Small molecule lysosome targeting chimera and application thereof
By designing small-molecular lysosome-targeted chimera (eHSPTACs), using the eHSP90-mediated lysosome pathway to degrade secreted proteins and membrane proteins, the problem of lack of targeting of existing PD-L1 inhibitors is solved, and targeted treatment for diseases such as tumors is achieved.
Patent Information
- Application Number
- CN202510507617.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-22
- Publication Date
- 2025-07-29
AI Technical Summary
The existing PD-L1 inhibitors lack targeting in tumor treatment and cannot effectively block the immune escape pathway of tumor cells, resulting in poor treatment effect.
A small molecule lysosome-targeted chimera (eHSPTACs) is developed to induce intracellular enrollment of secreted proteins or membrane proteins into lysosomes for degradation through eHSP90 mediation, and to achieve targeted selective treatment using eHSP90, which is highly expressed in tumor cells.
Targeted treatment of secreted proteins and membrane protein-related diseases such as tumors, immunity and inflammation is achieved, and target proteins are effectively degraded through the lysosomal pathway to improve the therapeutic effect.
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Figure CN120383570A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of pharmaceutical technology, and particularly relates to a small molecule lysosome-targeting chimera and its application. Background Art
[0002] Studies have found that secreted proteins and membrane proteins play crucial roles in the life activities of cells, and the abnormal expression of secreted proteins and membrane proteins is correlated with many diseases.
[0003] Taking tumor diseases as an example, the membrane protein PD-L1 is a protein expressed on the surface of tumor cells. It plays a negative regulatory role by binding to the PD-1 receptor on the surface of T cells. The overexpression of the membrane protein PD-L1 inhibits the activation, differentiation, and proliferation of T cells, enabling tumor cells to escape T cell killing, acquire immune escape, and affecting the treatment effect of tumor patients.
[0004] Therefore, during the clinical treatment of tumor diseases, PD-L1 inhibitors are also a common treatment drug; however, during the process of treating tumors with traditional PD-L1 inhibitors, due to the lack of targeting, the immune escape pathway of tumor cells cannot be effectively blocked, resulting in poor treatment effects.
[0005] In view of this, the inventor believes that for tumors, immune and inflammatory diseases related to secreted proteins or membrane proteins, how to develop a new type of targeted therapeutic drug is a technical problem that needs to be solved urgently by those skilled in the art at present.
[0006] The information disclosed in this background art section is only intended to increase the understanding of the overall background of the present invention, and should not be regarded as an admission or any form of implication that this information constitutes the prior art known to those of ordinary skill in the art. Summary of the Invention
[0007] In view of the above technical problems, the embodiments of the present invention provide a small molecule lysosome-targeting chimera and its application to solve the problems raised in the above background art.
[0008] The design concept of the present invention: The present invention has developed a small molecule lysosome-targeting chimera (eHSPTACs) mediated by eHSP90 (extracellular heat shock protein 90); through research, it has been found that the small molecule lysosome-targeting chimera can induce the formation of a ternary complex of eHSP90-small molecule lysosome-targeting chimera-target protein, and can promote the endocytosis of secreted proteins or membrane proteins, guide them into lysosomes, and then degrade them through the lysosome pathway; since eHSP90 is highly expressed outside tumor cells, this technology has certain tumor cell targeting selectivity, and this technology can be used to treat tumors, immune and inflammatory diseases related to targeted secreted proteins or membrane proteins.
[0009] A small molecule lysosome-targeting chimera, the structural formula of which is shown as follows:
[0010] L1-Linker-L2;
[0011] Wherein, L1 is an HSP90 ligand; L2 is a target protein ligand; Linker is a linker.
[0012] Preferably, the HSP90 ligand is selected from Figure 1 One of L1-1, L1-2, L2-3 in Chinese style.
[0013] Preferably, the target protein includes at least a secreted protein and a membrane protein.
[0014] Preferably, the membrane protein includes at least the membrane protein PD-L1.
[0015] Preferably, the membrane protein ligand is selected from Figure 2 One of L2-1, L2-2, L2-3, L2-4, L2-5 in Chinese style; the secreted protein ligand is selected from L2-6 of the following formula.
[0016] Specifically, L2-1, L2-2, L2-3, L2-4, L2-5 are ligands BMS-8 of PD-L1 and its derivatives; L2-6 is a ligand of an AF488-labeled α-DNP antibody of a secreted protein.
[0017] Preferably, the linker is selected from Figure 3 One of the structures shown in Chinese style;
[0018] Wherein, n is any integer from 0 to 3, m is any integer from 1 to 10, X is any integer from 1 to 10, and y is any integer from 1 to 10.
[0019] Preferably, the structural formula of the small molecule lysosome-targeting chimera is selected from Figure 4 One of the structures shown in Chinese style.
[0020] Use of a small molecule lysosome-targeting chimera as described above in the preparation of a targeted drug.
[0021] Preferably, the targeted drug is used for treating a drug for a disease related to a secreted protein or a membrane protein.
[0022] Preferably, the targeted drug includes a drug for treating tumors, immune diseases or inflammatory diseases.
