Compound for degrading Pol theta based on hydrophobic label as well as preparation method and application of compound

Abstract

The invention discloses a compound for degrading Pol theta based on a hydrophobic label as well as a preparation method and application of the compound. The structural formula of the compound for targeted degradation of Pol theta is as shown in a general formula (I). Compared with a Pol theta inhibitor, the compound provided by the invention can degrade Pol theta protein at a proper dosage. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to compounds, preparation methods and uses thereof, and in particular to a series of compounds or pharmaceutically acceptable salts thereof for degrading Polθ based on hydrophobic tags, preparation methods and uses thereof. Background Art

[0002] Synthetic lethality provides a new approach for the treatment of mutant genes that were previously considered untargetable in traditional genotype-targeted therapies. DNA polymerase θ (Polθ, encoded by the POLQ gene) plays an important role in the microhomology end-mediated repair pathway. Polθ consists of a C-terminal DNA polymerase domain, an N-terminal ATPase / helicase domain, and a disordered central domain. In homologous recombination-deficient cells, Polθ can perform error-prone DNA synthesis at sites of DNA damage through the microhomology end-mediated repair pathway and is synthetically lethal in homologous recombination (HR)-deficient cancers. Polθ is essentially not expressed in normal cells, but is upregulated in a variety of cancers, including breast cancer, lung cancer, colorectal cancer, ovarian cancer, and gastric cancer. In addition, increased Polθ expression is associated with a poor prognosis in breast cancer. The survival of cancer cells lacking HR, NHEJ, or ATM is highly dependent on the activity of Polθ. Therefore, Polθ is an attractive target for novel synthetic lethal therapies in cancers with DNA repair defects. Summary of the invention

[0003] Purpose of the invention: The purpose of the present invention is to provide a small molecule compound that degrades Polθ based on a hydrophobic tag, which utilizes a hydrophobic tag degradation strategy to degrade Polθ. Another purpose of the present invention is to provide a method for preparing the above-mentioned compound or a pharmaceutically acceptable salt thereof. Another purpose of the present invention is to provide a pharmaceutical composition. The last purpose of the present invention is to provide the use of the compound or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating or preventing cancer with homologous recombination deficiency.

[0004] Technical solution: The compound for degrading Polθ based on a hydrophobic tag or a pharmaceutically acceptable salt thereof according to the present invention, the structural formula of the compound for degrading Polθ is shown in general formula (I):

[0005]

[0006] in,

[0007] Linker does not exist or is selected from:

[0008] Unsubstituted or secondary amino-substituted 5- to 8-membered cycloalkyl, unsubstituted or -(CH2) 1～ 4-NH-substituted 5-8 membered nitrogen-containing heterocyclic group, unsubstituted or secondary amino-substituted 6-11 membered spiro ring containing nitrogen and carbon, unsubstituted or secondary amino-substituted 6-10 membered bridged ring containing nitrogen and carbon, 4-6 membered nitrogen heterocyclic ring-4-6 membered nitrogen heterocyclic ring;

[0009] Ra is selected from a saturated alkyl group having 0 to 8 carbon atoms and an alkoxy group having 4 to 6 carbon atoms;

[0010] Rb is selected from an alkoxy group having 1 to 6 carbon atoms;

[0011] Rc is selected from an alkoxy group having 1 to 6 carbon atoms;

[0012] HyT is:

[0013]

[0014] The compound for degrading Polθ based on the hydrophobic tag or a pharmaceutically acceptable salt thereof,

[0015] Linker does not exist or is selected from:

[0016] Unsubstituted or secondary amino-substituted 6-membered cycloalkyl, unsubstituted or -(CH2) 1～4 -NH-substituted piperazinyl, unsubstituted or secondary amino-substituted 6-11-membered spiro ring containing nitrogen and carbon, unsubstituted or secondary amino-substituted 6-10-membered bridged ring containing nitrogen and carbon, 4-membered nitrogen heterocycle-6-membered nitrogen heterocycle;

[0017] Ra is selected from a saturated alkyl group having 0 to 8 carbon atoms and an alkoxy group having 4 to 6 carbon atoms;

[0018] Rb is selected from an alkoxy group having 1 to 6 carbon atoms;

[0019] Rc is selected from an alkoxy group having 1 to 6 carbon atoms.

[0020] The compound for degrading Polθ based on a hydrophobic tag or a pharmaceutically acceptable salt thereof, the linker is absent or selected from:

[0021] Unsubstituted or secondary amino-substituted 6-membered cycloalkyl, unsubstituted or -(CH2) 1～4 -NH-substituted piperazinyl, unsubstituted or secondary amino-substituted 6-11 membered spiro ring containing nitrogen and carbon, unsubstituted or secondary amino-substituted 6-10 membered bridged ring containing nitrogen and carbon, azetidine-piperazinyl;

[0022] Ra is selected from a saturated alkyl group having 0 to 8 carbon atoms and an alkoxy group having 4 to 6 carbon atoms;

[0023] Rb is selected from an alkoxy group having 1 to 6 carbon atoms;

[0024] Rc is selected from an alkoxy group having 1 to 6 carbon atoms.

