NSD3 degradation agent and preparation method and application thereof

Abstract

The present invention discloses a NSD3 degrading agent and a preparation method and application thereof, belonging to the technical field of drug synthesis. The present invention provides a class of compounds shown in formula I, which have a good effect of degrading NSD3 protein in vitro and can inhibit the proliferation of squamous cell carcinoma cell line NCI-H1703, and is expected to be a new way to prevent and treat NSD3-related diseases.

Description

Technical Field

[0001] The present invention belongs to the technical field of drug synthesis, and in particular relates to a NSD3 degradation agent and a preparation method and application thereof. Background Art

[0002] NSD3 (Nuclear Receptor Binding SET Domain Protein 3) is a histone lysine methyltransferase and a member of the NSD family. NSD3 is involved in epigenetic regulation and mainly affects gene expression by methylating lysine residues in the histone tail. Recent studies have shown that NSD3 plays an important role in many types of cancer, especially in the occurrence, development and metastasis of cancer. Abnormal overexpression or enhanced function of NSD3 is closely related to the occurrence of tumors, such as lung cancer, breast cancer, and hematological malignancies. Therefore, NSD3 has become a potential target for cancer treatment, and the research and development of small molecule inhibitors and degraders for NSD3 has also received widespread attention.

[0003] PROTAC (Proteolysis-Targeting Chimera) technology for targeted protein degradation is an emerging drug design strategy that utilizes the inherent ubiquitin-proteasome system (UPS) in cells to achieve degradation of target proteins. Traditional drug design often focuses on inhibiting the functional activity of proteins, while PROTAC technology is designed by designing a special bifunctional small molecule compound, one end of which can specifically bind to the target protein and the other end can bind to the E3 ubiquitin ligase, thereby causing the target protein to be ubiquitinated and directed to the proteasome for degradation.

[0004] Therefore, developing a PROTAC degrader targeting NSD3 with a novel structure is of great significance for the prevention or treatment of NSD3-related diseases. Summary of the invention

[0005] The purpose of the present invention is to provide a class of quinoline-imidazole-benzene ring compounds, which can be used as NSD3 targeted degradation agents to achieve efficient prevention or treatment of diseases related to NSD3 function.

[0006] In a first aspect, the present invention provides a compound represented by formula I or a pharmaceutically acceptable salt thereof, wherein the structure of formula I is as follows:

[0007] ;

[0008] in:

[0009] X is a C1~C12 alkyl chain;

[0010] R1 is selected from H, (i.e. the dehydroxylated structure corresponding to arginine), (i.e. the dehydroxylated structure corresponding to histidine), (i.e. the dehydroxylated structure corresponding to glutamine), (i.e. the dehydroxylated structure of asparagine) or (That is, the dehydroxylated structure corresponding to glycine).

[0011] In some embodiments of the present invention, in the compound of formula I above, X is a C3-C10 alkyl chain.

[0012] In some embodiments of the present invention, in the above-mentioned compound of formula I, X is -(CH2) n -, n is selected from an integer of 1 to 12.

[0013] In some preferred embodiments of the present invention, in the compound of formula I above, n is selected from an integer of 3 to 10.

[0014] In some embodiments of the present invention, in the above-mentioned compound of formula I, when R1 is selected from , , , or When X is -(CH2)6-.

[0015] The present invention also provides some specific compounds, which are selected from:

[0016] , , , , , , , , or .

[0017] In a second aspect, the present invention also provides a method for preparing the compound represented by the above formula I, comprising:

[0018] When R1 is H, the method comprises the following steps:

[0019] A, 4-chloro-7-fluoroquinoline and N-methylimidazole are subjected to palladium-catalyzed coupling reaction to obtain intermediate 1; the reaction formula is as follows:

[0020] ;

[0021] B. Intermediate 1 and NBS undergo Wohl-Ziegler reaction to obtain intermediate 2; the reaction formula is as follows:

[0022] ;

[0023] C. 4-bromo-3,5-dimethylbenzaldehyde and biboric acid pinacol ester are subjected to Miyaura borylation reaction to obtain the corresponding biboric acid pinacol ester, namely intermediate 3; the reaction formula is as follows:

[0024] ;

[0025] D. Intermediate 2 and intermediate 3 are subjected to Suzuki reaction to obtain intermediate 4; the reaction formula is as follows:

[0026] ;

[0027] E. Intermediate 4 and raw material 5 are subjected to a reduction amination reaction with sodium triacetoxyborohydride to obtain intermediate 6; the reaction formula is as follows:

[0028] ;

[0029] F. Intermediate 6 is subjected to a Boc removal reaction to obtain a compound of formula I in which R1 is H; the reaction formula is as follows:

[0030] ;

[0031] When R1 is not H, based on steps A to F, the following steps are further included:

[0032] G, a compound of formula I in which R1 is H and an amino acid undergo a dehydration condensation reaction to obtain a compound of formula I in which R1 is not H; the amino acid is arginine, histidine, glutamine, asparagine or glycine; the reaction formula is as follows:

[0033] .

