A compound, its preparation and use

Abstract

The application discloses a compound and a preparation method and application thereof, and belongs to the technical field of biological medicines. The application provides a compound having a structure as shown in formula I or formula II, the compound has a significant inhibiting effect on NSD3 protein expression and a significant killing and growth inhibiting effect on lung squamous cell carcinoma cells. The compound provides a new scheme for treatment of lung squamous cell carcinoma and has a good application prospect in clinical treatment.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a compound, its preparation method, and its uses. Background Technology

[0002] In recent years, scientists have discovered that histone methyltransferase (NSD3) promotes the development of lung squamous cell cancer and head and neck squamous cell cancer through its enzymatic catalytic activity, providing a potential direction for the clinical treatment of lung squamous cell cancer.

[0003] Chromatin fragment amplification is a common genetic alteration in tumor cells, playing a crucial role in tumorigenesis. Amplification of the p11-12 segment of chromosome 8 (8p11AMP) is one of the most common genetic alterations in tumors, especially occurring frequently in squamous cell carcinoma of the lung. The NSD3 gene is a major gene with high-frequency amplification within the 8p11AMP segment and is also a key driver gene in the development of squamous cell carcinoma of the lung. NSD3 is a histone methyltransferase that specifically mediates the mono- and dimethylation of lysine residue 36 of histone H3 (H3K36me1 / 2). Research has revealed that, in addition to NSD3 gene amplification (derived from 8p11AMP), a mutant with enhanced NSD3 enzyme catalytic activity (NSD3T1232A) exists in squamous cell carcinoma of the lung, and it has been confirmed that the histone methyltransferase activity of NSD3 promotes the tumorigenesis of squamous cell carcinoma of the lung.

[0004] Given that NSD3 enzyme activity affects tumor development and progression, the development of small molecule inhibitors of NSD3 is of great significance for cancer treatment. Summary of the Invention

[0005] In order to solve the above-mentioned problems in the prior art, the purpose of this invention is to provide a compound, its preparation method and its use.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] This invention provides a compound having the structure shown in Formula I or Formula II:

[0008] ;

[0009] ;

[0010] Where m is 0, 1, or 2;

[0011] n can be 0, 1, 2, 3, 4, or 5.

[0012] Furthermore, the compound has a structure as shown in Formula III or Formula IV:

[0013] ;

[0014] .

[0015] The present invention also provides a method for preparing the compound shown in Formula I above, the method comprising the following steps:

[0016] (1) Compound B reacts with activators and organic bases;

[0017] (2) Add compound A to the reaction system of step (1), react, and obtain the compound shown in formula I;

[0018] Among them, compound A is m is 0, 1, or 2; compound B is ;

[0019] The molar ratio of compound A, compound B, activator, and organic base is (0.1~10):(0.1~10):(0.1~10):(0.1~10).

[0020] The solvent for the reaction in step (1) is an organic solvent; the reaction time is 0.1~2 h, and the reaction temperature is -10~10℃;

[0021] The reaction time in step (2) is 0.5 to 5 hours, and the reaction temperature is 15 to 50°C.

[0022] Further, the molar ratio of compound A, compound B, activator, and organic base is 1:1:1.5:3; the activator is 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, and the organic base is N,N-diisopropylethylamine;

[0023] The organic solvent in step (1) is N,N-dimethylformamide; the reaction time is 0.5 h, and the reaction temperature is -4~4℃;

[0024] The reaction time in step (2) is 2 hours, and the reaction temperature is 20~30℃.

[0025] Furthermore, compound A is .

[0026] The present invention also provides a method for preparing the compound shown in Formula II above, the method comprising the following steps:

[0027] (1) Compound B reacts with activators and organic bases;

[0028] (2) Add compound C to the reaction system of step (1), and react to obtain the compound shown in formula II;

[0029] Among them, compound C is n is 0, 1, 2, 3, 4, or 5; compound B is ;

[0030] The molar ratio of compound C, compound B, activator, and organic base is (0.1~10):(0.1~10):(0.1~10):(0.1~10).

[0031] The solvent for the reaction in step (1) is an organic solvent; the reaction time is 0.1~2 h, and the reaction temperature is -10~10℃;

[0032] The reaction time in step (2) is 0.5 to 5 hours, and the reaction temperature is 15 to 50°C.

[0033] Further, the molar ratio of compound C, compound B, activator, and organic base is 1:1:1.5:3; the activator is 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, and the organic base is N,N-diisopropylethylamine;

[0034] The organic solvent in step (1) is N,N-dimethylformamide; the reaction time is 0.5 h, and the reaction temperature is -4~4℃;

[0035] The reaction time in step (2) is 2 hours, and the reaction temperature is 20~30℃.

[0036] Furthermore, compound C is .

[0037] The present invention also provides the use of the above-mentioned compounds in the preparation of medicaments for the prevention and / or treatment of lung cancer.

[0038] Furthermore, the lung cancer mentioned is squamous cell carcinoma of the lung.

[0039] The present invention has achieved the following beneficial effects:

[0040] This invention provides a compound that significantly inhibits NSD3 protein expression and exhibits significant killing and growth-inhibiting effects on lung squamous cell carcinoma cells. This compound, as a small-molecule NSD3 degrader, offers a novel treatment option for lung squamous cell carcinoma and shows promising clinical application prospects.

[0041] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0042] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Attached Figure Description

[0043] Figure 1 The results of Western blot hybridization experiments show that the small molecule degrader 32649-1-1 inhibits the expression of NSD3 protein in H520 cells.

[0044] Figure 2 The results of Western blot hybridization experiments show that the small molecule degrader 32649-2-1 inhibits the expression of NSD3 protein in H1703 cells.

[0045] Figure 3 The results of Western blot hybridization experiments show that the small molecule degrader 32649-1-1 inhibits the expression of NSD3 protein in H1299 cells.

[0046] Figure 4 The results of Western blot hybridization experiments show that the small molecule degrader 32649-1-1 inhibits the expression of NSD3 protein in A549 cells.

[0047] Figure 5 The results of Western blot hybridization experiments show that the small molecule degrader 32649-2-1 inhibits the expression of NSD3 protein in H1299 cells.

[0048] Figure 6 The results of Western blot hybridization experiments show that the small molecule degrader 32649-2-1 inhibits the expression of NSD3 protein in A549 cells.

[0049] Figure 7 The results of Western blot hybridization experiments show that the small molecule degrader 32649-1-2 inhibits the expression of NSD3 protein in LK2 cells.