[0025] The compound for degrading Pol0 based on a hydrophobic tag or a pharmaceutically acceptable salt thereof, wherein the Linker does not exist or is selected from any one of the following structures:

[0026]

[0027] R1 is selected from:

[0028]

[0029] R2 is selected from: -CH2-, -CH2-CH2-.

[0030] The compound for degrading Pol0 based on a hydrophobic tag or a pharmaceutically acceptable salt thereof, wherein the Linker does not exist or is selected from any one of the following structures:

[0031]

[0032] A compound for degrading Polθ based on a hydrophobic tag or a pharmaceutically acceptable salt thereof, wherein the structural formula of the compound for degrading Polθ based on a hydrophobic tag is shown in any one of the following:

[0033]

[0034] The method for preparing the compound for degrading Polθ based on a hydrophobic tag comprises the following steps:

[0035] (1) (2-methoxyphenyl)boronic acid (Compound 1) and 3-bromoisonicotinic acid methyl ester (Compound 2) are subjected to Suzuki reaction to prepare 3-(2-methoxyphenyl)isonicotinic acid methyl ester (Compound 3);

[0036] (2) Compound 3 is hydrolyzed under alkaline conditions to obtain 3-(2-methoxyphenyl)isonicotinic acid (compound 4);

[0037] (3) 5-bromo-1,3,4-thiadiazol-2-amine (Compound 5) and methyl 4-hydroxymethylbenzoate (Compound 6) are subjected to substitution reaction to prepare methyl 4-(((5-amino-1,3,4-thiadiazol-2-yl)oxy)methyl)benzoate (Compound 7);

[0038] (4) Compound 4 and 7 are reacted with a condensing agent 2-(7-azobenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU) to produce methyl 4-(((5-(3-(2-methoxyphenyl)isonicotinamido)-1,3,4-thiadiazol-2-yl)oxy)methyl)benzoate (Compound 8);

[0039] (5) Compound 8 is hydrolyzed under alkaline conditions to obtain 4-(((5-(3-(2-methoxyphenyl)isonicotinamido)-1,3,4-thiadiazol-2-yl)oxy)methyl)benzoic acid (Compound 9);

[0040] (6) The linker modules 11b-11r are first reacted with adamantane acetic acid in the presence of a condensing agent 2-(7-azobenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU) to produce compounds 12b-12r;

[0041]

[0042] (7) Compound 12b-12r is subjected to a Boc protecting group removal reaction and then subjected to an amidation reaction with compound 9 to obtain a compound of formula (I).

[0043] Use of the compound for degrading Polθ based on the hydrophobic tag or a pharmaceutically acceptable salt thereof in the preparation of a drug targeted at degrading Polθ.

[0044] The use of the compound for degrading Polθ based on the hydrophobic tag or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating and / or preventing cancer. Preferably, the cancer includes breast cancer.

[0045] A pharmaceutical composition comprising the compound for degrading Polθ based on the hydrophobic tag or a pharmaceutically acceptable salt thereof.

[0046] Beneficial effects: Compared with Polθ inhibitors, the compounds of the present invention can degrade Polθ protein and are different from PROTAC technology in mechanism. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 is the degradation effect of the compound on Polθ protein in MDA-MB-436 cells;

[0048] Figure 2 This is a diagram of the degradation mechanism of compound HT-11. DETAILED DESCRIPTION

[0049] The NMR spectra of the final products and intermediates in the examples were measured using DMSO-d6 or CDCl3-d3 as solvent and TMS as internal standard, using a 300 MHz or 400 MHz NMR spectrometer from Bruker; high-resolution mass spectrometry (HRMS) was measured using an Agilent Q-TOF 6520 mass spectrometer.

[0050] The reagents used in the synthesis, purification and separation of the compounds are as follows: (1) Column chromatography silica gel: 200 or 300 mesh silica gel were purchased from Qingdao Ocean Chemical Industry; (2) HSGF254 TLC thin layer chromatography plate: purchased from Yantai Chemical Research Institute; (3) Conventional solvents used in column chromatography elution systems such as petroleum ether, dichloromethane, ethyl acetate, methanol, etc. and chemical reagents required for the reaction, unless otherwise specified, were all commercially available chemically pure or analytically pure products.

[0051] Example 1: Synthesis of connecting chain derivatives 12b-12r. Compound 10 (100 mg, 0.51 mmol) was dissolved in 5 ml of anhydrous DMF, and 2-(7-azobenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU, 194 mg, 0.51 mmol) and N,N-diisopropylethylamine (DIPEA, 276 mg, 2.13 mmol) were added in sequence. After reacting at room temperature for 20 min under nitrogen protection, amine compounds 11b-1r (1.0 mmol) of different lengths were added, and reacted at 50°C for 5 h under nitrogen protection. After the reaction was completed by TLC detection, the reaction solution was poured into water, extracted with ethyl acetate, the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain 12b-12r, which was directly added without purification.

[0052] Synthesis route:

[0053]

[0054] Example 2: Synthesis of N-(5-((4-((3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)propyl)carbamoyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14a)

[0055] Structural formula:

[0056]

[0057] Reference synthesis route:

[0058]

[0059] The synthesis method of step 1 to step 5 refers to the synthesis method of compound 9 in patent CN 118515661 B.