[0023] The small molecule lysosome-targeting chimera provided by the embodiments of the present invention and its applications have the following beneficial effects: The present invention has developed a small molecule lysosome-targeting chimera based on eHSP90-mediated, which can effectively achieve the lysosomal pathway degradation of secreted proteins and membrane proteins by coupling eHSP90 ligands and secreted protein / membrane protein ligands; this technology can be used to treat diseases related to targeted secreted proteins and membrane proteins, such as tumors, immunity, and inflammation, etc. BRIEF DESCRIPTION OF THE DRAWINGS
[0024] Figure 1 It is the structural formula of the HSP90 ligand in the present invention;
[0025] Figure 2 It is the structural formula of the membrane protein ligand and the secreted protein ligand in the present invention;
[0026] Figure 3 It is the structural formula of the linker in the present invention;
[0027] Figure 4 It is the structural formula of the small molecule lysosome-targeting chimera in the present invention;
[0028] Figure 5 It is the synthetic route diagram of Compound 11 in the present invention;
[0029] Figure 6 It is the synthetic route diagram of Compound dDNP-1 in the present invention;
[0030] Figure 7 It is the synthetic route diagram of Compound dDNP-2 in the present invention;
[0031] Figure 8 It is the synthetic route diagram of Compound dDNP-3 in the present invention;
[0032] Figure 9 It is the synthetic route diagram of Compound BMS-8 in the present invention;
[0033] Figure 10 It is the synthetic route diagram of Compound dPDL1-1 in the present invention;
[0034] Figure 11 It is the synthetic route diagram of Compound dPDL1-2 in the present invention;
[0035] Figure 12 It is the synthetic route diagram of Compound dPDL1-3 in the present invention;
[0036] Figure 13 It is the synthetic route diagram of Compound dPDL1-4 in the present invention;
[0037] Figure 14 It is the synthetic route diagram of Compound dPDL1-5 in the present invention;
[0038] Figure 15 This is the synthetic route diagram of compound dPDL1-6 in the present invention;
[0039] Figure 16 This is a schematic diagram of the mechanism by which small molecule eHSPTACs degrade secreted proteins and membrane proteins;
[0040] Among them, eHSPTACs specifically refer to the small molecule lysosome-targeting chimeras in the present invention;
[0041] Figure 17 This is the effect of dDNPs molecules on the endocytosis and degradation of AF488-labeled α-DNP antibody of secreted proteins;
[0042] Figure 18 This is the effect of dPDL1s molecules on the PD-L1 protein level in Hela, B16F10 and ID8 cells;
[0043] Figure 19 This is the effect of dPDL1-4 on the PD-L1 protein level in Hela cells at different concentrations and times. Specific embodiments
[0044] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative efforts belong to the protection scope of the present invention.
[0045] In view of the above technical problems, the embodiments of the present invention provide a small molecule lysosome-targeting chimera and its application to solve the problems raised in the above background technology.
[0046] I. Specific synthesis examples of compounds
[0047] Preparation Example 1: Preparation of compound dDNP-1
[0048] The specific synthetic route diagram of compound 11 is shown in detail in Figure 5 ; The specific synthetic route diagram of compound dDNP-1 is shown in detail in Figure 6 .
[0049] Synthesis of compound 2
[0050] Under an ice-water bath at 0 °C, phosphorus oxychloride (1.40 mL, 15.02 mmol, 2.5 equiv) was added dropwise to N,N-dimethylformamide (DMF, 6 mL), and stirring was continued for 10 min. This reaction solution was slowly added dropwise to a solution of compound 1 (4-isopropylresorcinol) (907 mg, 5.96 mmol, 1.0 equiv) in DMF (1.5 mL). The reaction was carried out at room temperature for 30 min, and then heated to 50 °C for 1 h. Subsequently, it was cooled to room temperature, and cold aqueous sodium hydroxide solution (2.88 g NaOH dissolved in 9 mL of water) was added. The mixture was stirred vigorously for 10 min and then heated to 70 °C for 15 min. After the reaction was cooled to room temperature, it was adjusted to pH 2 - 3 with concentrated hydrochloric acid and stirred for another 1 h. The reaction system was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation to obtain 815 mg of compound 2 as a white solid with a yield of 76%.
[0051] Synthesis of compound 3
[0052] At room temperature, benzyl bromide (BnBr, 0.79 mL, 6.65 mmol, 2.4 equiv) was slowly added dropwise to a solution of compound 2 (500 mg, 2.7 mmol, 1.0 equiv) and K2CO3 (1.15 g, 8.31 mmol, 3.0 equiv) in acetonitrile (4 mL). The reaction solution was heated to 75 °C and stirred overnight. Water was added to the reaction solution to precipitate a solid, which was filtered. The filter cake was rinsed with petroleum ether (PE), dried, and 633 mg of compound 3 as a yellow solid was obtained with a yield of 65%.
[0053] Synthesis of compound 5
[0054] Iron powder (Fe, 1.74 g, 31.11 mmol, 5.0 equiv) and ammonium chloride (NH4Cl, 2.33 g, 43.54 mmol, 7.0 equiv) were slowly added to a solution of compound 4 (2.00 g, 6.22 mmol, 1.0 equiv) in water / ethanol (6 mL H2O and 18 mL EtOH). The reaction mixture was stirred at 75 °C for 4 h. After the reaction was completed, it was cooled to room temperature. The mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine solution, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation to obtain 1.67 g of compound 5 as a light brown solid with a yield of 92%.
[0055] Synthesis of compound 6
[0056] At 0 °C, a solution of phenyl chloroformate (1.07 g, 6.84 mmol, 1.2 equiv) in dichloromethane (DCM, 1 mL) was added dropwise to a reaction solution of compound 5 (1.67 mg, 5.73 mmol, 1 equiv) and triethylamine (TEA, 2.67 g, 17.2 mmol, 3 equiv) in dichloromethane (DCM, 14 mL). Subsequently, the reaction was carried out at room temperature overnight. After the reaction was completed, the solvent was removed by rotary evaporation, and then purification was carried out by flash column chromatography (PE:EA, 1:1 - 1:3) to obtain 1.82 g of compound 6, a light yellow solid, with a yield of 42%.
[0057] Synthesis of Compound 7
[0058] At room temperature, hydrazine hydrate (971 mg, 19.42 mmol, 5.0 equiv) was added to a solution of compound 6 (1.6 g, 3.88 mmol, 1.0 equiv) in 1,4 - dioxane (15 mL). Then the reaction solution was stirred in an oil bath at 102 °C for 4 h. After the reaction was completed, extraction was carried out with dichloromethane, the organic phases were combined, the solvent was removed by rotary evaporation, and separation and purification were carried out by flash column chromatography (DCM:MeOH, 20:1 - 8:1) to obtain 651 mg of compound 7, a white solid, with a yield of 49%. 1 H NMR (400 MHz, DMSO - d6), δ 8.57 (s, 1H), 7.44 (d, J = 8.1 Hz, 2H), 7.34 (s, 1H), 7.13 (d, J = 8.2 Hz, 2H), 4.33 (s, 2H), 3.38 (s, 2H), 3.28 (t, J = 5.4 Hz, 4H), 2.27 (t, J = 4.9 Hz, 4H), 1.38 (s, 9H).
[0059] Synthesis of Compound 8
[0060] At room temperature, glacial acetic acid (AcOH, 32 mg, 0.54 mmol, 0.6 equiv) was added to a solution of compound 7 (313 mg, 0.90 mmol, 1.0 equiv.) and compound 3 (323 mg, 0.90 mmol, 1.0 equiv) in ethanol (EtOH, 6 mL). The reaction solution was stirred in an oil bath at 80 °C overnight. After the reaction was completed, extraction was carried out three times with ethyl acetate, the organic phases were combined, washed with saturated brine solution, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation to obtain 558 mg of compound 8, a light yellow solid, with a yield of 89%.
