A btk protein degrader with silicon-containing group as hydrophobic tag, preparation method, pharmaceutical composition and application thereof

Abstract

The application belongs to the field of chemical drugs, and discloses a BTK protein degrading agent with a silicon-containing group as a hydrophobic tag, a preparation method, a pharmaceutical composition and application thereof.The structure of the BTK protein degrading agent with the silicon-containing group as the hydrophobic tag is shown in formula I, the linker is a saturated aliphatic chain, an unsaturated aliphatic chain, a connecting structure composed of an aromatic ring and a saturated aliphatic chain, or a connecting structure composed of an aromatic ring and an unsaturated aliphatic chain; and R is a specific silicon-containing group as the hydrophobic tag.A BTK protein degrading agent with a silicon-containing group as a hydrophobic tag is obtained, the preparation process is simple and easy to implement, the obtained BTK protein degrading agent has high liver microsomal metabolic stability, has considerable oral bioavailability, shows high treatment efficiency in in-vivo experiments, has a certain inhibitory effect on the proliferation of various tumor cells, and is suitable for the development of cancer drugs such as leukemia.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of chemical drugs, and particularly relates to a BTK protein degrader with a silicon-containing group as a hydrophobic tag, a preparation method, a pharmaceutical composition and application thereof. BACKGROUND

[0002] Bruton's tyrosine kinase (BTK) is a member of the cytoplasmic non-receptor tyrosine kinase TEC family, which plays an important role in the growth, development, proliferation and differentiation of B cells, and is also a therapeutic target for B cell malignancies. Although the mainstream BTK inhibitors (Ibrutinib, Acalabrutinib, Zanubrutinib) have excellent therapeutic effects in clinical practice, like other targeted drugs, acquired drug resistance is gradually shortening the application range of these molecules, especially the mutation of the key site C481. The development of BTK degraders may be a solution.

[0003] The hydrophobic tag (Hyt) bifunctional molecule is composed of a target protein ligand, a linker and a hydrophobic group. By connecting a large and hydrophobic group to a small molecule that can bind to the target, such a double-headed molecule, after binding to the target, will be mistakenly considered by the intracellular protein repair mechanism as a misfolded part of the target protein, and then it can be folded by chaperone proteins and further degraded by proteasomes. The hydrophobic group in the Hyt molecule is often small in molecular weight, and thus may have higher solubility and drugability. The development of degraders based on hydrophobic tags is still in the exploratory stage. On the one hand, there are fewer reported hydrophobic tag fragments, and there is still a lot of room for optimization in terms of degradation activity and physicochemical properties. On the other hand, the exact degradation mechanism has not been clearly defined. Therefore, exploring more hydrophobic fragments with high activity and excellent physicochemical properties and clarifying their corresponding degradation mechanisms are crucial for the development of hydrophobic tag fragments based on clinical applications.

[0004] The hydrophobic fragments used in the currently reported hydrophobic tag degraders include adamantane, menthol, norbornene, Boc arginine, carborane, fluorenyl, etc. However, these conventional hydrophobic tag degraders have relatively low degradation effect, poor liver microsomal metabolic stability, poor solubility and are difficult to be orally administered, making them difficult to be used for further treatment and development in clinical practice. There is a need for the development of new hydrophobic tag degraders with better liver microsomal stability and oral administration to meet the clinical needs. SUMMARY

[0005] The present application aims to solve the problems of the prior art, and provides a BTK protein degrading agent with a silicon-containing group as a hydrophobic tag, which has high liver microsomal metabolic stability and can be orally taken, a preparation method, a pharmaceutical composition and an application thereof. Based on this, the first aspect of the present application provides a BTK protein degrading agent with a silicon-containing group as a hydrophobic tag, which has the structure shown in formula I:

[0006]

[0007] In the formula, the linker is a saturated aliphatic chain, an unsaturated aliphatic chain, a connecting structure composed of an aromatic ring and a saturated aliphatic chain, or a connecting structure composed of an aromatic ring and an unsaturated aliphatic chain; R is a trimethylsilyl group, a triethylsilyl group, a tri-n-propylsilyl group, a triisopropylsilyl group, an isopropyldimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, a cyclohexyldimethylsilyl group, a vinyl dimethylsilyl group, a trimethoxysilyl group, a triethoxysilyl group, a phenylvinylmethylsilyl group, or a triphenylsilyl group.

[0008] The present application provides a BTK protein degrading agent with a silicon-containing group as a hydrophobic tag. In the bifunctional molecule, the silicon-containing group is used as a hydrophobic tag, and the ibrutinib parent nucleus is used as a target protein ligand, thereby obtaining a protein degrading agent capable of effectively degrading BTK. The BTK protein degrading agent prepared by the present application can effectively induce the degradation of BTK in MOLM-13 (human acute myeloid leukemia cells) in a dose-dependent and time-dependent manner, has considerable liver microsomal stability and bioavailability, and exhibits excellent in vivo anticancer effect.

[0009] In the formula, the linker is a saturated aliphatic chain, an unsaturated aliphatic chain, a connecting structure composed of an aromatic ring and a saturated aliphatic chain, or a connecting structure composed of an aromatic ring and an unsaturated aliphatic chain. By selecting different types and lengths of ideal linkers, the purpose of not affecting the binding of the two proteins in space and maintaining their binding can be achieved. Based on this, in some more preferred embodiments, the structure of the BTK protein degrading agent with a silicon-containing group as a hydrophobic tag is shown in formula II or formula III:

[0010]

[0011] In the formula, m and n are independently selected from integers in the range of 1-10.

[0012] Further preferably, the structure of the BTK protein degrading agent with a silicon-containing group as a hydrophobic tag is shown in formula IV:

[0013]

[0014] The compound as shown in Formula IV can effectively induce degradation of BTK in human B lymphoma cell Ramos cell line and human diffuse large B cell lymphoma cell TMD8 cell line in a dose-dependent and time-dependent manner, has considerable liver microsomal stability and bioavailability, and exhibits excellent in vivo anticancer effect.