[0060] Step 6:

[0061] 4-(((5-(3-(2-methoxyphenyl)isonicotinamido)-1,3,4-thiadiazol-2-yl)oxy)methyl)benzoic acid (Compound 9, 100 mg, 0.22 mmol) was dissolved in anhydrous 3 ml DMF, and DIPEA (71 mg, 0.55 mmol) and 2-(7-azobenzotriazole)-tetramethyluronium hexafluorophosphate (84 mg, 0.22 mmol) were added in sequence. The mixture was stirred at room temperature for 20 min under N2 protection, and adamantane amine (Compound 13, 27.2 mg, 0.18 mmol) was added and reacted at 50°C for 5 h. After the reaction was completed as monitored by TCL, water was added, the mixture was extracted with ethyl acetate, washed with water, washed with saturated brine, dried over anhydrous Na2SO4, evaporated under reduced pressure, and column chromatography was performed to obtain 30 mg (14a) of a white solid. 1 H NMR (300MHz, DMSO-d6) δ12.88 (s, 1H), 8.71 (d, J = 5.0Hz, 1H), 8.61 (s, 1H), 7.80 (d, J = 8.0Hz, 2H), 7.62 (d, J = 4.8Hz, 2H), 7.52 (d .

[0062] Example 3: Synthesis of N-(5-((4-((2-(2-((3r,5r,7r)-adamantanamine-1-yl)acetylamino)ethyl)carbamoyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14b)

[0063] Structural formula:

[0064]

[0065] Reference synthesis route:

[0066]

[0067] The synthesis method of step 1 to step 5 refers to the synthesis method of compound 9 in patent CN118515661B.

[0068] Step 6

[0069] The connecting chain 12b (100 mg, 0.3 mmol) was dissolved in 1 mL of ethyl acetate, and 1 mL of 2.0 mol / L hydrochloric acid-ethyl acetate solution (Annaiji Chemical, W810184) was added. The reaction was allowed to react at room temperature for 3 h. After the reaction was completed as monitored by TLC, the mixture was concentrated under reduced pressure to obtain 100 mg of a yellow solid, which was used directly in the next step. 4-(((5-(3-(2-methoxyphenyl)isonicotinamido)-1,3,4-thiadiazol-2-yl)oxy)methyl)benzoic acid (Compound 9, 100 mg, 0.22 mmol) was dissolved in anhydrous 4 ml DMF, and DIPEA (143 mg, 1.1 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI, 42.18 mg, 0.22 mmol), and 1-hydroxybenzotriazole (HOBT, 29.73 mg, 0.22 mmol) were added in sequence. The mixture was stirred at room temperature for 20 min under N2 protection, and the above crude product was added. The mixture was reacted at 50°C overnight. After the reaction was completed under TCL monitoring, water was added, the mixture was extracted with ethyl acetate, washed with water, washed with saturated brine, dried over anhydrous Na2SO4, evaporated under reduced pressure, and column chromatography was performed to obtain 10 mg (12a) of a white solid. 1 H NMR (300MHz, DMSO-d6) δ12.89 (s, 1H), 8.71 (s, 1H), 8.61 (s, 1H), 8.49 (s, 1H), 7.86 (d, J = 8.7Hz, 3H), 7.59 (dd, J = 17.5, 6.4Hz, 3H), 7.36 (d, J=7.6Hz, 2H), 7.11-6.95 (m, 2H), 5.53 (s, 2H), 3.50 (s, 3H), 3.30 (s, 2H), 3.26-3.19 (m, 2H), 1.83-1.81 (m, 5H), 1.61-1.51 (m, 12H).

[0070] Example 4: Synthesis of N-(5-((4-((2-(2-(2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)carbamoyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14c)

[0071]

[0072] Refer to the synthesis method of Example 3. Replace compound 12b with tert-butyl (3-aminopropyl) carbamate, and keep other conditions unchanged. 1H NMR (300MHz, CDCl3) δ8.65 (d, J=5.0Hz, 1H), 8.57 (s, 1H), 7.84 (m, 3H), 7.59 (d, J=5.0Hz, 1H) , 7.41 (d, J=8.0Hz, 2H), 7.34 (td, J=7.9, 1.8Hz, 1H), 7.18 (dd, J=7.5, 1.7Hz, 1H), 7.00 (t, J=7 .4Hz, 1H), 6.86 (d, J=8.2Hz, 1H), 6.35 (t, J=6.4Hz, 1H), 5.36 (s, 2H), 3.56 (s, 3H), 3.46 (q, J =6.0Hz, 2H), 3.30 (q, J=6.0Hz, 2H), 1.94-1.92 (m, 5H), 1.71-1.69 (m, 5H), 1.59-1.54 (m, 9H).