[0061] Synthesis of Compound 9
[0062] At room temperature, potassium ferricyanide (K3Fe(CN)3, 800 mg, 2.43 mmol, 3.0 equiv) and sodium hydroxide (NaOH, 97 mg, 2.43 mmol, 3.0 equiv) were added to a solution of product 8 (558 mg, 0.81 mmol, 1.0 equiv) in ethanol (5 ml). The reaction solution was stirred in an oil bath at 95 °C for 5 h. After the reaction was completed, saturated ammonium chloride aqueous solution was added, and then extracted three times with ethyl acetate. The organic phases were combined, washed with saturated brine solution, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation. Then, it was separated and purified by flash column chromatography (PE:EA, 3:1 - 1:1) to obtain 437 mg of compound 9, a light yellow solid, with a yield of 78%. 1 1H NMR (400 MHz, DMSO-d6), δ 11.98 (s, 1H), 7.43–7.29 (m, 8H), 7.24–7.11 (m, 5H), 6.96 (d, J = 8.2 Hz, 2H), 5.08 (s, 2H), 4.81 (s, 2H), 3.45 (s, 2H), 3.28 (s, 4H), 3.14 - 3.19 (m, 1H), 2.27 (t, J = 5.0 Hz, 4H), 1.38 (s, 9H), 1.07 (d, J = 6.9 Hz, 6H).
[0063] Synthesis of compound 10
[0064] At room temperature, Pd / C (87.4 mg, 10% Pd content) was added to a solution of compound 9 (437 mg, 0.63 mmol, 1.0 equiv) in tetrahydrofuran (THF, 3 mL). Subsequently, it was placed under 1 atm of hydrogen (H2) and stirred at room temperature overnight. After the reaction was completed, it was separated and purified by flash column chromatography (DCM:MeOH, 30:1 - 10:1) to obtain 305 mg of compound 10, a white solid, with a yield of 95%.
[0065] Synthesis of compound 11
[0066] At room temperature, trifluoroacetic acid (TFA, 1 mL) was added to a solution of compound 10 (200 mg, 0.39 mmol) in dichloromethane (DCM, 2 mL). The reaction was stirred at room temperature for 1 h. After the reaction was completed, the solvent was removed by rotary evaporation, and then it was separated and purified by reverse-phase C18 flash chromatography (water containing 0.1% TFA and methanol as the mobile phase, methanol concentration gradient from 5% - 100% in 0 - 20 min) to obtain 197 mg of compound 11, in the form of TFA salt, a white solid, with a yield of 79%. 11H NMR (400 MHz, DMSO-d6) δ 11.96 (s, 1H), 9.65 (br, 3H), 9.33 (br, 1H), 7.62 (d, J = 7.9 Hz, 2H), 7.24 (d, J = 8.1 Hz, 2H), 6.92 (s, 1H), 6.31 (s, 1H), 4.34 (s, 2H), 3.80–3.57 (m, 4H), 3.53–3.35 (m, 4H), 3.01 (q, J = 6.8 Hz, 1H), 1.04 (d, J = 6.9 Hz, 6H).
[0067] Synthesis of Compound 12a
[0068] At room temperature, triethylamine (TEA, 41 mg, 0.40 mmol, 2.0 equiv) and 1-chloro-2,4-dinitrobenzene (49 mg, 0.24 mmol, 1.2 equiv) were added to a solution of tert-butyl 3-(2-aminoethoxy)propionate (38 mg, 0.20 mmol, 1.0 equiv) in 1,4-dioxane (2 mL). Then the reaction solution was stirred at room temperature for 16 h. After the reaction was completed, the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation. Then it was separated and purified by flash column chromatography (PE / EA, 4:1 - 2:1) to obtain 64 mg of Compound 12, a yellow oil, with a yield of 90%. 1 1H NMR (400 MHz, Chloroform-d) δ 9.15 (d, J = 2.7 Hz, 1H), 8.76 (br, 1H), 8.27 (dd, J = 9.4, 2.6 Hz, 1H), 6.95 (d, J = 9.5 Hz, 1H), 3.87–3.71 (m, 4H), 3.59 (q, J = 5.2 Hz, 2H), 2.53 (t, J = 6.3 Hz, 2H), 1.44 (s, 9H). HRMS (ESI-TOF) calcd for C 15 H 21 N3NaO7 + [M+Na] + 378.1272, found 378.1273.
[0069] Synthesis of Compound dDNP-1
[0070] At room temperature, TFA (0.5 mL) was added to a solution of compound 12a (8 mg, 0.022 mmol, 1.1 equiv.) in DCM (1 mL). The reaction solution was stirred at room temperature for 2 h and then concentrated under reduced pressure to obtain intermediate compound 13a without further purification. Further, intermediate compound 13a was dissolved in DMSO (1 mL), and compound 11 (10 mg, 0.02 mmol, 1.0 equiv), HATU (8 mg, 0.022 mmol, 1.1 equiv) and DIPEA (13 mg, 0.10 mmol, 5.0 equiv) were added successively. After the reaction solution was stirred at room temperature for 18 h, it was purified by reverse-phase C18 flash chromatography (using water containing 0.1% trifluoroacetic acid and methanol as the mobile phase, with a methanol concentration gradient of 5% - 100% from 0 - 20 min) to obtain 8 mg of compound dDNP-1 in the form of the TFA salt, as a yellow solid, with a two-step yield of 47%. 1 H NMR (400 MHz, Methanol-d4) δ 9.03 (d, J = 2.5 Hz, 1H), 8.28 (dd, J = 9.6, 2.6 Hz, 1H), 7.44 (d, J = 7.9 Hz, 2H), 7.29–7.23 (m, 2H), 7.19 (d, J = 9.6 Hz, 1H), 6.73 (s, 1H), 6.28 (s, 1H), 3.85–3.71 (m, 4H), 3.65 (t, J = 5.1 Hz, 2H), 3.63–3.54 (m, 6H), 3.02 (p, J = 6.9 Hz, 1H), 2.68 (t, J = 6.0 Hz, 2H), 2.50–2.39 (m, 4H), 0.91 (d, J = 6.9 Hz, 6H). HRMS (ESI-TOF) calcd for C 33 H 39 N8O9 + [M + H] + 691.2835, found 691.2843.
