Substituted aminophenyl furanopyridine compound and preparation method, pharmaceutical composition and application thereof

Abstract

The present invention belongs to the field of chemical medicine, and discloses a substituted aminophenyl furanopyridine compound and a preparation method, a pharmaceutical composition and an application thereof. The present invention obtains a novel substituted aminophenyl furanopyridine compound, as shown in Formula I. The preparation process is simple and easy, and the prepared compound has an efficient effect of inhibiting the deubiquitinating enzyme USP51, and has high selectivity, good safety, high solubility and high bioavailability. It can be used in the preparation of deubiquitinating enzyme USP51 inhibitors, and can be used to form a pharmaceutical composition, which has a certain inhibitory effect on the proliferation of various tumor cells and is suitable for the development of drugs for treating various cancers. Formula I.

Description

Technical Field

[0001] The invention belongs to the field of chemical medicines, and specifically relates to a substituted aminophenyl furanopyridine compound and a preparation method, a pharmaceutical composition and application thereof. Background Art

[0002] Malignant tumors are one of the major diseases that threaten human life. At present, the main treatments for malignant tumors are surgery, radiotherapy, and chemotherapy. Traditional anti-tumor drugs are hindered from further clinical application due to their toxic side effects. A large number of studies have shown that deubiquitinases play an extremely important physiological function by regulating protein interactions, localization, and enzyme activity, thereby affecting cellular processes, including transcription, DNA damage signals and DNA repair, cell cycle progression, oxidative stress, apoptosis, etc. It is closely related to the occurrence and development of tumors. Deubiquitinase inhibitors have also been found to play a significant anti-tumor effect in preclinical studies of various tumors, and have become a hot spot in the research and development of new anti-tumor drugs.

[0003] Epithelial-mesenchymal transition (EMT) and its reversal process and mesenchymal-epithelial transition (MET) play a key role in tumor metastasis. During the EMT process, epithelial cells lose the adhesion junctions and tight junctions that maintain close contact with neighboring cells. Activation of EMT by multiple transcription factors can induce the migration, invasion and metastasis of tumor cells. ZEB1, as a transcription factor that controls EMT induction, can promote tumor invasion, metastasis and therapeutic resistance. In many types of cancer, abnormal expression of ZEB1 is associated with invasive behavior, high tumor index, drug resistance, high metabolic plasticity and metastasis. For example, in breast cancer patients, high expression of ZEB1 was found in tumor cells of triple-negative / basal-like breast cancer.

[0004] USP51 is a deubiquitinase for ZEB1 (Zinc Finger E-Box BindingHomeobox 1), and targeting USP51 may also serve as an alternative approach to targeting the pro-oncogenic transcription factor ZEB1. Screening of a human deubiquitinase library revealed that USP51 is a ubiquitinase that can bind, deubiquitinate, and stabilize ZEB1. Loss of USP51 in mesenchymal-like breast cancer cells led to downregulation of ZEB1 protein and mesenchymal markers, upregulation of cadherin-E, and inhibition of cell invasion. Conversely, overexpression of USP51 in epithelial cells led to upregulation of ZEB1 and mesenchymal cell markers. In addition, USP51 was able to regulate the expression of ZEB1 target genes. Importantly, USP51 was overexpressed in breast cancer patients and associated with poor survival.

[0005] At present, the development of USP family inhibitors is mainly aimed at hot targets such as USP7, USP1, and USP9. As a relatively new deubiquitinase, USP51 has few studies and reports on its protease itself and its inhibitors. In addition, there are still many problems with the currently developed selective inhibitors of deubiquitinases: ① The relevant research on deubiquitinases is concentrated on USP family members, especially USP7, but most of the inhibitors reported on them show weak inhibitory activity, or contain undesirable chemical characteristics, or have poor selectivity for the currently known deubiquitinase family. ② At present, people's understanding of the protein structure and functional substrates of deubiquitinases is insufficient and unbalanced, and there is still a lack of exploration and cognition of the intrinsic mechanism related to deubiquitinases and tumorigenesis. This also leads to the defects of poor specificity of most current small molecule inhibitors of deubiquitinases. In addition, the original inhibitor screening methods and technologies also have defects such as poor compatibility with the actual physiological environment and easy false positives. Therefore, it is of great scientific significance to study and design and synthesize small molecule inhibitors targeting USP51 with novel structure, high inhibitory activity, high safety, and high bioavailability. Summary of the invention

