this compound has potential application value in the field of anti-tumor therapy.

[0008] A compound that targets and degrades HSP90α protein, as shown in the formula:

[0009] XYZ(Ⅰ-1)

[0010] Where X represents the ligand of HSP90α protein, Z represents the ligand of E3 ligase, and Y represents the chain connecting X and Z.

[0011] X is a compound of formula II-1, and Z is a compound of formula II-2.

[0012]

[0013]

[0014] Y is a compound represented by formula II-3.

[0015]

[0016] Each n is an independent integer between 2 and 7.

[0017]

[0018] Each n is an independent integer between 2 and 4. The compounds described above are as follows:

[0019]

[0020] Equation 1-1, n = 2 (X10c)

[0021]

[0022] Equation 1-2, n = 3 (X10d)

[0023]

[0024] Equation 1-3, n = 4 (X10e)

[0025]

[0026] Equation 1-4, n = 5 (X10f)

[0027]

[0028] Equation 1-5, n = 6 (X 10g)

[0029]

[0030] Equations 1-6, n = 7 (X10h)

[0031]

[0032] Equation 1-7, n = 2 (AP2)

[0033]

[0034] Equation 1-8, n = 3 (AP3)

[0035]

[0036] Equation 1-9, n = 4 (AP4).

[0037] A compound based on CRBN ligand-induced ubiquitination degradation of Hsp90α, the process is as follows:

[0038]

[0039] n = 2, 3, 4, 5, 6, 7

[0040]

[0041] Synthetic routes of x10 series compounds: (a) KI, K2CO3, MeCN, 14 h, 65 °C; (b) MeOH, Pb / C, H2, 12 h, 45 °C

[0042]

[0043] Synthetic routes of AP series compounds: (a) DIPEA, DMSO, 140℃, 1.5h; (b) TFA, DCM, rt14h; (c) HATU, DIPEA, DMF, rt14h; (d) Pb / C, H2, 45℃, 12h. Attached image description:

[0044] Figure 1 This diagram illustrates the degradation of HSP90α protein in the MCF-7 tumor cell line by the compound (X10f) shown in Formulas 1-4 according to the present invention. It demonstrates that compound X10f can significantly degrade the intracellular HSP90α protein level in MCF-7 breast cancer cells; a concentration of 0.2 μM X10f exhibits a degradative effect.

[0045] Figure 2 This diagram illustrates the degradation of HSP90α protein in the MCF-7 tumor cell line by the compound (X10g) shown in Formulas 1-5 of this invention and its related compounds. It demonstrates that compound X10g significantly degrades the intracellular HSP90α protein level in MCF-7 breast cancer cells. At an action time of 8 hours and a concentration of 0.5 μM, compared to other compounds in the same series, X10g exhibits a significant degradation effect on HSP90α while having no significant effect on HSP90β.

[0046] Figure 3This diagram illustrates the dose-response relationship and degradation mechanism of compound X10g (Formula 1-5) on HSP90α protein in the MCF-7 tumor cell line. It shows that compound X10g significantly degrades intracellular HSP90α in MCF-7 breast cancer cells at concentrations of 0.1 μM, 0.2 μM, 0.5 μM, 1 μM, 2 μM, and 5 μM. At an action time of 8 h and a concentration of 0.5 μM, it also exhibits strong degradation effects on total HSP90 protein, HSP90α, and its client protein survivin, while having no significant effect on HSP90β or its client protein CDK4. Furthermore, the competitive binding experiment shows that the selective degradation of HSP90α by X10g disappears in the presence of ligand AT13387, pomalidomide, and the proteasome inhibitor MG132 and E1 ubiquitin activator inhibitor MLN4924, indicating that the selective degradation of HSP90α by X10g is mediated through the ubiquitination proteasome pathway.

[0047] Figure 4 This diagram illustrates the in vivo pharmacokinetic parameters of the compounds (X10g) shown in Formulas 1-5 of this invention. It shows the in vivo half-life (t) of a single 15mg / kg X10g compound administered intraperitoneally to a rat. 1 / 2 The maximum absorbable concentration (C) is 150 min, and it is reached at 36 min. max The concentration was 302 ng / ml.

[0048] Figure 5 This diagram illustrates the in vivo antitumor activity of the compound (X10g) shown in Formulas 1-5 of this invention in a BALB / c mouse model of 4T-1 xenograft tumors. It demonstrates that after 20 days of administration of 60 mg / kg of compound X10g, the tumor inhibition rate reached 53.99%, superior to the AT13387 group; tumor weight decreased significantly compared to the solvent group, reflecting its good in vivo antitumor activity; furthermore, during the administration process, the body weight of mice in the X10g group was not significantly affected compared to the solvent group, while the body weight of mice in the AT13387 group showed a significant decreasing trend. Detailed Implementation

[0049] The embodiments of the present invention will be described in further detail below. These embodiments are exemplary and intended to explain the present invention, and should not be construed as limiting the present invention.

[0050] Example 1: Preparation of compound X9c

[0051] Compound X8 (120 mg, 0.209 mmol) was dissolved in 10 mL of acetonitrile, and potassium iodide (1.2 eq) and potassium carbonate (3 eq) were added. While stirring, a solution of compound 19Bc-h (125 mg, 0.307 mmol) in acetonitrile (5 mL) was added. The mixture was heated to 65 °C and reacted for 14 h. After the reaction was complete, the solid was removed by filtration, washed with dichloromethane / methanol, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain compound X9c as a yellow solid, 87 mg, yield 42.67%, mp: 147.2–148.7 °C. ESI-MS (m / z): 797.60 [M+H] + . 1 H NMR (600MHz, DMSO-D6) δ11.15(d,J=8.1Hz,1H),10.39(s,1H),8.51(d,J=5.4Hz,1H),7.78(t,J=6.6Hz,1H),7.57(d,J=5.1Hz,1H),7.45(t ,J=8.5Hz,3H),7.39(t,J=7.6Hz,3H),7.31(t,J=7.4Hz,1H),7.30–7.25(m,1H),7.20–7.13(m,2H),7.06(d,J=8.4Hz,1H),5.16(s,2H),5.1 2(dt,J=11.7,5.3Hz,1H),4.79(d,J=6.6Hz,2H),4.75(d,J=14.1Hz,2H),3.39(d,J=12.3Hz,2H),3.29–3.26(m,2H),2.93–2.85(m,1H),2.6 5–2.60(m,2H),2.57(d,J=7.8Hz,4H),2.53(d,J=5.6Hz,1H),2.51(d,J=6.0Hz,1H),2.39(s,4H),2.08–2.02(m,1H),1.17(d,J=6.9Hz,6H).

