Isoindoline-pyrrolidine derivative

Abstract

The invention provides an isoindoline-pyrrolidine derivative, and belongs to the technical field of chemical drugs. The structure of the isoindoline-pyrrolidine derivative is as shown in a formula I in the specification. The compound disclosed by the invention shows high selectivity and excellent degradation effect on TRPV1 in vitro and in vivo. In different animal pain models, the compound provided by the invention shows obvious analgesic activity. It is worthy of noting that the compound provided by the invention does not show adverse effects on body temperature, which are generally related to TRPV1 agonists or inhibitors, which indicates that the compound provided by the invention has higher safety. The compound provided by the invention is combined with a TRPV1 agonist or inhibitor for use, so that side effects caused by the TRPV1 agonist or inhibitor can be effectively relieved. The compound disclosed by the invention is the first PROTAC degradation device aiming at the ion channel at present, and the huge potential of the PROTAC technology in the field of ion channel modulation in the future is highlighted.

Description

Technical Field

[0001] The present invention belongs to the technical field of chemical drugs, and particularly relates to an isoindoline-pyrrolidine derivative. Background Art

[0002] Transient Receptor Potential Vanilloid 1 (TRPV1) is a non-selective cation-permeable ion channel with six transmembrane structures. It can be activated by capsaicin (Cap), inflammatory mediators, acids, endogenous cannabinoids, arachidonic acid, etc., and can also respond to temperature stimuli (>42°C). Due to its ability to respond to a wide variety of stimuli, the TRPV1 channel is thus called a polymodal receptor. The TRPV1 channel is widely present in various species from yeast to humans, mainly distributed in the brain tissue and primary sensory afferent nerve endings, and is also expressed in some non-neural tissues such as skin keratinocytes, bladder mucosal epithelial cells, gastrointestinal tract, stem cells, fibroblasts, T cells, alveolar wall cells, mast cells, and vascular smooth muscle cells. It plays an important role in pain transmission and modulation, and integration of various pain information. TRPV1 gene knockout mice show a significantly reduced or even absent response to heat. TRPV1 blockers can cause an increase in body temperature in animals and humans, indicating that TRPV1 is a biomolecule necessary for heat and pain sensation and participation in body temperature regulation.

[0003] Studies have found that the use of TRPV1 antagonists or agonists as drugs often causes side effects, such as high fever or burns, as well as hypothermia or a burning sensation, which limits the development of TRPV1 antagonists or agonists as systemic drugs. Therefore, it is of great significance to develop a drug that can effectively reduce the side effects caused by TRPV1 antagonists or agonists.

[0004] As bifunctional molecules, proteolysis-targeting chimeras (PROTACs) include a small molecule compound capable of binding to a target protein (protein of interest, POI), a linker group introduced at its appropriate position, and a small molecule compound capable of binding to an E3 ligase. As small molecule probes, PROTACs can bind to both the target protein and the E3 ligase simultaneously, thereby promoting the ubiquitination of the target protein and its recognition and degradation by the proteasome. Since the first PROTAC molecule was reported, this technology has attracted extensive attention in the academic and industrial communities, and several pharmaceutical companies have disclosed early clinical and / or Phase I / II clinical R & D projects. The PROTAC technology has been proven to be effective against a series of cytoplasmic and nuclear proteins, as well as several transmembrane proteins, such as α1A-adrenergic receptor, CC chemokine receptor type 9, receptor tyrosine kinase (RTK), and solute carrier family member A1 (SLC9A1). However, its applicability to ion channels remains unclear. Due to the inaccessible nature of proteins within the cell membrane, developing PROTAC degraders for transmembrane proteins presents unique difficulties. E3 ligases are usually present in the cytoplasm, while TRPV1 is typically located on the cell membrane, in sharp contrast to E3 ligases. This spatial difference poses extraordinary challenges to the formation of the target-PROTAC-ligase ternary complex. In addition, the binding sites of ion channels are usually located in the pockets of the transmembrane region of the cell membrane, which poses significant limitations to ligand recognition. These sites often exhibit different conformational changes during the dynamic process of channel opening and closing, thereby increasing the complexity of PROTAC design. Therefore, developing PROTACs capable of effectively degrading TRPV1 presents significant challenges. Summary of the Invention

[0005] The object of the present invention is to provide an isoindoline-pyrrolidine derivative, its preparation method and uses.

[0006] The present invention provides a compound, its stereoisomer or its salt, and the structure of the compound is shown in Formula I:

[0007]

[0008] Wherein, R1 is selected from C 1-6 alkyl; R2 is selected from C 1-6 alkyl; R3 is selected from C 1-6 alkyl;

[0009] m is an integer from 0 to 5;

[0010] L is selected from C 1-12 alkylene, O, S, O-C 1-12 alkylene, S-C 1-12 alkylene, 3-6 membered saturated heterocyclic group, 3-6 membered saturated cycloalkyl group;

[0011] Y is selected from O, S or none.

[0012] Furthermore, the structure of the compound is shown in Formula II:

[0013]

[0014] Wherein, L is selected from C 1-12 alkylene, O, S, O-C 1-12 alkylene, S-C 1-12 alkylene, 3- to 6-membered saturated heterocyclic group;

[0015] Y is selected from O, S or none.

[0016] Furthermore, the structure of the compound is shown in Formula III:

[0017]

[0018] Wherein, n is selected from an integer of 1-9.

[0019] Furthermore, the structure of the compound is shown in Formula III or Formula IV:

[0020]

[0021] Wherein, X is none or methylene;

[0022] a is 0 or 1; b is 0 or 1.

[0023] Furthermore, the compound is selected from:

[0024]

[0025]

[0026]

[0027] The present invention also provides a pharmaceutical composition, which is a preparation prepared from the above-mentioned compound, its stereoisomer or its salt as an active ingredient, plus a pharmaceutically acceptable auxiliary ingredient.

[0028] The present invention also provides the use of the above-mentioned compound, its stereoisomer or its salt in the preparation of a TRPV1 degrader.

[0029] As is well known to those skilled in the art, TRPV1 degrader can prevent and / or treat asthma, chronic obstructive pulmonary disease, idiopathic pulmonary fibrosis, chronic idiopathic cough, pruritus, esophageal cancer, cervical squamous cell carcinoma, type 2 diabetes.

[0030] The present invention also provides the use of the above-mentioned compound, its stereoisomer or its salt in the preparation of a medicament for preventing and / or treating asthma, chronic obstructive pulmonary disease, idiopathic pulmonary fibrosis, chronic idiopathic cough, pruritus, esophageal cancer, cervical squamous cell carcinoma, diabetes; preferably, the diabetes is type 2 diabetes.

[0031] The present invention also provides the use of the above-mentioned compound, its stereoisomer or its salt in the preparation of an analgesic medicament; preferably, the analgesic medicament is a medicament for treating chronic inflammatory pain, acute inflammatory pain, neuropathic pain or osteoarthritis pain.

[0032] The present invention also provides the use of the above-mentioned compound, its stereoisomer or its salt in combination with a TRPV1 agonist or a TRPV1 inhibitor in the preparation of a medicament for reducing the side effects caused by the TRPV1 agonist or the TRPV1 inhibitor; preferably, the TRPV1 agonist is sanshool, the TRPV1 inhibitor is AMG517, and the side effect is body temperature elevation.

[0033] The present invention proposes a programmable regulation strategy to achieve PROTAC degradation of ion channels through the process of "activation-endocytosis-degradation", and overcomes the challenges by using this strategy and develops a PROTAC that can effectively degrade TRPV1. The present invention constructs a compound using the TRPV1 agonist capsaicin as a ligand, and this compound has the dual functions of inducing endocytosis and RPOTAC.

[0034] The compound of the present invention shows high selectivity and excellent degradation effect on TRPV1 both in vitro and in vivo. In different animal pain models, the compound of the present invention shows significant analgesic activity. Notably, the compound of the present invention does not show the adverse effects on body temperature usually associated with TRPV1 agonists or inhibitors, which indicates that the compound of the present invention has higher safety. The compound of the present invention can effectively reduce the side effects caused by TRPV1 agonists or inhibitors when used in combination with TRPV1 agonists or inhibitors. The compound of the present invention is currently the first PROTAC degrader targeting ion channels, highlighting the great potential of PROTAC technology in the field of ion channel modulation in the future.