[0006] The purpose of the present invention is to solve the deficiencies of the prior art and provide substituted aminophenyl furanopyridine compounds and preparation methods, pharmaceutical compositions and applications thereof, specifically using the following technical solutions:

[0007] According to a first aspect of the present invention, there is provided a substituted aminophenyl furanopyridine compound, the structure of which is shown in Formula I:

[0008] Formula I;

[0009] Wherein, NR2R3 is selected from any one of a 3-9 membered saturated heterocyclic amino group and a dialkylamino group; the heterocyclic amino group contains at least one N atom and an O atom as ring atoms and is connected to the rest of the molecule through the amino part; the dialkylamino group is selected from any one of a dimethylamino group, a diethylamino group and a dipropylamino group; and n is an integer of 0-4.

[0010] Preferably, the substituted aminophenyl furanopyridine compound is one of Formula 1 to Formula 8, and the specific structural formula is as follows:

[0011] .

[0012] Preferably, the structure of the substituted aminophenyl furanopyridine compound is as shown in Formula II:

[0013] Formula II.

[0014] The substituted aminophenyl furanopyridine compound shown in formula II has the effect of highly selectively inhibiting USP51 enzyme activity and cardiotoxicity IC 50 The concentration of the drug is 10.39 μM, showing high safety; and it exhibits high clearance efficiency in multiple species, with low risk of metabolic clearance in vivo and high bioavailability.

[0015] According to the second aspect of the present invention, a preparation route of the above-mentioned substituted aminophenyl furanopyridine compound is also provided:

[0016] .

[0017] The specific steps are as follows: first, pyrrolopyrimidine A1 and 4-iodoanisole A2 undergo an aromatic CN coupling reaction under the action of cuprous iodide to obtain intermediate A, then 4-alkynylaniline B1 and 3-iodopyridone B2 undergo a tandem coupling reaction to obtain pyridofuran B3, which is then coupled with intermediate A to obtain intermediate B, and B undergoes a substitution reaction with different amino halides under the action of a strong base to obtain a series of target molecules.

[0018] According to the third aspect of the present invention, there is also provided the use of the above-mentioned substituted aminophenyl furanopyridine compound or a pharmaceutically acceptable salt thereof in the preparation of a deubiquitinating enzyme USP51 inhibitor. The compound or a pharmaceutically acceptable salt thereof has a highly effective inhibitory effect on the deubiquitinating enzyme USP51, has high selectivity, good safety, high solubility and high bioavailability.

[0019] Preferably, inhibitors of the deubiquitinase USP51 can be used to prepare drugs for treating diseases related to abnormalities of the deubiquitinase USP51. More preferably, the related diseases include breast cancer, ovarian cancer, lymphoma, lung cancer, colorectal cancer, prostate cancer, bladder cancer, liver cancer, taste cancer, neuroblastoma, and leukemia. The above-mentioned cancers include: gynecological cancers, such as ovarian cancer, cervical cancer, vaginal cancer, genital cancer, uterine / endometrial cancer, gestational trophoblastic tumor, fallopian tube cancer, uterine sarcoma; endocrine cancers, such as adrenal cortical carcinoma, pituitary cancer, pancreatic cancer, thyroid cancer, parathyroid cancer, thymic cancer, multiple endocrine tumors; bone cancers, such as osteosarcoma, Ewing's sarcoma, chondrosarcoma, etc.; lung cancers, such as small cell lung cancer and non-small cell lung cancer; brain and CN S tumors, such as neuroblastoma, acoustic neuroma, glioma and other brain tumors, spinal cord tumors, breast cancer, colorectal cancer, advanced colorectal adenocarcinoma; gastrointestinal cancers, such as liver cancer, extrahepatic bile duct cancer, gastrointestinal carcinoid tumors, gallbladder cancer, stomach cancer, esophageal cancer, small intestine cancer; genitourinary cancers, such as penile cancer, testicular cancer, prostate cancer; head and neck tumors, such as nose cancer, paranasal sinus cancer, nasopharyngeal cancer, oral cancer, lip cancer, salivary gland cancer, laryngeal cancer, lower Pharyngeal cancer, pharyngeal cancer; blood cancers, such as acute myeloid leukemia, acute lymphocytic leukemia, childhood leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, acute promyelocytic leukemia, plasma cell leukemia; bone marrow cancer blood diseases, such as myelodysplastic syndrome, myeloproliferative disorders, Fanwort anemia, aplastic anemia, idiopathic macroglobulinemia; lymphomas, such as Hodgkin's disease, non-Hodgkin's Lymphoma, peripheral T-cell lymphoma, cutaneous T-cell lymphoma, AIDS-related lymphoma; eye cancers, including retinoblastoma, uveal melanoma; skin cancers, such as melanoma, non-melanoma skin cancer, Merkel cell carcinoma; soft tissue sarcomas, such as Kaposi's sarcoma, childhood soft tissue sarcoma, adult soft tissue sarcoma, urinary system cancers, such as renal cancer Wilms' tumor, bladder cancer, urethral cancer and metastatic cell carcinoma. More preferably, breast cancer, colorectal cancer, ovarian cancer, prostate cancer, lung cancer.