[0071] Preparation Example 2: Preparation of Compound dDNP-2
[0072] For the specific synthetic route diagram of compound dDNP-2, see Figure 7 .
[0073] Synthesis of Compound 12b
[0074] According to the synthetic method of compound 12a, using tert-butyl 3-[2-(2-aminoethoxy)ethoxy]propionate (47 mg, 0.20 mmol) and 1-chloro-2,4-dinitrobenzene (49 mg, 0.24 mmol) as raw materials, 65 mg of compound 12b was obtained as a yellow oil, with a yield of 82%.1 H NMR (400 MHz, Chloroform-d) δ 9.15 (d, J=2.7 Hz, 1H), 8.80 (br, 1H), 8.27 (dd, J=9.5, 2.7 Hz, 1H), 6.95 (d, J=9.5 Hz, 1H), 3.83 (t, J=5.2 Hz, 2H), 3.77–3.63 (m, 6H), 3.59 (q, J=5.2 Hz, 2H), 2.51 (t, J=6.4 Hz, 2H), 1.44 (s, 9H). HRMS (ESI-TOF) calcd for C 17 H 25 N3NaO8 + [M+Na] + 422.1534, found 422.1542.
[0075] Synthesis of Compound dDNP-2
[0076] According to the synthesis method of compound dDNP-1, using compound 12b (9 mg, 0.022 mmol) as the raw material, the tert-butyl group was removed under acidic conditions and then coupled with compound 11 to obtain 8 mg of compound dDNP-2, a yellow solid, with a two-step yield of 48%. 1 H NMR (400 MHz, Methanol-d4) δ 9.02 (d, J=2.7 Hz, 1H), 8.27 (dd, J=9.6, 2.7 Hz, 1H), 7.46 (d, J=7.8 Hz, 2H), 7.27 (d, J=8.5 Hz, 2H), 7.19 (d, J=9.6 Hz, 1H), 6.72 (s, 1H), 6.27 (s, 1H), 3.80 (t, J=5.1 Hz, 2H), 3.75 (t, J=6.3 Hz, 2H), 3.72–3.54 (m, 12H), 3.01 (p, J=6.9 Hz, 1H), 2.66 (t, J=6.3 Hz, 2H), 2.53–2.40 (m, 4H), 0.91 (d, J=6.8 Hz, 6H). HRMS (ESI-TOF) calcd for C 35 H 43 N8O 10 + [M+H] + 735.3097, found 735.3110.
[0077] Preparation Example 3: Preparation of Compound dDNP-3
[0078] The specific synthetic route diagram of compound dDNP-3 is shown in detail in Figure 8 .
[0079] Synthesis of Compound 12c (DNP-ester)
[0080] According to the synthesis method of Compound 12a, using tert-butyl 3-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]propionate (56 mg, 0.20 mmol) and 1-chloro-2,4-dinitrobenzene (49 mg, 0.24 mmol) as raw materials, 54 mg of Compound 12c (DNP-ester) was obtained as a yellow oil, with a yield of 60%. 1 H NMR (400 MHz, Chloroform-d) δ 9.15 (d, J = 2.7 Hz, 1H), 8.80 (s, 1H), 8.27 (dd, J = 9.5, 2.7 Hz, 1H), 6.96 (d, J = 9.5 Hz, 1H), 3.83 (t, J = 5.2 Hz, 2H), 3.75–3.55 (m, 12H), 2.49 (t, J = 6.5 Hz, 2H), 1.44 (s, 9H). HRMS (ESI-TOF) calcd for C 19 H 29 N3NaO9 + [M+Na] + 466.1796, found 466.1797.
[0081] Synthesis of Compound dDNP-3
[0082] According to the synthesis method of Compound dDNP-1, using Compound 12c (10 mg, 0.022 mmol) as the raw material, the tert-butyl group was removed under acidic conditions and then coupled with Compound 11 to obtain 8 mg of Compound dDNP-3 as a yellow solid, with a two-step yield of 47%. 1 HNMR (400 MHz, Methanol-d4) δ 9.02 (d, J = 2.7 Hz, 1H), 8.27 (dd, J = 9.6, 2.7 Hz, 1H), 7.47 (d, J = 8.1 Hz, 2H), 7.27 (d, J = 8.2 Hz, 2H), 7.20 (d, J = 9.6 Hz, 1H), 6.73 (s, 1H), 6.27 (s, 1H), 3.80 (t, J = 5.2 Hz, 2H), 3.74 (t, J = 6.3 Hz, 2H), 3.71–3.54 (m, 16H), 3.02 (p, J = 6.9 Hz, 1H), 2.65 (t, J = 6.3 Hz, 2H), 2.53–2.41 (m, 4H), 0.92 (d, J = 6.9 Hz, 6H). HRMS (ESI-TOF) calcd for C 37 H 46 N8NaO 11 + [M+H]+ 801.3178, found 801.3189.
[0083] Preparation Example 4: Synthesis of Compound dPDL1-1
[0084] For the specific synthetic route diagram of Compound BMS-8, see Figure 9 ; for the specific synthetic route diagram of Compound dPDL1-1, see Figure 10 .