[0050] Figure 8 Western blot hybridization experiment to inhibit NSD3 protein expression in H520 cells using the small molecule degrader 32649-2-2.

[0051] Figure 9 The results of Western blot hybridization experiments show that the small molecule degrader 32649-1-3 inhibits the expression of NSD3 protein in H1703 cells.

[0052] Figure 10 The results of Western blot hybridization experiments show that the small molecule degrader 32649-2-3 inhibits the expression of NSD3 protein in H1703 cells.

[0053] Figure 11 The small molecule degrader 32649-1-1 was used to kill and inhibit the proliferation of H520 cells.

[0054] Figure 12 The small molecule degrader 32649-2-1 was used to kill and inhibit the proliferation of H1703 cells.

[0055] Figure 13 The small molecule degrader 32649-1-1 was used to kill and inhibit the proliferation of H1299 cells.

[0056] Figure 14 The small molecule degrader 32649-1-1 was used to kill and inhibit the proliferation of A549 cells.

[0057] Figure 15 The small molecule degrader 32649-2-1 was used to kill and inhibit the proliferation of H1299 cells.

[0058] Figure 16 The small molecule degrader 32649-2-1 was used to kill and inhibit the proliferation of A549 cells.

[0059] Figure 17 The small molecule degrader 32649-1-2 was used to kill and inhibit the proliferation of H1703 cells.

[0060] Figure 18 The small molecule degrader 32649-2-2 was used to kill and inhibit the proliferation of LK2 cells.

[0061] Figure 19 The small molecule degrader 32649-1-3 was used to kill and inhibit the proliferation of H520 cells.

[0062] Figure 20 The small molecule degrader 32649-2-3 was used to kill and inhibit the proliferation of H1703 cells. Detailed Implementation

[0063] The raw materials and equipment used in this invention are all known products, obtained by purchasing commercially available products.

[0064] The following experiments, where no temperature is specified, are reactions conducted under normal temperature conditions, which is room temperature, or 25±5℃.

[0065] 1. Preparation of intermediate 1 for CRBN ligand

[0066]

[0067] To a solution of 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione (1.0 g, 3.62 mmol, 1.0 eq) in DMSO (5 mL), triethylamine (1.51 mL, 10.9 mmol, 3.0 eq) and tert-butyl[2-[2-(2-aminoethoxy)ethoxy]ethyl]carbamate tert-butyl ester (1.08 g, 4.35 mmol, 1.2 eq) were added, followed by heating to 80 °C and stirring for 16 hours. TLC (petroleum ether:ethyl acetate = 1:1) showed complete reaction. The reaction mixture was diluted with H₂O, extracted with ethyl acetate, washed with saturated NaCl solution, dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 5:1 to 1:1) to give yellow bubbly intermediate 1 (1.13 g, 2.24 mmol, 62% yield).

[0068] The 1H NMR and HRMS data of intermediate 1 are as follows:

[0069] 1 H NMR (400 MHz, Chloroform- d δ 8.70 (s, 1H), 7.47 (dd, J = 8.5, 7.1Hz, 1H), 7.08 (d, J = 7.1 Hz, 1H), 6.89 (d, J = 8.5 Hz, 1H), 5.09 (s, 1H), 4.98 –4.84 (m, 1H), 3.70 (t, J = 5.3 Hz, 2H), 3.63 (s, 4H), 3.54 (t, J = 5.2 Hz, 2H), 3.45 (t, J = 5.3 Hz, 2H), 3.29 (t, J = 5.1 Hz, 2H), 2.84 – 2.66 (m, 3H), 2.14 –2.04 (m, 1H), 1.41 (s, 9H).

[0070] HRMS ( m / z ): [M + H] + Calculated for C 24 H 33 N4O8, 505.2298; found, 505.2288.

[0071] 2. Preparation of intermediate 2 for CRBN ligand

[0072]

[0073] Following the preparation method of intermediate 1 of CRBN ligand in step 1, intermediate 2 of CRBN ligand was prepared.

[0074] The 1H NMR and HRMS data of intermediate 2 are as follows:

[0075] 1 H NMR (400 MHz, Chloroform- d δ 8.72 (s, 1H), 7.46 (dd, J = 8.5, 7.1Hz, 1H), 7.06 (d, J = 7.0 Hz, 1H), 6.89 (d, J = 8.5 Hz, 1H), 6.46 (t, J = 5.6 Hz, 1H), 5.07 (t, J = 5.7 Hz, 1H), 4.92 – 4.85 (m, 1H), 3.70 (t, J = 5.4 Hz, 2H),3.66 – 3.56 (m, 8H), 3.50 (t, J = 5.2 Hz, 2H), 3.45 (q, J = 5.5 Hz, 2H), 3.27 (q, J = 5.5 Hz, 2H), 2.87 – 2.70 (m, 3H), 2.12 – 2.08 (m, 1H), 1.41 (s, 9H).

[0076] HRMS ( m / z ): [M + H] + Calculated for C 26 H 37 N4O9, 549.2561; found, 549.2553.

[0077] 3. Preparation of intermediate 3 for CRBN ligand

[0078]

[0079] Following the preparation method of intermediate 1 of CRBN ligand in step 1, intermediate 3 of CRBN ligand was prepared.

[0080] The 1H NMR and HRMS data of intermediate 3 are as follows:

[0081] 1 H NMR (400 MHz, Chloroform- d ) δ 8.56 (s, 1H), 7.47 (dd, J = 8.5, 7.1Hz, 1H), 7.08 (d, J = 7.1 Hz, 1H), 6.91 (d, J = 8.5 Hz, 1H), 6.48 (t, J = 5.7 Hz,1H), 5.12 (s, 1H), 4.95 – 4.83 (m, 1H), 3.70 (t, J = 5.4 Hz, 2H), 3.68 – 3.57(m, 12H), 3.52 (t, J = 5.2 Hz, 2H), 3.45 (q, J = 5.5 Hz, 2H), 3.29 (q, J = 5.4 Hz, 2H), 2.89 – 2.66 (m, 3H), 2.14 – 2.06 (m, 1H), 1.42 (s, 9H).

[0082] HRMS ( m / z ): [M + H] + Calculated for C 28 H 41 N4O 10 , 593.2823; found, 593.2815.