[0020] According to the fourth aspect of the present invention, a pharmaceutical composition is also provided, comprising the above-mentioned substituted aminophenyl furanopyridine compound or a pharmaceutically acceptable salt thereof. The pharmaceutical composition can be used to prepare drugs for treating breast cancer, ovarian cancer, lymphoma, lung cancer, colorectal cancer, prostate cancer, bladder cancer, liver cancer, palate cancer, neuroblastoma, and leukemia.

[0021] Preferably, the pharmaceutical composition comprises at least one of the excipients. More preferably, the excipient is at least one of gum arabic, syrup, lanolin, and starch. The excipient is stable in nature, has no incompatibility with the main drug, does not produce side effects, does not affect the efficacy, is not easy to deform, crack, mildew, or be worm-eaten at room temperature, is harmless to the human body, has no physiological effect, does not produce chemical or physical effects with the main drug, and does not affect the content determination of the main drug.

[0022] The beneficial effects of the present invention are as follows: the present invention obtains a novel substituted aminophenyl furanopyridine compound, the preparation process is simple and easy, the prepared compound has a highly effective inhibitory effect on the deubiquitinating enzyme USP51, has high selectivity, good safety, high solubility and high bioavailability. It can be used in the preparation of deubiquitinating enzyme USP51 inhibitors, and can be composed of pharmaceutical compositions, which have a certain inhibitory effect on the proliferation of various tumor cells and are suitable for the development of drugs for treating various cancers. DETAILED DESCRIPTION

[0023] The following will be combined with the embodiments to clearly and completely describe the concept and technical effects of the present invention, so as to fully understand the purpose, scheme and effect of the present invention. It should be noted that the embodiments and features in the embodiments of this application can be combined with each other without conflict.

[0024] Example 1

[0025] A substituted aminophenyl furanopyridine compound (denoted as compound 1), compound A, compound A1, compound A2, compound B1, compound B2, compound B3, and compound B, whose structures are shown below:

[0026] Compound 1 Compound A Compound A1

[0027] Compound A2 Compound B1 Compound B3 Compound B

[0028] The specific preparation method is:

[0029] 1) Preparation of compound A: Compound A1 (300 mg, 1.03 mmol) was dissolved in 1,4-dioxane (5 mL), and compound A2 (294 mg, 0.49 mmol) was added, and cuprous iodide (18 mg, 0.08 mmol), trans-1,2-cyclohexanediamine (120 mg, 0.70 mmol) and potassium phosphate (500 mg, 3.01 mmol) were added. The mixture was placed in an oil bath, heated to 110 °C, and refluxed for 3 hours. The mixture was filtered with diatomaceous earth, and the liquid in the bottle was concentrated with a rotary evaporator. The crude product was purified with silica gel column chromatography (petroleum ether: ethyl acetate = 5:1 to 3:1) to obtain compound A (white solid, 310 mg, yield: 70%).

[0030] Compound A was tested, and the test results were as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.69 (s, 1H), 7.67 (d, J = 14.3 Hz, 2H), 7.61-7.54 (m, 2H), 7.43 (s, 1H),7.16 (s,1H), 7.10 (d, J = 11.5 Hz, 2H), 7.01 (s, 2H), 6.52 (s, 1H), 3.85 (d, J = 14.3 Hz,3H), 2.31 (s, 3H). 13 C NMR (100 MHz,CDCl3) δ 158.1, 156.4, 151.8, 151.0, 137.9,131.0, 130.9,129.3, 129.3, 125.8, 124.9, 124.9, 118.6, 118.6, 114.4, 114.4,113.2, 101.3,55.6, 20.8.HRMS(ESI) calculated for C 20 H 19 N4O + [M+H] + : 331.1553,found: 331.1552.