[0085] Synthesis of Compound BMS-8
[0086] At 0 °C, triphenylphosphine (2.04 g, 7.78 mmol, 1.1 equiv), 3-(hydroxymethyl)-2-methylbiphenyl (1.42 g, 7.04 mmol, 1.0 equiv), and DIAD (1.47 mL, 7.78 mmol, 1.1 equiv) were successively added to a solution of 3-bromo-4-hydroxybenzaldehyde (1.40 g, 7.04 mmol, 1.0 equiv) in THF (80 mL). Subsequently, the reaction mixture was stirred at room temperature overnight, concentrated under reduced pressure, and purified by flash column chromatography (PE:EA, 20:1 - 15:1) to obtain 1.91 g of Compound 14 as a white solid in a yield of 71%. At room temperature, 2-piperidinecarboxylic acid (775 mg, 6 mmol, 3 equiv.) and acetic acid (10 drops of AcOH) were successively added to a solution of Compound 14 (763 mg, 2 mmol, 1.0 equiv) in DMF (10 mL). The reaction mixture was stirred at room temperature for 20 min, and then sodium cyanoborohydride (NaBH(CN)3, 377 mg, 6 mmol, 3 equiv) was added. Then, the reaction mixture was heated to 80 °C and stirred for 18 h. After the reaction was completed, it was concentrated under reduced pressure and purified by flash column chromatography (DCM:MeOH, 20:1 - 15:1) to obtain 277 mg of Compound BMS-8 as a white solid in a yield of 28%. 11H NMR (400 MHz, DMSO-d6) δ 7.59 (d, J = 1.9 Hz, 1H), 7.54 (dd, J = 7.6, 1.4 Hz, 1H), 7.50–7.43 (m, 2H), 7.42–7.35 (m, 1H), 7.35–7.25 (m, 5H), 7.21 (dd, J = 7.7, 1.5 Hz, 1H), 5.23 (s, 2H), 3.82 (d, J = 13.4 Hz, 1H), 3.46 (d, J = 13.4 Hz, 1H), 3.08 (dd, J = 8.1, 4.0 Hz, 1H), 2.91–2.83 (m, 1H), 2.25–2.21 (m, 4H), 1.86–1.76 (m, 1H), 1.76–1.64 (m, 1H), 1.57–1.42 (m, 3H), 1.38–1.31 (m, 1H). HRMS (ESI-TOF) calcd for C 27 H 28 BrNNaO3 + [M+Na] + 516.1145, found 516.1150.
[0087] Synthesis of Compound 15a
[0088] At room temperature, to a solution of compound 11 (21 mg, 0.05 mmol, 1.0 equiv) in DMSO (1 ml) were successively added 3-(2-((tert-butoxycarbonyl)amino)ethoxy)propanoic acid (14 mg, 0.06 mmol, 1.2 equiv), EDCI (19 mg, 0.10 mmol, 2.0 equiv), HOAt (10 mg, 0.075 mmol, 1.5 equiv) and NMM (25 mg, 0.25 mmol, 5.0 equiv). The reaction mixture was stirred at room temperature for 18 h and then purified by reverse-phase C18 flash chromatography (water and methanol containing 0.1% trifluoroacetic acid as the mobile phase, methanol concentration gradient from 5% to 100% in 0 - 20 min) to give 14 mg of compound 15a as the TFA salt, a white solid, in 45% yield. 11H NMR (400 MHz, Chloroform-d) δ 7.51–7.42 (m, 2H), 7.31–7.26 (m, 2H), 6.44 (s, 1H), 6.40 (s, 1H), 5.15 (s, 1H), 3.84–3.41 (m, 10H), 3.30 (s, 2H), 2.91 (p, J=7.1 Hz, 1H), 2.67–2.56 (m, 2H), 2.53–2.36 (m, 4H), 1.44 (s, 9H), 0.70 (d, J=6.7 Hz, 6H). HRMS (ESI-TOF) calcd for C 32 H 44 N6NaO7+ [M+Na] + 647.3164, found 647.3169.
[0089] Synthesis of Compound dPDL1-1
[0090] At room temperature, to a solution of compound 15a (10 mg, 0.016 mmol, 1.0 equiv) in MeOH (0.5 mL) was added a MeOH solution of 4 M HCl (0.5 mL). The reaction solution was stirred at room temperature for 4 h, and then concentrated under reduced pressure to obtain intermediate compound 16a without further purification. At room temperature, to the solution of the above-obtained intermediate compound 16a in DMSO (1 mL) were added BMS-8 (10 mg, 0.019 mmol, 1.2 equiv), EDCI (6 mg, 0.032 mmol, 2.0 equiv), HOAt (3 mg, 0.024 mmol, 1.5 equiv) and NMM (10 mg, 0.096 mmol, 6.0 equiv). The mixture was stirred at room temperature for 18 h, and then purified by reverse-phase C18 flash chromatography (using water containing 0.1% trifluoroacetic acid and methanol as the mobile phase, with a methanol concentration gradient from 5% to 100% in 0 - 20 min) to obtain 13 mg of compound dPDL1-1 in the form of the TFA salt, as a white solid, with a two-step yield of 68%. 11H NMR (400 MHz, Methanol-d4) δ 7.66 (d, J = 2.2 Hz, 1H), 7.46 (d, J = 8.1 Hz, 2H), 7.42–7.06 (m, 12H), 6.77 (s, 1H), 6.12 (s, 1H), 5.17 (s, 2H), 4.28 (s, 2H), 4.07–3.87 (m, 3H), 3.72–3.61 (m, 4H), 3.53–3.07 (m, 10H), 3.02–2.86 (m, 2H), 2.58 (t, J = 5.9 Hz, 2H), 2.15 (s, 3H), 2.05 (d, J = 14.0 Hz, 1H), 1.85–1.37 (m, 6H), 0.91 (d, J = 6.8 Hz, 6H). HRMS (ESI-TOF) calcd for C 54 H 62 BrN7NaO7 + [M+Na] + 1022.3786, found 1022.3790.
[0091] Preparation Example 5: Synthesis of Compound dPDL1-2
[0092] For the specific synthetic route diagram of Compound dPDL1-2, see Figure 11 .
[0093] Synthesis of Compound 15b
[0094] According to the synthetic method of Compound 15a, using Compound 11 (21 mg, 0.05 mmol, 1.0 equiv) and N-Boc-diglycolic acid (17 mg, 0.06 mmol, 1.2 equiv) as raw materials, Compound 15b was obtained as a white solid (17 mg, yield 50%). 1 1H NMR (400 MHz, Chloroform-d) δ 7.48 (d, J = 7.8 Hz, 2H), 7.31 (d, J = 7.8 Hz, 2H), 6.47 (s, 1H), 6.39 (s, 1H), 5.17 (s, 1H), 3.81 (t, J = 6.6 Hz, 2H), 3.69–3.42 (m, 12H), 3.39–3.21 (m, 2H), 2.92 (p, J = 6.8 Hz, 1H), 2.65 (t, J = 7.7 Hz, 2H), 2.53–2.35 (m, 4H), 1.44 (d, J = 4.7 Hz, 9H), 0.70 (d, J = 6.7 Hz, 6H). HRMS (ESI-TOF) calcd for C 34 H 48 N6NaO8 + [M+Na] +691.3426, found 691.3436.