[0083] 4. Preparation of intermediate 4 for VHL ligands

[0084]

[0085] 6-[(tert-Butoxycarbonyl)amino]hexanoic acid (1.0 g, 4.32 mmol, 1.0 eq) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.47 g, 6.49 mmol, 1.5 eq) were dissolved in 10 mL of dry DMF. N,N-diisopropylethylamine (2.14 mL, 13.0 mmol, 3.0 eq) was added dropwise under ice bath and stirred for 30 minutes. Then (2S,4R)-1-((S)-2-amino-3,3-dimethylbutyryl)-4-hydroxy-N-(4-(4-methylthiazolyl-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride (2.02 g, 4.32 mmol, 1.0 eq) was added and stirred at room temperature for 2 hours. TLC (dichloromethane:methanol = 10:1) showed that the reaction was complete. The reaction mixture was diluted with H2O, extracted with ethyl acetate, washed with saturated NaCl solution, dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, dichloromethane:methanol = 20:1 to 15:1) to give colorless bubbly intermediate 4 (2.25 g, 3.5 mmol, 81% yield).

[0086] The 1H NMR and HRMS data of intermediate 4 are as follows:

[0087] 1 H NMR (400 MHz, DMSO- d 6) δ 8.98 (s, 1H), 8.56 (t, J = 6.1 Hz, 1H), 7.84 (d, J = 9.3 Hz, 1H), 7.40 (q, J = 8.1 Hz, 4H), 6.73 (t, J = 5.7 Hz, 1H), 5.12 (d, J =3.5 Hz, 1H), 4.54 (d, J = 9.3 Hz, 1H), 4.49 – 4.39 (m, 2H), 4.35 (s, 1H), 4.21(dd, J = 15.9, 5.5 Hz, 1H), 3.66 (d, J = 4.0 Hz, 2H), 2.87 (q, J= 6.6 Hz, 2H), 2.44 (s, 3H), 2.30 – 1.84 (m, 4H), 1.53 – 1.42 (m, 2H), 1.36 (s, 11H), 1.21(q, J = 7.5 Hz, 2H), 0.93 (s, 9H).

[0088] HRMS ( m / z ): [M + H] + Calculated for C 33 H 50 N5O6S, 644.3482; found, 644.3471.

[0089] 5. Preparation of intermediate 5 for VHL ligands

[0090]

[0091] Following the preparation method of intermediate 4 of VHL ligand in step 4, intermediate 5 of VHL ligand is prepared.

[0092] The 1H NMR and HRMS data of intermediate 5 are as follows:

[0093] 1 H NMR (400 MHz, DMSO- d 6) δ 8.93 (s, 1H), 8.55 (t, J = 6.1 Hz, 1H), 7.81(d, J = 9.3 Hz, 1H), 7.41 (q, J = 8.1 Hz, 4H), 6.73 (t, J = 5.7 Hz, 1H), 5.12 (d, J =3.5 Hz, 1H), 4.55 (d, J = 9.2 Hz, 1H), 4.49 – 4.38 (m, 2H), 4.34 (s, 1H), 4.21(dd, J = 15.9, 5.4 Hz, 1H), 3.66 (d, J = 4.0 Hz, 2H), 2.88 (q, J= 6.6 Hz, 2H),2.44 (s, 3H), 2.30 – 1.83 (m, 4H), 1.53 – 1.32 (m, 4H), 1.36 – 1.24 (m, 13H),1.22 (q, J = 7.6 Hz, 2H), 0.95 (s, 9H).

[0094] HRMS ( m / z ): [M + H] + Calculated for C 35 H 54 N5O6S, 672.3795; found, 672.3789.

[0095] 6. Preparation of intermediate 6 for VHL ligands

[0096]

[0097] Following the preparation method of intermediate 4 of VHL ligand in step 4, intermediate 6 of VHL ligand is prepared.

[0098] The 1H NMR and HRMS data of intermediate 6 are as follows:

[0099] 1 H NMR (400 MHz, DMSO- d 6) δ 8.93 (s, 1H), 8.55 (t, J = 6.1 Hz, 1H), 7.81(d, J = 9.3 Hz, 1H), 7.41 (q, J = 8.1 Hz, 4H), 6.73 (t, J = 5.7 Hz, 1H), 5.12 (d, J =3.5 Hz, 1H), 4.55 (d, J = 9.2 Hz, 1H), 4.49 – 4.38 (m, 2H), 4.34 (s, 1H), 4.21(dd, J = 15.9, 5.4 Hz, 1H), 3.66 (d, J = 4.0 Hz, 2H), 2.88 (q, J= 6.6 Hz, 2H), 2.44 (s, 3H), 2.30 – 1.83 (m, 4H), 1.53 – 1.32 (m, 4H), 1.36 – 1.24 (m, 15H), 1.22 – 1.18 (m, 4H), 0.95 (s, 9H).

[0100] HRMS ( m / z ): [M + H] + Calculated for C 37 H 58 N5O6S, 700.4108; found,700.4112.

[0101] Example 1: Preparation of small molecule NSD3 degrader 32649-1-1

[0102]

[0103] Intermediate 1 (504 mg, 1.0 mmol, 1.0 eq) was dissolved in dichloromethane (5 mL), and 5 mL of 4M HCl / dioxane (20 mmol, 20 eq) was added. The mixture was reacted at room temperature for 2 hours and then concentrated under reduced pressure for later use (stored in a spare bottle). (4-oxo-6,7-dihydro-4H,5H-cyclopentadieno[4,5]thieno[2,3-D]pyrimidin-3-yl)-acetic acid (32649, 250 mg, 1.0 mmol, 1.0 eq) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (570 mg, 1.5 mmol, 1.5 eq) were dissolved in 2 mL of dry DMF. N,N-diisopropylethylamine (0.5 mL, 3.0 mmol, 3.0 eq) was added dropwise under ice bath conditions, and the mixture was stirred for 30 minutes. The solution was then transferred to a spare flask and stirred at room temperature for 2 hours. TLC (dichloromethane:methanol = 10:1) showed that the reaction was complete. The reaction mixture was diluted with H2O, extracted with ethyl acetate, washed with saturated NaCl solution, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, dichloromethane:methanol = 15:1 to 10:1) to give a yellow bubbly product 32649-1-1 (460 mg, 0.63 mmol, 63% yield).

[0104] The 1H NMR and HRMS data for 32649-1-1 are as follows:

[0105] 1 H NMR (400 MHz, Chloroform-d ) δ 8.70 (s, 1H), 7.97 (s, 1H), 7.47(dd, J = 8.5, 7.1 Hz, 1H), 7.08 (d, J = 7.1 Hz, 1H), 6.89 (d, J = 8.5 Hz, 1H), 5.09(s, 1H), 4.98 – 4.84 (m, 1H), 4.65 (s, 2H), 3.70 (t, J = 5.3 Hz, 2H), 3.63 (s, 4H), 3.54 (t, J = 5.2 Hz, 2H), 3.45 (t, J = 5.3 Hz, 2H), 3.29 (t, J = 5.1 Hz, 2H), 2.84 – 2.66 (m, 3H), 2.46 – 2.16 (m, 6H), 2.14 – 2.04 (m, 1H).