[0035] Obviously, based on the above content of the present invention, according to the common general technical knowledge and customary means in the art, without departing from the above basic technical idea of the present invention, various other forms of modifications, substitutions or changes can be made.

[0036] The following is a further detailed description of the above content of the present invention through specific embodiments in the form of examples. However, this should not be construed as limiting the scope of the above subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention fall within the scope of the present invention. Description of the Drawings

[0037] Figure 1 : Degradation effect of 20 μM compound on TRPV1 after 24 h (a); Degradation effect of different concentrations of compound 5a on TRPV1 after 24 h (b).

[0038] Figure 2 : Degradation effect of 10 μM compound on TRPV1 after 24 h (a); Degradation effect of different concentrations of compound TPD on TRPV1 after 24 h (b); Degradation effect of 10 μM compound TPD on TRPV1 at different times (c).

[0039] Figure 3 : Analgesic activity of the compound in CFA-induced chronic inflammatory pain model (a), formalin-induced acute inflammatory pain model (b), CCI-induced neuropathic pain model (c), STZ-induced diabetic neuropathic pain model (d), and MIA-induced osteoarthritis pain model (e).

[0040] Figure 4 : Effect of different concentrations of compound TPD on body temperature (a); Effect of drugs in each group on body temperature (b). Detailed Description of the Invention

[0041] The raw materials and equipment used in the present invention are all known products and are obtained by purchasing commercially available products.

[0042] Compound 5a-5i was synthesized according to the following Route A, and compound 10a-a0l was synthesized according to the following Route B.

[0043] Route A:

[0044]

[0045] Route B:

[0046]

[0047] The following are examples of synthesizing specific compounds of the present invention.

[0048] Example 1: (E)-N-(4-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)propoxy)-3-methoxybenzyl)-8-methylnon-6-enamide (5a)

[0049]

[0050] A mixture of intermediate 3a (0.1 g, 234.52 μmol), 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (61.1 mg, 228.21 μmol) and potassium carbonate (94.27 mg, 703.57 μmol) was dissolved in 15 mL of N,N-dimethylformamide. The mixture was heated at 45 °C overnight and then cooled to room temperature. The reaction was quenched with water and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine and then concentrated by evaporation under vacuum. The crude product was purified by silica gel chromatography using a petroleum ether / dichloromethane gradient to give the desired product as a white solid (5a, 20% yield). 1 H NMR (400 MHz, CDCl3) δ 8.11 (s, 1H), 7.67 (t, J = 7.8 Hz, 1H), 7.45 (d, J = 7.8 Hz, 1H), 7.28 (s, 1H), 6.91 (d, J = 8.6 Hz, 1H), 6.78 (s, 2H), 5.68 (s, 1H), 5.33 (dd, J = 7.8, 8.6 Hz, 2H), 4.94 (m, 1H), 4.43–4.25 (m, 6H), 3.81 (s, 3H), 2.94–2.67 (m, 3H), 2.43–2.33 (m, 2H), 2.25–1.94 (m, 6H), 1.64 (d, J = 7.2 Hz, 2H), 1.44–1.29 (m, 2H), 0.94 (d, J = 6.6 Hz, 4H), 0.85 (d, J = 6.6 Hz, 2H). 13 C NMR (101 MHz, CDCl3) δ 172.82, 170.81, 168.00, 156.52, 149.60, 147.59, 138.09, 136.52, 133.74, 126.49, 120.22, 119.05, 115.93, 113.75, 111.59, 77.33, 77.22, 77.01, 76.70, 65.79, 65.22, 55.89, 49.10, 43.36, 36.72, 32.23, 31.39, 30.96, 29.29, 28.99, 25.26, 22.65. HRMS m / z (ESI) calcd for C 34 H 41 N3O8 620.2894; found, 620.2965.

[0051] Compounds 5b - 5i were synthesized by referring to the synthetic method of reference compound 5a.

[0052] Example 2: (E)-N-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)butoxy)-3-methoxybenzyl)-8-methylnon-6-enamide (5b)

[0053] White solid, 19% yield.

[0054]

[0055] 1 H NMR (400 MHz, CDCl3) δ 8.12 (s, 1H), 7.65 (t, J = 7.8 Hz, 1H), 7.46 (d, J = 7.2 Hz, 1H), 7.27 (d, J = 4.0 Hz, 1H), 6.92 (d, J = 8.6 Hz, 1H), 6.77 (s, 2H), 5.66 (s, 1H), 5.48–4.95 (m, 2H), 4.93 (d, J = 5.0 Hz, 1H), 4.46–4.24 (m, 6H), 3.80 (s, 3H), 2.95–2.68 (m, 3H), 2.42–1.87 (m, 8H), 1.64 (d, J = 7.2 Hz, 2H), 1.44–1.26 (m, 4H), 0.94 (d, J = 6.7 Hz, 4H), 0.85 (d, J = 6.5 Hz, 2H). 13 C NMR (101 MHz, CDCl3). 13 C NMR (101 MHz, CDCl3) δ 172.89, 170.81, 168.00, 165.56, 156.52, 149.61, 147.60, 138.09, 136.53, 133.93, 131.51, 126.49, 120.22, 119.05, 117.38, 115.94, 113.75, 111.59, 65.79, 65.22, 55.89, 49.11, 43.37, 38.98, 38.39, 36.72, 32.23, 31.40, 30.97, 29.30, 28.99, 27.89, 27.02, 25.26, 22.64. HRMS m / z (ESI) calcd for C 35 H 43 N3O8 634.3050; found, 634.3121.

[0056] Example 3: (E)-N-(4-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)pentyl)oxy)-3-methoxybenzyl)-8-methylnon-6-enamide (5c)

[0057] White solid, 15% yield.

[0058]

[0059] 1 H NMR (400 MHz, CDCl3) δ 8.13 (s, 1H), 7.67 (t, J = 7.6 Hz, 1H), 7.46 (d, J = 7.2 Hz, 1H), 7.28 (d, J = 4.4 Hz, 1H), 6.91 (d, J = 7.2 Hz, 3H), 6.79 (s, 1H), 5.66 (s, 2H), 5.30 (m, 1H), 4.98 (s, 1H), 4.44–4.24 (m, 6H), 3.80 (s, 3H), 2.95–2.67 (m, 3H), 2.43–2.32 (m, 2H), 2.32–1.87 (m, 6H), 1.71–1.59 (m, 3H), 1.25 (m, 5H), 0.96 (d, J = 6.6 Hz, 4H), 0.82 (d, J = 6.6 Hz, 2H). 13 C NMR (101 MHz, CDCl3) δ 173.21, 172.89, 172.57, 170.81, 168.00, 167.06, 165.56, 156.52, 149.60, 147.59, 141.02, 138.09, 136.52, 133.63, 131.80, 126.49, 120.22, 119.05, 115.93, 113.75, 111.59, 93.80, 65.79, 65.22, 55.87, 49.11, 43.36, 38.38, 36.72, 32.23, 31.39, 30.96, 29.29, 28.99, 25.26, 22.62. HRMS m / z (ESI) calcd for C 36 H 45 N3O8 648.3207; found, 648.3283.

[0060] Example 4: (E)-N-(4-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)hexyl)oxy)-3-methoxybenzyl)-8-methylnon-6-enamide (5d)

[0061] White solid, 12% yield.

[0062]

[0063] 11H NMR (400 MHz, CDCl3) δ 8.04 (s, 1H), 7.61 (t, J = 7.8 Hz, 1H), 7.43 (d, J = 7.2 Hz, 1H), 7.19 (d, J = 8.4 Hz, 1H), 6.82 (q, J = 8.4 Hz, 3H), 5.71 (s, 1H), 5.41–5.29 (m, 2H), 4.92 (dd, J = 10.0, 5.2 Hz, 1H), 4.34 (d, J = 6.4 Hz, 2H), 4.16 (t, J = 6.4 Hz, 2H), 4.02 (t, J = 6.6 Hz, 2H), 3.83 (s, 3H), 2.91–2.61 (m, 3H), 2.32–1.80 (m, 10H), 1.71–1.62 (m, 2H), 1.56 - 1.47 (s, 6H), 0.99 (d, J = 6.6 Hz, 4H), 0.84 (d, J = 6.6 Hz, 2H). 13 13C NMR (101 MHz, CDCl3) δ 172.83, 170.87, 167.98, 167.07, 165.67, 156.71, 149.57, 148.00, 138.08, 136.49, 133.83, 131.00, 126.50, 120.10, 118.93, 117.13, 115.73, 112.93, 111.66, 69.47, 69.04, 56.00, 53.43, 49.09, 43.43, 38.95, 36.89, 36.72, 32.23, 31.39, 30.97, 29.62, 29.37, 29.29, 29.05, 29.00, 28.78, 27.94, 27.24, 25.87, 25.83, 25.28, 22.65, 22.63, 14.20. HRMS m / z (ESI) calcd for C 37 H 47 N3O8 662.3363; found, 662.3441

[0064] Example 5: (E)-N-(4-((7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)heptyl)oxy)-3-methoxybenzyl)-8-methylnon-6-enamide (5e)

[0065] White solid, yield 19%.