[0031] 2) Preparation of compound B3: Under argon protection, compound B1 (p-alkynylaniline, 197 mg, 1.70 mmol) was dissolved in tetrahydrofuran (7 mL), and compound B2 (3-iodo-2-hydroxypyridine, 250 mg, 1.13 mmol), cuprous iodide (21.5 mg, 0.113 mmol), triethylamine (0.46 mL) and bistriphenylphosphine palladium dichloride (80.3 mg, 0.113 mmol) were added. After refluxing overnight, the liquid in the bottle was concentrated by rotary evaporator, the reaction solution was washed with saturated NaCl solution, extracted with ethyl acetate (3×100 mL), the organic phase was dried with anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4: 1) to obtain compound B3 (white solid, 185 mg, yield: 78%).

[0032] Compound B3 was tested, and the test results were as follows: 1 H NMR (400 MHz, Chloroform- d ) δ8.30 – 8.13 (m, 1H), 7.82(dd, J = 7.6, 1.8 Hz, 1H), 7.78 – 7.64 (m, 2H), 7.17(dt, J =6.7, 3.2 Hz, 1H), 6.85 – 6.64 (m, 3H), 3.87 (s, 2H).

[0033] 3) Preparation of compound B: Compound A (130 mg, 0.41 mmol) was dissolved in 1,4-dioxane (4 mL), and compound B3 (130 mg, 0.62 mmol) was added, palladium acetate (6 mg, 0.02 mmol), 1,1'-binaphthyl-2,2'-bis(diphenylphosphine) (35 mg, 0.02 mmol) and cesium carbonate (260 mg, 0.73 mmol). The mixture was placed in an oil bath, heated to 110 °C, refluxed for 8 hours, filtered with diatomaceous earth, and the liquid in the bottle was concentrated with a rotary evaporator. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1 to 1:1) to obtain compound B (white solid, 140 mg, yield: 79%).

[0034] Compound B was tested, and the test results are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ8.74 (s, 1H), 8.25 (dd, J= 4.9, 1.6 Hz, 1H), 7.89 – 7.80 (m, 5H), 7.70 – 7.64(m, 2H), 7.51 (s, 1H),7.23 (d, J = 3.6 Hz, 1H), 7.19 (dd, J = 7.6, 4.9 Hz, 1H),7.12 –7.06 (m, 2H), 6.88 (s, 1H), 6.57 (d, J = 3.7 Hz, 1H), 3.92 (s, 3H). 13 CNMR(101 MHz, CDCl3) δ 161.8, 158.4, 156.1, 155.7, 151.5, 151.0,143.1, 141.5,130.7, 128.9, 126.5, 126.1, 125.1, 122.6, 121.9, 119.4, 118.1,114.5, 113.8,101.3, 98.2, 55.7. HRMS (ESI): m / z calcd for C 26 H 20 N2O2 + [M+H] + : 434.1612 ; found434.1614.

[0035] 4) Preparation of compound 1: Compound B (70.0 mg, 0.162 mmol) was dissolved in tetrahydrofuran (1 mL) at 0 °C, and sodium hydride (7.14 mg, 0.178 mmol, 60%) was added. After 10 minutes, N,N-dimethylaminoethyl bromide hydrobromide (44 mg, 0.178 mmol) was added. The reaction was allowed to react at room temperature for 1 hour, and water was added to quench the reaction. After the tetrahydrofuran was spin-dried, water (10 mL) and ethyl acetate (3×10 mL) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain compound 1 (white solid, 58.1 mg, yield: 63%).

[0036] Compound 1 was tested, and the test results were as follows: 1H NMR (400 MHz, Chloroform-d) δ8.64 (s, 1H), 8.28 (dd, J = 5.0, 1.7Hz, 1H), 7.98 – 7.91 (m, 2H), 7.89 (dd, J= 7.6, 1.7 Hz, 1H), 7.65 – 7.59 (m,2H), 7.50 – 7.44 (m, 2H), 7.21 (dd, J =7.6, 4.9 Hz, 1H), 7.19 (d, J = 3.7 Hz,1H), 6.98 (d, J = 2.2 Hz, 1H), 6.96 (d,J = 3.5 Hz, 2H), 4.30 (t, J = 7.5 Hz,2H), 3.82 (s, 3H), 2.80 (t, J = 7.5 Hz,2H), 2.37 (s, 6H). 13 C NMR (100 MHz, Chloroform-d) δ 161.9, 158.0, 155.6, 152.1, 150.8, 145.9, 143.6, 131.0, 129.3, 126.8, 126.2, 126.0, 125.8, 124.7, 121.7, 119.5, 114.3, 113.0, 101.2, 99.6, 56.4, 55.6, 47.9, 45.2. HRMS (ESI): m / z calcd forC 31 H 31 N6O2 + [M+H] + :505.2347 ; found 505.2344.