[0106] HRMS ( m / z ): [M + H] + Calculated for C 30 H 33 N6O8S, 637.2081; found, 637.2085.

[0107] Example 2: Preparation of small molecule NSD3 degrader 32649-2-1

[0108]

[0109] The preparation method of Example 1 is used as a reference, except that intermediate 1 is replaced with intermediate 4 to prepare 32649-2-1.

[0110] The 1H NMR and HRMS data for 32649-2-1 are as follows:

[0111] 1 H NMR (400 MHz, Chloroform- d ) δ 8.66 (s, 1H), 7.97 (s, 1H), 7.35 –7.27 (m, 4H), 7.21 (dd, J = 14.1, 5.1 Hz, 1H), 6.58 (d, J= 8.9 Hz, 1H), 4.76 –4.68 (m, 1H), 4.63 – 4.46 (m, 5H), 4.30 (d, J = 15.1 Hz, 1H), 4.13 (d, J = 11.4Hz, 1H), 3.64 (dd, J = 11.3, 3.3 Hz, 1H), 3.21 – 3.14 (m, 2H), 2.95 (ddd, J =13.0, 10.5, 6.8 Hz, 4H), 2.49 (s, 3H), 2.46 – 2.38 (m, 3H), 2.26 – 2.12 (m,3H), 1.63 – 1.38 (m, 6H), 0.98 (s, 9H).

[0112] HRMS ( m / z ): [M + H] + Calculated for C 39 H 50 N7O6S2, 776.3264; found,776.3258.

[0113] The following is a method for preparing the control sample of this invention.

[0114] Comparative Example 1: Preparation of small molecule NSD3 degrading agent 32649-1-2

[0115]

[0116] The preparation method of Example 1 is used, except that intermediate 1 is replaced with intermediate 2 to prepare 32649-1-2.

[0117] The 1H NMR and HRMS data for 32649-1-2 are as follows:

[0118] 1 H NMR (400 MHz, Chloroform- d ) δ 8.72 (s, 1H), 7.97 (s, 1H), 7.46(dd, J = 8.5, 7.1 Hz, 1H), 7.06 (d, J = 7.0 Hz, 1H), 6.89 (d, J = 8.5 Hz, 1H), 6.46(t, J = 5.6 Hz, 1H), 5.07 (t,J = 5.7 Hz, 1H), 4.92 – 4.85 (m, 1H), 4.65 (s, 2H), 3.70 (t, J = 5.4 Hz, H), 3.66 – 3.56 (m, 8H), 3.50 (t, J = 5.2 Hz, 2H), 3.45 (q, J = 5.5 Hz, 2H), 3.27 (q, J = 5.5 Hz, 2H), 2.87 – 2.70 (m, 3H), 2.46 – 2.15 (m, 6H), 2.12 – 2.08 (m, 1H).

[0119] HRMS ( m / z ): [M + H] + Calculated for C 32 H 37 N6O9S, 681.2343; found, 681.2349.

[0120] Comparative Example 2: Preparation of small molecule NSD3 degrading agent 32649-1-3

[0121]

[0122] The preparation method of Example 1 is used, except that intermediate 1 is replaced with intermediate 3 to prepare 32649-1-3.

[0123] The 1H NMR and HRMS data for 32649-1-3 are as follows:

[0124] 1 H NMR (400 MHz, Chloroform- d ) δ 8.56 (s, 1H), 7.97 (s, 1H), 7.47(dd, J = 8.5, 7.1 Hz, 1H), 7.08 (d, J = 7.1 Hz, 1H), 6.91 (d, J = 8.5 Hz, 1H), 6.48(t, J = 5.7 Hz, 1H), 5.12 (s, 1H), 4.95 – 4.83 (m, 1H), 4.65 (s, 2H), 3.70 (t, J= 5.4 Hz, 2H), 3.68 – 3.57 (m, 12H), 3.52 (t, J = 5.2 Hz, 2H), 3.45 (q, J = 5.5Hz, 2H), 3.29 (q, J = 5.4 Hz, 2H), 2.89 – 2.66 (m, 3H), 2.47 – 2.17 (m, 6H)2.14– 2.06 (m, 1H).

[0125] HRMS ( m / z ): [M + H] + Calculated for C 34 H 41 N6O 10 S, 725.2605; found, 725.2611.

[0126] Comparative Example 3: Preparation of small molecule NSD3 degrader 32649-2-2

[0127]

[0128] The preparation method of Example 1 is used, except that intermediate 1 is replaced with intermediate 5 to prepare 32649-2-2.

[0129] The 1H NMR and HRMS data for 32649-2-2 are as follows:

[0130] 1 H NMR (400 MHz, Chloroform- d ) δ 8.63 (s, 1H), 7.95 (s, 1H), 7.35 –7.27 (m, 4H), 7.21 (dd, J = 14.2, 5.2 Hz, 1H), 6.58 (d, J = 8.8 Hz, 1H), 4.76 –4.68 (m, 1H), 4.63 – 4.46 (m, 5H), 4.30 (d, J = 15.2 Hz, 1H), 4.13 (d, J = 11.4Hz, 1H), 3.64 (dd, J = 11.2, 3.3 Hz, 1H), 3.21 – 3.14 (m, 2H), 2.95 (ddd, J=13.0, 10.5, 6.8 Hz, 4H), 2.49 (s, 3H), 2.46 – 2.38 (m, 3H), 2.26 – 2.11 (m,5H), 1.63 – 1.33 (m, 8H), 0.98 (s, 9H).

[0131] HRMS ( m / z ): [M + H] + Calculated for C 41 H 54 N7O6S2, 804.3577; found, 804.3582

[0132] Comparative Example 4: Preparation of small molecule NSD3 degrading agent 32649-2-3

[0133]

[0134] The preparation method of Example 1 is used, except that intermediate 1 is replaced with intermediate 6 to prepare 32649-2-3.