[0015] The second aspect of the present application provides a preparation method of the above-mentioned BTK protein degrader with a silicon-containing group as a hydrophobic tag, when the structure of the above-mentioned BTK protein degrader with a silicon-containing group as a hydrophobic tag is as shown in Formula II, the preparation method is shown in Route I; when the structure of the above-mentioned BTK protein degrader with a silicon-containing group as a hydrophobic tag is as shown in Formula III, the preparation method is shown in Route II.

[0016] Route I: tert-butyl diphenylchlorosilane (compound B) is reacted with azidoethanol (compound A) in the presence of imidazole to obtain intermediate C, and ibutinib nucleus (compound D) is condensed with alkyne acids of different lengths in the presence of N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate and N,N-diisopropylethylamine to obtain intermediates E1-E6, and click reaction of intermediate C and intermediates E1-E6 obtains final products 1-6.

[0017]

[0018] Route II: tert-butyl diphenylchlorosilane (compound B) is reacted with brominated alcohols (compounds F1-F4) of different lengths in the presence of imidazole to obtain intermediates G1-G4, and then substitution reaction with compound D in the presence of cesium carbonate to obtain final products 7-10.

[0019]

[0020] The preparation method of the present application is simple in route, the raw materials used are cheap and easy to obtain, and the overall yield of the reaction is high.

[0021] The above-mentioned BTK protein degrader with a silicon-containing group as a hydrophobic tag or a pharmaceutically acceptable salt thereof can be used for preparing a medicament for treating related cancers with BTK abnormalities. The above-mentioned related cancers include leukemia, lung cancer, gastric cancer, breast cancer, colorectal cancer, and pancreatic cancer. More preferably, the related cancer is leukemia.

[0022] The third aspect of the present application provides a pharmaceutical composition comprising the BTK protein degrader with a silicon-containing group as a hydrophobic tag or a pharmaceutically acceptable salt thereof (or, the BTK protein degrader with a silicon-containing group as a hydrophobic tag or a pharmaceutically acceptable salt thereof as the main active ingredient). In some preferred embodiments, the above-mentioned pharmaceutical composition can further comprise at least one of an excipient, a solvent and a pharmaceutical carrier. Among them, the excipient is at least one of gum arabic, sugar syrup, lanolin and starch; there is no incompatibility with the main drug, no side effects, no influence on the curative effect, not easy to deform, dry, mold, insect damage at room temperature, harmless to the human body, no physiological effect, no chemical or physical action with the main drug, no influence on the content determination of the main drug. The solvent can be water, glycerol or ethanol.

[0023] The present application has the following beneficial effects: the present application obtains a BTK protein degrader with a silicon-containing group as a hydrophobic tag, the preparation process is simple and easy to operate, the obtained BTK protein degrader has high liver microsomal metabolic stability, has considerable oral bioavailability, shows high therapeutic effect in in vivo experiments, has certain inhibitory effect on the proliferation of various tumor cells, and is suitable for the development of cancer drugs such as leukemia. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 The figure shows the effect of compound 2 on the concentration-dependent degradation of BTK in the MOLM-13 cell line; (A) is a Western Blot diagram, and (B) is a DC 50 FIG.

[0025] Figure 2 The figure shows the effect of compound 2 on the time-dependent degradation of BTK in the MOLM-13 cell line. DETAILED DESCRIPTION

[0026] The concept and technical effects of the present application will be described clearly and completely in combination with the embodiments and the drawings below, so as to fully understand the purpose, scheme and effect of the present application. It should be noted that the embodiments in the present application and the features in the embodiments can be combined with each other without conflict.

[0027] Example 1

[0028] A BTK protein degrader with a silicon-containing group as a hydrophobic tag (denoted as compound 1-6), the structures of which are respectively as follows:

[0029]

[0030]

[0031] The structures of the intermediate compounds A, compound B, compound C, compound D, compound alkyne acid, compounds E1-E6 used are as follows, respectively:

[0032]

[0033] The specific preparation method is as follows:

[0034] 1) Preparation of compound C:

[0035] Compound A (174 mg, 2.00 mmol) was dissolved in dichloromethane (12 mL) under ice bath, compound B (825 mg, 3.00 mmol) and imidazole (408 mg, 6.00 mmol) were added, respectively, after 30 minutes, the ice bath was removed, and after stirring overnight at room temperature, the solid was removed by filtration, the remaining organic phase was dried with anhydrous sodium sulfate, concentrated, and then compound C was obtained by silica gel flash column chromatography (petroleum ether: ethyl acetate = 60:1).

[0036] Compound C was detected, and the detection results are as follows: 1 H NMR (400 MHz, CDCl3) δ 7.72-7.66 (m, 4H), 7.41 (tt, J = 8.1, 5.7 Hz, 6H), 3.81 (dd, J = 5.5, 4.6 Hz, 2H), 3.29 (t, J = 5.0 Hz, 2H), 1.07 (s, 9H). 13 C NMR (100 MHz, CDCl3) δ 135.6, 133.1, 129.9, 127.8, 63.3, 53.2, 26.7, 19.1.

[0037] 2) Preparation of compounds E1-E6:

[0038] Preparation of compound E1:

[0039] Compound 3-butynoic acid (168 mg, 1.50 mmol) was dissolved in N,N- dimethylformamide (3 mL), N-N-diisopropylethylamine (330 μL, 2.00 mmol) was added, the reaction system was cooled to 0 °C, then N,N,N',N'-tetramethyl-0-(7- azabenzotriazol-1-yl) uronium hexafluorophosphate (570 mg, 1.50 mmol) was added. Stirring at 0 °C for 10 min, then compound D (463 mg, 1.20 mmol) was added, the reaction was carried overnight, after the reaction was completed, the reaction was extracted with ethyl acetate (3 x 10 mL), then the organic phase was combined and washed with saturated sodium bicarbonate solution (10 mL), 1 N hydrochloric acid solution (10 mL), saturated NaCl solution (10 mL), respectively, the organic phase was dried with anhydrous sodium sulfate, concentrated, the obtained crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain compound E1.