[0073] Example 5: Synthesis of N-(5-((4-((4-(2-((3r,5r,7r)-adamantan-1-yl)acetylamino)butyl)carbamoyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14d)

[0074]

[0075] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl (4-aminobutyl) carbamate, and other conditions remained unchanged. 1 H NMR (300MHz, DMSO-d6) δ12.89 (s, 1H), 8.71 (d, J = 4.9Hz, 1H), 8.61 (s, 1H), 8.51 (t, J = 5.7Hz, 1H), 7.86 (d, J=8.2Hz, 2H), 7.68 (t, J=5.7Hz, 1H), 7.63 (d, J=5.0Hz, 1H), 7.56 (d, J=8.2Hz, 2H), 7.40- 7.35 (m, 2H), 7.09-7.05 (m, 1H), 7.02-6.94 (m, 1H), 5.53 (s, 2H), 3.51 (s, 3H), 3.26 (q, J=6.5Hz, 2 H), 3.04 (q, J=6.4Hz, 2H), 1.91-1.84 (m, 3H), 1.80 (s, 2H), 1.67-1.49 (m, 14H), 1.46-1.36 (m, 2H).

[0076] Example 6: Synthesis of N-(5-((4-((5-(2-((3r,5r,7r)-adamantyl-1-yl)acetylamino)pentyl)carbamoyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14e)

[0077]

[0078] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl (5-aminopentyl) carbamate, and other conditions remained unchanged. 1 H NMR (300MHz, DMSO-d6) δ12.88 (s, 1H), 8.71 (d, J=5.0Hz, 1H), 8.60 (s, 1H), 8.47 (t, J=5.7Hz, 1H), 7. 86 (d, J=8.0Hz, 2H), 7.64 (t, J=6.0Hz, 2H), 7.56 (d, J=8.0Hz, 2H), 7.37 (t, J=7.6Hz, 2H), 7.06 (t, J= 7.4Hz, 1H), 6.98 (d, J=8.2Hz, 1H), 5.53 (s, 2H), 3.51 (s, 3H), 3.24 (q, J=6.6Hz, 2H), 3.01 (q, J=6.4H z, 2H), 1.88 (s, 3H), 1.79 (s, 2H), 1.66-1.49 (m, 14H), 1.40 (q, J=7.1Hz, 2H), 1.29 (t, J=7.9Hz, 2H).

[0079] Example 7: Synthesis of N-(5-((4-((6-(2-((3r,5r,7r)-adamantanamine-1-yl)acetylamino)hexyl)carbamoyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14f)

[0080]

[0081] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl (6-aminohexyl) carbamate, and other conditions remained unchanged. 1H NMR (300MHz, CDCl3) δ11.85 (s, 1H), 8.64 (d, J=5.OHz, 1H), 8.56 (s, 1H), 7.79 (d, J=8.1Hz, 2H), 7.58 (d, J=5. 0Hz, 1H), 7.39 (d, J=8.1Hz, 2H), 7.33 (dd, J=7.8, 1.8Hz, 1H), 7.16 (dd, J=7.5, 1.7Hz, 1H), 7.00 (q, J=6.5Hz, 2H), 6.86 (d, J=8.3Hz, 1H), 5.77 (t, J=5.9Hz, 1H), 5.36 (s, 2H), 3.56 (s, 3H), 3.40 (q, J=6.7Hz, 2H), 3.19 (q, J=6.6Hz, 2H), 1.92 (t, J=3.2Hz, 3H), 1.89 (s, 2H), 1.72-1.54 (m, 14H), 1.52-1.41 (m, 3H), 1.40-1.28 (m, 4H).

[0082] Example 8: Synthesis of N-(5-((4-((8-(2-((3r,5r,7r)-adamantyl-1-yl)acetylamino)octyl)carbamoyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14 g)

[0083]

[0084] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl (8-aminooctyl) carbamate, and other conditions remained unchanged. 1 H NMR (300MHz, CDCl3) δ11.14 (s, 1H), 8.69 (d, J = 5.1Hz, 1H), 8.59 (s, 1H), 7.76 (d, J = 7.9Hz, 2H), 7.62 (d, J=5.1Hz, 1H), 7.43 (d, J=8.2Hz, 2H), 7.37 (d, J=7.7Hz, 1H), 7.19 (d, J=7.4Hz, 1H), 7.04 (t, J=7.4Hz, 1H) , 6.91 (d, J = 8.3Hz, 1H), 6.51 (t, J = 5.7Hz, 1H), 5.41 (s, 2H), 3.63 (s, 3H), 3.43 (q, J = 6.8Hz, 2H), 3.21 (q , J=6.7Hz, 2H), 1.95 (s, 3H), 1.89 (s, 2H), 1.67-1.57 (m, 14H), 1.47 (t, J=6.6Hz, 2H), 1.36-1.25 (m, 8H).