[0034] Wherein, in the above preparation method, in step A, the amount of materials used in the palladium-catalyzed coupling reaction is: 4-chloro-7-fluoroquinoline, N-methylimidazole, palladium acetate, potassium acetate and N -The molar volume ratio of methylpyrrolidone is 1 mmol: 1~3 mmol: 0.01~0.1mmol: 1~3 mmol: 5~30 mL.

[0035] Wherein, in the above preparation method, in step A, the conditions for the palladium-catalyzed coupling reaction are: reaction at 90-120° C. under inert gas protection.

[0036] Wherein, in the above preparation method, in step B, the amount of materials used in the Wohl-Ziegler reaction is: the molar volume ratio of intermediate 1, NBS and acetonitrile is 1 mmol: 1~1.5 mmol: 5~30 mL.

[0037] Wherein, in the above preparation method, in step C, the amount of materials used in the Miyaura borylation reaction is: the molar volume ratio of 4-bromo-3,5-dimethylbenzaldehyde, diboronic acid pinacol ester, Pd(dppf)Cl2, potassium acetate and 1,4-dioxane is 1 mmol: 2~4 mmol: 0.01~0.1 mmol: 1~3 mmol: 5~30 mL.

[0038] Wherein, in the above preparation method, in step C, the conditions for the Miyaura borylation reaction are: reaction at 80-100° C. under inert gas protection.

[0039] Wherein, in the above preparation method, in step D, the amount of materials used in the Suzuki reaction is: the molar volume ratio of intermediate 2, intermediate 3, triphenylphosphine palladium, cesium carbonate, 1,4-dioxane and water is 1 mmol: 1~3 mmol: 0.01~0.1 mmol: 2~6mmol: 20~60 mL: 5~15 mL.

[0040] Wherein, in the above preparation method, in step D, the conditions for the Suzuki reaction are: reaction at 80-100° C. under inert gas protection.

[0041] Wherein, in the above preparation method, in step E, the amount of materials used in the reductive amination reaction is: the molar volume ratio of intermediate 4, raw material 5, sodium triacetoxyborohydride, dichloromethane and acetic acid is 1 mmol: 1~3 mmol: 2~4 mmol: 50~200 mL: 0.5~2 mL.

[0042] In the above preparation method, in step F, the amount of materials used in the de-Boc reaction is: the molar volume ratio of intermediate 6, trifluoroacetic acid and dichloromethane is 1 mmol: 2~5 mL: 2~5 mL.

[0043] Wherein, in the above preparation method, in step G, the amount of materials used in the dehydration condensation reaction is: the molar volume ratio of the compound of formula I in which R1 is H, the amino acid, TBTU, DIEA and N,N-dimethylformamide is 1 mmol: 1~2 mmol: 1~2 mmol: 2~4mmol: 50~200 mL.

[0044] In a third aspect, the present invention provides a pharmaceutical composition comprising the compound of formula I or a pharmaceutically acceptable salt thereof as an active ingredient, and a pharmaceutically acceptable carrier or excipient.

[0045] A further object of the present invention is to provide a method for preparing the pharmaceutical composition of the present invention, which comprises combining any compound of Formula I or a pharmaceutically acceptable salt thereof, or a mixture thereof, with one or more pharmaceutically acceptable carriers or excipients.

[0046] In a fourth aspect, the present invention provides use of the aforementioned compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present invention, in the preparation of an NSD3 degrading agent.

[0047] In a fifth aspect, the present invention provides use of the aforementioned compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present invention, in the preparation of a medicament for preventing and / or treating squamous cell carcinoma of the lung.

[0048] Terminology explanation:

[0049] In the present invention, unless otherwise specified, "C1-C12 alkyl chain" refers to a saturated alkane chain containing 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms in a straight or branched form. For example, methylene, -(CH2)2-, -CH(CH3)-, -(CH2)3-, -C(CH3)2-, -CH(CH3)CH2-, -(CH2)9-, -CH(CH3)(CH2)7-, -(CH2) 12 - etc. Similarly, "C3-C10 alkyl chain" refers to a saturated alkane chain containing 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms in a straight or branched form.

[0050] Beneficial effects of the present invention:

[0051] The present invention designs and synthesizes a series of quinoline-imidazole-benzene ring compounds with novel structures. In vitro experiments prove that the compounds of the present invention have a good NSD3 degradation effect on cells, and some compounds also have a concentration gradient NSD3 degradation effect; in addition, the compounds of the present invention also have a good proliferation inhibition effect on NCI-H1703 cells; the compounds of the present invention are expected to be used as a new way to prevent and treat NSD3-related diseases and have good medical application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 Western blotting of the effects of 10 μM compounds ND1-ND10 on NSD3 degradation at the cellular level.