[0135] The 1H NMR and HRMS data for 32649-2-3 are as follows:

[0136] 1 H NMR (400 MHz, Chloroform- d ) δ 8.66 (s, 1H), 7.97 (s, 1H), 7.35 –7.27 (m, 4H), 7.21 (dd, J = 14.1, 5.1 Hz, 1H), 6.58 (d, J = 8.9 Hz, 1H), 4.76 –4.68 (m, 1H), 4.63 – 4.46 (m, 5H), 4.30 (d, J = 15.1 Hz, 1H), 4.13 (d, J = 11.4Hz, 1H), 3.64 (dd, J = 11.3, 3.3 Hz, 1H), 3.21 – 3.14 (m, 2H), 2.95 (ddd, J =13.0, 10.5, 6.8 Hz, 4H), 2.49 (s, 3H), 2.46 – 2.38 (m, 3H), 2.26 – 2.05 (m,7H), 1.65 – 1.33 (m, 10H), 0.98 (s, 9H).

[0137] HRMS ( m / z ): [M + H] + Calculated for C 43 H 58 N7O6S2, 832.3890; found, 832.3898.

[0138] The following experimental examples demonstrate the beneficial effects of the present invention.

[0139] In the following experiments, H1703 cells, LK2 cells, and H520 cells were lung squamous cell carcinoma cells, while H1299 cells and A549 cells were lung adenocarcinoma cells.

[0140] Experimental Example 1: Inhibition of NSD3 Protein Expression by Small Molecule Degraders

[0141] 1. Inhibition of NSD3 protein expression in H520 cells by the small molecule degrader 32649-1-1

[0142] The following are the experimental steps for Western blot analysis of proteins using small molecule degrading agents:

[0143] 1. After H520 cells adhered to the culture vessel, they were cultured in complete culture medium containing different concentrations (0, 0.1 µM, 1 µM, 2 µM) of the small molecule degrader 32649-1-1. After 48 hours in a 37°C incubator, the cells were removed to collect proteins.

[0144] II. Preparation of Protein Lysis Buffer

[0145] 1. Preparation of protein lysis buffer 2×sample buffer 40ml system: 80% glycerol 10ml, sterile water 1ml, 10% SDS 24ml, 1M Tris-HCl (pH 6.8) 5ml. Mix thoroughly and store at room temperature for later use.

[0146] 2. Preparation of protein loading buffer: Mix β-mercaptoethanol and bromophenol blue powder thoroughly in a 1.5 ml EP tube and store at -20°C for later use.

[0147] III. Total Protein Extraction

[0148] 1) Remove the cell culture dish from the 37℃ constant temperature incubator and assess the cell density under a microscope;

[0149] 2) Place the culture dish on ice (the entire operation should be performed on ice), aspirate the upper culture medium, and wash twice with cooled PBS;

[0150] 3) Add 20-50 μl of PBS and protein lysis buffer to each well at a 1:1 ratio according to the cell density. After the liquid covers the surface of the culture dish, immediately use a cell scraper to thoroughly mix and lyse the added solution with the cells attached to the surface of the culture dish, and collect the solution into an EP tube.

[0151] 4) Heat in a 100℃ constant temperature metal bath for 10 minutes to denature the protein, then briefly centrifuge at 12000 rpm and store at 4℃ for short-term use.

[0152] IV. Protein Concentration Quantification (BCA Method)

[0153] 1) Before the experiment, mix PBS and BSA stock solution quantitatively to prepare BSA standard with a concentration of 0.5 mg / ml, and store on ice or at 4°C for later use;

[0154] 2) Preparation of BCA mixture: Based on the number of test samples and corresponding gradient standards set in the experiment, prepare a light green BCA working solution at a ratio of BCA reagent:Cu reagent = 50:1. After thorough mixing, store at room temperature for short-term use.

[0155] 3) Starting with the first well, add the standard in decreasing volumes from 20 μl to 0 μl to the protein standard well, and then add PBS to make up the liquid in each well to 20 μl, so that the concentration of the standard in each well forms a series of concentration gradients;

[0156] 4) The absorbance was detected using a microplate reader at a wavelength of 562 nm, the OD value was recorded, and a protein standard curve was plotted.

[0157] 5) Sample protein processing: Take a 96-well plate and the prepared protein sample, add 18 μl PBS and 2 μl sample to each well, and add 200 μl BCA mixture. React in a 37℃ metal bath or incubator for 30 minutes.

[0158] 6) After taking the sample after the reaction and detecting the OD value, calculate the sample protein concentration according to the completed protein standard curve, and calculate the sample volume during electrophoresis according to the protein loading amount. The specific calculation formula is: protein loading volume = protein loading amount / sample protein concentration.

[0159] 7) Mix the sample protein with the prepared protein loading buffer at a ratio of 20:1, incubate at 12,000 rpm and store at -20°C for later use.

[0160] V. SDS-PAGE gel electrophoresis

[0161] 1) Preparation of separating gel: Different concentrations of separating gel were selected according to the molecular weight of the proteins to be detected. The required separating gel concentrations for detecting proteins RNase1, p-ALK (Y1604), and p-ALK (Y1282 / 1283) in this experiment were 10%, 8%, and 8%, respectively. The separating gel system is shown in Table 1. The gel casting rack was installed on a horizontal table. The mixture (5 ml) was prepared according to the separating gel system (as shown in Table 1) and quickly, carefully, and steadily poured into the pre-reserved gap between the casting plates. Then, 100-200 μl of isopropanol or anhydrous ethanol was slowly poured horizontally from left to right along the edge of the casting plate to flatten the gel surface. One side of the casting rack was slightly lifted and shaken to remove any potential air bubbles. Finally, the gel was left to stand on a horizontal table at room temperature for 30-60 minutes to allow the separating gel to solidify.

[0162] Table 1. Separating gel preparation system (5 ml)

[0163]

[0164] 2) Preparation of the stacking gel: After the separating gel has solidified, carefully remove the isopropanol or anhydrous ethanol and allow it to air dry under natural ventilation. Then, prepare the top layer gel (2 ml) according to the stacking gel system (as shown in Table 2), and quickly and steadily pour it into the gaps of the gel casting plate until liquid overflows. Slowly and steadily insert a 15-well comb. Let it stand on a horizontal table at room temperature for 30-60 minutes.

[0165] Table 2. Stacking gel preparation system (2ml)

[0166]

[0167] 3) After the upper layer of concentrated gel has solidified, carefully remove the gel plate from the gel preparation rack and secure it tightly in the electrophoresis tank. Carefully, slowly and vertically pull out the comb to leave the sample loading hole, and pour an appropriate amount of electrophoresis solution into the electrophoresis tank according to the number of gel plates.

[0168] 4) Protein loading: Remove the protein samples stored at -20℃ and allow them to thaw to room temperature. Load the samples sequentially in equal volumes according to the calculated loading volume, and reserve 1-2 wells for 5μl of protein loading indicator (marker) as needed for the experiment.