[0040] Compound E1 was detected, and the detection results were as follows: 1 H NMR (400 MHz, CDC13) δ 8.30 (s, 1H), 7.83 - 7.70 (m, 2H), 7.41 - 7.29 (m, 2H), 7.25 (d, J = 8.1 Hz, 1H), 7.19 (d, J = 8.0 Hz, 1H), 7.1

[0041] 5 - 7.06 (m, 4H), 7.03 - 6.96 (m, 3H), 4.88 (dt, J = 5.4, 2.7 Hz, 1H), 3.88 (d, J = 2.8 Hz, 2H), 3.65 - 3.44 (m, 5H), 2.26 (t, J = 3.0 Hz, 1H), 2.18 - 1.99 (m, 1H), 2.00 - 1.82 (m, 4H).

[0042] Preparation of compound E2:

[0043] Compound 3-butynoic acid was replaced with compound 4-pentynoic acid (147 mg, 1

[0044] .50 mmol) according to the same method as preparing compound E1 to obtain compound E2.

[0045] Compound E2 was detected, and the detection results were as follows: 1 H NMR (400 MHz, CDC13) δ 8.34 (d, J = 13

[0046] H NMR (400 MHz, CDC13) δ 8.35 (d, J = 9.4 Hz, 1H), 7.64 (dd, J = 8.6, 3.0 Hz, 2H), 7.47 - 7.33 (m, 2H), 7.16 (ddd, J = 12.1, 8.0, 2.9 Hz, 3H), 7.08 (d, J = 8.1 Hz, 2H), 5.69 (s, 2H), 4.98 - 4.45 (m, 2H), 4.15 - 3.84 (m, 1H), 3.72 - 3.25 (m, 1H), 3.19 - 2.74 (m, 1H), 2.69 - 2.17 (m, 5H), 2.06 - 1.78 (m, 6H).

[0047] Preparation of compound E3:

[0048] Compound 3-butynoic acid was replaced with compound 5-hexynoic acid (168 mg, 1.50 mmol) according to the same method as for the preparation of compound El to obtain compound E3.

[0049] Compound E3 was tested and the results of the test are as follows:

[0050] Compound E3 was tested and the results of the test are as follows: 1 H NMR (400 MHz, CDC13) δ 8.35 (d, J = 9.4 Hz, 1H), 7.64 (dd, J = 8.6, 3.0 Hz, 2H), 7.47 - 7.33 (m, 2H), 7.16 (ddd, J = 12.1, 8.0, 2.9 Hz, 3H), 7.08 (d, J = 8.1 Hz, 2H), 5.69 (s, 2H), 4.98 - 4.45 (m, 2H), 4.15 - 3.84 (m, 1H), 3.72 - 3.25 (m, 1H), 3.19 - 2.74 (m, 1H), 2.69 - 2.17 (m, 5H), 2.06 - 1.78 (m, 6H).

[0051] Preparation of compound E4:

[0052] Compound 3-butynoic acid was replaced with compound 6-heptynoic acid (189 mg, 1.50 mmol) according to the same method as for the preparation of compound El to obtain compound E4.

[0053] Compound E4 was tested and the results of the test are as follows: 1H NMR (400 MHz, CDC13) δ 8.35 (d, J = 11.1 Hz, 1H), 7.64 (dd, J = 8.7, 2.9 Hz, 2H), 7.46 - 7.33 (m, 2H), 7.15 (dt, J = 8.4, 3.5 Hz, 3H), 7.08 (d, J = 8.0 Hz, 2H), 5.71 (s, 2H), 4.99 - 4.49 (m, 2H), 4.17 - 3.82 (m, 1H), 3.78 - 3.22 (m, 1H), 3.18 - 2.72 (m, 1H), 2.40 (tq, J = 11.7, 4.4, 3.8 Hz, 2H), 2.35 - 2.28 (m, 1H), 2.21 (dtd, J = 13.9, 7.1, 2.7 Hz, 3H), 1.97 - 1.88 (m, 3H), 1.77 (tdd, J = 12.7, 8.2, 5.9 Hz, 2H), 1.65 - 1.51 (m, 2H).

[0054] Preparation of compound E5:

[0055] Compound 7-octynoic acid (210 mg, 1.50 mmol) was used to replace compound 3-butynoic acid according to the same method for preparing compound El to obtain compound E5.

[0056] Compound E5 was detected, and the detection results are as follows: 1 H NMR (400 MHz, CDC13) δ 8.35 (d, J = 11.1 Hz, 1H), 7.64 (dd, J = 8.7, 2.9 Hz, 2H), 7.46 - 7.33 (m, 2H), 7.15 (dt, J = 8.4, 3.5 Hz, 3H), 7.08 (d, J = 8.0 Hz, 2H), 5.71 (s, 2H), 4.99 - 4.49 (m, 2H), 4.17 - 3.82 (m, 1H), 3.78 - 3.22 (m, 1H), 3.18 - 2.72 (m, 1H), 2.40 (tq, J = 11.7, 4.4, 3.8 Hz, 2H), 2.35 - 2.28 (m, 1H), 2.21 (dtd, J = 13.9, 7.1, 2.7 Hz, 3H), 1.97 - 1.88 (m, 3H), 1.77 (tdd, J = 12.7, 8.2, 5.9 Hz, 2H), 1.65 - 1.51 (m, 2H).

[0057] Preparation of compound E6:

[0058] Compound 8-nonynoic acid (231 mg, 1.50 mmol) was used to replace compound 3-butynoic acid according to the same method for preparing compound El to obtain compound E6.