[0053] Figure 2 Western blotting of the effects of compounds ND1 and ND10 on NSD3 in 293T cells transfected with V5-NSD3. DETAILED DESCRIPTION

[0054] The scheme of the present invention will be explained below in conjunction with embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are followed.

[0055] Example 1

[0056] N 1 -(4-(5-(7-fluoroquinolin-4-yl)-1-methyl-1 H -imidazol-4-yl)-3,5-dimethylbenzyl)propane-1,3-diamine

[0057]

[0058] (1) Using 20 mL of N-methylpyrrolidone as solvent, add 1 mmol of 4-chloro-7-fluoroquinoline, 2 mmol of N-methylimidazole, 0.1 mmol of palladium acetate and 3 mmol of potassium acetate in a three-necked flask and react at 100°C overnight under nitrogen protection. The formation of the reaction product, namely intermediate 1 (R f =0.7). EA was used for extraction, and the organic phase was concentrated and purified by column (DCM: MeOH = 20: 1) to obtain intermediate 1 with a yield of 48%.

[0059] (2) 20 mL of acetonitrile was used as solvent, 330 mg of intermediate 1 was added, and 285 mg of NBS was added under ice bath. After the solution was completely dissolved, the mixture was transferred to room temperature for reaction. The reaction was monitored by TLC. After the reaction was complete, column purification (DCM: MeOH = 30: 1) was performed to obtain intermediate 2 with a yield of 56%.

[0060] (3) With 0.05 mmol of Pd(dppf)Cl2 as catalyst, 3 mmol of anhydrous potassium acetate, 1 mmol of 4-bromo-3,5-dimethylbenzaldehyde and 3 mmol of diboric acid pinacol ester were added, and the mixture was heated to 80°C in 20 mL of anhydrous 1,4-dioxane under nitrogen protection. The reaction was monitored by TLC. After the reaction was completed, intermediate 3 was purified by column purification (PE:EA=3:1) with a yield of 38%.

[0061] (4) 20 mL of a mixed solution of 1,4-dioxane and 5 mL of water was used as solvent, 3 mmol of cesium carbonate, 0.5 mmol of intermediate 2, 1 mmol of intermediate 3 and 0.05 mmol of triphenylphosphine palladium were added, and the mixture was reacted at 100°C under nitrogen protection. The reaction was monitored by TLC. After the reaction was completed, intermediate 4 was purified by column purification (DCM: MeOH = 20:1) with a yield of 40%.

[0062] (5) 10 mL of dichloromethane was used as solvent, 0.05 mmol of intermediate 4 and 0.05 mmol of N -tert-Butyloxycarbonyl-1,3-propanediamine and 0.1 mmol of sodium triacetoxyborohydride, add 0.05 mL of acetic acid to induce the reaction, and react at room temperature for 5 h. The reaction was monitored by TLC, and after the reaction was complete, column purification (DCM: MeOH = 20: 1) was performed to obtain intermediate 6 with a yield of 60%.

[0063] (6) 3 mmol of intermediate 6 was dissolved in 10 mL of dichloromethane, followed by 10 mL of trifluoroacetic acid. The reaction was allowed to react at room temperature for 4 hours and monitored by TLC. After the reaction was complete, the product was purified by column chromatography (DCM: MeOH = 20:1) and dried in vacuo to obtain the target compound ND1. The product was a light yellow solid with a yield of 80% in the last step. 1 H NMR (400 MHz, DMSO- d 6) δ 8.89 (d, J = 4.4 Hz, 1H), 8.08 (s, 1H), 7.80 (dd, J = 10.2, 2.7 Hz, 1H), 7.75 – 7.63 (m,1H), 7.60 – 7.47 (m, 1H), 7.27 (d, J = 4.4 Hz, 1H), 7.17 (s, 1H), 7.07 (s,1H), 3.87 (s, 2H), 3.48 (s, 3H), 2.86 (q, J = 6.9 Hz, 4H), 2.08 (s, 3H), 1.95(dd, J = 17.4, 10.2 Hz, 2H), 1.87 (s, 3H). Molecular Weight: 417.5324.