[0052] Compound X9d was prepared in Example 1 using compound X8 (120 mg, 0.256 mmol) and 19Bd (129 mg, 0.307 mmol) to give 76 mg of a yellow solid (X9d), yield 36.65%, mp: 146.1–146.9 °C. ESI-MS (m / z): 811.60 [M+H] + . 1H NMR (400MHz, DMSO-D6) δ11.18(s,1H),9.72(s,1H),8.48(d,J=8.5Hz,1H),7.84(t,J=7.7Hz,1H),7.63(d,J=7.3Hz ,2H),7.46(dt,J=21.1,7.5Hz,6H),7.38–7.16(m,4H),7.11(d,J=8.6Hz,1H),5.20(s,2H),5.15(dd,J=12.4,5.2H z,1H),4.81(d,J=16.1Hz,4H),3.33–3.25(m,2H),2.90(t,J=14.3Hz,1H),2.61(d,J=18.8Hz,2H),2.55(s,1H),2. 43–2.23(m,8H),2.18(s,1H),2.07(d,J=7.0Hz,2H),1.72–1.69(m,1H),1.53–1.47(m,2H),1.21(d,J=6.9Hz,6H).

[0053] Compound X9e was synthesized from Example 1 using X8 (120 mg, 0.256 mmol) and 19Bf (138 mg, 0.307 mmol) via general method B to obtain 96 mg of a yellow solid (X9f), yield 44.73%, mp: 134.5–135.2 °C. ESI-MS (m / z): 839.60 [M+H] + . 1H NMR (400MHz, DMSO-D6) δ11.17(s,1H),9.71(s,1H),8.47(d,J=8.4Hz,1H),7.84(t,J=7.9Hz,1H),7.62(d,J=7.4Hz,1H),7.49(t,J=7.2H z,4H),7.44(t,J=7.4Hz,2H),7.35(dd,J=15.2,7.8Hz,2H),7.21(d,J=8.5Hz,2H),7.11(d,J=8.6Hz,1H),5.20(s,2H),5.15(dd,J=12.8, 5.4Hz,1H),4.82(d,J=14.4Hz,4H),3.48(s,2H),3.30(d,J=6.3Hz,2H),2.93–2.86(m,1H),2.63(d,J=3.3Hz,1H),2.58(s,1H),2.54(d, J=4.4Hz,1H),2.49–2.17(m,8H),2.09–2.03(m,1H),1.67–1.59(m,2H),1.58–1.44(m,2H),1.34(d,J=8.7Hz,2H),1.21(d,J=6.9Hz,6H).

[0054] Compound X9f was synthesized in Example 1 using compound X8 (120 mg, 0.256 mmol) and 19Bf (138 mg, 0.307 mmol) to produce 96 mg of a yellow solid (X9f), yield 44.73%, mp: 134.5–135.2 °C. ESI-MS (m / z): 839.60 [M+H] + . 1H NMR (400MHz, DMSO-D6) δ11.17(s,1H),9.71(s,1H),8.47(d,J=8.4Hz,1H),7.84(t,J=7.9Hz,1H),7.62(d,J=7.4Hz,1H),7.49(t,J=7.2H z,4H),7.44(t,J=7.4Hz,2H),7.35(dd,J=15.2,7.8Hz,2H),7.21(d,J=8.5Hz,2H),7.11(d,J=8.6Hz,1H),5.20(s,2H),5.15(dd,J=12.8, 5.4Hz,1H),4.82(d,J=14.4Hz,4H),3.48(s,2H),3.30(d,J=6.3Hz,2H),2.93–2.86(m,1H),2.63(d,J=3.3Hz,1H),2.58(s,1H),2.54(d, J=4.4Hz,1H),2.49–2.17(m,8H),2.09–2.03(m,1H),1.67–1.59(m,2H),1.58–1.44(m,2H),1.34(d,J=8.7Hz,2H),1.21(d,J=6.9Hz,6H).

[0055] Compound X9 g was synthesized from Compound X8 (120 mg, 0.256 mmol) and 19 B g (142 mg, 0.307 mmol) in Example 1 via general method B to yield 98 mg of a yellow solid (X9 g), with a yield of 44.91%, at a temperature of 133.8–136.4 °C. ESI-MS (m / z): [M+H] + . 1H NMR (400MHz, DMSO-D6) δ11.18(s,1H),9.71(s,1H),8.47(d,J=8.4Hz,1H),7.84(t,J=7.9Hz,1H),7.63(d,J=7.2Hz,1H),7.49(t,J=7.1Hz,4 H),7.44(t,J=7.4Hz,2H),7.38–7.32(m,2H),7.23(d,J=9.3Hz,2H),7.11(d,J=8.6Hz,1H),5.20(s,2H),5.15(dd,J=12.8,5.4Hz,1H),4.83 (d,J=15.1Hz,4H),4.03(q,J=7.1Hz,1H),3.51(d,J=25.1Hz,2H),3.33(d,J=6.8Hz,2H),2.90(td,J=16.9,15.4,5.3Hz,4H),2.64(s,1H),2 .59(s,1H),2.54(d,J=4.4Hz,1H),2.48–2.25(m,4H),2.11–2.02(m,1H),1.68–1.55(m,4H),1.36(d,J=25.2Hz,6H),1.21(d,J=6.9Hz,6H).