[0059] Compound E6 was detected, and the detection results are as follows: 1H NMR (400 MHz, CDC13) δ 8.30 (s, 1H), 7.91 - 7.56 (m, 2H), 7.41 - 7.31 (m, 2H), 7.25 (d, J = 8.1 Hz, 1H), 7.19 (d, J = 8.0 Hz, 1H), 7.16 - 7.06 (m, 4H), 7.04 - 6.98 (m, 3H), 4.91 (dt, J = 5.4, 2.7 Hz, 1H), 3.82 (d, J = 2.7 Hz, 2H), 3.64 - 3.41 (m, 3H), 2.38 (t, J = 7.9 Hz, 3H), 2.12 (td, J = 6.1, 3.1 Hz, 3H), 2.08 - 1.99 (m, 2H), 2.00 - 1.86 (m, 4H), 1.63 - 1.52 (m, 2H), 1.52 - 1.41 (m, 3H), 1.42 - 1.31 (m, 4H).

[0060] 3) Preparation of compound 1-6:

[0061] Preparation of compound 1:

[0062] E1 (104 mg, 0.230 mmol) and compound C (91.1 mg, 0.280 mmol) were dissolved in a mixed solvent of t-BuOH / H20 (5 mL, 1:1), CuS04·5H20 (18.6 mg, 0.116 mmol), sodium L-ascorbate (38.4 mg, 0.194 mmol) were added, and the reaction was allowed to proceed at room temperature overnight. After completion of the reaction, CuS04·5H20 was removed by filtration, and the filtrate was concentrated in vacuo, and compound 1 was isolated by silica gel flash column chromatography (dichloromethane:methanol = 50:1).

[0063] Compound 1 was detected, and the detection results thereof were as follows: 1 H NMR (400 MHz, CDC13) δ 8.30 (s, 1H), 7

[0064] .84 - 7.75 (m, 3H), 7.65 (s, 1H), 7.64 - 7.58 (m, 5H), 7.41 - 7.32 (m, 11H), 7.25 (d, J = 8.1 Hz, 1H), 7.19 (d, J = 8.1 Hz, 1H), 7.15 - 7.07 (m, 4H), 7.03 - 6.95 (m, 3H), 4.90 (dt, J = 5.6, 2.7 Hz, 1H), 4.31 (td, J = 4.1, 1.1 Hz, 3H), 4.18 (t, J = 4.1 Hz, 3H), 3.90 - 3.77 (m, 3H), 3.67 (d, J = 2.4 Hz, 3H), 3.56 - 3.45 (m, 2H), 2.16 - 1.82 (m, 5H), 1.03 (s, 9H).

[0065] Preparation of compound 2:

[0066] Compound E1 was replaced with compound E2 (107 mg, 0.230 mmol) according to the same method as for the preparation of compound 1 to obtain compound 2.

[0067] Compound 2 was tested and the results of the test are as follows: 1 H NMR (400 MHz, CDC13) δ 8.34 (s, 1H), 7

[0068] .63 (d, J = 8.2 Hz, 2H), 7.51 (q, J = 7.6, 5.8 Hz, 4H), 7.38 (dp, J = 15.6, 7.2 Hz, 9H), 7.16 (dd, J = 13.2, 7.7 Hz, 3H), 7.08 (d, J = 8.0 Hz, 2H), 5.67 (s, 2H), 4.88 - 4.55 (m, 2H), 4.43 (dt, J = 16.8, 5.3 Hz, 2H), 4.14 - 3.91 (m, 3H), 3.70 - 3.22 (m, 1H), 3.08 (q, J = 7.0 Hz, 2H), 2.85 - 2.68 (m, 2H), 2.34 - 2.17 (m, 2H), 1.95 (s, 2H), 1.66 (s, 1H), 0.99 (d, J = 14.0 Hz, 9H).

[0069] 2.68 (m, 2H), 2.34 - 2.17 (m, 2H), 1.95 (s, 2H), 1.66 (s, 1H), 0.99 (d, J = 14.0 Hz, 9H). 13 CNMR (100 MHz, CDC13) δ 170.6, 158.7, 157.9, 156.4, 155.9, 155.8, 154.3, 146.8, 135.5, 132.8, 132.8, 130.1, 127.9, 124.2, 124.2, 123.0, 122.9, 119.7, 119.7, 119.3, 62.8, 53.5, 52.7, 52.4, 45.9, 33.0, 30.4, 26.8, 25.2, 24.2, 21.3.

[0070] Preparation of compound 3:

[0071] Compound E1 was replaced with compound E3 (111 mg, 0.230 mmol) according to the same method as for the preparation of compound 1 to obtain compound 3.

[0072] Compound 3 was tested and the results of the test are as follows: 1H NMR (400 MHz, CDC13) δ 8.35 (d, J = 7.0 Hz, 1H), 7.63 (dd, J = 8.6, 3.5 Hz, 2H), 7.53 - 7.47 (m, 4H), 7.44 - 7.32 (m, 9H), 7.15 (dt, J = 8.5

[0073] , 3.3 Hz, 3H), 7.08 (d, J = 8.0 Hz, 2H), 5.70 (s, 2H), 4.94 - 4.55 (m, 2H), 4.43 (dt, J = 18.7, 5.2 Hz, 2H), 4.12 - 3.81 (m, 3H), 3.66 - 3.20 (m, 1H), 3.13 - 2.65 (m, 3H), 2.43 (dp, J = 24.3, 9.1, 8.2 Hz, 2H), 2.12 - 1.88 (m, 5H), 1.67 (ddt, J = 19.6, 15.3, 7.6 Hz, 1H), 0.99 (d, J = 11.6 Hz, 9H). 13 C NMR (100 MHz, CDC13) δ 171.3, 158.6, 158.5, 157.9, 156.3, 155.8, 155.6, 154.3, 147.3, 144.1, 143.9, 135.4, 132.7, 130.0, 127.9, 124.1, 124.1, 122.3, 119.6, 119.2, 62.7, 53.5, 52.4, 52.3, 44.9, 41.1, 32.6, 30.3, 26.7, 25.2, 25.0, 19.1.