[0085] Example 9: Synthesis of N-(5-((4-((2-(2-(2-((3r,5r,7r)-adamantan-1-yl)acetylamino)ethoxy)ethyl)carbamoyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14h)

[0086]

[0087] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl (2-(2-(2-aminoethoxy)ethoxy)carbamate, and other conditions remained unchanged. 1 H NMR (300MHz, CDCl3) δ10.92 (s, 1H), 8.72 (d, J = 5.0Hz, 1H), 8.62 (s, 1H), 7.81 (d, J = 1.7Hz, 1H), 7.79 (d, J = 1.9Hz, 1H), 7.64 ( d, J=5.0Hz, 1H), 7.47 (d, J=1.8Hz, 1H), 7.45 (d, J=1.8Hz, 1H), 7.43-7.38 (m, 1H), 7.23 (dd, J=7.5, 1.7Hz, 1H), 7.06 (td, J=7 .5, 1.0Hz, 1H), 6.92 (dd, J=8.4, 1.0Hz, 1H), 6.87 (s, 1H), 5.84 (t, J=5.7Hz, 1H), 5.42 (s, 2H), 3.67 (d, J=2.6Hz, 4H), 3.65 (s , 3H), 3.63-3.59 (m, 4H), 3.53 (t, J=5.1Hz, 2H), 3.41 (q, J=5.3Hz, 2H), 1.96-1.91 (m, 3H), 1.89 (s, 2H), 1.70-1.56 (m, 12H).

[0088] Example 10: Synthesis of N-(5-((4-(4-(2-((3r,5r,7r)-adamantan-1-yl)acetylamino)ethyl)piperazine-1-carbonyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14i)

[0089]

[0090] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl 4-(2-aminoethyl)piperidine-1-carboxylate, and other conditions remained unchanged. 1H NMR (300MHz, DMSO-d6) δ12.85 (s, 1H), 8.68 (d, J = 5.0Hz, 1H), 8.58 (s, 1H), 7.62 (d, J = 5.4 Hz, 2H), 7.55 (d, J=7.8Hz, 2H), 7.41 (d, J=8.1Hz, 2H), 7.35 (d, J=8.2Hz, 2H), 7.05 (t, J=7. 4Hz, 1H), 6.98 (d, J=8.2Hz, 1H), 5.50 (s, 2H), 3.59 (s, 2H), 3.51 (s, 3H), 3.32-3.22 (m, 4H) , 3.15-3.11(m, 2H), 2.36(t, J=6.7Hz, 4H), 1.90(s, 3H), 1.80(s, 2H), 1.68-1.53(m, 12H).

[0091] Example 11: Synthesis of N-(5-((4-((1R,5S)-6-(2-((3R,5R,7R)-adamantan-1-yl)acetyl)-3,6-diazabicyclo[3.1.1]heptane-3-carbonyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14j)

[0092]

[0093] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl 3,6-diazabicyclo[3.1.1]heptane-3-carboxylate, and other conditions remained unchanged. 1 H NMR (300MHz, DMSO-d6) δ12.89 (s, 1H), 8.71 (d, J = 5.0Hz, 1H), 8.61 (s, 1H), 7.63 (d, J = 5.0H) z, 1H), 7.56 (d, J=8.0Hz, 2H), 7.47 (d, J=7.9Hz, 1H), 7.42-7.35 (m, 3H), 7.06 (t, J=7.4Hz, 1H), 6.99 (d, J=8.3Hz, 1H), 5.52 (s, 1H), 4.55-4.12 (m, 2H), 3.95 (dd, J=13.6, 3.8Hz, 1H), 3.82-3.57(m, 2H), 3.51(s, 3H), 3.48-3.38(m, 1H), 2.03-1.72(m, 5H), 1.72-1.49(m, 14H).

[0094] Example 12: Synthesis of N-(5-((4-((1R,5S)-3-(2-((3R,5R,7R)-adamantan-1-yl)acetyl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14k)

[0095]

[0096] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate, and other conditions remained unchanged. 1 H NMR (300MHz, DMSO-d6) δ12.90 (s, 1H), 8.71 (d, J = 5.0Hz, 1H), 8.61 (s, 1H), 7.63 (d, J = 5.0Hz, 1H), 7 .59-7.53(m, 4H), 7.41-7.35(m, 2H), 7.11-7.03(m, 1H), 6.99(d, J=8.2Hz, 1H), 5.54(s, 2H), 4.65( s, 1H), 4.31-3.82 (m, 3H), 3.52 (s, 3H), 3.31-3.23 (m, 1H), 3.22-3.10 (m, 1H), 2.79 (d, J=12.9Hz, 1 H), 2.22 (d, J=13.6Hz, 1H), 2.10-1.95 (m, 1H), 1.90 (s, 3H), 1.83-1.80 (m, 2H), 1.66-1.55 (m, 13H).

[0097] Example 13: Synthesis of N-(5-((4-(2-(2-((3r,5r,7r)-adamantan-1-yl)acetyl)-2,7-diazaspiro[3.5]nonane-7-carbonyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (141)

[0098]

[0099] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate, and other conditions remained unchanged. 1H NMR (300MHz, DMSO-d6) δ12.89 (s, 1H), 8.71 (d, J = 5.0Hz, 1H), 8.60 (s, 1H), 7.63 (d, J=5.0Hz, 1H), 7.55 (d, J=8.0Hz, 2H), 7.43-7.34 (m, 4H), 7.10-7.03 (m, 1H), 6.99 (d, J=8.1Hz, 1H), 5.51 (s, 2H), 3.83 (s, 2H), 3.56 (s, 4H), 3.5 1(s, 3H), 3.23(s, 2H), 1.93-1.87(m, 3H), 1.78(s, 2H), 1.73-1.53(m, 16H).