[0056] Compound X9h was synthesized in Example 1 from compound X8 (120 mg, 0.256 mmol) and 19Bh (146 mg, 0.307 mmol) to obtain 89 mg of yellow solid (X9h), yield 40.13%, mp: 130.1–132.4 °C%. ESI-MS (m / z): 1H NMR (400MHz, DMSO-D6) δ11.18(s,1H),9.70(s,1H),8.47(d,J=8.4Hz,1H),7.83(t,J=7.9Hz,1H),7.62(d,J=7.2Hz,1H),7.49(t,J=7.0Hz, 4H),7.44(t,J=7.4Hz,2H),7.39–7.32(m,2H),7.22(d,J=8.2Hz,2H),7.11(d,J=8.7Hz,1H),5.20(s,2H),5.15(dd,J=12.8,5.4Hz,1H),4. 83(d,J=14.9Hz,4H),3.49(d,J=13.7Hz,2H),3.30(s,2H),2.96–2.86(m,1H),2.63(d,J=3.7Hz,1H),2.59(d,J=5.6Hz,1H),2.54(d,J=4.5 Hz,1H),2.51–2.19(m,8H),2.10–2.03(m,1H),1.61(d,J=9.3Hz,2H),1.50(d,J=7.5Hz,2H),1.30(d,J=6.2Hz,8H),1.21(d,J=6.9Hz,6H).

[0057] Example 2, Preparation of the compound shown in Formula 1-1

[0058]

[0059] Equation 1-1, n = 2(X10c)

[0060] Compound X9c (87 mg, 0.109 mmol) was dissolved in 10 mL of methanol, and 10% palladium on carbon was added. The mixture was purged with nitrogen three times, followed by purging with hydrogen at 45 °C overnight. After the reaction, the palladium on carbon was removed by diatomaceous earth filtration, the solvent was removed by vacuum evaporation, and the target compound was separated by silica gel column chromatography to obtain 32 mg of the compound, yield 41.56%, mp: 178.2–179.8 °C. HPLC: 96.132%. HRMS (ESI) m / z: [M+H] + calcd for C 39 H 42 N6O7 706.3115, found 707.2219. 1H NMR (400MHz, DMSO) δ11.20(s,1H),10.43(s,1H),9.82(s,1H),8.55(d,J=8.5Hz,1H),7.83(t,J=7.9Hz,1H),7.74–7.6 6(m,3H),7.62(d,J=7.3Hz,1H),7.39(s,1H),7.36–7.28(m,2H),7.21(d,J=8.6Hz,2H),6.85(d,J=8.2Hz,1H),5.16(d d,J=12.7,5.4Hz,1H),4.82(s,4H),4.16–4.12(m,2H),3.51–3.41(m,2H),3.24(q,J=6.9Hz,1H),2.99–2.86(m,1H),2 .75–2.52(m,8H),2.44(s,2H),2.37–2.31(m,1H),2.09(d,J=6.3Hz,1H),1.62(q,J=6.3Hz,1H),1.19(d,J=6.9Hz,6H).

[0061]

[0062] Equation 1-2, n = 3(X10d)

[0063] The compounds shown in Formulas 1-2 were synthesized from compound X9d (76 mg, 0.094 mmol) in Example 2 to obtain 23 mg of a yellow solid, with a yield of 33.97%, mp: 177.4–178.9 °C. HPLC: 97.711%. HRMS (ESI) m / z: [M+H] + calcdfor C 40 H 44 N6O7 720.3271, found 721.3378. 1H NMR (400MHz, DMSO-D6) δ11.18(s,1H),9.83(s,1H),9.71(s,1H),8.48(d,J=8.6Hz,1H),7.84(t,J=7.9Hz,1H),7.62(d, J=7.3Hz,1H),7.39(s,1H),7.32(dd,J=8.2,2.3Hz,1H),7.29–7.13(m,3H),6.84(d,J=8.3Hz,1H),5.15(dd,J=12.7,5.4 Hz,1H),4.80(d,J=9.7Hz,4H),3.49(d,J=19.8Hz,2H),3.22(p,J=6.9Hz,1H),2.90(t,J=13.3Hz,1H),2.63(s,1H),2.5 8(s,1H),2.55(s,2H),2.47(s,2H),2.41–2.19(m,8H),2.08–2.03(m,1H),1.77(d,J=5.4Hz,2H),1.19(d,J=6.9Hz,6H).

[0064]

[0065] Equation 1-3, n = 4(X10e)

[0066] The compounds shown in Formulas 1-3 were synthesized from compound X9e (86 mg, 0.104 mmol) in Example 2 to obtain 35 mg of a yellow solid, yield 41.90%, mp: 176.9–177.2 °C. HPLC: 95.541%. HRMS (ESI) m / z: [M+H] + calcdfor C 41 H 46 N6O7 found 734.3428, found 735.3539. 1H NMR (400MHz, DMSO-D6) δ11.18(s,1H),9.83(s,1H),9.71(s,1H),8.46(d,J=8.4Hz,1H),7.83(t,J=7.9Hz,1H),7.62(d,J=7.2Hz,1H ),7.39(s,1H),7.35–7.28(m,2H),7.20(d,J=5.6Hz,2H),6.84(d,J=8.3Hz,1H),5.15(dd,J=12.7,5.4Hz,1H),4.81(d,J=7.0Hz,4H ),3.50–3.43(m,2H),3.22(q,J=6.9Hz,1H),2.94–2.85(m,1H),2.63(d,J=3.5Hz,1H),2.59(d,J=5.0Hz,1H),2.54(d,J=4.5Hz,2H) ,2.48–2.19(m,8H),2.06(p,J=7.4,6.7Hz,1H),1.66–1.58(m,2H),1.53(d,J=6.7Hz,2H),1.30–1.21(m,2H),1.19(d,J=6.9Hz,6H).