[0066]

[0067] 11H NMR (400 MHz, CDCl3) δ 8.06 (s, 1H), 7.67 (t, J = 7.8 Hz, 1H), 7.45 (d, J = 7.2 Hz, 1H), 7.21 (d, J = 8.5 Hz, 1H), 6.80 (q, J = 8.4 Hz, 3H), 5.70 (s, 1H), 5.41–5.28 (m, 2H), 4.93 (dd, J = 12.0, 5.4 Hz, 1H), 4.37 (d, J = 5.4 Hz, 2H), 4.18 (t, J = 6.4 Hz, 2H), 4.00 (t, J = 6.4 Hz, 2H), 3.84 (s, 3H), 2.80 (m, 3H), 2.20 (t, J = 7.2 Hz, 3H), 2.11 (d, J = 10.4 Hz, 1H), 2.06–1.96 (m, 2H), 1.94–1.80 (m, 4H), 1.70–1.63 (m, 2H), 1.56–1.32 (m, 9H), 0.95 (d, J = 6.6 Hz, 4H), 0.85 (d, J = 6.6 Hz, 2H). 13 13C NMR (101 MHz, CDCl3) δ 172.83, 170.87, 167.98, 167.07, 156.71, 149.57, 148.00, 138.08, 136.49, 133.83, 131.00, 126.50, 120.10, 118.93, 117.13, 115.73, 112.93, 111.66, 69.47, 69.04, 56.00, 53.43, 49.09, 43.43, 38.95, 36.89, 36.72, 32.23, 30.97, 29.62, 28.78, 27.94, 25.87, 25.83, 25.28, 22.65, 22.63, 14.21. HRMS m / z (ESI) calcd for C 38 H 49 N3O8 676.3520; found, 676.3582.

[0068] Example 6: (E)-N-(4-((8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octyl)oxy)-3-methoxybenzyl)-8-methylnon-6-enamide (5f)

[0069] White solid, 19% yield.

[0070]

[0071] 11H NMR (400 MHz, CDCl3) δ 8.07 (s, 1H), 7.68 (t, J = 7.8 Hz, 1H), 7.46 (d, J = 7.2 Hz, 1H), 7.22 (d, J = 8.4 Hz, 1H), 6.81 (q, J = 8.4 Hz, 3H), 5.70 (s, 1H), 5.41–5.28 (m, 2H), 4.93 (dd, J = 12.0, 5.2 Hz, 1H), 4.37 (d, J = 5.4 Hz, 2H), 4.18 (t, J = 6.4 Hz, 2H), 4.02 (t, J = 6.6 Hz, 2H), 3.85 (s, 3H), 2.82 (m, 3H), 2.27–1.78 (m, 10H), 1.72–1.62 (m, 3H), 1.55–1.31 (m, 9H), 0.96 (d, J = 6.6 Hz, 4H), 0.85 (d, J = 6.6 Hz, 2H). 13 13C NMR (101 MHz, CDCl3) δ 172.83, 170.87, 167.98, 167.07, 165.67, 156.71, 149.57, 148.00, 138.08, 136.49, 133.83, 131.00, 126.50, 120.10, 118.93, 117.13, 115.73, 112.93, 111.66, 69.47, 69.04, 56.00, 53.43, 49.09, 43.43, 38.95, 36.89, 36.72, 32.23, 31.39, 30.97, 29.62, 29.37, 29.29, 29.05, 29.00, 28.78, 27.94, 27.24, 25.87, 25.83, 25.28, 22.65, 22.64, 14.23. HRMS m / z (ESI) calcd for C 39 H 51 N3O8 690.3676; found, 690.3752.

[0072] Example 7: (E)-N-(4-((9-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)nonyl)oxy)-3-methoxybenzyl)-8-methylnon-6-enamide (5 g)

[0073] White solid, 11% yield.

[0074]

[0075] 11H NMR (400 MHz, CDCl3) δ 8.12 (s, 1H), 7.67 (t, J = 7.6 Hz, 1H), 7.44 (d, J = 6.8 Hz, 1H), 7.21 (d, J = 8.2 Hz, 1H), 6.81 (s, 3H), 5.71 (s, 1H), 5.61–4.99 (m, 2H), 4.99 (s, 1H), 4.37 (d, J = 4.6 Hz, 2H), 4.17 (s, 2H), 3.99 (s, 2H), 3.84 (s, 3H), 2.95–2.64 (m, 3H), 2.16 (m, 4H), 2.04–1.74 (m, 6H), 1.63 (s, 11H), 1.37 (s, 5H), 0.95 (d, J = 6.6 Hz, 4H), 0.85 (d, J = 6.6 Hz, 2H). 13 13C NMR (101 MHz, CDCl3) δ 172.83, 170.90, 168.01, 167.09, 165.67, 156.74, 149.56, 148.00, 138.08, 136.47, 133.82, 130.96, 126.50, 120.09, 118.90, 117.12, 115.68, 112.87, 111.65, 69.43, 69.10, 56.00, 49.08, 43.43, 38.95, 36.88, 36.72, 32.23, 31.38, 30.96, 29.62, 29.31, 29.18, 29.12, 28.86, 27.94, 27.24, 25.87, 25.71, 25.27, 22.63, 14.20. HRMS m / z (ESI) calcd for C 40 H 53 N3O8 704.3833; found, 704.3898.

[0076] Example 8: (E)-N-(4-((10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)decyl)oxy)-3-methoxybenzyl)-8-methylnon-6-enamide (5h)

[0077] White solid, 15% yield.

[0078]

[0079] 11H NMR (400 MHz, CDCl3) δ 8.13 (s, 1H), 7.65 (t, J = 7.6 Hz, 1H), 7.43 (d, J = 6.8 Hz, 1H), 7.20 (d, J = 8.4 Hz, 1H), 6.79 (s, 3H), 5.31 (m, 2H), 4.98–4.88 (m, 1H), 4.35 (d, J = 4.8 Hz, 2H), 4.16 (s, 2H), 3.99 (s, 2H), 3.83 (s, 3H), 2.95–2.65 (m, 3H), 2.16 (m, 4H), 2.02–1.77 (m, 6H), 1.66 (s, 5H), 1.27 (m, 11H), 0.94 (d, J = 6.6 Hz, 4H), 0.86 (d, J = 6.6 Hz, 2H). 13 13C NMR (101 MHz, CDCl3) δ 172.94, 172.83, 170.92, 168.05, 167.10, 165.70, 156.75, 149.58, 148.04, 138.08, 136.48, 133.82, 130.96, 126.50, 120.11, 118.92, 117.10, 115.68, 112.92, 111.68, 69.49, 69.14, 56.01, 53.44, 49.09, 43.44, 38.96, 36.88, 36.72, 32.23, 31.40, 30.97, 29.62, 29.49, 29.44, 29.36, 29.29, 29.26, 29.15, 28.90, 27.94, 27.24, 25.93, 25.79, 25.28, 22.65, 22.63, 14.20. HRMS m / z (ESI) calcd for C 41 H 55 N3O8 718.3989; found, 718.3898.

[0080] Example 9: (E)-N-(4-((11-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)undecyl)oxy)-3-methoxybenzyl)-8-methylnon-6-enamide (5i)

[0081] White solid, 13% yield.