[0074] Preparation of compound 4:

[0075] Compound E1 was replaced with compound E4 (114 mg, 0.230 mmol) according to the same method as preparing compound 1 to obtain compound 4.

[0076] Compound 4 was detected, and the detection results are as follows: 1H NMR (400 MHz, CDC13) δ 8.35 (d, J = 8.8 Hz, 1H), 7.64 (dd, J = 8.5, 3.5 Hz, 2H), 7.51 (t, J = 7.0 Hz, 4H), 7.45 - 7.33 (m, 9H), 7.16 (dd, J = 13.2, 7.7 Hz, 3H), 7.08 (d, J = 8.0 Hz, 2H), 5.66 (s, 2H), 4.92 - 4.52 (m, 2H), 4.43 (dt, J = 13.9, 5.2 Hz, 2H), 4.15 - 3.49 (m, 4H), 3.28 - 3.11 (m, 1H), 2.75 (dt, J = 15.4, 6.2 Hz, 2H), 2.39 (q, J = 8.9, 8.3 Hz, 2H), 2.28 (dt, J = 33.8, 8.1 Hz, 2H), 1.88 - 1.61 (m, 6H), 1.00 (d, J = 7.4 Hz, 9H). 13 C NMR (100 MHz, CDC13) δ 171.6, 158.7, 158.6, 158.0, 157.9, 156.4, 155.9, 155.8, 154.4, 147.7, 135.5, 132.8, 130.1, 128.0, 124.2, 124.2, 122.3, 122.2, 119.7, 119.3, 62.9, 53.6, 52.4, 50.0, 41.8, 33.2, 30.4, 30.1, 29.4, 26.8, 25.6, 24.9, 19.2.

[0077] Preparation of compound 5:

[0078] Compound E1 was replaced with compound E5 (117 mg, 0.230 mmol) according to the same method as for the preparation of compound 1 to obtain compound 5.

[0079] Compound 5 was tested, and the results of the test are as follows: 1 H NMR (400 MHz, CDC13) δ 8.35 (d, J = 7.7 Hz, 1H), 7.63 (dd, J = 8.5, 3.8 Hz, 2H), 7.50 (t, J = 7.4 Hz, 4H), 7.42 (d, J = 14.9 Hz, 3H), 7.39

[0080] -7.30 (m, 6H), 7.15 (t, J = 6.8 Hz, 3H), 7.07 (d, J = 8.0 Hz, 2H), 5.79 (s, 2H), 4.87 - 4.55 (m

[0081] ,2H),4.43(dt,J=10.9,5.1Hz,2H),3.96(dt,J=9.5,5.3Hz,2H),3.87–3.60(m,1H),3.16(dt,J=60.2,12.4Hz,1H),2.76–2.69(m,2H),2.34(t ,J=7.5Hz,2H),2.28–2.18(m,2H),1.94(d,J=13.9Hz,2H),1.69(dt,J=14.9,7.7Hz,5H),1.40(dt,J=22.2,7.6Hz,2H),0.99(d,J=11.4Hz,9H). 13 C NMR (100MHz, CDCl3) δ171.8,158.6,158.5,157.9,156.4,155.8,155.6,154.3,154.1,147.9,144.1,135.4,132.6,130.0,12 7.9,124.1,124.1,122.1,119.6,119.2,62.8,53.5,52.3,49.9,45.7,41.7,33.4,29.4,29.1,26.7,25.5,25.1,24.1,19.1.

[0082] Preparation of compound 6:

[0083] Compound 6 was obtained by replacing Compound E1 with Compound E6 (120 mg, 0.230 mmol) according to the same method as for preparing Compound 1.

[0084] Compound 6 was tested, and the test results were as follows: 1 H NMR(400MHz,CDCl3)δ8.30(s,1H),7

[0085] .83–7.74(m,3H),7.67–7.57(m,5H),7.48(s,1H),7.41–7.30(m,11H),7.25(d,J=8.1Hz,1H),7.19(d,J=8.1Hz,1H),7.15–7.05(m, 4H),7.04–6.95(m,3H),4.92(dq,J=5.4,2.8Hz,1H),4.31(td,J=4.2,0.9Hz,3H),4.18(t,J=4.1Hz,3H),3.82(d,J=2.7Hz,2H),3.5

[0086] 8 - 3.33 (m, 3H), 2.66 (t, J = 8.0 Hz, 2H), 2.38 (t, J = 7.9 Hz, 2H), 2.13 - 1.88 (m, 5H), 1.79 (p, J = 8.1 Hz, 3H), 1.63 - 1.51 (m, 3H), 1.48 - 1.27 (m, 6H), 1.03 (s, 9H).

[0087] Example 2

[0088] A BTK protein degrader with silicon-containing group as hydrophobic tag (denoted as compounds 7-10) whose structures are shown as follows, respectively:

[0089]

[0090] The structures of the intermediate compounds F1, F2, F3, F4, B, G1, G2, G3, G4, D used are shown as follows, respectively:

[0091]

[0092] The specific preparation method is as follows:

[0093] 1) Preparation of compounds G1-G4:

[0094] Preparation of compound G1:

[0095] Compound F1 (278 mg, 2.00 mmol) was used to replace compound A in the same method as that for preparing compound C in Example 1 to obtain compound G1.

[0096] Compound G1 was detected, and the detection results are as follows: 1 H NMR (400 MHz, CDC13) δ 7.74 - 7.49 (m, 4H), 7.46 - 7.29 (m, 6H), 3.83 (t, J = 5.4 Hz, 2H), 3.44 (t, J = 4.9 Hz, 2H), 2.03 (p, J = 5.2 Hz, 2H), 1.03 (s, 9H).

[0097] Preparation of compound G2:

[0098] Compound F2 (306 mg, 2.00 mmol) was used to replace compound A in the same method as that for preparing compound C in Example 1 to obtain compound G2.