[0037] The chemical formula of N,N-dimethylaminoethyl bromide hydrobromide used in this embodiment is as follows:

[0038] .

[0039] Example 2

[0040] A substituted aminophenyl furanopyridine compound (denoted as compound 2) has the following structure:

[0041] Compound 2

[0042] The specific preparation method is: using 4-(2-bromoethyl)morpholine hydrobromide to obtain compound 2 according to the synthesis steps of compound 1 described in Example 1.

[0043] Compound 2 was tested, and the test results were as follows: 1 H NMR (400 MHz,Chloroform-d) δ 8.64(s, 1H), 8.36 – 8.22 (m, 1H), 7.91 (dd, J = 12.8, 7.9 Hz,3H), 7.62 (d, J =8.6 Hz, 2H), 7.48 (d, J = 8.3 Hz, 2H), 7.22 (dd, J = 7.6, 5.0Hz, 1H), 7.18(d, J = 3.6 Hz, 1H), 6.97 (t, J = 4.5 Hz, 3H), 6.51 (d, J = 3.6Hz, 1H), 4.23(t, J = 7.3 Hz, 2H), 3.83 (s, 3H), 3.63 (t, J = 4.5 Hz, 4H), 2.70(t, J = 7.3Hz, 2H), 2.46 (t, J = 4.5 Hz, 4H). 13 C NMR (100 MHz, Chloroform-d) δ 161.9,158.1, 157.9, 155.7, 152.2, 150.7, 146.3, 143.6, 131.0, 129.3, 127.0, 126.1, 125.9 , 125.7 , 124.8 , 121.7 , 119.5 , 114.2 ,112.8 , 101.2 , 99.5, 67.0 , 56.3 , 55.5 , 54.0 , 47.8 . HRMS (ESI): m / z calcdfor C 33 H 33 N6O3 + [M+H] + :547.2452 ; found 547.2455.

[0044] The chemical formula of 4-(2-bromoethyl)morpholine hydrobromide used in this example is as follows:

[0045] .

[0046] Example 3

[0047] A substituted aminophenyl furanopyridine compound (denoted as compound 3) has the following structure:

[0048] Compound 3

[0049] The specific preparation method is: using 4-(2-bromoethyl)piperazine hydrobromide to obtain compound 3 according to the synthesis steps of compound 1 described in Example 1.

[0050] Compound 3 was tested, and the test results were as follows: 1 H NMR (400 MHz, Chloroform-d) δ 8.62(s, 1H), 8.28 (dd, J = 5.0, 1.5 Hz, 1H), 7.90 (t, J = 8.9Hz, 3H), 7.61 – 7.56(m, 2H), 7.45 (d, J = 8.3 Hz, 2H), 7.21 (dd, J = 7.7, 4.9Hz, 1H), 7.17 (d, J= 3.7 Hz, 1H), 7.01 – 6.95 (m, 3H), 6.51 (d, J = 3.7 Hz, 1H), 4.22 (t, J = 7.1Hz, 2H), 3.84 (s, 3H), 2.75 (t, J = 7.0 Hz, 2H), 2.67 (t,8H), 2.43 (s, 3H). 13 CNMR (100 MHz, Chloroform-d) δ 161.9 , 158.1 ,158.0 , 155.6 , 152.1 , 150.7 ,146.1 , 143.6 , 130.9 , 129.4 , 127.0 , 126.2 HRMS (ESI): m / z calcd for C 33 H 33 N6O3 + [M+H] + : 560.2768; found 560.2767.

[0051] The chemical formula of 4-(2-bromoethyl)piperazine hydrobromide used in this embodiment is as follows:

[0052] .

[0053] Example 4

[0054] A substituted aminophenyl furanopyridine compound (denoted as compound 4) has the following structure:

[0055] Compound 4

[0056] The specific preparation method is: using 4-(2-bromoethyl)tetrahydropyrrole hydrobromide to obtain compound 4 according to the synthesis steps of compound 1 described in Example 1.