[0082]

[0083] 11H NMR (400 MHz, CDCl3) δ 8.14 (s, 1H), 7.67 (t, J = 7.6 Hz, 1H), 7.44 (d, J = 6.8 Hz, 1H), 7.21 (m, 3H), 6.81 (s, 1H), 5.70 (s, 1H), 5.33 (m, 2H), 5.01–4.89 (m, 1H), 4.36 (d, J = 4.8 Hz, 2H), 4.17 (s, 2H), 3.99 (s, 2H), 3.84 (s, 3H), 2.95–2.66 (m, 3H), 2.16 (dd, J = 9.8, 8.6 Hz, 4H), 2.05–1.79 (m, 6H), 1.66 (s, 5H), 1.28 (m, 13H), 0.95 (d, J = 6.6 Hz, 4H), 0.85 (d, J = 6.6 Hz, 2H). 13 13C NMR (101 MHz, CDCl3) δ 172.95, 172.83, 170.92, 168.05, 167.10, 165.70, 156.77, 149.58, 148.03, 138.08, 136.48, 133.82, 130.96, 126.50, 120.11, 118.92, 117.10, 115.68, 112.92, 111.67, 69.49, 69.14, 56.01, 53.44, 49.09, 43.44, 38.96, 36.88, 36.72, 32.23, 31.40, 30.97, 29.63, 29.48, 29.44, 29.36, 29.29, 29.26, 29.15, 28.90, 27.94, 27.24, 25.93, 25.79, 25.28, 22.66, 22.64, 14.20. HRMS m / z (ESI) calcd for C 42 H 57 N3O8 732.4161; found, 732.4219.

[0084] Example 10: (E)-N-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)azetidin-3-yl)oxy)-3-methoxybenzyl)-8-methylnon-6-enamide (10a)

[0085] A mixture of intermediate 9ab (0.2 g, 544.79 μmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (168.57 mg, 610.27 μmol) and DIPEA (0.3 ml, 1.66 mmol) was dissolved in 8 mL of DMSO. The mixture was heated to 100 °C for 2 h and then cooled to room temperature. The reaction was quenched with water and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine and then concentrated by evaporation under vacuum. The crude product was purified by silica gel chromatography using a petroleum ether / dichloromethane gradient to give the desired product as a yellow solid (10a, 46% yield).

[0086]

[0087] 1 H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 8.30 (s, 1H), 7.73 (d, J = 8.2 Hz, 1H), 6.95 (d, J = 8.2 Hz, 2H), 6.86–6.74 (m, 3H), 5.62–5.25 (m, 2H), 5.20 (s, 2H), 4.59–4.48 (m, 2H), 4.26 (d, J = 5.0 Hz, 2H), 4.08 (d, J = 7.0 Hz, 2H), 3.81 (s, 3H), 2.94 (m, 1H), 2.72–2.60 (m, 1H), 2.33–1.96 (m, 5H), 1.57 (d, J = 6.8 Hz, 2H), 1.33 (d, J = 6.8 Hz, 4H), 0.99 (d, J = 6.4 Hz, 4H), 0.90 (d, J = 6.4 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.28, 172.49, 170.56, 167.91, 167.63, 155.12, 149.46, 145.02, 137.84, 134.27, 134.08, 127.07, 125.30, 119.69, 117.85, 115.12, 114.07, 112.09, 105.43, 67.49, 58.92, 55.88, 49.20, 42.16, 38.90, 35.84, 35.69, 32.15, 31.45, 30.84, 29.53, 29.17, 27.85, 27.17, 25.83, 25.37, 23.01, 22.66. HRMS m / z (ESI) calcd for C 34 H 40 N4O7 [M + H] +617.2897; found, 617.2964.

[0088] Referring to the method of synthetic compound 10a, synthesize 10b - 10l.

[0089] Example 11: (E)-N-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)azetidin-3-yl)oxy)-3-methoxybenzyl)-8-methylnon-6-enamide (10b)

[0090] Yellow solid, 25% yield.

[0091]

[0092] 1 H NMR (400 MHz, DMSO-d6) δ 11.07 (s, 1H), 8.24 (t, J = 5.6 Hz, 1H), 7.65–7.56 (m, 1H), 7.17 (d, J = 7.0 Hz, 1H), 6.94–6.84 (m, 2H), 6.75 (q, J = 5.6 Hz, 2H), 5.42–5.23 (m, 2H), 5.06 (dd, J = 12.8, 5.2 Hz, 2H), 4.66–4.57 (m, 2H), 4.24–4.09 (m, 4H), 3.76 (s, 3H), 2.93–2.80 (m, 1H), 2.62–2.53 (m, 1H), 2.26–1.90 (m, 6H), 1.59–1.26 (m, 5H), 0.93 (d, J = 6.6 Hz, 4H), 0.84 (d, J = 6.6 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.26, 172.54, 172.49, 170.46, 167.64, 166.97, 149.43, 147.92, 144.98, 137.84, 135.54, 133.85, 133.73, 127.07, 120.61, 119.74, 113.90, 112.63, 112.15, 111.09, 67.37, 61.14, 55.91, 49.13, 42.18, 38.89, 35.84, 35.69, 32.14, 31.41, 30.84, 29.53, 29.16, 27.85, 27.17, 25.82, 25.36, 23.00, 22.55. HRMS m / z (ESI) calcd for C 34 H 40 N4O7 [M + H] +617.2897; found, 617.2963.

[0093] Example 12: E)-N-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)pyrrolidin-3-yl)oxy)-3-methoxybenzyl)-8-methylnon-6-enamide (10c)

[0094] Yellow solid, 29% yield.

[0095]

[0096] 1 1H NMR (400 MHz, DMSO-d6) δ 11.08 (s, 1H), 8.24 (t, J = 5.6 Hz, 1H), 7.66 (d, J = 8.4 Hz, 1H), 6.97 (d, J = 7.4 Hz, 2H), 6.92–6.84 (m, 2H), 6.77 (d, J = 8.0 Hz, 1H), 5.45–5.10 (m, 2H), 5.10 (s, 2H), 4.20 (d, J = 5.8 Hz, 2H), 3.78–3.67 (m, 4H), 3.57 (t, J = 9.2 Hz, 3H), 2.98–2.82 (m, 1H), 2.66–2.52 (m, 2H), 2.30–1.88 (m, 8H), 1.60–1.42 (m, 2H), 1.36–1.25 (m, 2H), 0.92 (d, J = 6.6 Hz, 4H), 0.84 (d, J = 6.6 Hz, 2H). 13 13C NMR (101 MHz, DMSO-d6) δ 173.30, 172.57, 172.51, 170.61, 168.13, 167.70, 152.29, 150.43, 145.19, 137.83, 134.50, 134.22, 127.06, 125.46, 119.78, 116.82, 116.45, 115.95, 112.27, 106.13, 77.77, 55.91, 54.11, 49.16, 46.53, 42.16, 38.89, 35.84, 35.69, 32.14, 31.46, 30.96, 30.83, 29.52, 29.15, 27.84, 27.16, 25.82, 25.36, 22.99, 22.71. HRMS m / z (ESI) calcd for C 35 H 42 41N4O7 632.3053; found, 631.3118.

[0097] Example 13: (E)-N-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pyrrolidin-3-yl)oxy)-3-methoxybenzyl)-8-methylnon-6-enamide (10d)

[0098] Yellow solid, 37% yield.

[0099]

[0100] 1 1H NMR (400 MHz, DMSO-d6) δ 11.07 (s, 1H), 8.23 (s, 1H), 7.60 (t, J = 7.8 Hz, 1H), 7.16 (d, J = 7.8 Hz, 2H), 6.99–6.84 (m, 2H), 6.75 (d, J = 7.9 Hz, 1H), 5.47–5.07 (m, 2H), 5.09 (d, J = 4.9 Hz, 2H), 4.25–4.04 (m, 3H), 3.81 (dd, J = 17.5, 8.9 Hz, 1H), 3.68 (s, 3H), 3.55 (d, J = 7.3 Hz, 2H), 3.00–2.81 (m, 1H), 2.51 (s, 3H), 2.33–1.81 (m, 8H), 1.51 (dd, J = 14.8, 7.4 Hz, 2H), 1.37–1.23 (m, 3H), 0.92 (d, J = 6.7 Hz, 4H), 0.84 (d, J = 6.6 Hz, 2H). 13 13C NMR (101 MHz, DMSO-d6) δ 173.28, 172.55, 172.49, 170.54, 167.56, 167.52, 167.07, 150.41, 146.38, 145.29, 137.83, 135.46, 134.43, 134.39, 134.11, 127.06, 121.84, 119.76, 116.65, 112.25, 110.78, 77.78, 57.63, 55.87, 49.19, 42.15, 38.89, 35.83, 35.68, 32.13, 31.41, 30.90, 30.83, 29.51, 29.15, 27.84, 27.16, 25.81, 25.36, 22.99, 22.55. HRMS m / z (ESI) calcd for C 35 H 42 N4O7 632.3053; found, 631.3210.