[0064] Example 2

[0065] N 1-(4-(5-(7-fluoroquinolin-4-yl)-1-methyl-1 H -imidazol-4-yl)-3,5-dimethylbenzyl)butane-1,4-diamine

[0066]

[0067] The synthesis steps are the same as in Example 1, except N -tert-Butyloxycarbonyl-1,3-propylenediamine N -tert-Butyloxycarbonyl-1,4-butanediamine to obtain the target compound ND2. White solid, the final step yield is 84%. 1 H NMR (400 MHz, DMSO- d 6) δ8.89 (d, J = 4.4 Hz, 1H), 8.10 (s, 1H), 7.83 – 7.78 (m, 1H), 7.74 – 7.68 (m,1H), 7.56 – 7.52 (m, 1H), 7.28 (d, J = 4.5 Hz, 1H), 7.21 (s, 1H), 7.11 (s,1H), 3.93 (s, 2H), 3.48 (s, 3H), 2.78 (dt, J = 22.5, 7.4 Hz, 4H), 2.08 (s,3H), 1.87 (s, 3H), 1.74 – 1.65 (m, 2H), 1.64 – 1.56 (m, 2H).

[0068] Example 3

[0069] N 1 -(4-(5-(7-fluoroquinolin-4-yl)-1-methyl-1 H -imidazol-4-yl)-3,5-dimethylbenzyl)pentane-1,5-diamine

[0070]

[0071] The synthesis steps are the same as in Example 1, except N -tert-Butyloxycarbonyl-1,3-propanediamine was replaced with mono-BOC-pentanediamine to obtain the target compound ND3. White solid, the final step yield was 86%. 1 H NMR (400 MHz, DMSO-d 6) δ 8.88 (d, J =4.4 Hz, 1H), 8.06 (s, 1H), 7.83 – 7.76 (m, 1H), 7.72 – 7.66 (m, 1H), 7.55 –7.48 (m, 1H), 7.27 (d, J = 4.5 Hz, 1H), 7.00 (s, 1H), 6.95 – 6.88 (m, 1H), 3.64 (s, 2H), 3.48 (s, 3H), 2.73 – 2.65 (m, 2H), 2.05 (s, 3H), 1.86 (s, 3H), 1.57 – 1.42 (m, 4H), 1.36 – 1.30 (m, 2H), 1.25 – 1.22 (m, 2H).

[0072] Example 4

[0073] N 1 -(4-(5-(7-fluoroquinolin-4-yl)-1-methyl-1 H -imidazol-4-yl)-3,5-dimethylbenzyl)hexane-1,6-diamine

[0074]

[0075] The synthesis steps are the same as in Example 1, except N -tert-Butyloxycarbonyl-1,3-propylenediamine N -(6-aminohexyl)carbamic acid tert-butyl ester to obtain the target compound ND4 as a white solid with a final step yield of 80%. 1 H NMR (400 MHz, Methanol- d 4) δ 8.85 (d, J = 4.5 Hz, 1H), 8.61 (s, 1H), 7.86 – 7.80 (m, 1H), 7.74 (dd, J = 9.8, 2.6 Hz, 1H), 7.53 – 7.46 (m, 1H), 7.38 (d, J= 4.5 Hz,1H), 7.19 (s, 1H), 7.09 (s, 1H), 4.05 (s, 2H), 3.66 (s, 3H), 3.53 – 3.44 (m,1H), 2.98 – 2.89 (m, 4H), 2.22 (s, 3H), 2.02 (s, 3H), 1.67 (q, J = 7.6 Hz, 4H), 1.43 – 1.38 (m, 4H), 1.30 – 1.26 (m, 2H).

[0076] Example 5

[0077] N 1 -(4-(5-(7-fluoroquinolin-4-yl)-1-methyl-1 H -imidazol-4-yl)-3,5-dimethylbenzyl)heptane-1,7-diamine

[0078]

[0079] The synthesis steps are the same as in Example 1, except N -tert-Butyloxycarbonyl-1,3-propanediamine was replaced with 1-tert-Butyloxycarbonyl-1,7-diaminoheptane to obtain the target compound ND5. A light yellow solid with a final step yield of 82%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.88 (d, J = 4.4 Hz, 1H), 8.06 (s, 1H), 7.82 – 7.76 (m, 1H), 7.72 –7.66 (m, 1H), 7.54 – 7.47 (m, 1H), 7.27 (d, J = 4.4 Hz, 1H), 6.98 (s, 1H), 6.91 (s, 1H), 3.64 (s, 2H), 3.48 (s, 3H), 2.76 – 2.64 (m, 2H), 2.03 (s, 3H), 1.86 (s, 3H), 1.59 – 1.36 (m, 4H), 1.39 – 1.01 (m, 8H).