[0067]

[0068] Equation 1-4, n = 5(X10f)

[0069] The compounds shown in Formulas 1-5 were synthesized from compound X9f (86 mg, 0.115 mmol) in Example 2 to obtain 38 mg of a yellow solid, with a yield of 43.34%, at 165.5–167.7 °C. HPLC: 96.560%. HRMS (ESI) m / z: [M+H] + calcd forC 42 H 48 N6O7 748.3584, found 749.3691. 1H NMR (400MHz, DMSO-D6) δ11.17(s,1H),9.82(s,1H),9.71(s,1H),8.47(d,J=8.4Hz,1H),7.83(t,J=7.9Hz,1H),7.62(d,J=7.3Hz,1 H),7.39(s,1H),7.32(dd,J=8.3,2.3Hz,2H),7.25–7.17(m,2H),6.85(d,J=8.2Hz,1H),5.15(dd,J=12.7,5.5Hz,1H),4.81(d,J=8 .5Hz,4H),3.47(s,2H),3.23(p,J=6.9Hz,1H),2.94–2.85(m,1H),2.63(d,J=3.8Hz,1H),2.60–2.57(m,1H),2.54(d,J=4.7Hz,2H) ,2.50–2.19(m,8H),2.10–2.02(m,1H),1.63(q,J=7.4Hz,2H),1.49(d,J=5.3Hz,2H),1.28(d,J=8.6Hz,4H),1.19(d,J=6.9Hz,6H). 13 C NMR(101MHz,DMSO)δ173.23,172.38,170.25,169.82,168.17,167.12,156.61,137.5 6,136.99,136.59,135.69,134.41,131.91,129.00,128.71,127.43,126.73,126.48, 126.20,123.76,123.04,118.80,117.42,114.75,61.72,56.98(2C),55.40,54.83,52 .26(2C),51.28,49.38,36.78,31.42,29.50,26.82,26.42,24.94,22.83(2C),22.47.

[0070]

[0071] Equation 1-5, n = 6 (X 10g)

[0072] The compounds shown in Formulas 1-5 were synthesized from compound X9 g (98 mg, 0.115 mmol) in Example 2 to produce 35 mg of a yellow solid, with a yield of 39.92%, mp: 163.1–164.7 °C. HPLC: 96.728%. HRMS (ESI) m / z: [M+H] + calcdfor C 43 H 50N6O7 762.3741, found 763.3843. 1 H NMR (400MHz, DMSO-D6) δ11.18(s,1H),9.83(s,1H),9.71(s,1H),8.47(d,J=8.4Hz,1H),7.83(t,J=7.9Hz,1H),7.62(d,J=7.2H z,1H),7.39(s,1H),7.36–7.29(m,2H),7.20(s,2H),6.84(d,J=8.2Hz,1H),5.15(dd,J=12.8,5.3Hz,1H),4.81(d,J=10.6Hz,4 H),3.47(s,2H),3.23(p,J=6.9Hz,1H),2.94–2.86(m,1H),2.63(d,J=3.3Hz,1H),2.58(s,1H),2.54(d,J=4.2Hz,2H),2.47–2. 24(m,8H),2.10–2.03(m,1H),1.61(d,J=7.7Hz,2H),1.48–1.39(m,2H),1.37–1.27(m,6H),1.23(s,2H),1.19(d,J=6.8Hz,6H).

[0073]

[0074] Equations 1-6, n = 7(X10h)

[0075] The compounds shown in Formulas 1-6 were synthesized from Compound X9h (89 mg, 0.103 mmol)C in Example 2 to obtain 37 mg of a yellow solid, yield 46.27%, mp: 160.1–161.5 °C. HPLC: 95.361%. 1H NMR (400MHz, DMSO-D6) δ11.18(s,1H),9.83(s,1H),9.70(s,1H),8.47(d,J=8.4Hz,1H),7.83(t,J=7.9Hz,1H),7.62(d,J=7.3Hz,1 H),7.39(s,1H),7.32(dd,J=8.2,2.2Hz,2H),7.22(s,2H),6.84(d,J=8.3Hz,1H),5.15(dd,J=12.8,5.4Hz,1H),4.81(d,J=9.7Hz, 4H),3.47(s,2H),3.23(p,J=6.8Hz,1H),2.95–2.85(m,1H),2.63(d,J=3.8Hz,1H),2.61–2.57(m,1H),2.54(d,J=4.4Hz,2H),2.50 –2.14(m,8H),2.06(dd,J=9.6,4.4Hz,1H),1.66–1.57(m,2H),1.44(d,J=7.4Hz,2H),1.27(d,J=6.5Hz,8H),1.19(d,J=6.9Hz,6H). 13 C NMR (101MHz, DMSO) δ173.23,172.46,170.25,169.82,168.19,167.12,156.63,137.84,137. 02,136.85,136.59,134.41,131.90,129.12,128.78,127.40,126.68,126.49,126.19,123. 83,123.08,118.76,117.38,114.75,67.86,61.46,56.72,55.40,54.81,53.95,52.45,51.9 3,50.68,49.38,36.94,31.42,30.26,28.77,26.82,26.60,25.11,23.71,22.83(2C),22.47.

[0076] Example 3 Preparation of compound Pa2

[0077] 2-(2,6-dioxopyridin-3-yl)4-fluoroisoindole-1,3-dione (100 mg 0.36 mmol), NH2–PEG2-COOtBu (109.73 mg 0.47 mmol), and DIPEA (1.5 mmol) were stirred in DMSO at 145 °C for 1.5 h. The mixture was then poured into water, and the organic phase was extracted with ethyl acetate. After drying with anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. Purification by column chromatography yielded compound Pa2 as a yellow-green oil, 100 mg, yield 56.78%. ESI-MS (m / z): 512.30 [M+Na]+; 528.30 [M+K]+. 1HNMR (600MHz, DMSO-D6) δ11.10 (s, 1H), 7.58 (dd, J=8.5, 7.1Hz, 1H), 7.14 (d, J=8.6Hz, 1H), 7.04(d,J=7.0Hz,1H),6.60(t,J=5.8Hz,1H),5.05(dd,J=12.8,5.5Hz,1H),3.61(t,J=5.5Hz ,4H),3.58(t,J=6.2Hz,6H),3.58–3.51(m,5H),3.54–3.46(m,4H),3.46(q,J=5.6Hz,3H),2. 91–2.84(m,1H),2.63–2.50(m,7H),2.39(t,J=6.2Hz,2H),2.07–1.97(m,1H),1.37(s,11H).