[0099] Compound G2 was detected, and the detection results are as follows: 1H NMR (400 MHz, CDC13) δ 7.77 - 7.51 (m, 4H), 7.50 - 7.24 (m, 6H), 3.71 (t, J = 5.9 Hz, 2H), 3.44 (t, J = 4.5 Hz, 2H), 1.98 - 1.55 (m, 4H), 1.03 (s, 9H).

[0100] Preparation of compound G3:

[0101] Compound G3 was obtained by replacing compound A with compound F3 (334 mg, 2.00 mmol) according to the same method as that for preparing compound C in Example 1.

[0102] Compound G3 was detected, and the detection results thereof are as follows: 1 H NMR (400 MHz, CDC13) δ 7.75 - 7.54 (m, 4H), 7.50 - 7.27 (m, 6H), 3.68 (t, J = 6.0 Hz, 2H), 3.45 (t, J = 4.6 Hz, 2H), 1.87 - 1.67 (m, 2H), 1.64 - 1.56 (m, 2H), 1.52 (qd, J = 7.0, 1.2 Hz, 2H), 1.03 (s, 9H).

[0103] Preparation of compound G4:

[0104] Compound G4 was obtained by replacing compound A with compound F4 (362 mg, 2.00 mmol) according to the same method as that for preparing compound C in Example 1.

[0105] Compound G4 was detected, and the detection results thereof are as follows: 1 H NMR (400 MHz, CDC13) δ 7.84 - 7.57 (m, 4H), 7.51 - 7.23 (m, 6H), 3.68 (t, J = 6.0 Hz, 2H), 3.45 (t, J = 4.6 Hz, 2H), 1.84 (tt, J = 7.6, 4.6 Hz, 2H), 1.57 (dtd, J = 7.1, 6.5, 5.8 Hz, 2H), 1.50 - 1.26 (m, 4H), 1.03 (s, 9H).

[0106] 2) Preparation of compound 7-10:

[0107] Preparation of compound 7:

[0108] Compound D (155 mg, 0.400 mmol) was dissolved in acetonitrile (5 mL), followed by the addition of compound G1 (226 mg, 0.600 mmol), cesium carbonate (391 mg, 1.20 mmol), and the reaction was refluxed overnight, followed by cooling to room temperature and concentration, diluted with water (20 mL) and extracted with ethyl acetate (3 x 10 mL), followed by combining the organic phase, drying the organic phase using anhydrous sodium sulfate, concentration, and purifying the obtained crude product using silica gel column chromatography (petroleum ether: ethyl acetate = 40: 1) to obtain compound 7.

[0109] Compound 7 was tested, and the test results thereof were as follows: 1 H NMR (400 MHz, CDC13) δ 8.30 (s, 1H), 7.85 - 7.75 (m, 3H), 7.65 - 7.55 (m, 5H), 7.41 - 7.32 (m, 11H), 7.25 (d, J = 8.1 Hz, 1H), 7.19 (d, J = 8.1 Hz, 1H), 7.15 - 7.07 (m, 4H), 6.99 (dd, J = 7.8, 1.4 Hz, 2H), 4.72 (dddd, J = 5.5, 4.2, 3.1, 1.4 Hz, 1H), 3.74 (t, J = 6.5 Hz, 3H), 2.96 (dd, J = 12.4, 1.3 Hz, 1H), 2.84 (dd, J = 12.5, 4.0 Hz, 1H), 2.68 - 2.56 (m, 3H), 2.03 (dddd, J = 12.3, 7.7, 6.3, 3.0 Hz, 1H), 1.93 (ddt, J = 12.3, 8.3, 5.7 Hz, 1H), 1.86 - 1.78 (m, 2H), 1.73 (pd, J = 6.6, 3.7 Hz, 3H), 1.03 (s, 9H).

[0110] Preparation of compound 8:

[0111] Compound G1 was replaced with compound G2 (235 mg, 0.600 mmol) according to the same method as in the preparation of compound 7 to obtain compound 8.

[0112] Compound 8 was tested, and the test results thereof were as follows: 1H NMR (400 MHz, CDC13) δ 8.30 (s, 1H), 7.90 - 7.73 (m, 2H), 7.67 - 7.56 (m, 5H), 7.42 - 7.31 (m, 11H), 7.25 (d, J = 8.1 Hz, 1H), 7.19 (d, J = 8.1 Hz, 1H), 7.17 - 7.06 (m, 4H), 7.03 - 6.92 (m, 3H), 4.76 (dddd, J = 5.5, 4.1, 3.0, 1.3 Hz, 1H), 3.85 - 3.54 (m, 3H), 2.96 (dd, J = 12.4, 1.3 Hz, 1H), 2.84 (dd, J = 12.4, 3.9 Hz, 1H), 2.72 - 2.56 (m, 3H), 2.55 - 2.45 (m, 2H), 2.03 (dddd, J = 12.3, 7.7, 6.3, 3.0 Hz, 1H), 1.93 (ddt, J = 12.3, 8.3, 5.7 Hz, 1H), 1.70 - 1.50 (m, 5H), 1.03 (s, 9H).

[0113] Preparation of compound 9:

[0114] Compound G1 was replaced with compound G3 (243 mg, 0.600 mmol) according to the same method as for preparing compound 7 to obtain compound 9.

[0115] Compound 9 was detected, and the detection results are as follows: 1 H NMR (400 MHz, CDC13) δ 8.30 (s, 1H), 7.90 - 7.73 (m, 2H), 7.67 - 7.56 (m, 5H), 7.42 - 7.31 (m, 11H), 7.25 (d, J = 8.1 Hz, 1H), 7.19 (d, J = 8.1 Hz, 1H), 7.17 - 7.06 (m, 4H), 7.03 - 6.92 (m, 3H), 4.76 (dddd, J = 5.5, 4.1, 3.0, 1.3 Hz, 1H), 3.85 - 3.54 (m, 3H), 2.96 (dd, J = 12.4, 1.3 Hz, 1H), 2.84 (dd, J = 12.4, 3.9 Hz, 1H), 2.72 - 2.56 (m, 3H), 2.55 - 2.45 (m, 2H), 2.03 (dddd, J = 12.3, 7.7, 6.3, 3.0 Hz, 1H), 1.93 (ddt, J = 12.3, 8.3, 5.7 Hz, 1H), 1.70 - 1.50 (m, 5H), 1.03 (s, 9H).