[0057] Compound 4 was tested, and the test results are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ8.67(s, 1H), 8.29 (d, J = 4.9 Hz, 1H), 7.92 (t, J = 8.3 Hz, 3H), 7.55(d, J = 8.4Hz, 2H), 7.45 (d, J = 8.2 Hz, 2H), 7.23 (dd, J = 7.7, 5.0 Hz, 1H), 7.18 (d, J = 3.7Hz, 1H), 7.04 – 6.97 (m, 3H), 6.55(d, J = 3.6 Hz, 1H), 4.51 (t, J = 7.5 Hz, 2H),3.85 (s, 3H), 3.26(t, J = 7.6 Hz, 2H), 3.11 (m, 4H), 1.93 (d, J = 6.3 Hz, 4H). 13 CNMR (100 MHz, Chloroform- d ) δ 161.9, 158.4, 157.5, 155.3, 152.1, 151.1,145.0, 143.7, 130.6, 129.5, 126.8, 126.7, 126.6, 126.2, 125.7, 121.6, 119.6,114.4, 113.2, 101.2, 99.9, 55.7, 53.9, 52.7, 46.9, 23.5. HRMS (ESI): m / zcalcdfor C 33 H 33 N6O2 + [M+H] +:531.2503 ; found 531.2503.

[0058] The chemical formula of 4-(2-bromoethyl)pyrrole hydrobromide used in this example is as follows:

[0059] .

[0060] Example 5

[0061] A substituted aminophenyl furanopyridine compound (denoted as compound 5) has the following structure: Compound 5

[0062] The specific preparation method is: using 4-(2-bromoethyl)tetrahydropyrrole hydrobromide to obtain compound 5 according to the synthesis steps of compound 1 described in Example 1.

[0063] Compound 5 was tested, and the test results are as follows: 1 H NMR (400 MHz, Chloroform-d) δ 8.64(d, J = 1.8 Hz, 1H), 8.31 – 8.24 (m, 1H), 7.95 – 7.87 (m,3H), 7.60 (d, J =8.5 Hz, 2H), 7.47 – 7.41 (m, 2H), 7.24 – 7.17 (m, 2H), 7.03 –6.96 (m, 3H), 6.55 – 6.49 (m, 1H), 4.17 (t, J = 7.0 Hz, 2H), 3.83 (s, 3H), 2.76(t, J = 7.6Hz, 2H), 2.43 (s, 6H), 2.11 (p, J = 6.4 Hz, 2H). 13 C NMR(100 MHz, Chloroform-d)δ 161.9, 158.1, 158.0, 155.5, 152.0, 150.7, 145.7, 143.6, 130.9,129.4, 126.8, 126.2, 126.0 , 125.9 , 124.7 , 121.7 , 119.6, 114.4 , 113.0, 101.2 , 99.7 , 56.5 , 55.6 , 48.3 , 43.8 , 24.5. HRMS (ESI): m / z calcd forC 31 H 31 N6O2 + [M+H] +: 519.2503; found 519.2507.

[0064] The chemical formula of 4-(2-bromoethyl)pyrrole hydrobromide used in this example is as follows:

[0065] .

[0066] Example 6

[0067] A substituted aminophenyl furanopyridine compound (denoted as compound 6) has the following structure: Compound 6

[0068] The specific preparation method is: using 4-(2-bromopropyl)morpholine hydrobromide according to the synthesis steps of compound 1 described in Example 1 to obtain compound 6.

[0069] Compound 6 was tested, and the test results were as follows: 1 H NMR (400 MHz, Chloroform-d) δ 8.64(s, 1H), 8.29 (dd, J = 4.9, 1.7 Hz, 1H), 7.94 – 7.88 (m,3H), 7.65 – 7.61 (m,2H), 7.49 – 7.45 (m, 2H), 7.22 (dd, J = 7.6, 4.9 Hz, 1H),7.19 (d, J = 3.7 Hz,1H), 6.99 (d, J = 2.3 Hz, 1H), 6.97 (d, J = 1.3 Hz, 2H),6.52 (d, J = 3.7 Hz,1H), 4.16 – 4.10 (m, 2H), 3.84 (s, 3H), 3.68 (t, J = 4.7Hz, 4H), 2.49 – 2.38(m, 6H), 1.99 (p, J = 7.3 Hz, 3H). 13C NMR (100MHz, Chloroform-d) δ 161.9 ,158.2 , 157.8 , 155.7 , 152.1 , 150.7 , 146.3 ,143.6 , 131.1 , 129.3 , 126.7, 125.9 , 125.6 , 124.4 , 121.7 , 119.5 , 114.3 ,112.9 , 101.2 , 99.4 , 66.7 , 56.5 , 55.6 , 53.6 , 48.9 , 24.8 . HRMS (ESI):m / z calcd for C 33 H 33 N6O3 + [M+H] + :561.2609 ; found 561.2607.