[0078] Compound Pa3, synthesized in Example 3 via 2-(2,6-dioxopyridin-3-yl)4-fluoroisoindole-1,3-dione (100 mg 0.36 mmol) and NH2–PEG3-COOtBu (130.28 mg 0.47 mmol), was a yellow-green oily substance, 130 mg, yield 67.72%. ESI-MS (m / z): 556.340 [M+Na] + 572.30[M+K] +. 1H NMR (600MHz, DMSO-D6) δ11.10(s,1H),7.58(dd,J=8.6,7.1Hz,1H),7.14(d,J=8.6Hz,1H),7.04( d,J=7.0Hz,1H),6.60(t,J=5.8Hz,1H),5.05(dd,J=12.9,5.4Hz,1H),3.61(t,J=5.5Hz,2H),3.5 6(d,J=6.2Hz,2H),3.55(t,J=2.6Hz,2H),3.53–3.51(m,2H),3.50–3.47(m,3H),3.47–3.45(m,3 H),2.94–2.82(m,1H),2.61–2.52(m,2H),2.39(t,J=6.2Hz,2H),2.05–1.98(m,1H),1.37(s,9H).

[0079] Compound Pa4 was synthesized from Example 3 using 2-(2,6-dioxopyridin-3-yl)4-fluoroisoindole-1,3-dione (100 mg 0.36 mmol) and NH2–PEG4-COOtBu (150.97 mg 0.47 mmol) as starting materials via general synthetic method A. Intermediate Pa4 was a yellow-green oil, 150 mg, yield 73.98%. ESI-MS (m / z): 1H NMR (600MHz, DMSO-D6) δ11.20(s,1H),7.68(dd,J=8.3,7.3Hz,1H),7.25(d,J=8.6Hz,1H),7.14( dd,J=7.1,0.6Hz,1H),6.70(t,J=5.9Hz,1H),5.16(dd,J=12.9,5.5Hz,1H),3.72(t,J=5.5Hz,2H ),3.68–3.65(m,4H),3.64–3.62(m,2H),3.60–3.58(m,4H),3.61–3.53(m,11H),3.27(d,J=5.3H z,1H),3.05–2.93(m,1H),2.75–2.65(m,1H),2.65–2.58(m,2H),2.18–2.08(m,1H),1.48(s,9H).

[0080] Example 4: Synthesis of compound Pb2

[0081] Compound Pa2 (100 mg 0.20 mmol) was added to 10 mL of DCM solution, followed by 2 mL of LTFA. The resulting mixture was stirred at room temperature for 14 hours. The solvent was removed under reduced pressure and concentrated under vacuum. Purification was performed by column chromatography to give compound Pb2 as a yellow oil, 85 mg, in a yield of 96.00%. ESI-MS (m / z): 1 H NMR (600MHz, DMSO-D6) δ11.10(s,1H),7.59(dd,J=8.5,7.1Hz,1H),7.15(d,J =8.6Hz,1H),7.04(d,J=7.0Hz,1H),6.61(s,1H),5.06(dd,J=12.9,5.4Hz,1H) ,3.65–3.56(m,6H),3.55(dd,J=5.8,3.3Hz,2H),3.51(t,J=4.5Hz,3H),3.47( q,J=5.6Hz,3H),2.63–2.54(m,2H),2.42(t,J=6.4Hz,2H),2.07–1.98(m,1H).

[0082] Compound Pb3 was synthesized from compound Pa3 (130 mg 0.24 mmol) as a starting material in Example 3. Compound Pb3 was a yellow oily substance, 100 mg, yield 85.96%. ESI-MS (m / z): 1 H NMR (600MHz, DMSO-D6) δ11.09(s,1H),7.60–7.55(m,1H),7.14(d,J=8.7Hz,1H),7.04(d, J=7.0Hz,1H),6.60(t,J=5.9Hz,1H),5.05(dd,J=12.9,5.5Hz,1H),3.62(t,J=5.5Hz,2H) ,3.59–3.55(m,4H),3.53–3.50(m,2H),3.49(dd,J=5.6,3.1Hz,2H),3.46(dd,J=6.4,3.2 Hz, 4H), 2.91–2.84 (m, 1H), 2.62–2.52 (m, 2H), 2.42 (t, J = 6.3Hz, 2H), 2.05–2.00 (m, 1H).

[0083] Compound Pb4 was synthesized in Example 3 from compound Pa4 (100 mg 0.18 mmol). Compound Pb4 was a yellow oil, 85 mg, yield 94.40%. ESI-MS (m / z): 600.35 [M+Na] + 616.30 [M+K] + . 1H NMR (600MHz, DMSO-D6) δ11.10(s,1H),7.58(dd,J=8.6,7.0Hz,1H),7.15(d,J=8.6Hz, 1H),7.04(d,J=7.0Hz,1H),6.61(t,J=5.8Hz,1H),5.05(dd,J=12.9,5.5Hz,1H),3.62( t,J=5.5Hz,2H),3.60–3.55(m,5H),3.54–3.51(m,2H),3.51–3.49(m,3H),3.49–3.45( m,9H),2.88(ddd,J=17.0,13.9,5.4Hz,1H),2.42(t,J=6.3Hz,2H),2.06–2.00(m,1H).