[0080] Example 6

[0081] ( S )-2-amino- N-(6-((4-(5-(7-fluoroquinolin-4-yl)-1-methyl-1 H -imidazol-4-yl)-3,5-dimethylbenzyl)amino)hexyl)-5-guanidinopentanamide

[0082]

[0083] The synthesis steps are the same as those in Example 1. After obtaining Example 4, a condensation reaction is performed. The specific steps are as follows:

[0084] (1) 0.055 mmol of L-arginine was dissolved in 10 mL of N,N-dimethylformamide (DMF), and 0.06 mmol of TBTU was added to activate the mixture at room temperature for 20 minutes. Subsequently, 0.05 mmol of ND4 obtained in Example 4 and 0.165 mmol of DIEA were added, and the reaction was continued at room temperature overnight.

[0085] (2) The next day, the reaction progress was monitored by TLC. After the reaction of the raw materials was completed, 100 mL of H2O and 20 mL of EA were added for extraction three times, the polar phases were combined, and the aqueous phase was discarded. The organic phase was then washed with H2O, saturated sodium bicarbonate solution, and saturated NaCl aqueous solution in turn. The organic solvent was then removed by reduced pressure distillation, and the reaction was monitored by TLC. After the reaction was complete, it was purified by column purification (DCM: MeOH = 20:1) and vacuum dried to obtain the target compound ND6. The yield was 52%. 1 H NMR (400 MHz, Methanol- d 4) δ9.36 (s, 1H), 8.98 – 8.92 (m, 1H), 7.94 – 7.88 (m, 1H), 7.84 – 7.78 (m, 1H), 7.59 – 7.53 (m, 1H), 7.52 – 7.45 (m, 1H), 7.07 – 6.99 (m, 1H), 6.96 – 6.88(m, 1H), 3.92 – 3.87 (m, 1H), 3.76 (s, 3H), 3.33 – 3.31 (m, 1H), 3.23 – 3.14(m, 4H), 3.03 – 2.85 (m, 2H), 2.24 (s, 3H), 2.00 (s, 3H), 1.96 – 1.80 (m,4H), 1.76 – 1.63 (m, 4H), 1.55 – 1.42 (m, 4H), 1.31 – 1.24 (m, 4H).

[0086] Example 7

[0087] 2-amino- N -(6-((4-(5-(7-fluoroquinolin-4-yl)-1-methyl-1 H -imidazol-4-yl)-3,5-dimethylbenzyl)amino)hexyl)-3-(1 H -imidazol-4-yl)propanamide

[0088]

[0089] The synthesis steps are the same as those in Example 1. After obtaining Example 4, a condensation reaction is performed. The specific steps are as follows:

[0090] (1) 0.055 mmol of L-histidine was dissolved in 10 mL of N,N-dimethylformamide (DMF), and 0.06 mmol of TBTU was added to activate the mixture at room temperature for 20 minutes. Subsequently, 0.05 mmol of ND4 obtained in Example 4 and 0.165 mmol of DIEA were added, and the reaction was continued at room temperature overnight.

[0091] (2) The reaction progress was monitored by TLC on the next day. After the reaction of the raw materials was completed, 100 mL of H2O and 20 mL of EA were added for extraction three times, the polar phases were combined, and the aqueous phase was discarded. The organic phase was then washed with H2O, saturated sodium bicarbonate solution, and saturated NaCl aqueous solution in turn. The organic solvent was then removed by vacuum distillation, and the reaction was monitored by TLC. After the reaction was complete, it was purified by column purification (DCM: MeOH = 20:1) and vacuum dried to obtain the target compound ND7. The yield was 42%. 1 H NMR (400 MHz, Methanol- d 4) δ9.33 (s, 1H), 8.98 – 8.80 (m, 3H), 7.90 (t, J = 7.4 Hz, 1H), 7.84 – 7.75 (m,1H), 7.55 (t, J = 8.6 Hz, 1H), 7.50 – 7.46 (m, 1H), 7.43 (d, J= 10.1 Hz,2H), 7.09 – 6.99 (m, 1H), 6.97 – 6.86 (m, 1H), 4.78 – 4.71 (m, 2H), 4.53 (s,1H), 4.23 – 4.16 (m, 1H), 3.75 (s, 3H), 3.28 – 3.20 (m, 2H), 3.20 – 3.15 (m,2H), 2.31 – 2.20 (m, 3H), 2.06 – 2.01 (m, 2H), 2.01 (s, 3H), 1.47 – 1.34 (m,4H), 1.35 – 1.25 (m, 4H).