[0070] The chemical formula of 4-(2-bromopropyl)morpholine hydrobromide used in this example is as follows:

[0071] .

[0072] Example 7

[0073] A substituted aminophenyl furanopyridine compound (denoted as compound 7) has the following structure: Compound 7

[0074] The specific preparation method is: using 4-(2-bromopropyl)piperazine hydrobromide to obtain compound 7 according to the synthesis steps of compound 1 described in Example 1.

[0075] Compound 7 was tested, and the test results are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ8.63(s, 1H), 8.28 (dd, J = 4.9, 1.7 Hz, 1H), 7.97 – 7.87 (m, 3H), 7.63 –7.54(m, 2H), 7.46 – 7.37 (m, 2H), 7.22 (dd, J = 7.6, 4.9 Hz, 1H), 7.18(d, J = 3.7 Hz,1H), 7.06 – 7.00 (m, 2H), 6.98 (s, 1H), 6.52 (d, J = 3.7 Hz, 1H), 4.20 (t, J=6.6 Hz, 2H), 3.85 (s, 3H), 3.12 – 3.03 (m, 2H), 2.69 (s, 8H), 2.57 (s, 3H), 2.35 – 2.23 (m, 2H). 13 C NMR (100MHz, Chloroform- d ) δ 161.9, 158.2, 158.1,155.4, 151.9, 150.8, 145.3,143.7, 130.8, 129.5, 126.8, 126.6, 126.2, 126.1,125.0, 121.6, 119.6, 114.4,113.1, 101.3, 99.8, 55.7, 53.7, 53.4, 47.8, 24.7,23.4. HRMS (ESI): m / z calcdfor C 33 H 33 N6O3 + [M+H] + :574.2925 ; found 574.2927.

[0076] The chemical formula of 4-(2-bromopropyl)piperazine hydrobromide used in this embodiment is as follows:

[0077] .

[0078] Example 8

[0079] A substituted aminophenyl furanopyridine compound (denoted as compound 8) has a structure as shown below: Compound 8

[0080] The specific preparation method is: using 4-(2-bromopropyl)tetrahydropyrrole hydrobromide according to the synthesis steps of compound 1 described in Example 1 to obtain compound 8.

[0081] Compound 8 was tested, and the test results are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ8.63(s, 1H), 8.28 (dd, J = 4.9, 1.7 Hz, 1H), 7.96 – 7.91 (m, 2H), 7.90 (dd, J =7.6, 1.6 Hz, 1H), 7.61 – 7.55 (m, 2H), 7.44 – 7.39 (m, 2H), 7.22 (dd, J= 7.6,4.9 Hz, 1H), 7.18 (d, J = 3.7 Hz, 1H), 7.04 – 7.00 (m, 2H), 6.98(s, 1H), 6.52(d, J = 3.7 Hz, 1H), 4.20 (t, J = 6.6 Hz, 2H), 3.85(s, 3H), 3.60 (s, 2H), 3.10 –3.03 (m, 2H), 2.52 (s, 2H), 2.33 – 2.24 (m, 2H),2.12 – 1.86 (m, 4H). 13 C NMR (101 MHz, CDCl3) δ 161.9,158.2, 158.1, 155.4, 151.9, 150.8, 145.3, 143.7,130.8, 129.5, 126.8, 126.6,126.2, 126.1, 125.0, 121.6, 119.6, 114.4, 113.1,101.3, 99.8, 55.7, 53.7, 53.4,47.8, 24.7, 23.4. HRMS (ESI): m / z calcd forC 33 H 33 N6O2 + [M+H] + : 545.2660; found 545.2663.

[0082] The chemical formula of 4-(2-bromopropyl)pyrrolidine hydrobromide used in this embodiment is as follows:

[0083] .

[0084] Example 9

[0085] In this example, the compounds prepared in Examples 1-8 were tested for USP51 in vitro kinase activity using the Ub-AMC (C-terminal-7-amido-labeled ubiquitin protein-4-methylcoumarin label) method.