[0084] Example 5, Synthesis of compound APb2

[0085] Compound Pb2 (100 mg 0.23 mmol) was dissolved in 5 mL of LMF solvent. HATU (114 mg 0.299 mmol) was added, and the mixture was stirred at room temperature for half an hour. Then, compound X8 (107.93 mg 0.23 mmol) was added, followed by 0.5 mL of DIPEA. The reaction was carried out at room temperature for 14 hours. After the reaction was complete, water was added to quench the reaction. The mixture was extracted with ethyl acetate, concentrated, and purified by silica gel column chromatography to give compound APb2 as an orange-yellow solid, 123 mg, in a yield of 60.47%. ESI-MS (m / z): 885.50 [M+H] + . 1H NMR (600MHz, DMSO-D6) δ11.06(s,1H),7.55–7.51(m,1H),7.44(dd,J=13.5,7.5Hz,4H),7.39(t,J=7.7Hz,2H),7.32–7.25(m,2H) ,7.18–7.14(m,2H),7.08(dd,J=17.2,8.7Hz,2H),7.01–6.97(m,1H),6.56(d,J=6.4Hz,1H),5.15(s,2H),5.00(dd,J=13.0,5.4Hz ,1H),4.80–4.72(m,4H),3.57(d,J=4.7Hz,4H),3.50(d,J=4.9Hz,2H),3.46(s,4H),3.41(d,J=5.7Hz,4H),3.28(d,J=7.0Hz,1H) ,2.87–2.79(m,1H),2.55(s,1H),2.52(s,1H),2.50(s,2H),2.25(d,J=18.6Hz,4H),1.97(d,J=5.7Hz,1H),1.17(d,J=6.9Hz,6H).

[0086] Compound APb3 was synthesized from compound Pb3 (100 mg 0.21 mmol) and compound X8 (98.55 mg 0.21 mmol) as raw materials in Example 4. The resulting compound APb3 was an orange-yellow solid, 100 mg, with a yield of 51.29%. 1H NMR (600MHz, DMSO-D6) δ11.08(s,1H),7.59–7.53(m,1H),7.47(ddd,J=13.0,6.3,1.5Hz,4H),7.44–7.40(m,2H),7.36–7.27(m,2H),7.19(q,J=8.6 ,7.9Hz,2H),7.14–7.08(m,2H),7.02(d,J=7.0Hz,1H),6.61–6.55(m,1H) ,5.19(s,2H),5.04(dd,J=12.9,5.4Hz,1H),4.82(d,J=4.9Hz,2H),4.78(d ,J=11.1Hz,2H),4.10(q,J=5.3Hz,1H),3.59(dq,J=17.4,6.3,5.8Hz,4H) ,3.54(q,J=2.9Hz,2H),3.53–3.50(m,2H),3.47(d,J=13.0Hz,8H),3.42–3 .37(m,4H),2.91–2.83(m,1H),2.59(s,1H),2.55(d,J=16.1Hz,1H),2.52 (s,2H),2.29(d,J=29.3Hz,4H),2.04–1.96(m,1H),1.20(d,J=6.9Hz,6H).

[0087] Compound APb4 was synthesized from compound Pb4 (150 mg 0.29 mmol) and compound X8 (136.09 mg 0.29 mmol) as starting materials, yielding an orange-yellow solid of 130 mg in a yield of 46.10%. ESI-MS (m / z): 973.45 [M+H] + 995.40 [M+Na] + . 1 H NMR (600MHz, DMSO-D6) δ11.10 (s, 1H), 7.57 (t, J = 7.6Hz, 1H), 7.49–7.46 (m, 4H), 7.42(t,J=7.6Hz,2H),7.36–7.30(m,2H),7.20(q,J=9.5,8.9Hz,2H),7.13(dd,J= 8.7,2.6Hz,1H),7.10(d,J=8.7Hz,1H),7.03(d,J=7.0Hz,1H),6.59(d,J=6.3Hz, 1H),5.19(s,2H),5.05(dd,J=12.9,5.4Hz,1H),4.83(d,J=5.4Hz,2H),4.78(d,J=

[0088] 11.7Hz,2H),3.61–3.57(m,4H),3.55(q,J=3.5,3.1Hz,3H),3.52(q,J=4.5,4.1Hz,3H),3.47(d,J=18.4Hz,14H),3.4 1(s,4H),2.93–2.81(m,1H),2.58(d,J=18.5Hz,1H),2.34–2.22(m,5H),2.00(t,J=9.1Hz,2H),1.20(d,J=7.0Hz,6H).

[0089]

[0090] Equation 1-7, n = 2 (AP2)

[0091] The compounds shown in Formulas 1-7 were synthesized from APb2 (50 mg 0.057 mmol) as a starting material in Example 5. The resulting compound AP2 was a pale yellow solid, 27 mg, with a yield of 60.12%. ESI-MS (m / z): 795.50 [M+H] + 833.45 [M+K] + . 1 H NMR (600MHz, DMSO-D6) δ11.09(s,1H),9.64(s,1H),7.56(dd,J=8.6,7.1Hz,1H),7.23(d,J=51.0Hz,3H),7.13(d,J=8.6Hz,1H),7 .05–7.01(m,2H),6.59(t,J=5.9Hz,1H),6.39(s,1H),5.04(dd,J=12.9,5.4Hz,1H),4.75(s,4H),3.60(q,J=6.6,5.9Hz,4H),3.53 (dd,J=5.8,3.2Hz,2H),3.49(t,J=2.6Hz,3H),3.44(d,J=5.6Hz,7H),3.08(p,J=6.9Hz,1H),2.90–2.83(m,1H),2.59(t,J=3.6Hz, 1H), 2.56 (t, J = 3.3Hz, 1H), 2.52 (s, 1H), 2.47 (d, J = 4.4Hz, 1H), 2.28 (d, J = 24.0Hz, 4H), 2.03–1.98 (m, 1H), 1.13 (d, J = 6.9Hz, 6H).

[0092]

[0093] Equation 1-8, n = 3 (AP3)

[0094] The compounds shown in Formulas 1-8 were synthesized from APb3 (60 mg 0.06 mmol) as a starting material in Example 5. The resulting compound AP3 was a pale yellow solid, 33 mg, with a yield of 60.91%. 1 H NMR (600MHz, DMSO-D6) δ11.09(s,1H),9.80(s,1H),7.60–7.55(m,1H),7.38(s,1H),7.34–7.29(m,2H),7.20(d,J=6.8Hz,2H),7.1 3(d,J=8.6Hz,1H),7.03(d,J=7.0Hz,1H),6.83(d,J=8.3Hz,1H),6.59(t,J=5.8Hz,1H),5.04(dd,J=12.9,5.5Hz,1H),4.86–4.75( m,4H),3.62–3.54(m,6H),3.53–3.50(m,2H),3.48(s,2H),3.46(d,J=3.5Hz,6H),3.41(s,4H),3.22(p,J=7.1Hz,1H),2.91–2.84( m,1H),2.62–2.59(m,1H),2.58–2.53(m,1H),2.52(s,2H),2.29(d,J=28.2Hz,4H),2.04–1.98(m,1H),1.18(d,J=6.9Hz,6H).HPLC purity: 97.89%.