[0101] Example 14: (E)-N-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)oxy)-3-methoxybenzyl)-8-methylnon-6-enamide (10e)

[0102] Yellow solid, 32% yield.

[0103]

[0104] 1 H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.26 (s, 1H), 7.72 (d, J = 8.6 Hz, 1H), 7.40 (s, 1H), 7.32 (d, J = 8.2 Hz, 1H), 7.03 (d, J = 8.2 Hz, 1H), 6.93 (s, 1H), 6.80 (d, J = 7.8 Hz, 1H), 5.52–5.25 (m, 2H), 5.12 (dd, J = 12.8, 5.2 Hz, 1H), 4.55 (s, 1H), 4.24 (d, J = 5.6 Hz, 2H), 3.86 (d, J = 12.8 Hz, 2H), 3.78 (s, 3H), 3.22 (d, J = 5.2 Hz, 2H), 2.99–2.86 (m, 1H), 2.70–2.58 (m, 2H), 2.17 (t, J = 7.2 Hz, 2H), 1.99 (d, J = 6.0 Hz, 4H), 1.72 (d, J = 8.4 Hz, 2H), 1.55 (m, 3H), 1.41–1.29 (m, 3H), 0.97 (d, J = 6.6 Hz, 4H), 0.89 (d, J = 6.6 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.29, 172.47, 170.57, 168.07, 167.42, 155.14, 150.79, 145.29, 137.83, 134.53, 134.05, 127.08, 125.52, 119.77, 118.20, 117.64, 112.39, 108.35, 73.83, 56.03, 49.21, 49.07, 44.96, 42.19, 38.9, 35.68, 32.15, 31.44, 30.84, 30.24, 29.15, 27.86, 27.17, 25.37, 23.01, 22.65. HRMS m / z (ESI) calcd for C 36 H 44 N4O7 [M+H] + 645.3210; found, 645.3276.

[0105] Example 15: (E)-N-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidin-4-yl)oxy)-3-methoxybenzyl)-8-methylnon-6-enamide (10f)

[0106] Yellow solid, 39% yield.

[0107]

[0108] 1 H NMR (400 MHz, DMSO-d6) δ 11.15 (s, 1H), 8.29 (s, 1H), 7.75 (t, J = 7.6 Hz, 1H), 7.47–7.37 (m, 2H), 7.05 (d, J = 8.2 Hz, 1H), 6.95 (s, 1H), 6.81 (d, J = 7.8 Hz, 1H), 5.50–5.29 (m, 2H), 5.16 (dd, J = 12.6, 5.0 Hz, 1H), 4.51 (s, 1H), 4.26 (d, J = 5.2 Hz, 2H), 3.81 (s, 3H), 3.62 (s, 2H), 3.29–3.21 (m, 2H), 2.93 (dd, J = 21.9, 9.2 Hz, 1H), 2.63 (dd, J = 17.8, 10.7 Hz, 2H), 2.28–2.04 (m, 6H), 2.04–1.81 (m, 4H), 1.62–1.53 (m, 2H), 1.41–1.32 (m, 2H), 0.99 (d, J = 6.6 Hz, 4H), 0.90 (d, J = 6.6 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.29, 172.55, 172.49, 170.50, 167.56, 166.79, 150.84, 150.26, 145.42, 137.83, 136.28, 134.13, 134.03, 127.07, 124.45, 119.78, 117.77, 116.98, 115.11, 112.41, 73.77, 56.05, 49.25, 48.57, 42.20, 38.90, 35.84, 35.69, 32.15, 31.43, 31.19, 30.84, 29.53, 29.15, 27.85, 27.18, 25.83, 25.37, 23.00, 22.52. HRMS m / z (ESI) calcd for C 36 H 44 N4O7 [M + H] + 645.3210; found, 645.3278.

[0109] Example 16: (E)-N-(4-((1-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)azetidin-3-yl)methoxy)-3-methoxybenzyl)-8-methylnon-6-enamide (10 g)

[0110] Yellow solid, 20% yield.

[0111]

[0112] 1 H NMR (400 MHz, DMSO-d6) δ 11.08 (s, 1H), 8.23 (s, 1H), 7.65 (d, J = 8.2 Hz, 1H), 6.95 (d, J = 8.2 Hz, 1H), 6.86 (s, 1H), 6.82 (s, 1H), 6.75 (d, J = 8.2 Hz, 1H), 6.68 (d, J = 8.2 Hz, 1H), 5.53–5.11 (m, 2H), 5.06 (dd, J = 9.6, 5.8 Hz, 1H), 4.18 (t, J = 7.2 Hz, 6H), 3.88 (dd, J = 8.0, 5.8 Hz, 2H), 3.72 (s, 3H), 3.25–3.15 (m, 1H), 2.94–2.83 (m, 1H), 2.64–2.52 (m, 2H), 2.28–1.87 (m, 5H), 1.50 (dq, J = 10.8, 7.4 Hz, 2H), 1.30 (dd, J = 10.8, 7.4 Hz, 2H), 0.93 (d, J = 6.6 Hz, 4H), 0.84 (d, J = 6.6 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.29, 172.47, 170.58, 167.98, 167.66, 155.61, 149.58, 147.24, 137.82, 134.27, 133.54, 127.07, 125.26, 119.79, 117.26, 114.62, 114.53, 112.06, 104.90, 71.17, 55.95, 54.46, 49.18, 42.15, 38.89, 35.84, 35.68, 32.14, 31.45, 30.83, 29.52, 29.35, 29.14, 27.85, 27.17, 25.82, 25.37, 23.00, 22.68. HRMS m / z (ESI) calcd for C 35 H 42 N4O7 [M + H] +631.3053; found, 631.3120.

[0113] Example 17: (E)-N-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)azetidin-3-yl)methoxy)-3-methoxybenzyl)-8-methylnon-6-enamide (10h)

[0114] Yellow solid, 22% yield.

[0115]

[0116] 1 1H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.29 (d, J = 5.6 Hz, 1H), 7.63 (dd, J = 8.2, 7.6 Hz, 1H), 7.17 (d, J = 7.0 Hz, 1H), 6.98 (s, 1H), 6.93–6.85 (m, 2H), 6.80 (d, J = 8.2 Hz, 1H), 5.46–5.12 (m, 2H), 5.13 (s, 1H), 4.39 (s, 2H), 4.22 (dd, J = 10.6, 6.8 Hz, 4H), 4.09 (s, 2H), 3.77 (s, 3H), 3.16 (td, J = 10.4, 6.8 Hz, 1H), 2.93 (m, 1H), 2.69–2.57 (m, 2H), 2.17 (s, 6H), 1.56 (dd, J = 10.2, 7.4 Hz, 2H), 1.41–1.30 (m, 2H), 0.98 (d, J = 6.6 Hz, 4H), 0.89 (d, J = 6.6 Hz, 2H). 13 13C NMR (101 MHz, DMSO-d6) δ 173.28, 172.54, 172.48, 170.50, 167.71, 166.98, 149.55, 148.46, 147.26, 137.82, 135.39, 133.74, 133.46, 127.06, 120.35, 119.78, 114.42, 112.12, 112.01, 110.47, 71.26, 56.97, 55.92, 49.10, 42.16, 38.89, 35.84, 35.69, 32.13, 31.42, 30.83, 29.52, 29.14, 27.85, 27.16, 25.82, 25.37, 22.99, 22.58. HRMS m / z (ESI) calcd for C 35 H 42 N4O7 [M+H] +631.3053; found, 631.3119.

[0117] Example 18: (E)-N-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)pyrrolidin-3-yl)methoxy)-3-methoxybenzyl)-8-methylnon-6-enamide (10i)

[0118] Yellow solid, yield 17%.