[0169] 5) Electrophoresis: After loading the sample, start electrophoresis at a constant voltage of 80V. When you see bead-like bubbles appearing at the bottom of the gel plate, it indicates that electrophoresis has started. Then you can see the bromophenol blue in the protein sample being compressed into a straight line. After the protein reaches a clearly distinguishable transparent separating gel interface according to the marker, adjust the voltage to a constant voltage of 120V. When the bromophenol blue indicator drops to the bottom edge of the gel plate, stop electrophoresis and start the membrane transfer.

[0170] 6) Transfer: Prepare the necessary filter paper, methanol cartridge for activation, transfer clamps, PVDF membrane, etc., and pre-cool the transfer buffer at 4°C before transfer. Place a piece of filter paper in a horizontal container and pour in an appropriate amount of transfer buffer, making it slightly higher than the thickness of the filter paper. Wet and activate the PVDF membrane with methanol, then place it on the filter paper. Take out the electrophoresis gel, peel the protein gel from the gel, trim the edges appropriately, and place it on the PVDF membrane. Cover it with another layer of filter paper, continuously removing any air bubbles generated during the process. Finally, following the "black gel, white membrane" principle, place the transparent side of the transfer clamp as the bottom layer, cover it with a sponge pad, and horizontally place the previously clamped filter paper and PVDF membrane on top. Cover it with another sponge pad, align the black sides of the transfer clamp, and fasten the transfer clamp clips. Secure the clamped transfer clamp in the transfer tank, add transfer buffer until it submerges the transfer indicator line, place the transfer tank in a suitable container, and begin the transfer process. Add ice water to the container for cooling. Using a constant current of 300mA for membrane transfer, the appropriate transfer time was selected according to the size of the target protein. The transfer times for R1 and p-ALK proteins were 1 and 2 hours, respectively.

[0171] 7) Blocking: Prepare 5% skim milk and an appropriate amount of PBST for washing beforehand. After the transfer is complete, open the transfer clamp and carefully remove the PVDF membrane. Wash the membrane in PBST 2-3 times, 2 minutes each time. Then transfer it to 5% skim milk prepared with PBST, place it on a shaker, and block for 1 hour.

[0172] 8) Primary antibody incubation: Prepare primary antibodies using primary antibody dilution buffer, 5% milk, or 5% BSA according to the specified ratio. The concentration of primary antibodies in this experiment was 1:1000 (all antibodies in this experiment were self-made monoclonal mouse antibodies). Each antibody preparation yielded 4 ml. Before incubation, prepare the antibody, pour it into the corresponding band cell of the incubation chamber, and place it on ice to maintain antibody activity at a low temperature. Remove the PVDF membrane from the 5% skim milk and rinse it briefly with PBST to remove residual milk. Based on the size of the protein bands detected in the experiment, use the colorimetric marker as a scale to divide the membrane into bands, and then place it into the corresponding antibody cell. After sealing, place it on a shaker at 4°C and shake gently for 12 hours.

[0173] 9) Recovery of primary antibody: The primary antibody was recovered 12 hours later;

[0174] 10) Secondary antibody incubation: Use horseradish enzyme-labeled goat anti-mouse IgG (H+L) secondary antibody (Zhongshan Jinqiao, catalog number: ZB-2305). Prepare the appropriate secondary antibody dilution solution in advance using 5% skim milk prepared with PBST at a ratio of 1:5000. Rinse the PVDF membrane at least 3 times with PBST, 5-10 minutes each time, and discard the liquid after rinsing. Then pour the secondary antibody dilution solution into the corresponding species of PVDF membrane compartment and shake gently on a shaker for 1 hour.

[0175] 11) Recovery of secondary antibody: The secondary antibody was recovered and the PVDF membrane was rinsed again with PBST 3 times, 5-10 minutes each time;

[0176] 12) ECL Development: Prepare the ultrasensitive ECL developer (UltraSignal ultrasensitive ECL chemiluminescent substrate, Sizhengbai, catalog number: 4AW011-100) in advance. Mix equal volumes of ECL solutions A and B in an EP tube and store at low temperature in the dark. Before development, place the PVDF membrane on a foam plate, slightly wipe off any remaining PBST, and then drop the prepared ultrasensitive ECL developer onto the membrane according to its size. After both sides are completely wetted, expose the membrane in a Bio-rad imaging instrument.

[0177] The experimental results are shown in Figure 1 The small molecule degrader 32649-1-1 significantly inhibited the expression of NSD3 protein in H520 cells.

[0178] 2. Inhibition of NSD3 protein expression in H1703 cells by the small molecule degrader 32649-2-1

[0179] The protein immunoblotting hybridization experiment was performed in step 1, with the only difference being that the small molecule degrader 32649-1-1 was replaced with 32649-2-1, and H520 cells were replaced with H1703 cells.

[0180] The experimental results are shown in Figure 2 The small molecule degrader 32649-2-1 significantly inhibited the expression of NSD3 protein in H1703 cells.

[0181] 3. Inhibition of NSD3 protein expression in H1299 cells by the small molecule degrader 32649-1-1

[0182] The protein immunoblotting hybridization experiment (Western blot) procedure in step 1 is followed, except that H520 cells are replaced with H1299 cells.

[0183] The experimental results are shown in Figure 3 The small molecule degrader 32649-1-1 had no significant inhibitory effect on NSD3 protein expression in H1299 cells.

[0184] 4. Inhibition of NSD3 protein expression in A549 cells by the small molecule degrader 32649-1-1

[0185] The protein immunoblotting hybridization experiment in step 1 is followed, except that H520 cells are replaced with A549 cells.

[0186] The experimental results are shown in Figure 4 The small molecule degrader 32649-1-1 had no significant inhibitory effect on NSD3 protein expression in A549 cells.

[0187] 5. Inhibition of NSD3 protein expression in H1299 cells by the small molecule degrader 32649-2-1

[0188] The protein immunoblotting hybridization experiment was performed in step 1, with the only difference being that the small molecule degrader 32649-1-1 was replaced with 32649-2-1, and H520 cells were replaced with H1299 cells.

[0189] The experimental results are shown in Figure 5 The small molecule degrader 32649-2-1 had no significant inhibitory effect on NSD3 protein expression in H1299 cells.

[0190] 6. Inhibition of NSD3 protein expression in A549 cells by the small molecule degrader 32649-2-1

[0191] The protein immunoblotting hybridization experiment was performed in step 1, with the only difference being that the small molecule degrader 32649-1-1 was replaced with 32649-2-1, and H520 cells were replaced with A549 cells.