[0092] Example 8

[0093] N 1 -(4-(5-(7-fluoroquinolin-4-yl)-1-methyl-1 H -imidazol-4-yl)-3,5-dimethylbenzyl)octane-1,8-diamine

[0094]

[0095] The synthesis steps are the same as in Example 1, except N -tert-Butyloxycarbonyl-1,3-propanediamine was replaced with 1-tert-Butyloxycarbonyl-1,8-diaminooctane to obtain the target compound ND8. A light yellow solid with a final step yield of 80%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.89 (d, J = 4.5 Hz, 1H), 8.10 (s, 1H), 7.80 (dd, J = 10.1, 2.7 Hz,1H), 7.69 (dd, J = 9.3, 6.1 Hz, 1H), 7.52 (td, J = 8.8, 2.7 Hz, 1H), 7.29 (d, J= 4.5 Hz, 1H), 7.17 (s, 1H), 7.09 (s, 1H), 3.91 (s, 2H), 3.49 (s, 3H), 2.97– 2.56 (m, 4H), 2.06 (s, 3H), 1.89 (s, 3H), 1.66 – 1.41 (m, 4H), 1.33 – 1.16 (m, 8H).

[0096] Example 9

[0097] N 1 -(4-(5-(7-fluoroquinolin-4-yl)-1-methyl-1 H -imidazol-4-yl)-3,5-dimethylbenzyl)nonane-1,9-diamine

[0098]

[0099] The synthesis steps are the same as in Example 1, except N -tert-Butyloxycarbonyl-1,3-propylenediamine N -BOC-1,9-nonanediamine, to obtain the target compound ND9. Light yellow solid, the final step yield is 83%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.88(d, J = 4.4 Hz, 1H), 8.07 (s, 1H), 7.83 – 7.75 (m, 1H), 7.69 (dd, J = 9.3,6.2 Hz, 1H), 7.55 – 7.46 (m, 1H), 7.28 (d, J = 4.4 Hz, 1H), 7.05 (s, 1H), 6.97 (s, 1H), 3.74 (s, 2H), 3.48 (s, 3H), 2.72 (t, J = 7.6 Hz, 2H), 2.63 –2.53 (m, 2H), 2.04 (s, 3H), 1.88 (s, 3H), 1.58 – 1.46 (m, 4H), 1.27 – 1.15(m, 11H).

[0100] Example 10

[0101] N 1-(4-(5-(7-fluoroquinolin-4-yl)-1-methyl-1 H -imidazol-4-yl)-3,5-dimethylbenzyl)decane-1,10-diamine

[0102]

[0103] The synthesis steps are the same as in Example 1, except N -tert-Butyloxycarbonyl-1,3-propanediamine was replaced with 1-BOC-1,10-diaminodecane to obtain the target compound ND10. Pale yellow solid, the final step yield was 85%. 1 H NMR (400 MHz, DMSO- d 6) δ8.88 (d, J = 4.4 Hz, 1H), 8.06 (s, 1H), 7.83 – 7.74 (m, 1H), 7.74 – 7.66 (m,1H), 7.51 (td, J = 8.8, 2.7 Hz, 1H), 7.27 (d, J = 4.4 Hz, 1H), 6.93 (s, 1H), 6.86 (s, 1H), 3.56 (s, 2H), 3.48 (s, 3H), 2.71 (t, J = 7.6 Hz, 2H), 2.46 –2.38 (m, 2H), 2.03 (s, 3H), 1.86 (s, 3H), 1.58 – 1.47 (m, 2H), 1.43 – 1.34(m, 2H), 1.26 – 1.16 (m, 12H).

[0104] Embodiment 11

[0105] CCK8 was used to detect the proliferation of NCI-H1703 cells after 72 hours of drug treatment, with Sorafenib as the positive drug. The results are shown in Table 1. The specific operation is as follows:

[0106] (1) Collect cells and count and plate: Remove the upper layer of culture medium from NCI-H1703 cells in the logarithmic growth phase in a 10 cm culture dish with a pipette, wash twice with sterile phosphate buffered saline (PBS), and then add 1 mL of 0.25% trypsin to digest. Incubate in an incubator at 37°C and 5% carbon dioxide for 1 minute, then remove and add fresh RPMI 1640 complete culture medium (containing 10% fetal bovine serum, 100 U / mL penicillin and 100 mg / ml streptomycin) to terminate digestion. Then transfer the mixed cell suspension to a 15 mL centrifuge tube and centrifuge at 400 g for 3 minutes at room temperature. Discard the supernatant, add 4 ml of culture medium to resuspend, disperse the cells into a single cell suspension, and take 20 μL of the cell suspension, add trypan blue solution, mix and dilute, and count to obtain the cell density. Calculate the total number of cells required based on the number of 3000 cells per well in a 96-well plate. Take out the liquid volume corresponding to the required number of cells from the single-cell suspension prepared above, and make up the taken-out cell suspension to the corresponding total volume according to the 100 μL cell suspension volume per well of the 96-well plate. Then mix it evenly again, use a sample pipette to evenly and quickly add the cell suspension to the 96-well plate, add 200 μL PBS solution to the side wells to prevent excessive evaporation of the culture medium volume in the center well, and then quickly transfer it to a 37°C, 5% carbon dioxide incubator for overnight culture.