[0119]

[0120] 1 1H NMR (400 MHz, DMSO-d6) δ 11.07 (s, 1H), 8.23 (d, J = 5.4 Hz, 1H), 7.65 (d, J = 8.4 Hz, 1H), 6.89 (d, J = 7.4 Hz, 5H), 5.53–5.07 (m, 2H), 5.08 (s, 1H), 4.18 (d, J = 5.8 Hz, 2H), 3.98 (p, J = 9.2 Hz, 2H), 3.74 (s, 3H), 3.66–3.39 (m, 4H), 2.95–2.78 (m, 2H), 2.63–2.53 (m, 2H), 2.12 (s, 8H), 1.56–1.45 (m, 2H), 1.36–1.25 (m, 2H), 0.93 (d, J = 6.6 Hz, 4H), 0.84 (d, J = 6.6 Hz, 2H). 13 13C NMR (101 MHz, DMSO-d6) δ 178.13, 177.11, 177.02, 174.23, 171.31, 170.15, 153.34, 151.34, 150.45, 141.51, 140.22, 139.79, 137.11, 132.12, 128.55, 124.54, 118.93, 116.67, 115.14, 114.24, 75.47, 60.75, 59.44, 55.98, 53.72, 46.39, 44.34, 43.73, 41.19, 40.33, 37.78, 36.12, 35.18, 34.88, 33.89, 33.29, 32.71, 31.67, 30.97, 29.22, 27.56, 26.34. HRMS m / z (ESI) calcd for C 36 H 44 N4O7 [M+H] + 644.3210; found, 645.3280.

[0121] Example 19: (E)-N-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pyrrolidin-3-yl)methoxy)-3-methoxybenzyl)-8-methylnon-6-enamide (10j, also known as TPD)

[0122] Yellow solid, 28% yield.

[0123]

[0124] 1 H NMR (400 MHz, DMSO-d6) δ 11.06 (s, 1H), 8.22 (t, J = 5.6 Hz, 1H), 7.58 (dd, J = 8.4, 7.2 Hz, 1H), 7.13 (t, J = 7.2 Hz, 2H), 6.93 (d, J = 8.2 Hz, 1H), 6.86 (s, 1H), 6.74 (d, J = 8.2 Hz, 1H), 5.46–5.12 (m, 2H), 5.07 (dd, J = 12.8, 5.4 Hz, 1H), 4.18 (d, J = 5.8 Hz, 2H), 4.04–3.93 (m, 2H), 3.73 (s, 4H), 3.63 (t, J = 6.6 Hz, 2H), 3.58–3.50 (m, 1H), 2.88 (s, 1H), 2.73 (m, 1H), 2.64–2.53 (m, 2H), 2.26–1.78 (m, 8H), 1.57–1.44 (m, 2H), 1.37–1.25 (m, 2H), 0.93 (d, J = 6.6 Hz, 4H), 0.84 (d, J = 6.6 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 178.04, 177.25, 177.20, 175.29, 172.34, 171.75, 154.23, 152.11, 151.17, 142.57, 140.07, 139.17, 138.03, 131.82, 126.58, 124.54, 118.93, 116.83, 116.72, 115.34, 75.47, 60.75, 59.54, 55.56, 53.94, 46.90, 43.64, 43.23, 40.59, 40.43, 36.88, 36.18, 35.58, 34.27, 33.89, 33.08, 32.60, 31.91, 30.57, 30.12, 27.74, 27.37. HRMS m / z (ESI) calcd for C 36 H 44 N4O7 [M+H] +6454.3210; found, 645.3276.

[0125] Example 20: (E)-N-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methoxy)-3-methoxybenzyl)-8-methylnon-6-enamide (10k)

[0126] Yellow solid, 19% yield.

[0127]

[0128] 1 1H NMR (400 MHz, DMSO-d6) δ 11.14 (s, 1H), 8.27 (s, 1H), 7.75 (t, J = 7.8 Hz, 1H), 7.43–7.28 (m, 2H), 7.03–6.88 (m, 2H), 6.80 (d, J = 7.8 Hz, 1H), 5.60–5.24 (m, 2H), 5.13 (m, 1H), 4.20 (dd, J = 6.8, 9.2 Hz, 4H), 3.96–3.82 (m, 2H), 3.82 (s, 3H), 3.07 (t, J = 9.2 Hz, 2H), 3.00–2.88 (m, 1H), 2.63 (dd, J = 6.8, 7.6 Hz, 2H), 2.35–1.85 (m, 9H), 1.57 (t, J = 7.6 Hz, 2H), 1.49–1.32 (m, 4H), 0.99 (d, J = 6.6 Hz, 4H), 0.90 (d, J = 6.6 Hz, 2H). 13 13C NMR (101 MHz, DMSO-d6) δ 173.29, 172.49, 172.43, 170.58, 168.11, 167.44, 155.44, 149.42, 147.50, 137.82, 134.52, 132.91, 127.07, 125.48, 119.77, 118.13, 117.92, 113.74, 112.01, 108.28, 73.15, 55.98, 49.20, 47.48, 42.16, 38.89, 35.77, 35.69, 32.14, 31.45, 30.84, 29.53, 29.14, 28.21, 27.85, 27.17, 25.83, 25.37, 23.00, 22.66. HRMS m / z (ESI) calcd for C 37 H 46 N4O7 [M+H] + 659.3366; found, 659.3434.

[0129] Example 21: (E)-N-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidin-4-yl)methoxy)-3-methoxybenzyl)-8-methylnon-6-enamide (10l)

[0130] Yellow solid, 25% yield.

[0131]

[0132] 1 H NMR (400 MHz, DMSO-d6) δ 11.09 (s, 1H), 8.22 (s, 1H), 7.69 (t, J = 8.0 Hz, 1H), 7.39–7.30 (m, 2H), 6.95–6.83 (m, 2H), 6.75 (d, J = 8.0 Hz, 1H), 5.46–5.22 (m, 2H), 5.10 (dd, J = 9.8, 5.6 Hz, 1H), 4.19 (d, J = 5.6 Hz, 2H), 3.86 (d, J = 5.8 Hz, 2H), 3.75 (s, 5H), 2.98–2.81 (m, 3H), 2.64–2.54 (m, 2H), 2.07 (m, 9H), 1.51 (d, J = 7.8 Hz, 4H), 1.35–1.26 (m, 2H), 0.93 (d, J = 6.6 Hz, 4H), 0.85 (d, J = 6.6 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 173.29, 172.44, 170.51, 167.58, 166.78, 150.64, 149.47, 147.57, 137.82, 136.23, 134.14, 132.90, 127.08, 124.45, 119.80, 116.89, 114.95, 113.78, 112.06, 73.39, 56.02, 51.20, 49.24, 42.17, 35.69, 35.45, 32.14, 31.43, 30.84, 29.53, 29.14, 29.07, 27.86, 25.38, 23.01, 22.54. HRMS m / z (ESI) calcd for C 37 H 46 N4O7 [M + H] + 659.3366; found, 659.3437.

[0133] Example 22: (E)-N-(4-((1-(2-(1-ethyl-2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pyrrolidin-3-yl)methoxy)-3-methoxybenzyl)-8-methylnon-6-enamide (10m)

[0134]

[0135] To a solution of 10j (0.2 g, 319.20 μmol) in THF (10 mL) was slowly added NaH (60%, 20 mg), and the mixture was stirred at 0 °C for 10 minutes. CH3CH2I (72.57 mg, 465.28 μmol) was added dropwise with a syringe and the reaction was carried out at 25 °C for 2 h. The reaction residue was extracted with ethyl acetate and aqueous HCl solution (1 N, 20 mL). The organic layer was washed with water, dried over Na2SO4, and evaporated in vacuo. The crude product was purified by column chromatography to give 10m (65 mg, 32%) as a yellow solid.

[0136] 1 1H NMR (400 MHz, DMSO-d6) δ 11.65–11.60 (m, 1H), 8.28–8.15 (m, 1H), 7.58 (dd, J = 8.4, 7.2 Hz, 1H), 7.19–7.10 (m, 2H), 6.92 (d, J = 8.2 Hz, 1H), 6.85 (s, 1H), 6.73 (d, J = 8.4 Hz, 1H), 5.44–5.12 (m, 2H), 5.21–5.03 (m, 1H), 4.17 (d, J = 5.8 Hz, 2H), 3.96 (d, J = 2.8 Hz, 2H), 3.79–3.49 (m, 9H), 2.96 (d, J = 2.8 Hz, 1H), 2.79–2.67 (m, 2H), 2.51 (s, 2H), 2.24–1.80 (m, 7H), 1.54–1.46 (m, 2H), 1.36–1.26 (m, 2H), 1.01 (t, J = 7.0 Hz, 3H), 0.92 (d, J = 6.6 Hz, 4H), 0.83 (d, J = 6.6 Hz, 2H). 1313C NMR (101 MHz, DMSO-d6) δ 172.47, 172.41, 171.85, 169.80, 167.58, 167.02, 149.49, 147.36, 146.44, 137.81, 135.32, 134.41, 133.32, 127.07, 121.83, 119.77, 114.21, 112.09, 110.60, 70.73, 55.98, 54.81, 50.76, 49.78, 42.14, 38.89, 38.52, 38.44, 35.84, 35.68, 35.11, 32.14, 31.65, 30.84, 29.52, 29.14, 28.26, 27.85, 27.17, 25.82, 25.37, 22.99, 21.90, 13.37.