[0095] Synthesis of Compound dPDL1-2
[0096] According to the synthesis method of compound dPDL1-1, using compound 15b (11 mg, 0.016 mmol) as the raw material, the Boc protecting group was removed under acidic conditions, and then coupled with compound BMS-8 to obtain 15 mg of compound dPDL1-2 in the form of TFA salt, a white solid, with a two-step yield of 73%. 1 H NMR (400 MHz, Methanol-d4) δ 7.66 (d, J = 2.2 Hz, 1H), 7.47 (d, J = 8.0 Hz, 2H), 7.42–7.07 (m, 12H), 6.78 (s, 1H), 6.12 (s, 1H), 5.17 (s, 2H), 4.28 (d, J = 4.6 Hz, 2H), 3.93 (d, J = 13.1 Hz, 1H), 3.80–3.41 (m, 14H), 3.39–3.07 (m, 6H), 3.03–2.85 (m, 2H), 2.53 (t, J = 6.2 Hz, 2H), 2.15 (s, 3H), 2.05 (d, J = 14.1 Hz, 1H), 1.86–1.34 (m, 6H), 0.91 (d, J = 6.9 Hz, 6H). HRMS (ESI-TOF) calcd for C 56 H 66 BrN7NaO8 + [M+Na] + 1066.4048, found 1066.4052.
[0097] Preparation Example 6: Synthesis of Compound dPDL1-3
[0098] The specific synthetic route diagram of compound dPDL1-3 is shown in detail in Figure 12 .
[0099] Synthesis of Compound 15c
[0100] According to the synthesis method of compound 15a, using compound 11 (21 mg, 0.05 mmol, 1.0 equiv) and 2,2-dimethyl-4-oxo-3,8,11,14-tetraoxa-5-azapentadecanoic acid (19 mg, 0.06 mmol, 1.2 equiv) as raw materials, compound 15c was obtained as a white solid of 21 mg with a yield of 52%. 11H NMR (400 MHz, Methanol-d4) δ 7.58 (d, J = 8.1 Hz, 2H), 7.39 (d, J = 8.1 Hz, 2H), 6.90 (s, 1H), 6.22 (s, 1H), 4.40 (s, 2H), 3.76 (t, J = 6.0 Hz, 2H), 3.68–3.55 (m, 12H), 3.49 (t, J = 5.7 Hz, 2H), 3.38–3.25 (m, 4H), 3.19 (t, J = 5.7 Hz, 2H), 3.08 (p, J = 6.9 Hz, 1H), 2.70 (t, J = 6.1 Hz, 2H), 1.43 (s, 9H), 1.03 (d, J = 6.9 Hz, 6H). HRMS (ESI-TOF) calcd for C 36 H 52 N6NaO9+ [M+Na] + 735.3688, found 735.3696.
[0101] Synthesis of Compound dPDL1-3
[0102] According to the synthesis method of compound dPDL1-1, using compound 15c (11 mg, 0.013 mmol) as the raw material, the Boc protecting group was removed under acidic conditions and then coupled with compound BMS-8 to obtain 12 mg of compound dPDL1-3 in the form of TFA salt, a white solid, with a two-step yield of 69%. 1 1H NMR (400 MHz, Methanol-d4) δ 7.81 (d, J = 2.2 Hz, 1H), 7.62 (d, J = 8.1 Hz, 2H), 7.57–7.37 (m, 7H), 7.37–7.22 (m, 5H), 6.93 (s, 1H), 6.28 (s, 1H), 5.33 (s, 2H), 4.42 (s, 2H), 4.08 (d, J = 13.2 Hz, 1H), 3.99–3.23 (m, 24H), 3.19–3.02 (m, 2H), 2.72 (t, J = 5.8 Hz, 2H), 2.31 (s, 3H), 2.20 (d, J = 14.0 Hz, 1H), 2.03–1.53 (m, 6H), 1.07 (d, J = 6.9 Hz, 6H). HRMS (ESI-TOF) calcd for C 58 H 71 BrN7O9 + [M+H] + 1088.4491, found 1088.4498.
[0103] Preparation Example 7: Synthesis of Compound dPDL1-4
[0104] The synthetic route diagram of compound dPDL1-4 is specifically shown in Figure 13 .
[0105] Synthesis of compound 15d
[0106] According to the synthesis method of compound 15a, using compound 11 (21 mg, 0.05 mmol, 1.0 equiv) and 2,2-dimethyl-4-oxo-3,8,11,14,17-pentaoxa-5-azadocosane-20-carboxylic acid (22 mg, 0.06 mmol, 1.2 equiv) as raw materials, compound 15d was obtained as a white solid (26 mg, yield 68%). 1 H NMR (400 MHz, Methanol-d4) δ 7.46 (d, J = 8.0 Hz, 2H), 7.26 (d, J = 8.1 Hz, 2H), 6.71 (s, 1H), 6.28 (s, 1H), 3.74 (t, J = 6.2 Hz, 2H), 3.66–3.55 (m, 18H), 3.50 (t, J = 5.6 Hz, 2H), 3.22 (t, J = 5.6 Hz, 2H), 3.02 (p, J = 6.9 Hz, 1H), 2.66 (t, J = 6.2 Hz, 2H), 2.50 (t, J = 4.9 Hz, 2H), 2.44 (t, J = 5.3 Hz, 2H), 1.43 (s, 9H), 0.91 (d, J = 6.9 Hz, 6H). HRMS (ESI-TOF) calcd for C 38 H 56 N6NaO 10 + [M + Na] + 779.3950, found 779.3954.
[0107] Synthesis of compound dPDL1-4
[0108] According to the synthesis method of compound dPDL1-1, using compound 15d (26 mg, 0.034 mmol) as the raw material, the Boc protecting group was removed under acidic conditions and then coupled with compound BMS-8 to obtain 26 mg of compound dPDL1-4 in the form of TFA salt, as a white solid, with a two-step yield of 67%. 11H NMR (400 MHz, DMSO-d6) δ 11.96 (s, 1H), 9.63 (s, 1H), 9.44 (s, 1H), 7.84–7.76 (m, 1H), 7.61 (s, 1H), 7.54 (d, J = 7.6 Hz, 1H), 7.50–7.13 (m, 13H), 6.79 (s, 1H), 6.31 (s, 1H), 5.23 (s, 2H), 3.71 (d, J = 13.1 Hz, 1H), 3.61 (t, J = 6.7 Hz, 2H), 3.55–3.40 (m, 21H), 3.27–3.16 (m, 1H), 3.08–2.94 (m, 2H), 2.73 (dd, J = 22.8, 10.6 Hz, 2H), 2.57–2.52 (m, 2H), 2.38–2.32 (m, 2H), 2.32–2.26 (m, 2H), 2.25 (s, 3H), 1.88 (t, J = 11.3 Hz, 1H), 1.75 (d, J = 12.3 Hz, 1H), 1.66 (d, J = 12.0 Hz, 1H), 1.59–1.48 (m, 2H), 1.37 (d, J = 12.1 Hz, 1H), 1.22 (q, J = 8.8, 6.7 Hz, 1H), 0.96 (d, J = 6.9 Hz, 6H). HRMS (ESI-TOF) calcd for C 60 H 74 BrN7NaO[[ID=…]] 10 + [M+Na] + 1154.4573, found 1154.4580.