[0107] (2) On the second day, after observing that the cells have attached to the wall, the compound was prepared into a 10 mM stock solution, and the compound was serially diluted using RPMI 1640 complete medium to obtain 100 μM, 20 μM and 4 μM compound solutions. 100 μL of the above concentrations of compound solution was added to each well of the 96-well plate, and there were 3 parallel wells for each concentration. 3 wells were used as the Control group, with only cells and no drug treatment, and 3 wells were used as the Blank group, with only culture medium without cells or drugs. As the Blank group, these 6 wells were added with an equal volume of fresh RPMI 1640 complete medium when the drug was added. After 72 hours of compound treatment, 20 μL of CCK8 solution (Baoguang Bio, BG0025) was added to each well. The wells were incubated in an incubator at 37°C and 5% carbon dioxide until the solution turned yellow. The absorbance (OD) of the sample was measured at a wavelength of 450 nm using an enzyme reader (BioTek Cytation 3). The cell proliferation inhibition rate was calculated according to the following formula:

[0108] Inhibition rate (%) = (OD Control -OD Compound ) / (OD Control -OD Blank ) × 100%;

[0109] Among them, OD Control : Absorbance reading of wells containing cells, culture medium, CCK-8 solution, but no drug; OD Compound : Absorbance readings of wells containing cells, culture medium, CCK-8 solution, and compound solution; OD Blank : Absorbance readings of wells containing culture medium and CCK-8 solution, but not containing cells or drugs.

[0110] Table 1 The results of the inhibition of NCI-H1703 cell proliferation by the compounds of the present invention

[0111]

[0112] a All experiments were repeated at least 3 times.

[0113] As shown in Table 1, ND1, ND2, ND4, ND5, ND8~ND10 all have good proliferation inhibition effects on NCI-H1703 cells, especially ND8~ND10, the inhibition rate of cells at 10 μM exceeds 50%.

[0114] Example 12

[0115] Western blotting was used to detect the degradation effect of the compound on NSD3 in 293T cells overexpressing NSD3. The specific operation is as follows:

[0116] (1) 293T cells in the logarithmic growth phase were digested with 0.25% trypsin and then terminated with DMEM complete medium (containing 10% fetal bovine serum, 100 U / mL penicillin and 100 mg / ml streptomycin). 2×10 6 Cells were plated into one well of a 6-well plate, and transfection was performed after the cells were cultured overnight and adhered to the wall and the cell density reached 60%.

[0117] (2) Change the cell culture medium 1 hour in advance by adding fresh DMEM complete medium solution containing chloroquine (final concentration is 25 mM).

[0118] (3) Prepare transfection system: Prepare solution A and solution B according to Table 2. The whole process should be performed in a clean bench. First, add pure water to a 1.5 ml sterile microcentrifuge tube. Then, use a pipette to accurately transfer the Plvx-V5-NSD3 plasmid solution and CaCl2 solution to the centrifuge tube. Mix well with a vortexer to obtain solution A. Accurately transfer solution B to a new 1.5 ml sterile microcentrifuge tube. Then, place solution B on a vortexer at 1500 rpm to vortex solution B. Evenly add solution A to solution B at a rate of 2 drops / second to prepare DNA-calcium phosphate complex. Remove cells, evenly add DNA-calcium phosphate complex to culture medium, gently mix well, and place in an incubator. After 10 hours, remove cells, replace with fresh culture medium, and continue to culture overnight.

[0119] Table 2 Plasmid transfection system

[0120]

[0121] (4) 24 hours after transfection, add culture medium containing the corresponding concentration of compound: prepare the compound into 10 mM stock solution and then use DMEM complete culture medium solution for gradient dilution to prepare a mixed solution containing a final concentration of 10 μM or 5 μM compound, and continue to culture for 24 hours. Then collect the cell sample, use cell lysis buffer (containing protease and phosphatase inhibitors) for lysis on ice, collect the supernatant after centrifugation for 10 minutes, add protein loading buffer and mix well, denature at 100 degrees Celsius for 10 minutes, and use protein immunoblotting to detect the degradation of NSD3 protein. The results are as follows: Figure 1 and Figure 2 shown.

[0122] Figure 1 The figure is a protein immunoblot diagram of the degradation effect of 10 μM ND1-ND10 compound on NSD3 at the cellular level. BI-9321 is a probe small molecule targeting NSD3, which has a strong affinity binding effect on NSD3. The protein immunoblot experiment results of the present invention use this molecule as a positive control. Figure 1 It can be seen that when the action concentration is also 10 μM, the NSD3 bands of ND1~ND6 basically disappear, indicating that ND1~ND6 have better degradation effects on the long isomer (represented by NSD3-L) and short isomer (represented by NSD3-S) of NSD3 protein than BI-9321; the NSD3 band of ND10 is lighter, and it has a degradation effect comparable to BI-9321; ND7~9 has a degradation effect comparable to BI-9321 on the long isomer of NSD3 protein. It can be seen that the compounds of the present invention have a good degradation effect on NSD3.