[0100] Example 14: Synthesis of N-(5-((4-((7-(2-((3r,5r,7r)-adamantan-1-yl)acetylamino)heptyl)carbamoyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14m)

[0101]

[0102] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl (7-aminoheptyl) carbamate, and other conditions remained unchanged. 1 H NMR (300MHz, DMSO-d6) δ12.89 (s, 1H), 8.71 (d, J = 5.0Hz, 1H), 8.61 (s, 1H), 8.48 (t, J = 5.7Hz, 1H) , 7.86 (d, J=8.0Hz, 2H), 7.63 (t, J=4.9Hz, 2H), 7.56 (d, J=7.9Hz, 2H), 7.40-7.35 (m, 2H), 7.06 (t, J=7.4Hz, 1H), 6.98 (d, J=8.3Hz, 1H), 5.53 (s, 2H), 3.51 (s, 3H), 3.24 (q, J=6.5Hz, 2H), 3.00 (q, J= 6.3Hz, 2H), 1.87(s, 3H), 1.79(s, 2H), 1.65-1.47(m, 14H), 1.39-1.35(m, 2H), 1.30-1.22(m, 6H).

[0103] Example 15: Synthesis of N-(5-((4-(9-(2-((3r,5r,7r)-adamantan-1-yl)acetyl)-3,9-diazaspiro[5.5]undecane-3-carbonyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14n)

[0104]

[0105] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl 3,9-diazaspiro[5.5]undecane-3-carboxylate, and other conditions remained unchanged. 1 H NMR (300MHz, DMSO-d6) δ12.89 (s, 1H), 8.71 (d, J=5.0Hz, 1H), 8.61 (s, 1H), 7.63 (d , J=5.0Hz, 1H), 7.55 (d, J=7.8Hz, 2H), 7.39 (dd, J=15.2, 7.7Hz, 4H), 7.06 (t, J=7. 4Hz, 1H), 6.99 (d, J=8.2Hz, 1H), 5.51 (s, 2H), 3.60 (s, 2H), 3.51 (s, 3H), 3.44 (s, 4 H), 3.27(s, 2H), 2.08(s, 2H), 1.89(s, 3H), 1.67-1.54(m, 13H), 1.49-1.33(m, 7H).

[0106] Example 16: Synthesis of N-(5-((4-((2-(2-((3r,5r,7r)-adamantanamine-1-yl)acetylamino)ethoxy)ethyl)carbamoyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14o)

[0107]

[0108] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl (2-(2-aminoethoxy)ethyl)carbamate, and other conditions remained unchanged. 1 H NMR (300MHz, CDCl3) δ11.99 (s, 1H), 8.62 (d, J = 5.0Hz, 1H), 8.54 (s, 1H), 7.80 (d, J = 8.0Hz, 2H), 7. 57 (d, J=5.0Hz, 1H), 7.38 (d, J=8.2Hz, 2H), 7.35-7.27 (m, 2H), 7.16 (dd, J=7.6, 1.7Hz, 1H), 6.98 ( t, J=7.4Hz, 1H), 6.84 (d, J=8.3Hz, 1H), 6.22 (t, J=5.6Hz, 1H), 5.34 (s, 2H), 3.59 (s, 4H), 3.54 (s, 3H), 3.51 (t, J=5.1Hz, 2H), 3.37 (q, J=5.3Hz, 2H), 1.87 (s, 2H), 1.85 (s, 3H), 1.61-1.48 (m, 12H).

[0109] Example 17: Synthesis of N-(5-((4-((4-(2-((3r,5r,7r)-adamantan-1-yl)acetylamino)cyclohexyl)carbamoyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14p)

[0110]

[0111] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl (4-aminocyclohexyl) carbamate, and other conditions remained unchanged. 1 H NMR (300MHz, DMSO-d6) δ12.89 (s, 1H), 8.68 (d, J = 5.0Hz, 1H), 8.58 (s, 1H), 8.25 (dd, J =40.5, 7.2Hz, 1H), 7.87 (d, J = 8.1Hz, 2H), 7.67-7.52 (m, 4H), 7.40-7.32 (m, 2H), 7.05 ( t, J=7.4Hz, 1H), 6.98 (d, J=8.2Hz, 1H), 5.51 (s, 2H), 3.75 (d, J=23.9Hz, 2H), 3.51 (s, 3 H), 1.88 (d, J=10.9Hz, 5H), 1.82-1.76 (m, 3H), 1.68-1.50 (m, 15H), 1.45-1.26 (m, 2H).