[0116] Preparation of compound 10:

[0117] Compound G1 was replaced with compound G4 (252 mg, 0.600 mmol) following the same procedure as for the preparation of compound 7 to obtain compound 10.

[0118] Compound 10 was tested and the results of the testing are as follows: 1 H NMR (400 MHz, CDC13) δ 8.30 (s, 1H), 7.82 - 7.72 (m, 3H), 7.67 - 7.55 (m, 5H), 7.41 - 7.32 (m, 11H), 7.25 (d, J = 8.1 Hz, 1H), 7.19 (d, J = 8.1 Hz, 1H), 7.16 - 7.07 (m, 4H), 7.04 - 6.95 (m, 3H), 4.76 (dddd, J = 5.5, 4.1, 3.0, 1.3 Hz, 1H), 3.68 (t, J = 6.0 Hz, 3H), 2.96 (dd, J = 12.4, 1.3 Hz, 1H), 2.84 (dd, J = 12.4, 3.9 Hz, 1H), 2.70 - 2.54 (m, 3H), 2.49 (td, J = 6.4, 2.8 Hz, 3H), 2.03 (dddd, J = 12.3, 7.7, 6.3, 3.0 Hz, 1H), 1.98 - 1.88 (m, 1H), 1.88 - 1.70 (m, 2H), 1.65 - 1.43 (m, 5H), 1.43 - 1.24 (m, 6H), 1.03 (s, 9H).

[0119] Comparative Example 1

[0120] Compound 11 is a non-silicon-containing group hydrophobic tag comparative. Its structure is shown below:

[0121]

[0122] The synthetic route is as follows:

[0123]

[0124] The structures of the intermediate compounds H, I, J, E2 used are shown below:

[0125]

[0126] The specific preparation method is as follows:

[0127] 1) Preparation of compound J:

[0128] Compound I (456 mg, 3.00 mmol) was dissolved in tetrahydrofuran (15 mL) at 0°C, followed by the addition of sodium hydride (180 mg, 4.50 mmol, the content of sodium hydride used was 60%), after stirring for ten minutes at the temperature, compound H (1.45 g, 6.00 mmol) was added, and the reaction was allowed to proceed overnight at room temperature, after the completion of the reaction, ethyl acetate (40 mL) was added, and the organic phase was washed with saturated brine (20 mL), the organic phase was dried using anhydrous sodium sulfate, and concentrated, and the obtained crude product was purified using a silica gel column (petroleum ether: ethyl acetate = 20: 1) to obtain compound J.

[0129] Compound J was tested, and the test results thereof were as follows: 1 H NMR (400 MHz, CDCl3) δ 3.68 (t, J = 6.8 Hz, 2H), 3.41 (t, J = 6.7 Hz, 2H), 2.32 (dt, J = 10.8, 5.3 Hz, 3H), 2.11 (d, J = 5.1 Hz, 6H), 1.70 (t, J = 5.6 Hz, 6H).

[0130] 2) Preparation of compound 11:

[0131] Compound C was replaced with compound J (62.0 mg, 0.280 mmol) according to the same method as in the preparation of compound 1 to obtain compound 11.

[0132] Compound 11 was tested, and the test results thereof were as follows: 1 H NMR (400 MHz, CDCl3) δ 3.68 (t, J = 6.8 Hz, 2H), 3.41 (t, J = 6.7 Hz, 2H), 2.32 (dt, J = 10.8, 5.3 Hz, 3H), 2.11 (d, J = 5.1 Hz, 6H), 1.70 (t, J = 5.6 Hz, 6H).

[0133] Compound performance test

[0134] 1) Western blotting was used to evaluate the ability of the above BTK protein degraders with silicon-containing groups as hydrophobic tags (BTK protein degraders 1-10 prepared in Examples 1-2) and BTK protein degraders with non-silicon-containing groups as hydrophobic tags (BTK protein degrader 11 prepared in Comparative Example 1) to degrade BTK in MOLM-13 (human acute myeloid leukemia cells), and the concentration of each compound used was 5 μM. The degradation efficiency results are shown in Table 1, and the IC 50 The evaluation results are shown in Table 1.

[0135] Table 1. BTK degradation efficiency and IC 50

[0136]

[0137] As can be seen from Table 1, each compound has a certain ability to degrade BTK, and is better than the BTK protein degrader comparative compound 11 with non-silicon-containing groups as hydrophobic tags, and compound 2 is the most excellent. Compound 2 was selected for further degradation effect evaluation, and MOLM-13 cell lines were selected and set at eight concentration gradients of 0, 0.15, 0.3, 0.6, 1.25, 2.5, 5, and 10 μM to determine whether the compound can degrade BTK in a concentration-dependent manner. The results are shown in Figure 1 The results show that compound 2 can degrade BTK in MOLM-13 cell lines in a concentration-dependent manner, and the degradation DC 50 is 2.78 μM. MOLM-13 cell lines were selected and set at six time gradients of 0, 3, 6, 9, 12, and 24 h to determine whether the preferred compounds can degrade BTK in a time-dependent manner. The results are shown in Figure 2 The results show that compound 2 can degrade BTK in MOLM-13 cell lines in a time-dependent manner.

[0138] 2) Compound 2 was evaluated for liver microsomal metabolic stability, and non-silicon-containing group hydrophobic tag compound 11 was used as a comparison, and Diclofenac was used as a positive control.