[0109] Preparation Example 8: Synthesis of Compound dPDL1-5
[0110] The specific synthetic route diagram of Compound dPDL1-5 is shown in detail in Figure 14 .
[0111] Synthesis of Compound 15e
[0112] According to the synthetic method of Compound 15a, using Compound 11 (21 mg, 0.05 mmol, 1.0 equiv) and 2,2-dimethyl-4-oxo-3,8,11,14,17,20-hexaoxa-5-azatricosane-23-carboxylic acid (25 mg, 0.06 mmol, 1.2 equiv) as raw materials, Compound 15e was obtained as a white solid (30 mg, yield 66%). 11H NMR (400 MHz, Methanol-d4) δ 7.49 (d, J = 8.2 Hz, 2H), 7.30 (d, J = 8.3 Hz, 2H), 6.80 (s, 1H), 6.12 (s, 1H), 4.31 (s, 2H), 3.77–3.34 (m, 24H), 3.24–3.06 (m, 6H), 2.98 (p, J = 6.6 Hz, 1H), 2.65–2.57 (m, 2H), 1.33 (s, 9H), 0.93 (d, J = 6.9 Hz, 6H).
[0113] Synthesis of Compound dPDL1-5
[0114] According to the synthesis method of compound dPDL1-1, using compound 15e (17 mg, 0.02 mmol) as the raw material, the Boc protecting group was removed under acidic conditions and then coupled with compound BMS-8 to obtain 16 mg of compound dPDL1-5 in the form of TFA salt, a white solid, with a two-step yield of 67%. 1 1H NMR (400 MHz, Methanol-d4) δ 7.66 (d, J = 2.2 Hz, 1H), 7.48 (d, J = 8.4 Hz, 2H), 7.42–7.06 (m, 12H), 6.79 (s, 1H), 6.12 (s, 1H), 5.17 (s, 2H), 4.37–4.20 (m, 3H), 3.98–3.41 (m, 29H), 3.38–3.10 (m, 4H), 3.05–2.92 (m, 2H), 2.60–2.53 (m, 2H), 2.16 (s, 3H), 2.05 (d, J = 14.2 Hz, 1H), 1.83–1.38 (m, 5H), 0.92 (d, J = 6.9 Hz, 6H). HRMS (ESI-TOF) calcd for C 62 H 79 BrN7O 11 + [M + H] + 1178.4995, found 1178.5016.
[0115] Preparation Example 9: Synthesis of Compound dPDL1-6
[0116] The specific synthetic route diagram of compound dPDL1-6 is shown in detail in Figure 15 .
[0117] Synthesis of Compound 15f
[0118] According to the synthesis method of compound 15a, using compound 11 (21 mg, 0.05 mmol, 1.0 equiv) and 21-(Boc-amino)-4,7,10,13,16,19-hexaoxaheneicosanoic acid (27 mg, 0.06 mmol, 1.2 equiv) as starting materials, compound 15f was obtained as a white solid (32 mg, yield 66%). 1 H NMR (400 MHz, Methanol-d4) δ 7.49 (d, J = 8.3 Hz, 2H), 7.30 (d, J = 8.4 Hz, 2H), 6.80 (s, 1H), 6.12 (s, 1H), 4.31 (s, 2H), 3.83–3.34 (m, 28H), 3.31–3.07 (m, 6H), 2.98 (p, J = 6.9 Hz, 1H), 2.66–2.54 (m, 2H), 1.33 (s, 9H), 0.93 (d, J = 6.9 Hz, 6H).
[0119] Synthesis of compound dPDL1-6
[0120] According to the synthesis method of compound dPDL1-1, using compound 15f (17 mg, 0.019 mmol) as the starting material, the Boc protecting group was removed under acidic conditions and then coupled with compound BMS-8 to obtain 11 mg of compound dPDL1-6 in the form of TFA salt, as a white solid, with a two-step yield of 48%. 1 H NMR (400 MHz, Methanol-d4) δ 7.66 (d, J = 2.2 Hz, 1H), 7.48 (d, J = 8.4 Hz, 2H), 7.42–7.07 (m, 12H), 6.79 (s, 1H), 6.12 (s, 1H), 5.17 (s, 2H), 4.35–4.21 (m, 3H), 4.02–3.40 (m, 33H), 3.39–3.07 (m, 4H), 3.03–2.86 (m, 2H), 2.60–2.53 (m, 2H), 2.16 (s, 3H), 2.05 (d, J = 14.2 Hz, 1H), 1.86–1.37 (m, 5H), 0.92 (d, J = 6.9 Hz, 6H). HRMS (ESI-TOF) calcd for C 64 H 83 BrN7O 12 + [M + H] + 1220.5278, found 1220.5280.
[0121] II. Examples of Pharmacological Activity Experiments of Compounds
[0122] Steps of Biological Activity Experiments
[0123] Fluorescence microscopy experiment
[0124] Mix 10 μM of DNP-ester (12c), dDNP-1, dDNP-2, and dDNP-3 with AF488-α-DNP antibody (final concentration 100 nM) respectively, and incubate at 37 °C for 6 hours to form binary complexes. Add the above mixture into the culture wells of Hela cells and culture at 37 °C for 12 h. When verifying the effect of lysosome inhibitor chloroquine (CQ) on protein degradation, co-incubate the cells with the above binary complexes and a solution containing CQ (concentration 50 μM) for 12 h. To detect the co-localization of α-DNP antibody with lysosome markers, after removing the cell culture medium, wash the cells once with PBS, and then incubate the cells in a medium containing LysoTracker Red for 30 min. Stain the cell nuclei with Hoechst 33258 for 15 min. Remove the cell culture medium, wash the cells with PBS, and fix with 4% paraformaldehyde (PFA) at room temperature for 30 min. After fixation, wash the cells again with PBS, and permeabilize with a PBS solution containing 0.5% Triton X-100 for 5 min. Then block the cells with Tris buffer containing 0.2% Tween20 and 3% BSA at room temperature for 1 h. Finally, analyze by photographing with a Zeiss LSM900 confocal microscope.