[0192] The experimental results are shown in Figure 6 The small molecule degrader 32649-2-1 had no significant inhibitory effect on NSD3 protein expression in A549 cells.

[0193] 7. Inhibition of NSD3 protein expression in H1703 cells by the small molecule degrader 32649-1-2

[0194] The protein immunoblotting hybridization experiment was performed in step 1, with the only difference being that the small molecule degrader 32649-1-1 was replaced with 32649-1-2, and H520 cells were replaced with H1703 cells.

[0195] The experimental results are shown in Figure 7 The small molecule degrader 32649-1-2 had no significant inhibitory effect on NSD3 protein expression in H1703 cells.

[0196] 8. Inhibition of NSD3 protein expression in LK2 cells by small molecule degrader 32649-2-2

[0197] The protein immunoblotting hybridization experiment was performed in step 1, with the only difference being that the small molecule degrader 32649-1-1 was replaced with 32649-2-2, and the H520 cells were replaced with LK2 cells.

[0198] The experimental results are shown in Figure 8 The small molecule degrader 32649-2-2 had no significant inhibitory effect on NSD3 protein expression in LK2 cells.

[0199] 9. Inhibition of NSD3 protein expression in H520 cells by the small molecule degrader 32649-1-3

[0200] The protein immunoblotting hybridization experiment is performed in step 1, with the only difference being that the small molecule degrader 32649-1-1 is replaced with 32649-1-3.

[0201] The experimental results are shown in Figure 9 The small molecule degrader 32649-1-3 had no significant inhibitory effect on NSD3 protein expression in H520 cells.

[0202] 10. Inhibition of NSD3 protein expression in H1703 cells by the small molecule degrader 32649-2-3

[0203] The protein immunoblotting hybridization experiment was performed in step 1, with the only difference being that the small molecule degrader 32649-1-1 was replaced with 32649-2-3, and H520 cells were replaced with H1703 cells.

[0204] The experimental results are shown in Figure 10 The small molecule degrader 32649-2-3 had no significant inhibitory effect on NSD3 protein expression in H1703 cells.

[0205] The above results indicate that the small molecule degraders 32649-1-1 and 32649-2-1 provided in the embodiments of the present invention can effectively inhibit the expression of NSD3 protein in lung squamous cell carcinoma cells, but have no significant inhibitory effect on the expression of NSD3 protein in lung adenocarcinoma cells; the small molecule degraders 32649-1-2, 32649-1-3, 32649-2-2, and 32649-2-3 provided in the comparative examples of the present invention have no significant inhibitory effect on the expression of NSD3 protein in both lung squamous cell carcinoma cells and lung adenocarcinoma cells.

[0206] Experimental Example 2: Cell killing and proliferation inhibition effects of small molecule degrading agents

[0207] 1. Killing and proliferation-inhibiting effects of small molecule degrader 32649-1-1 on H520 cells

[0208] The killing and proliferation-inhibiting effects of the small molecule degrader 32649-1-1 on H520 cells were detected using the MTT assay.

[0209] I. Experimental Procedure

[0210] (1) When the cell density reaches 80-90% or more, remove the cell (usually use a 10cm dish), discard the culture medium, and wash the cell dish twice with PBS.

[0211] (2) Add 0.5-1 ml of trypsin and place in a 37°C incubator to digest the cells. After 5-10 minutes, when the cells no longer adhere to the wall, add complete culture medium to stop the digestion.

[0212] (3) Collect cells, centrifuge at 800 rpm for 3 min, discard the supernatant, add 3 ml of complete culture medium and mix well to make a cell suspension, then use the cell suspension to count cells, and re-prepare the cell suspension required for the experiment (3000 cells per 100 µl).

[0213] (4) After preparing the cell suspension, mix it gently, add 100µl to each well, and evenly spread an equal amount of cells into the 96-well plate.

[0214] Note: Because the cells will continue to settle after mixing, it is necessary to mix repeatedly during the seeding process, such as mixing once after every 6 wells, to ensure that the seeded cell density is exactly the same in each well. This is crucial for the MTT results.

[0215] (5) Place the inoculated cell culture plate in a 37°C incubator and culture until the cells adhere to the plate. Then add the drug at different concentration gradients (0, 0.315 µM, 1.25 µM, 5 µM, 20 µM) (i.e., small molecule degrading agent). Prepare the complete culture medium containing different concentrations of the drug in EP tubes. Add 100 µl of culture medium containing different concentrations of the drug to each well (Note: The prepared concentrations are all 2x, so that when added to the 96-well plate, they are exactly the same as the 100 µl of culture medium liquid in the previous 96-well plate, ensuring the accuracy of the drug concentration). Set 3 to 6 replicates for each concentration gradient.

[0216] (6) Incubate at 37℃ for 72 hours, during which the effects of the drug can be observed under an inverted microscope.

[0217] (7) After 72 hours, remove the well, add 20µl of MTT solution to each well, and then place it in a 37°C incubator for 4 hours.

[0218] (8) Remove the plate to terminate the culture, discard the supernatant, place several sheets of filter paper on the table, and then gently invert the 96-well plate so that the supernatant is absorbed by the filter paper, reducing the loss of results. Then add 200 µl of dimethyl sulfoxide (DMSO) to each well and shake on a shaker at low speed for 10 min to fully dissolve the crystals. Measure the absorbance of each well at OD562 nm using an ELISA reader.

[0219] The experimental results are shown in Figure 11The small molecule degrader 32649-1-1 has a significant effect on killing and inhibiting the proliferation of H520 cells.

[0220] 2. Killing and proliferation-inhibiting effects of small molecule degrader 32649-2-1 on H1703 cells

[0221] The experimental procedure is the same as in step 1, except that the small molecule degrader 32649-1-1 is replaced with 32649-2-1 and the H520 cells are replaced with H1703 cells.

[0222] The experimental results are shown in Figure 12 The small molecule degrader 32649-2-1 has a significant effect on killing and inhibiting the proliferation of H1703 cells.

[0223] 3. Killing and proliferation-inhibiting effects of small molecule degrader 32649-1-1 on H1299 cells

[0224] The experimental procedure is the same as in step 1, except that H520 cells are replaced with H1299 cells.

[0225] The experimental results are shown in Figure 13 The small molecule degrader 32649-1-1 had no significant effect on killing or inhibiting the proliferation of H1299 cells.

[0226] 4. Killing and proliferation-inhibiting effects of small molecule degrader 32649-1-1 on A549 cells.

[0227] The experimental procedure is the same as in step 1, except that H520 cells are replaced with A549 cells.