[0123] Figure 2The figure is a Western blotting of the effects of ND1 and ND10 compounds on NSD3 in 293T cells transfected with V5-NSD3. Figure 2 It can be seen that the protein imprinting experiment confirmed that 10 μM ND1 and ND10 compounds had significant degradation effects on long and short isomers of NSD3 protein, which was much better than the DMSO treatment group and significantly better than 5 μM, indicating that compounds such as ND1 and ND10 of the present invention have a concentration gradient NSD3 protein degradation effect, and the degradation effect is not caused by toxicity to cells.

Claims

1. A compound represented by formula I or a pharmaceutically acceptable salt thereof, characterized in that: The structure of Formula I is as follows: ； in: X is -(CH2) n -, n is an integer selected from 3 to 10; R1 is H.

2. The method for preparing the compound represented by formula I according to claim 1, characterized in that: The following steps are involved: A, 4-chloro-7-fluoroquinoline and N-methylimidazole are subjected to palladium-catalyzed coupling reaction to obtain intermediate 1; the reaction formula is as follows: ； B. Intermediate 1 and NBS undergo Wohl-Ziegler reaction to obtain intermediate 2; the reaction formula is as follows: ； C. 4-bromo-3,5-dimethylbenzaldehyde and biboric acid pinacol ester are subjected to Miyaura borylation reaction to obtain the corresponding biboric acid pinacol ester, namely intermediate 3; the reaction formula is as follows: ； D. Intermediate 2 and intermediate 3 are subjected to Suzuki reaction to obtain intermediate 4; the reaction formula is as follows: ； E. Intermediate 4 and raw material 5 are subjected to a reduction amination reaction with sodium triacetoxyborohydride to obtain intermediate 6; the reaction formula is as follows: ； F. Intermediate 6 is subjected to a Boc removal reaction to obtain a compound of formula I in which R1 is H; the reaction formula is as follows: 。 3. The preparation method according to claim 2, characterized in that: At least one of the following is met: In step A, the amount of materials used in the palladium-catalyzed coupling reaction is: the molar volume ratio of 4-chloro-7-fluoroquinoline, N-methylimidazole, palladium acetate, potassium acetate and N-methylpyrrolidone is 1 mmol: 1-3 mmol: 0.01-0.1 mmol: 1-3 mmol: 5-30 mL; In step A, the conditions for the palladium-catalyzed coupling reaction are: reaction at 90-120° C. under inert gas protection; In step B, the amount of materials used in the Wohl-Ziegler reaction is: the molar volume ratio of intermediate 1, NBS and acetonitrile is 1 mmol: 1-1.5 mmol: 5-30 mL; In step C, the amount of materials used in the Miyaura borylation reaction is: the molar volume ratio of 4-bromo-3,5-dimethylbenzaldehyde, diboronic acid pinacol ester, Pd(dppf)Cl2, potassium acetate and 1,4-dioxane is 1 mmol: 2-4 mmol: 0.01-0.1 mmol: 1-3 mmol: 5-30 mL; In step C, the conditions for the Miyaura borylation reaction are: reaction at 80-100° C. under inert gas protection; In step D, the amount of materials used in the Suzuki reaction is: the molar volume ratio of intermediate 2, intermediate 3, triphenylphosphine palladium, cesium carbonate, 1,4-dioxane and water is 1 mmol: 1-3 mmol: 0.01-0.1 mmol: 2-6 mmol: 20-60 mL: 5-15 mL; In step D, the conditions for the Suzuki reaction are: reaction at 80-100° C. under inert gas protection; In step E, the amount of materials used in the reductive amination reaction is: the molar volume ratio of intermediate 4, raw material 5, sodium triacetoxyborohydride, dichloromethane and acetic acid is 1 mmol: 1-3 mmol: 2-4 mmol: 50-200 mL: 0.5-2 mL; In step F, the amount of materials used in the de-Boc reaction is: the molar volume ratio of intermediate 6, trifluoroacetic acid and dichloromethane is 1 mmol: 2~5 mL: 2~5 mL.

4. A pharmaceutical composition, characterized in that: The invention uses the compound described in claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient, and is supplemented with a pharmaceutically acceptable carrier or excipient.

5. Use of the compound or pharmaceutically acceptable salt thereof according to claim 1 or the pharmaceutical composition according to claim 4 in the preparation of a NSD3 degrading agent.

6. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 4 in the preparation of a medicament for preventing and treating lung squamous cell carcinoma.

Images