[0112] Example 18: Synthesis of N-(5-((4-(6-(2-((3r,5r,7r)-adamantan-1-yl)acetylamino)-2-azapyridine[3.3]heptane-2-carbonyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14q)

[0113]

[0114] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl 6-amino-2-azaspiro[3.3]heptane-2-carboxylate, and other conditions remained unchanged. 1H NMR (300MHz, DMSO-d6) δ18.87 (s, 1H), 8.71 (d, J=5.0Hz, 1H), 8.60 (s, 1H), 7.92 (t, J=6.4 Hz, 1H), 7.66-7.62 (m, 3H), 7.55 (d, J=7.9Hz, 2H), 7.40-7.35 (m, 2H), 7.06 (t, J=7.4Hz, 1H ), 6.99 (d, J = 8.2Hz, 1H), 5.52 (s, 2H), 4.28 (d, J = 33.1Hz, 2H), 4.12-3.92 (m, 3H), 3.51 (s, 3H), 2.49-2.43(m, 2H), 2.08-2.02(m, 2H), 1.89(s, 3H), 1.76(s, 2H), 1.67-1.48(m, 12H).

[0115] Example 19: Synthesis of N-(5-((4-(3-(4-(2-((3r, 5r, 7r)-adamantan-1-yl)acetyl)piperazin-1-yl)azetidin-1-carbonyl)benzyl)oxy)-1,3,4-thiadiazol-2-yl)-3-(2-methoxyphenyl)isonicotinamide (14r)

[0116]

[0117] Referring to the synthesis method of Example 3, compound 12b was replaced with tert-butyl 3-(piperazin-1-yl)azetidine-1-carboxylate, and other conditions remained unchanged. 1 H NMR (300MHz, DMSO-d6) 612.89 (s, 1H), 8.70 (d, J = 5.0Hz, 1H), 8.59 (s, 1H), 7.67 (d, J = 7.9Hz, 2H) , 7.62 (d, J=4.9Hz, 1H), 7.55 (d, J=8.0Hz, 2H), 7.35 (d, J=7.8Hz, 2H), 7.06 (t, J=7.5Hz, 1H), 6.98 (d, J=8.1Hz, 1H), 5.52 (s, 2H), 4.35-4.25 (m, 1H), 4.16-4.02 (m, 2H), 3.90-3.85 (m, 1H), 3.51-3 .47(m, 8H), 3.18-3.11(m, 1H), 2.33-2.22(m, 4H), 2.08(s, 2H), 1.90(s, 3H), 1.66-1.55(m, 12H).

[0118] Embodiment 20:

[0119] Activity test

[0120] (I) Western blot analysis of the effects of compounds on Polθ expression in MDA-MB-436 cells

[0121] 1. Experimental Methods

[0122] After the MDA-MB-436 cells were treated with the compound, the culture medium was discarded, and the cells were washed with PBS for 2-3 times. Protease inhibitors and RIPA lysis buffer were added in sequence to completely lyse the cells, and then centrifuged (4°C, 12,000g, 10min). The upper layer was the total protein solution obtained. Referring to the instructions of the kit, 7.5% separation gel and 5% concentration gel were prepared according to the molecular weight of the protein to be tested, and SDS-PAGE gel was cast. After adding an appropriate amount of pre-cooled 1× electrophoresis buffer, the previously biomarked sample or total protein extract in the cell was added to the lane (pre-stained protein Maker and sample). 90V stabilized electrophoresis for about 30 minutes. After the sample enters the separation gel, adjust the voltage to 120V and continue electrophoresis. When the target band reaches the appropriate position (refer to the position of the pre-stained protein Maker), the electrophoresis is terminated. Electrophoresis can be terminated when bromophenol blue just runs out, and the mold is transferred. The band of the target protein was stripped out, attached to a PVDF membrane, transferred to the PVDF membrane by electrophoresis, and then blocked with 5% skim milk on a decolorizing shaker for 1 hour. Add the primary antibody, incubate overnight at 4°C, and then wash three times with TBST, each washing time is 10 minutes. Add the secondary antibody, incubate at room temperature for 2 hours, and then wash three times with TBST, each washing time is 10 minutes. Prepare the ECL mixed solution in the dark room in advance according to the ratio of ECLA:ECLB=1:1, then place the treated PVDF membrane face up in the exposure box, add the prepared ECL mixed solution, react for 1-2 minutes, and then discard the reaction solution, adjust the exposure conditions according to the luminescence intensity of the developer, and start exposure.

[0123] 2. Experimental results

[0124] like Figure 1 As shown, the compounds 14a, 14b, 14c, 14e, 14g, 14i, 14k, and 141 disclosed in the present invention showed a concentration-dependent degradation effect on Polθ, and showed a certain degree of degradation effect on Polθ when the administration concentration was 15 μM and the administration time was 24 h. Figure 2 As shown, compound 14k treated MDA-MB-436 cells and showed a Polθ degradation effect. After the cells were pre-treated with a proteasome inhibitor for 4 hours and then treated with compound 14k, the results showed that the degradation effect of compound 14k disappeared, indicating that compound 14k degrades Polθ protein through the proteasome pathway.