[0228] The experimental results are shown in Figure 14 The small molecule degrader 326491-1 had no significant effect on killing or inhibiting the proliferation of A549 cells.

[0229] 5. Killing and proliferation-inhibiting effects of small molecule degrader 32649-2-1 on H1299 cells.

[0230] The experimental procedure is the same as in step 1, except that the small molecule degrader 32649-1-1 is replaced with 32649-2-1 and the H520 cells are replaced with H1299 cells.

[0231] The experimental results are shown in Figure 15 The small molecule degrader 32649-2-1 had no significant effect on killing or inhibiting the proliferation of H1299 cells.

[0232] 6. Killing and proliferation-inhibiting effects of small molecule degrader 32649-2-1 on A549 cells.

[0233] The experimental procedure is the same as in step 1, except that the small molecule degrader 32649-1-1 is replaced with 32649-2-1 and the H520 cells are replaced with A549 cells.

[0234] The experimental results are shown in Figure 16 The small molecule degrader 32649-2-1 had no significant effect on killing or inhibiting the proliferation of A549 cells.

[0235] 7. Killing and proliferation-inhibiting effects of small molecule degrader 32649-1-2 on H1703 cells

[0236] The experimental procedure is the same as in step 1, except that the small molecule degrader 32649-1-1 is replaced with 32649-1-2 and the H520 cells are replaced with H1703 cells.

[0237] The experimental results are shown in Figure 17 The small molecule degrader 32649-1-2 had no significant effect on killing or inhibiting the proliferation of H1703 cells.

[0238] 8. Killing and proliferation-inhibiting effects of small molecule degrader 32649-2-2 on LK2 cells

[0239] The experimental procedure is the same as in step 1, except that the small molecule degrader 32649-1-1 is replaced with 32649-2-2 and the H520 cells are replaced with LK2 cells.

[0240] The experimental results are shown in Figure 18 The small molecule degrader 32649-2-2 had no significant effect on killing or inhibiting the proliferation of LK2 cells.

[0241] 9. Killing and proliferation-inhibiting effects of small molecule degrader 32649-1-3 on H520 cells

[0242] The experimental procedure is the same as in step 1, except that the small molecule degrader 32649-1-1 is replaced with 32649-1-3.

[0243] The experimental results are shown in Figure 19 The small molecule degrader 32649-1-3 had no significant effect on killing or inhibiting the proliferation of H520 cells.

[0244] 10. Killing and proliferation-inhibiting effects of small molecule degrader 32649-2-3 on H1703 cells.

[0245] The experimental procedure is the same as in step 1, except that the small molecule degrader 32649-1-1 is replaced with 32649-2-3 and the H520 cells are replaced with H1703 cells.

[0246] The experimental results are shown in Figure 20 The small molecule degrader 32649-2-3 had no significant effect on killing or inhibiting the proliferation of H1703 cells.

[0247] The above results indicate that the small molecule degrading agents 32649-1-1 and 32649-2-1 provided in the embodiments of the present invention have significant killing and proliferation-inhibiting effects on lung squamous cell carcinoma cells, but no significant killing and proliferation-inhibiting effects on lung adenocarcinoma cells; the small molecule degrading agents 32649-1-2, 32649-1-3, 32649-2-2, and 32649-2-3 provided in the comparative examples of the present invention have no significant killing and proliferation-inhibiting effects on lung squamous cell carcinoma cells and lung adenocarcinoma cells.

[0248] In summary, this invention provides a small-molecule NSD3 degrading agent that significantly inhibits NSD3 protein expression and exhibits significant killing and growth-inhibiting effects on lung squamous cell carcinoma cells. This small-molecule NSD3 degrading agent offers a novel treatment option for lung squamous cell carcinoma and shows promising clinical application prospects.

Claims

1. A compound, characterized in that, The compound has a structure as shown in Formula I or Formula II: ； ； Where m is 0, 1, or 2; n can be 0, 1, 2, 3, 4, or 5.

2. The compound according to claim 1, characterized in that, The compound has a structure as shown in Formula III or Formula IV: ； 。 3. A method for preparing the compound of formula I according to claim 1, characterized in that, The method includes the following steps: (1) Compound B reacts with activators and organic bases; (2) Add compound A to the reaction system of step (1), react, and obtain the compound shown in formula I; Among them, compound A is m is 0, 1, or 2; compound B is ; The molar ratio of compound A, compound B, activator, and organic base is (0.1~10):(0.1~10):(0.1~10):(0.1~10). The solvent for the reaction in step (1) is an organic solvent; the reaction time is 0.1~2 h, and the reaction temperature is -10~10℃; The reaction time in step (2) is 0.5 to 5 hours, and the reaction temperature is 15 to 50°C.

4. The method according to claim 3, characterized in that, The molar ratio of compound A, compound B, activator, and organic base is 1:1:1.5:3; the activator is 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, and the organic base is N,N-diisopropylethylamine. The organic solvent in step (1) is N,N-dimethylformamide; the reaction time is 0.5 h, and the reaction temperature is -4~4℃; The reaction time in step (2) is 2 hours, and the reaction temperature is 20~30℃.

5. The method according to claim 3 or 4, characterized in that, The compound A is .

6. A method for preparing the compound of formula II according to claim 1, characterized in that, The method includes the following steps: (1) Compound B reacts with activators and organic bases; (2) Add compound C to the reaction system of step (1), and react to obtain the compound shown in formula II; Among them, compound C is n is 0, 1, 2, 3, 4, or 5; compound B is ; The molar ratio of compound C, compound B, activator, and organic base is (0.1~10):(0.1~10):(0.1~10):(0.1~10). The solvent for the reaction in step (1) is an organic solvent; the reaction time is 0.1~2 h, and the reaction temperature is -10~10℃; The reaction time in step (2) is 0.5 to 5 hours, and the reaction temperature is 15 to 50°C.

7. The method according to claim 6, characterized in that, The molar ratio of compound C, compound B, activator, and organic base is 1:1:1.5:3; the activator is 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, and the organic base is N,N-diisopropylethylamine. The organic solvent in step (1) is N,N-dimethylformamide; the reaction time is 0.5 h, and the reaction temperature is -4~4℃; The reaction time in step (2) is 2 hours, and the reaction temperature is 20~30℃.

8. The method according to claim 6 or 7, characterized in that, The compound C is .

9. Use of the compound of claim 1 or 2 in the preparation of a medicament for the prevention and / or treatment of lung cancer.

10. The use according to claim 9, characterized in that, The lung cancer mentioned is squamous cell carcinoma of the lung.

Images