[0137] HRMS m / z (ESI) calcd for C 38 H 48 N4O7 673.3523; found, 673.3566.

[0138] The beneficial effects of the present invention are demonstrated by the following experimental examples.

[0139] Experimental Example 1, Degradation Activity against TRPV1

[0140] 1. Experimental Method

[0141] Cells were lysed with DDM (n-dodecyl-D-maltoside) lysis buffer containing protease inhibitors. The Minute Total Protein Extraction Kit (AT-022, Invent Biotechnologies, Inc.) was used to homogenize tissues. In the assay, cells were lysed in lysis buffer (0.025 M Tris, 0.15 M NaCl, 0.001 M EDTA, 1% NP-40, 5% glycerol, pH 7.4). The whole lysate was incubated with 10 μg of antibody or normal IgG. The bound proteins were eluted with 0.1 M glycine (pH 2.5) and then neutralized with 1 M Tris buffer to prevent interference with the heavy chain (about 55 kDa). The concentration was determined using G250. Proteins were separated by SDS-PAGE gel and transferred to a polyvinylidene difluoride membrane (Millipore). The membrane was blocked with 5% skim milk for 1 hour and incubated with specific primary antibody overnight at 4°C. The next day, the membrane was incubated with secondary antibody for 1 hour at 37°C. TM

[0142] 2. Experimental Results

[0143] The results are as Figure 1 and​Figure 2 As shown, it can be seen that the compounds of the present invention can effectively degrade TRPV1. Among them, the half degradation concentration (DC 50 ) of compound 5a is 9.9 μM, and the DC 50 of compound 10j (TPD) is 5.9 μM.

[0144] Experimental Example 2, Analgesic Activity

[0145] 1. Experimental Method

[0146] (1) Chronic Inflammatory Pain Model Induced by Complete Freund's Adjuvant (CFA)

[0147] In this experiment, C57 mice were randomly divided into 6 groups, namely the solvent group, different dose groups of TPD (25 mg / kg, 50 mg / kg, 100 mg / kg, 200 mg), and the positive control indomethacin (indo) group. The administration dose of indomethacin was 10 mg / kg, and the solvent group was given an equal volume of PEG200. Oral administration was performed, and the administration volume was 0.2 mL. One day before modeling, the baseline of mechanical stimulation response threshold and thermal stimulation response threshold of the mice was measured. After measuring the baseline, 20 μL of CFA solution was subcutaneously injected into the plantar surface of the left hind paw of the mice. On the first day after modeling, the pain thresholds before administration (recorded as 0 min) and at 0, 20, 40, 60, 90, 120, 180, 240, 300, 360, 420, 480, 600 min after administration were measured. The results are as Figure 3 shown in a.

[0148] (2) Acute Inflammatory Pain Model Induced by Formalin

[0149] C57 mice were randomly divided into 3 groups, namely the solvent group, the TPD group, and the indomethacin group, with 8 mice in each group. The administration dose of the candidate compound was 100 mg / kg. The administration dose of indomethacin was 10 mg / kg, and the solvent group was given an equal volume of PEG200. The administration method was oral, and the administration volume was 0.2 mL. Analytical pure formaldehyde was diluted with physiological saline to the required concentration for the experiment (20 μL or 50 μL of 5% formalin diluted with saline). The activities of the mice were restricted using a transparent and clean resin box, which was placed on a metal rack with mesh holes. An animal behavior observation mirror was placed under the metal rack to observe and record the reactions of the mice. All mice needed to adapt to the environment for at least 30 minutes before the experiment started. After the experiment ended, the mice in each group were administered drugs according to the aforementioned grouping plan. 20 minutes after drug administration, 20 μL of 5% formalin solution was subcutaneously injected into the plantar surface of the left hind paw of the mice. Within 30 minutes after the formalin solution was injected into the mice, the time of the mice shaking and licking their left hind paws was recorded by video. After the experiment ended, a stopwatch was used to count the reaction time of the mice. During the analysis, the 0 - 5 minutes after the formalin solution was given was recorded as Phase I, and the 15 - 30 minutes after the formalin solution was given was recorded as Phase II. The experiment was conducted using a blind method, and the results were uniformly judged by trained observers without knowing the specific drug administration grouping. The results are as Figure 3 shown in

[0150] (3) Neuropathic pain model induced by chronic constriction injury of the sciatic nerve (CCI)

[0151] In this experiment, C57 mice were randomly divided into 3 groups, namely the solvent group, the TPD group, and the positive control pregabalin (PGB) group. The administration dose of TPD was 100 mg / kg, and the administration dose of pregabalin was 30 mg / kg. The solvent group was given an equal volume of intralipid. The administration method was intraperitoneal injection, and the administration volume was 0.2 mL. One day before modeling, the baseline of the mechanical stimulation response threshold and the cold stimulation response threshold of the mice were measured. After measuring the baseline, a modeling operation was performed. With the assistance of a small animal anesthesia machine, the mice were anesthetized with isoflurane (induction concentration: 3-4%, maintenance concentration: 1-1.5%). The mice were placed in the left lateral position, and the surface projection position of the sciatic nerve was determined between the greater trochanter of the femur and the ischial tuberosity. The skin was prepared, and the surgical site was disinfected with iodophor. An incision was made along the course of the sciatic nerve, and the skin and fascia were bluntly separated to expose the thick white nerve, which was the main trunk of the sciatic nerve. The sciatic nerve was separated with a glass microprobe and ligated with chromic catgut. The tightness should be such that the hind paw of the mouse showed a slight tremor. Two surgical knots were tied in sequence, with a distance of 1 mm between the two knots. The sciatic nerve was replaced, and the fascia and skin were sutured with 4-0 silk thread in sequence. After disinfection again, the mice were placed on a small animal warming blanket and waited for anesthesia to wear off. After the mice woke up, they were put back into the breeding cage. One week after the operation, the incision site and general condition of the mice were closely observed, and penicillin was given to prevent infection. On the 7th day after modeling, the pain thresholds of each group of mice were measured before administration (recorded as 0 min) and at 0, 20, 40, 60, 90, 120, 180, 240, 300, 360, 420, 480, 600 min after administration, and every half hour thereafter. The results are as Figure 3 shown in

[0152] (4) Diabetic Neuropathic Pain Model Induced by Streptozocin (STZ)

[0153] In this experiment, C57 mice were randomly divided into 3 groups, namely the solvent group, the TPD group, and the positive control pregabalin group. The administration dose of TPD was 100 mg / kg, the administration dose of pregabalin was 30 mg / kg, and the solvent group was given an equal volume of fat emulsion. The administration route was oral, and the administration volume was 0.2 mL. One day before modeling, the baseline of the mechanical stimulation response threshold and the cold stimulation response threshold of the mice were measured. After measuring the baseline, the mice were fasted overnight. On the morning of the second day after fasting, the fasting blood glucose of the mice was measured, and the mice with fasting blood glucose levels between 3.7 and 6.9 mmol / L were included in the study. The STZ powder was dissolved in sodium citrate buffer to prepare a 1 g / mL solution, and the pH value was adjusted to 4.5. The mice were intraperitoneally injected with 200 mg / kg of STZ to induce diabetes. One week after STZ induction, the mice were fasted overnight again. On the morning of the second day after fasting, the fasting blood glucose of the mice was measured again, and the mice with fasting blood glucose levels higher than 16.7 mmol / L were recorded as successfully diabetes-induced mice. The pain thresholds of the successfully diabetes-induced mice in each group were measured before administration (recorded as 0 min) and at 0, 20, 40, 60, 90, 120, 180, 240, 300, 360, 420, 480, and 600 min after administration. The results are as Figure 3 shown in

[0154] (5) Osteoarthritis pain model induced by monoiodoacetate (MIA)