Claims

1. A compound for degrading Polθ based on a hydrophobic tag or a pharmaceutically acceptable salt thereof, characterized in that: The structural formula of the compound for degrading Polθ is shown in general formula (I): in, Linker does not exist or is selected from: Unsubstituted or secondary amino-substituted 5- to 8-membered cycloalkyl, unsubstituted or -(CH2) 1～4 -NH-substituted 5-8 membered nitrogen-containing heterocyclic group, unsubstituted or secondary amino-substituted 6-11 membered spiro ring containing nitrogen and carbon, unsubstituted or imino-substituted 6-10 membered bridged ring containing nitrogen and carbon, 4-6 membered nitrogen heterocyclic ring-4-6 membered nitrogen heterocyclic ring; Ra is selected from a saturated alkyl group having 0 to 8 carbon atoms and an alkoxy group having 4 to 6 carbon atoms; Rb is selected from an alkoxy group having 1 to 6 carbon atoms; Rc is selected from an alkoxy group having 1 to 6 carbon atoms; HyT is:

2. The compound for degrading Polθ based on a hydrophobic tag according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Linker does not exist or is selected from: Unsubstituted or secondary amino-substituted 6-membered cycloalkyl, unsubstituted or -(CH2) 1～4 -NH-substituted piperazinyl, unsubstituted or secondary amino-substituted 6-11-membered spiro ring containing nitrogen and carbon, unsubstituted or secondary amino-substituted 6-10-membered bridged ring containing nitrogen and carbon, 4-membered nitrogen heterocycle-6-membered nitrogen heterocycle; Ra is selected from a saturated alkyl group having 0 to 8 carbon atoms and an alkoxy group having 4 to 6 carbon atoms; Rb is selected from an alkoxy group having 1 to 6 carbon atoms; Rc is selected from an alkoxy group having 1 to 6 carbon atoms.

3. The compound for degrading Polθ based on a hydrophobic tag according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Linker does not exist or is selected from: Unsubstituted or secondary amino-substituted 6-membered cycloalkyl, unsubstituted or -(CH2) 1～4 -NH-substituted piperazinyl, unsubstituted or secondary amino-substituted 6-11 membered spiro ring containing nitrogen and carbon, unsubstituted or secondary amino-substituted 6-10 membered bridged ring containing nitrogen and carbon, azetidine-piperazinyl; Ra is selected from a saturated alkyl group having 0 to 8 carbon atoms and an alkoxy group having 4 to 6 carbon atoms; Rb is selected from an alkoxy group having 1 to 6 carbon atoms; Rc is selected from an alkoxy group having 1 to 6 carbon atoms.

4. The compound for degrading Polθ based on a hydrophobic tag according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The Linker does not exist or is selected from any of the following structures: R1 is selected from: R2 is selected from: -CH2-, -CH2-CH2-.

5. The compound for degrading Polθ based on a hydrophobic tag according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The Linker does not exist or is selected from any of the following structures:

6. A compound for degrading Polθ based on a hydrophobic tag or a pharmaceutically acceptable salt thereof, characterized in that: The structural formula of the compound for degrading Polθ based on the hydrophobic tag is shown in any one of the following:

7. A method for preparing a compound for degrading Polθ based on a hydrophobic tag according to any one of claims 1 to 6, characterized in that: The steps include: (1) (2-methoxyphenyl)boronic acid and 3-bromoisonicotinate methyl ester are subjected to Suzuki reaction to prepare 3-(2-methoxyphenyl)isonicotinate methyl ester; (2) 3-(2-methoxyphenyl)isonicotinic acid methyl ester is hydrolyzed under alkaline conditions to obtain 3-(2-methoxyphenyl)isonicotinic acid; (3) 5-bromo-1,3,4-thiadiazol-2-amine and methyl 4-hydroxymethylbenzoate are subjected to substitution reaction to prepare methyl 4-(((5-amino-1,3,4-thiadiazol-2-yl)oxy)methyl)benzoate; (4) 3-(2-methoxyphenyl)isonicotinic acid and methyl 4-(((5-amino-1,3,4-thiadiazol-2-yl)oxy)methyl)benzoate are reacted with 2-(7-azobenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU) as a condensing agent to produce methyl 4-(((5-(3-(2-methoxyphenyl)isonicotinamido)-1,3,4-thiadiazol-2-yl)oxy)methyl)benzoate; (5) methyl 4-(((5-(3-(2-methoxyphenyl)isonicotinamido)-1,3,4-thiadiazol-2-yl)oxy)methyl)benzoate is hydrolyzed under alkaline conditions to give 4-(((5-(3-(2-methoxyphenyl)isonicotinamido)-1,3,4-thiadiazol-2-yl)oxy)methyl)benzoic acid; (6) The linker modules 11b-11r are first reacted with adamantane acetic acid in the presence of a condensing agent 2-(7-azobenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate to produce compounds 12b-12r; (7) Compound 12b-12r is deprotected from the Boc protecting group and then undergoes an amidation reaction with 4-(((5-(3-(2-methoxyphenyl)isonicotinamido)-1,3,4-thiadiazol-2-yl)oxy)methyl)benzoic acid to obtain a compound of the general formula (I).

8. Use of the compound for degrading Polθ based on a hydrophobic tag or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6 in the preparation of a drug for targeted degradation of Polθ.

9. Use of the compound for degrading Polθ based on a hydrophobic tag or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6 in the preparation of a drug for treating and / or preventing cancer.

10. A pharmaceutical composition, characterized in that A compound for degrading Polθ based on a hydrophobic tag according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof.

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