[0095]

[0096] Equation 1-9, n = 4 (AP4)

[0097] The compounds shown in Formulas 1-9 were synthesized from APb4 (66 mg 0.07 mmol) in Example 5. Compound AP4 was a pale yellow solid, 30 mg, with a yield of 50.09%. ESI-MS (m / z): 883.50 [M+H]+; 905.45 [M+Na]+. 1H NMR (600MHz, DMSO-D6) δ11.09(s,1H),9.83(s,1H),7.59–7.55(m,1H),7.38(s,1H),7.31(dd,J=8.3,2.3Hz,2H),7.20(t,J=7.7Hz,2H),7.13( d,J=8.6Hz,1H),7.03(d,J=7.0Hz,1H),6.83(d,J=8.3Hz,1H),6.59(t,J=5.8Hz,1H),5.04(dd,J=12.9,5.5Hz,1H),4.85–4.76(m,4H),3.62–3 .57(m,4H),3.55(d,J=4.0Hz,2H),3.52(d,J=4.2Hz,2H),3.49(d,J=6.4Hz,2H),3.45(d,J=4.8Hz,14H),3.22(p,J=6.9Hz,1H),2.92–2.81(m, 1H), 2.59 (t, J = 3.1Hz, 1H), 2.57 (t, J = 3.3Hz, 1H), 2.52 (d, J = 4.3Hz, 2H), 2.29 (d, J = 29.9Hz, 4H), 2.01 (q, J = 5.3Hz, 1H), 1.18 (d, J = 6.9Hz, 6H).

[0098] Example 6: Half-maximal inhibitory rate against tumor cell lines from different sources

[0099] Weigh an appropriate amount of the test compound powder, dissolve it in DMSO to make a final concentration of 10 mM, 10 μL / tube, store at -20℃ protected from light, thaw before use, and dilute to the required concentration.

[0100] Human breast cancer cells MDA-231, MDA-468, and MCF-7 were seeded at a density of 4000 cells / well, and MX-1 cells at a density of 3000 cells / well in 96-well plates, 180 μL per well. The plates were incubated overnight at 37°C, 5% CO2, and saturated humidity. After cell attachment, 20 μL of the target compound (maximum concentration 200 μM, half-diluted) was added to each experimental group. The negative control group received no drug, while the positive controls included AT13387, pomalidomide, and lenalidomide. Each group had three replicates, and incubation lasted 72 hours. 20 μL of 5 mg / mL MTT solution was added to each well, and the plates were incubated for another 4 hours. After culturing, the supernatant was discarded, and 180 μL of LDMSO was added to dissolve the cells. The plates were then incubated at 37°C, 5% CO2, and saturated humidity for 20 minutes. The absorbance (OD value) was measured at 570 nm using a microplate reader. The cell growth inhibition rate is calculated based on absorbance.

[0101] The formula for calculating cell inhibition rate is as follows:

[0102] Cell growth inhibition rate (%) = (OD control group - OD experimental group value) / OD control group × 100%.

[0103] Table 1 shows the half-maximal inhibitory rates of the compounds obtained in this invention against four types of tumor cells. Compounds X10f and X10g have strong proliferative inhibitory activity.

[0104] Table 1. Inhibitory effect of the compounds obtained in this invention on tumor cell proliferation.

[0105]

[0106] The X10 series compounds exhibited significantly weaker inhibitory activity against the proliferation of four breast cancer cell lines compared to AT13387. A possible reason is that, compared to AT13387, the X10 series PROTAC compounds have only one phenolic hydroxyl group at the HSP90α protein terminus, resulting in poorer solubility. Furthermore, the absence of the phenolic hydroxyl group at position 2 increases selectivity for the α isoform while reducing its effect on other isoforms, which may be a significant reason for the decreased activity.

[0107] X10c showed poor inhibitory effects on all four breast cancer cell lines, while compounds X10f and X10g exhibited relatively good proliferative inhibitory activity.

[0108] The AP series showed significantly higher activity than the X10 series, especially in MCF-7 cells, where its activity was close to or comparable to that of AT13387.

[0109] Example 7: Degradation activity test of the compounds of the present invention against HSP90α

[0110] Preparation of the compound: Dissolve in DMSO to prepare a stock solution of 20 mmol / L and store at -20℃. Thaw before use and dilute with culture medium to the required concentration.

[0111] Total protein extraction: Collect cells, wash twice with PBS, centrifuge at 2000 rpm for 5 minutes, and discard the supernatant by pipette tip. Add an appropriate volume of lysis buffer to the cell pellet and lyse on ice for 30 minutes, vortexing every 10 minutes to ensure complete cell lysis. Centrifuge at 12000 rpm for 15 minutes at 4°C (centrifuge should be pre-cooled), and transfer the supernatant to a new EP tube. Prepare an appropriate amount of BCA working solution (180 μL / sample, A:B = 50:1) and mix thoroughly. Add 120 μL of BSA standard to a 96-well plate and dilute eight times with 60 μL ddH2O to create nine concentration gradients. Dilute the protein sample 10-fold (6 μL + 54 μL ddH2O). Set up two replicates for each standard and sample, adding 180 μL of working solution to each replicate. Next, add 20 μL of standard or sample to the corresponding working solution replicates and incubate at 37°C for 15 min. Measure the absorbance at 570 nm using a microplate reader. Calculate the protein concentration of each sample based on the standard protein curve derived from the standard concentration and absorbance values. Then add 1 / 3 of the sample volume of 4× loading buffer, denature at 98°C for 5 min, aliquot, and store at -20°C.