[0139] The specific method for evaluating the stability of liver microsomal metabolism is as follows: the liver microsomes are taken out from the-80°C refrigerator and placed on a 37°C water bath constant temperature oscillator for pre-incubation for 3 min, and then thawed for use. Then a certain amount of NADPH is weighed, and a proper amount of magnesium chloride solution is added to dissolve into a 2 mM solution for use. The incubation system mixture solution (without NADPH) is prepared according to the composition ratio of the experimental incubation system (3 mM MgCl2-PB solution, 1 μM test compound, 0.5 mg / mL liver microsomes), and is divided into 40 μL / tube.

[0140] The 0 min sample is added with 240 μL of internal standard working precipitant, and then 40 μL of NADPH solution (40 μL of magnesium chloride solution is added to the negative control group), and the other samples are added with 40 μL of NADPH solution to start the reaction (40 μL of magnesium chloride solution is added to the negative control group), and then incubated in a 37°C water bath for 5 min, 15 min, 30 min and 60 min, and then 240 μL of internal standard precipitant is added, and 40 μL of NADPH solution is added to the positive control group to start the reaction, and then incubated in a 37°C water bath for 5 min and 15 min, and then 240 μL of internal standard precipitant is added. All the samples are vortexed and centrifuged to take 150 μL of supernatant, and then vortexed after 150 μL of water is added, and then analyzed by LC-MS / MS. It is found by analysis and calculation that compound 2 shows clearance efficiency in two species of humans and mice. And it is significantly stable in the BTK protein degrader comparative example 11 with a non-silicon group as a hydrophobic tag. The results are shown in Table 2.

[0141] Table 2 Stability of liver microsomal metabolism

[0142]

[0143] The oral bioavailability of compound 2 is evaluated.

[0144] The specific method for evaluating the oral bioavailability is as follows: after the rats are fasted overnight, three are orally administered (10 mg / kg), and three are administered via the tail vein (1 mg / kg), and then blood is taken at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h and 8 h, and then placed in pre-heparin sodium tubes, and then vortexed to mix the blood and heparin sodium, and then centrifuged (4°C, 3000 rpm, 10 min) to obtain plasma, and then 50 μL of the plasma sample is taken, and then 50 μL of diluent (50% methanol / water) and 250 μL of methanol precipitant are added, and then vortexed and centrifuged (4°C, 12000 rpm, 10 min), and then the supernatant is filtered and stored for LC-MS / MS detection. The results (Table 3) show that compound 2 has good oral bioavailability of 17.9%.

[0145] Table 3 Kinetic parameters

[0146]

[0147] From Table 2 and the results of the evaluation test of the oral bioavailability of Compound 2, it can be seen that the present application is optimized in terms of hydrophobic tag type and linker type, and the selected Compound 2 can exert higher metabolic stability and in vivo bioavailability.

[0148] 3) In vivo pharmacodynamics PD experiment was evaluated in a xenograft model, and the mice received a single oral dose of Compound 2 (10 mg / kg). The mice were euthanized at the designated time points (0 hours, 6 hours, 12 hours, 24 hours, 48 hours and 72 hours) after oral administration, and the tumor tissues were harvested for Western blotting analysis. The data are shown in Table 4, and the results show that Compound 2 significantly reduces BTK expression within 6 hours, and the effect lasts up to 48 hours. These findings indicate that the degrading agent 2 can maintain BTK degradation in vivo for a long time.

[0149] Table 4 PD data

[0150]

[0151] 4) In vivo pharmacodynamic evaluation of Compound 2, MOLM-13 cells were selected to establish a xenograft model.

[0152] The specific method is as follows: MOLM-13 cells were injected into NOD / SCID mice. Two days after cell injection, the mice were divided into three groups, Compound 2 oral administration group 1 (5mpk, 6 mice, daily administration), Compound 2 oral administration group 2 (10mpk, 6 mice, daily administration), and control group (6 mice).

[0153] After 18 days, the body weight of the mice in the administration group and the body weight of the mice in the control group did not fluctuate significantly and the mice were not significantly abnormal, indicating that Compound 2 was safe and well tolerated, and the tumor weight and tumor volume of the mice in the administration group were significantly lower than those in the control group, and the tumor inhibition rate ((1-administration group tumor weight / control group tumor weight)*100%) is shown in Table 5. The results show that Compound 2 has a strong in vivo therapeutic effect.

[0154] Table 5 TGI of MOLM-13 cell xenograft model

[0155]

[0156] The above is only a preferred embodiment of the present application, and the present application is not limited to the above-described embodiments, as long as the same means achieve the technical effects of the present application, it should belong to the protection scope of the present application. The technical solutions and / or embodiments within the protection scope of the present application can have various modifications and changes.

Claims

1. A BTK protein degrader with a silicon-containing group as a hydrophobic tag, characterized in that: Its structure is shown in Formula II or Formula III: Formula II; Formula III; wherein m and n are independently selected from integers ranging from 1 to 10.

2. The BTK protein degrader with a silicon-containing group as a hydrophobic tag according to claim 1, characterized in that Its structure is shown in Formula IV: Formula IV.

3. A method for preparing a BTK protein degrader with a silicon-containing group as a hydrophobic tag according to claim 1, characterized in that: When the structure of the BTK protein degrader with a silicon-containing group as a hydrophobic tag is shown in Formula II, the preparation method thereof is shown in Scheme 1; when the structure of the BTK protein degrader with a silicon-containing group as a hydrophobic tag is shown in Formula III, the preparation method thereof is shown in Scheme 2; Route 1: ； Route 2: 。 4. Use of the BTK protein degrader according to claim 1 or 2, which has a silicon-containing group as a hydrophobic tag, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating cancers associated with BTK abnormalities.

5. The use according to claim 4, characterized in that The related cancer is leukemia.

6. A pharmaceutical composition, characterized in that The BTK protein degrader comprising the silicon-containing group as a hydrophobic tag according to claim 1 or 2, or a pharmaceutically acceptable salt thereof.

7. The pharmaceutical composition according to claim 6, characterized in that Pharmaceutically acceptable excipients are also included.

8. The pharmaceutical composition according to claim 7, characterized in that The excipient is at least one of gum arabic, syrup, lanolin and starch.

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