[0155] In this experiment, C57 mice were randomly divided into 3 groups, namely the solvent group, the TPD group, and the positive control celecoxib (Cele) group. The administration dose of TPD was 100 mg / kg, and the administration dose of celecoxib was 30 mg / kg. The solvent group was given an equal volume of PEG200. Oral administration was performed, and the administration volume was 0.2 mL. One day before modeling, the baseline mechanical stimulation response threshold and thermal stimulation response threshold of the mice were measured. After measuring the baseline, the modeling surgery was performed. With the assistance of a small animal anesthesia machine, the mice were anesthetized with isoflurane (induction concentration: 3 - 4%, maintenance concentration: 1 - 1.5%). The hair on the knees of the mice was shaved off to expose the knee joint field of view, and it was disinfected with iodophor. The knee joint of the mouse was fixed in the flexed position, and the patellar tendon was located. The needle of a micro syringe was inserted into the space below the patella and advanced perpendicularly to the tibia. There was no resistance during insertion. After penetrating into the joint cavity, a sense of falling through was felt. At this time, 1 mg / 10 μl of sodium monoiodoacetate was injected. After withdrawing the needle, the knee joint of the mouse was massaged to make the drug evenly distributed, and it was disinfected again. After the mouse woke up, it was put back into the breeding cage. The knee joints of the mice were observed for one week after that, and anti-infection treatment was given if necessary. On the 7th day after modeling, the pain thresholds before administration (recorded as 0 min) and at 0, 20, 40, 60, 90, 120, 180, 240, 300, 360, 420, 480, 600 min after administration were measured. The time point when the analgesic effect reached the peak was selected for threshold determination after administration on days 14, 21, and 28. The results are as Figure 3 shown in

[0156] In the above experimental method, a radiant heat source (model 37370; Ugo - Basile) was used to evaluate the sensitivity of the hind paw to noxious heat stimulation. The stimulation intensity was set to produce an initial latency of approximately 10 seconds, and the cut - off time was set to 20 seconds to avoid accidental damage. The mice were allowed to acclimate in a glass - bottomed plexiglass chamber for 3 days, and the paw - withdrawal latency of each mouse was measured at a 5 - minute stimulation interval. To determine the withdrawal latency, the results of three trials were averaged.

[0157] 2. Experimental Results

[0158] The results are as Figure 3 shown. It can be seen that in the chronic inflammatory pain model induced by CFA, the acute inflammatory pain model induced by formalin, the neuropathic pain model induced by CCI, the neuropathic pain model induced by STZ, and the osteoarthritis pain model induced by MIA, 100 mg / kg of TPD has obvious advantages in analgesic effect compared with the positive compound.

[0159] Experimental Example 3. Effect on Body Temperature

[0160] 1. Experimental Method

[0161] (1) Experiment on the effect of TPD on the change of mouse body temperature

[0162] In this experiment, C57 mice were randomly divided into 5 groups, namely the solvent group, and the TPD (25 mg / kg, 50 mg / kg, 100 mg / kg, 200 mg / kg) groups. The oral administration method was used, and the administration volume was 0.2 mL. The test was conducted by inserting the thermistor probe of a digital thermometer (TH-212) 1 cm deep into the rectum of the animal lubricated with vaseline. Rectal temperature was recorded before the administration of the test dose (0 minute) and at 0, 20, 40, 60, 90, 120, 180, 240, 300, 360, 420, 480, and 600 minutes after the administration of the test dose. The ambient temperature was 23.4 ± 4 °C. The results are as Figure 4 shown in

[0163] (2) Experiment on the perturbation of TPD on the body temperature of mice by the TRPV1 agonist capsaicin (Cap) and the TRPV1 inhibitor AMG517

[0164] In this experiment, normal C57 mice were randomly divided into 5 groups, namely the solvent group, the group that was orally administered 100 mg / kg of TPD in advance and then subcutaneously administered 1 mg / kg of capsaicin and orally administered 1 mg / kg of AMG517, the group that was only subcutaneously administered 1 mg / kg of capsaicin alone, and the experimental group that was orally administered AMG517. The administration volume was 0.2 mL. There was also a group of TRPV1 - / - (C57 group with TRPV1 completely knocked out), a total of 6 groups. Rectal temperature was recorded before the administration of the test dose (50 minutes, 0 minute) and at 0, 20, 40, 60, 90, 120, 180, 240, 300, 360, 420, 480, and 600 minutes after the administration of the test dose. The ambient temperature was 23.4 ± 4 °C. The results are as Figure 4 shown in

[0165] 2. Experimental results

[0166] The results are as Figure 4 shown. It can be seen that after the administration of different doses of TPD, the body temperature is not affected, and at the same time, it can weaken the tendency of the TRPV1 agonist Cap to lower the body temperature and the TRPV1 inhibitor AMG517 to increase the body temperature, indicating that the combined use of TPD with the TRPV1 agonist Cap or the TRPV1 inhibitor AMG517 can effectively reduce the side effects caused by the TRPV1 agonist Cap or the TRPV1 inhibitor AMG517.

[0167] In summary, the present invention provides an isoindoline-pyrrolidine derivative represented by Formula I. The compounds of the present invention exhibit high selectivity and excellent degradation effect on TRPV1 both in vitro and in vivo. In different animal pain models, the compounds of the present invention all exhibit significant analgesic activity. Notably, the compounds of the present invention do not exhibit the adverse effects on body temperature usually associated with TRPV1 agonists or inhibitors, indicating that the compounds of the present invention have higher safety. The compounds of the present invention can effectively reduce the side effects caused by TRPV1 agonists or inhibitors when used in combination with TRPV1 agonists or inhibitors. The compounds of the present invention are currently the first PROTAC degrader targeting ion channels, highlighting the great potential of PROTAC technology in the field of ion channel modulation in the future.

Claims

1. A compound, its stereoisomer or its salt, characterized in that, The structure of the said compound is shown in Formula I: Among them, R1 is selected from C 1-6 alkyl; R2 is selected from C 1-6 alkyl; R3 is selected from C 1-6 alkyl; m is an integer selected from 0 to 5; L is selected from C 1-12 alkylene, O, S, O-C 1-12 alkylene, S-C 1-12 alkylene, 3-6 membered saturated heterocyclic group, 3-6 membered saturated cycloalkyl group; Y is selected from O, S or none.

2. The compound, stereoisomer or salt thereof according to claim 1, characterized in that, The structure of the said compound is shown in Formula II: Among them, L is selected from C 1-12 alkylene, O, S, O-C 1-12 alkylene, S-C 1-12 alkylene, 3- to 6-membered saturated heterocyclic group; Y is selected from O, S or none.

3. The compound, stereoisomer or salt thereof according to claim 2, characterized in that, The structure of the said compound is shown in Formula III: wherein, n is an integer selected from 1 to 9.

4. The compound, its stereoisomer or its salt according to claim 2, characterized in that, The structure of the said compound is shown in Formula III or Formula IV: wherein, X is none or methylene; a is 0 or 1; b is 0 or 1.

5. The compound, its stereoisomer or its salt according to claim 1, characterized in that, The said compound is selected from:

6. A pharmaceutical composition, characterized in that, The said pharmaceutical composition is a preparation prepared with the compound, its stereoisomer or its salt according to any one of claims 1-5 as the active ingredient, plus a pharmaceutically acceptable auxiliary ingredient.

7. Use of the compound, its stereoisomer or its salt according to any one of claims 1-5 in the preparation of a TRPV1 degrader.

8. The use according to claim 7, wherein, The said TRPV1 degrader is a drug for preventing and / or treating asthma, chronic obstructive pulmonary disease, idiopathic pulmonary fibrosis, chronic idiopathic cough, pruritus, esophageal cancer, cervical squamous cell carcinoma, diabetes; preferably, the said diabetes is type 2 diabetes.

9. Use of the compound, its stereoisomer or its salt according to any one of claims 1-5 in the preparation of an analgesic drug; preferably, the said analgesic drug is a drug for treating chronic inflammatory pain, acute inflammatory pain, neuropathic pain or osteoarthritis pain.

10. Use of the compound, its stereoisomer or its salt according to any one of claims 1-5 in combination with a TRPV1 agonist or a TRPV1 inhibitor in the preparation of a drug for reducing the side effects caused by the TRPV1 agonist or the TRPV1 inhibitor; preferably, the said TRPV1 agonist is capsaicin, the said TRPV1 inhibitor is AMG517, and the said side effect is body temperature increase.

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