[0125] Western blot experiment
[0126] Culture the cells in a 12-well plate until the density reaches 70%. PD-L1 activity screening ( Figure 18 ) Treat different cells with 10 μM of dPDL1-1, dPDL1-2, dPDL1-3, and dPDL1-4 for 12 h. Concentration-dependent experiment of dPDL1-4 on PD-L1 degradation ( Figure 19 A in), treat HeLa cells with different concentrations of dPDL1-4 for 12 h. Time-dependent experiment of dPDL1-4 on PD-L1 degradation ( Figure 19In part B), HeLa cells were treated with 10 μM d PDL1-4 for different times. Then the cells were collected and lysed on ice for 30 min in urea buffer containing protease inhibitor mixture (TRANSDI111). For protein extraction from in vivo tumor tissues, the homogenized tissues were lysed in urea buffer containing protease inhibitor mixture. The lysate was centrifuged at 12,000 rpm for 10 min, and then the concentration of the protein lysate was determined using Bradford reagent (Sigma-Aldrich, B6916) and boiled for 5 min together with 6× loading buffer. Equal amounts of protein were separated by SDS-PAGE (Epizyme Biotech PG111) and transferred onto a 0.45 μm polyvinylidene difluoride membrane (Millipore). Then the membrane was blocked with 5% non-fat milk (biosharp BS102) in PBS-T buffer at room temperature for 1 h, and subsequently incubated overnight at 4 °C with primary antibodies (PD-L1 antibody, Proteintech 66248-1-Ig, 1:1000; GAPDH antibody, Proteintech 60004-1-Ig, 1:10,000; HSP90 antibody, Proteintech 60318-1-Ig, 1:1000; ATP1A1 antibody, Proteintech 14418-1-AP, 1:4000). The membrane was incubated with horseradish peroxidase (HRP)-labeled anti-rabbit IgG antibody (diluted 1:2000) and anti-mouse IgG antibody (diluted 1:2000) at room temperature for 1 h, and then imaged using Tanon 5200 (Tanon Science & Technology). The degradation level of PD-L1 was analyzed using Image J software.
[0127] Biological data description
[0128] Figure 17 The results showed that:
[0129] In the experiment with HeLa cells, DNP-ester (12c) failed to induce the endocytosis of extracellular AF488-α-DNP antibody. AF488 is a fluorescent label attached to the α-DNP antibody to observe the endocytosis of the antibody. Since no endocytosis occurred, there was no obvious change in the intracellular fluorescence signal, indicating that DNP-ester (12c) has no promoting effect on the endocytosis of this antibody.
[0130] However, dDNP-1 and dDNP-3 enhanced the intracellular fluorescence signal, which means that more fluorescently labeled α-DNP antibodies were taken up by the cells, demonstrating that dDNP-1 and dDNP-3 can effectively promote the endocytosis of extracellular α-DNP antibody.
[0131] In contrast, the cytoplasmic fluorescence intensity of the dDNP-2 treatment group was weak, presumably related to the insufficient efficiency of ternary complex formation.
[0132] Co-incubation with CQ (chloroquine) further enhanced the cytoplasmic fluorescence intensity. CQ is a substance that can inhibit lysosomal function. Normally, lysosomes degrade the α-DNP antibody that enters the cell. When lysosomal function is inhibited (using CQ), antibody degradation decreases and accumulates in the cell, thereby increasing the intracellular fluorescence intensity. And obvious co-localization of the signal with the lysosomal marker LysoTracker was observed, that is, the fluorescence signal of the antibody coincides with the position of the lysosome, which further proves that the α-DNP antibody is degraded through the lysosomal pathway after entering the cell.
[0133] Figure 18 The experimental results showed that in HeLa, B16F10 and ID8 cells, the dPDL1s molecule could promote the degradation of the membrane protein PD-L1, while the PD-L1 inhibitor BMS-8 could not. In ID8 cells, when the cells were treated with 10 μM dPDL1-4 for 12 h, PD-L1 could be degraded to about 30%.
[0134] Figure 19 The experimental results showed that in HeLa cells, dPDL1-4 could degrade the membrane protein PD-L1 in a time-dependent and concentration-dependent manner.
[0135] The embodiments described above are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention should fall within the protection scope determined by the claims of the present invention.
Claims
1. A small molecule lysosome-targeting chimera, characterized in that, The structural formula is shown below: L1-Linker-L2; Among them, L1 is the HSP90 ligand; L2 is the target protein ligand; Linker is the connector.
2. The small molecule lysosome-targeting chimera according to claim 1, wherein The HSP90 ligand is selected from one of the following formulas L1-1, L1-2, and L2-3:
3. The small molecule lysosome-targeting chimera according to claim 1, wherein Target proteins include at least secretory proteins and membrane proteins.
4. The small molecule lysosome targeting chimera according to claim 3, characterized in that The membrane protein includes at least the membrane protein PD-L1.
5. The small molecule lysosome targeting chimera according to claim 3, characterized in that The membrane protein ligand is selected from one of the following formulas L2-1, L2-2, L2-3, L2-4, and L2-5; the secretory protein ligand is selected from the following formula L2-6; 6. The small molecule lysosome-targeting chimera according to claim 1, wherein The linker is selected from one of the following structures: Wherein, n is any integer from 0 to 3, m is any integer from 1 to 10, x is any integer from 1 to 10, and y is any integer from 1 to 10.
7. The small molecule lysosome-targeting chimera according to claim 1, wherein The structural formula of the small molecule lysosome-targeting chimera specifically includes one of the following structures:
8. Use of the small molecule lysosome targeting chimera according to any one of claims 1 to 7 in the preparation of targeted drugs.
9. Use of the small molecule lysosome targeting chimera according to claim 8 in the preparation of targeted drugs or targeted probes, characterized in that: The targeted drug is used to treat diseases associated with secretory proteins or membrane proteins.
10. Use of the small molecule lysosome-targeting chimera according to claim 8 in the preparation of a targeted drug or a targeted probe, characterized in that, The targeted drugs include drugs for treating tumors, immune diseases or inflammatory diseases.