[0112] Immunoblot analysis: Prepare electrophoresis gels with the appropriate percentage based on protein molecular weight. After solidification, assemble the plate onto the electrophoresis clamp, place it in the electrophoresis tank, add 1× electrophoresis buffer, and carefully remove the comb vertically. Calculate the solution volume for 30 μg of protein based on the protein content; this is the loading amount. Thaw and mix the samples, then add the protein markers and each sample group sequentially. Electrophoresis is performed at a constant voltage of 80V. After the protein markers separate, electrophoresis is performed at a constant voltage of 100V. Stop electrophoresis when the bromophenol blue dye reaches the end of the gel. Then, place the transfer clamp, two sponge pads, and two filter papers in a dish containing pre-cooled transfer buffer. Cut the target protein fraction according to the marker and place it in the transfer clamp. Cut a PVDF membrane to the appropriate size according to the size of the gel strip and immerse it in methanol for at least 30 seconds. Assemble the membrane in the following order: negative electrode clamp – sponge – filter paper – gel block – PVDF membrane – filter paper – sponge – positive electrode clamp, ensuring no air bubbles between each layer. Place the membrane in the transfer tank and fill it with transfer buffer. Place ice outside the tank to cool it down. Transfer at a constant current of 200mA for 120 minutes. After transfer, label the PVDF membrane and place it in 5% skim milk blocking buffer, protein side up, and gently shake on a shaker at room temperature for 1 hour.

[0113] Remove the PVDF membrane strip from the blocking solution and wash it three times with 1×TBST for 5 min each time. Then, prepare a sealing bag according to the size of the strip, place the strip in the bag, add the corresponding primary antibody incubation solution, and incubate overnight at 4°C. Incubate with secondary antibody. After the primary antibody incubation is complete, remove the strip, wash it three times with 1×TBST for 5 min each time, and then incubate it in the secondary antibody incubation solution for 1 h. Prepare Smart-ECL Enhanced (a 1:1 mixture of solutions A and B) developing solution and develop it on a Clinx Science Instruments chemiluminescence analyzer.

[0114] Compound X10f significantly degraded HSP90α protein in MCF-7 cell lines after 4 and 6 hours of treatment, but had almost no effect on HSP90β. The degradation effect decreased after prolonged treatment to 8 and 12 hours, which may be related to compensatory synthesis of HSP90α protein within the cells. After 24 hours of treatment, significant degradation was observed at a concentration of 0.2 μM. Figure 1 This indicates that compound X10f has a significant selective degradation effect on HSP90α.

[0115] Compound X10g significantly degraded HSP90α at a concentration of 0.5 μM after 8 hours of treatment with MCF-7 cell line, and the total amount of HSP90 protein was also affected. The degradation effect was most pronounced at a concentration of 0.2 μM after 24 hours of treatment. Figures 2-5 ).

[0116] Both X10f and X10g exhibited a "hook effect" after 24 hours of treatment. This is likely due to the competitive formation of POI-PROTAC or E3-PROTAC binary complexes at lower or higher concentrations of PROTAC compounds, rather than the formation of degradable POI-PROTAC-E3 ternary complexes.

[0117] Example 8: Pharmacokinetic parameters and in vivo antitumor activity detection of the compounds of the present invention

[0118] Weigh an appropriate amount of the test compound powder and dissolve it in a mixed solvent (DMSO:PEG300:Tween80:NaCl = 10:40:5:45) to make a final concentration of 5 mg / mL.

[0119] Rats were administered a single intraperitoneal injection of 15 mg / kg x 10 g. Following administration, blood samples (300 μL each) were collected from the canthus of the rat at 5, 10, 15, 30, 45, 60, 90, 120, 180, 240, and 480 min. The plasma samples were centrifuged at 8000 rpm and 4℃ for 5 min, and the supernatant (100 μL each) was collected. The supernatant was then diluted with 5 times its volume of methanol and centrifuged at 12000 rpm and 4℃ for 15 min to obtain the final sample. The concentration of x 10 g in the plasma samples was determined by LC-MS.

[0120] To establish a first-generation xenograft tumor model, 1×10⁻⁶ cells were injected into the right axilla of BALB / c mice. 6 Four T-1 cells were injected. When the tumor volume was large enough, the tumor tissue was removed and evenly injected into the right axilla of 24 BALB / c mice using an inoculation needle. Mice with a tumor volume of 50 mm3 were randomly divided into four groups (n=6): saline group (ip), solvent control group (ip), AT13387 group (50 mg / kg, ip, twice a week), and X10g group (60 mg / kg, ip, every day), and the drug administration process lasted for 20 days.

[0121] When a single dose of 15 mg / kg x 10 g is administered intraperitoneally to a rat, its in vivo half-life (t) 1 / 2 The maximum absorbable concentration (C) is 150 min, and it is reached at 36 min. max ) is 302 ng / ml ( Figure 4 After 20 days of drug treatment, both the AT13387 group and the X10g group showed inhibitory effects on tumor growth. Figure 5 The tumor inhibition rate in the X10g group was 53.99%, which was superior to the 43.24% in the AT13387 group. Figure 5 In addition, tumor weight decreased significantly. Figure 5 However, the body weight of mice in the AT13387 group showed a significant decreasing trend, while the body weight of mice in the X10g group remained relatively stable. Figure 5 Furthermore, the mice survived well during the administration process, with no mice dying.

Claims

1. A compound based on CRBN ligand-induced ubiquitination degradation of Hsp90α, characterized in that, The compounds are those represented by formulas 1-1 to 1-9: ； Formula 1-1 ； Formula 1-2 ； Formula 1-3 ； Formula 1-4 ； Formula 1-5 ； Formula 1-6 ； Formula 1-7 ； Formula 1-8 ； Equations 1-9.

2. The method for preparing the compound based on CRBN ligand-induced ubiquitination degradation of Hsp90α according to claim 1, characterized in that, The process is as follows: ； 。 3. The use of the compound of claim 1, which is based on CRBN ligand-induced ubiquitination and degradation of Hsp90α, for the preparation of a drug for treating or preventing tumors, wherein the tumor is breast cancer.

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