[0086] Detect the direct interaction ability of the above-mentioned small molecule drugs with protein kinases. These substrates are effectively cleaved or hydrolyzed by various deubiquitinating enzymes, releasing a highly fluorescent moiety. The fluorescent substrate AMC does not emit fluorescence when bound to the ubiquitin chain. When USP cleaves the fluorescent substrate AMC from the ubiquitin chain, AMC emits fluorescence, and the inhibitory degree of USP is analyzed by detecting the fluorescence intensity. At the same time, choose dihydromyricetin (DIH) and epigallocatechin (EPI) as positive controls. Add the fluorescent substrate AMC and each drug molecule into the buffer respectively, then add USP51 protein, and centrifuge and mix the samples at 4 °C and 12,000 rmp, and then detect the fluorescence intensity by machine.

[0087] To more intuitively compare the differences in compound enzyme activities, the IC 50 is divided into four categories: 100 nM < IC 50 < 1 μM (+), 50 nM < IC50 < 100 nM (++), 10 nM < IC50 < 50 nM (+++), IC50 < 10 nM (++++). The test results are shown in Table 1 below:

[0088] Table 1

[0089] Compound No. IC50 (μM) Compound No. IC50 (μM) DHI + 4 +++ EPI + 5 ++++ 1 ++++ 6 ++++ 2 ++++ 7 +++ 3 ++++ 8 +++

[0090] From the results of the above table, it can be seen that Compounds 1-8 all show excellent activities in inhibiting USP51, far exceeding those of the positive controls DHI and EPI. Among them, the activity of Compound 2 is stronger than that of other analogs, and it is expected to be developed into a potential small molecule inhibitor of USP51.

[0091] Example 10

[0092] In this example, the hERG cardiac toxicity of the preferred Compound 2 was evaluated. HEK293 cells stably transfected and expressing the human cardiac HERG ion channel were used. The cultured cells were placed under an inverted microscope, and the recording electrode was brought into contact with the cell surface through a micromanipulator. Subsequently, membrane capacitance compensation and series resistance compensation were performed to smooth the current trace for subsequent testing. An 8-channel perfusion drug delivery system controlled by a solenoid valve was used. The prepared mother liquor of Compound 2 was diluted to 30 μM, 10 μM, 3 μM, 1 μM, and 0.3 μM respectively according to the ratio and added to the perfusion drug delivery system. The flow rate was controlled by gravity to continuously perfuse the cells. The drug delivery electrode was connected and adjusted to the upper left of the cell. After the current of the cell was recorded, perfusion with the control group (without Compound 2) was carried out. After the current was stable, perfusion with Compound 2 was switched to observe its effect. Through analysis with Clampfit software, the cardiac toxicity IC 50It is 10.39 μM, showing high safety.

[0093] Embodiment 11

[0094] In this example, the stability of the preferred compound 2 was evaluated in 5 different species of liver microsomes. The liver microsomes were taken out from the -80 ° C refrigerator, placed in a 37 ° C water bath constant temperature oscillator for pre-incubation for 3 min, and melted for use. Then a certain amount of NADPH was weighed, and an appropriate amount of magnesium chloride solution was added to dissolve it into a 2 mM solution for use. According to the composition ratio of the experimental incubation system (3Mm MgCl2-PB solution, 1 μM compound 2, 0.5 mg / mL liver microsomes), a mixed solution of the incubation system (excluding NADPH) was prepared and packaged in 40 μL / tube. 240 μL of internal standard working precipitant was added to the sample at 0 min, and then 40 μL of NADPH solution was added (40 μL of magnesium chloride solution was added to the negative control group), and 40 μL of NADPH solution was added to other samples to start the reaction (40 μL of magnesium chloride solution was added to the negative control group), and 240 μL of internal standard precipitant was added after incubation in a 37 ℃ water bath for 5 min, 15 min, 30 min, and 60 min. The positive control group was added with 40 μL of NADPH solution to start the reaction, incubated in a 37 ℃ water bath for 5 min, and 240 μL of internal standard precipitant was added after 15 min. All samples were vortexed and centrifuged to obtain 150 μL of supernatant, vortexed after adding 150 μL of water, and analyzed by LC-MS / MS injection. Through analysis and calculation, it was found that compound 2 showed medium clearance efficiency in multiple species and had a low risk of metabolic clearance in vivo. The test results are shown in Table 2 below:

[0095] Table 2

[0096] Species Remaining(T=60 min, %) Remaining(NCF=60 min, %) T1 / 2(min) InvitroCLint (μL / min / mg) Mouse 52.1% 90.5% 65.4 21.2 Human 93.9% 103% >60 1.4 Rat 70.7% 86.9% 128 10.8 Dog 54.1% 90.0% 72.2 19.2 Monkey 15.0% 115% 22.2 62.4

[0097] *When the final remaining volume (%) is greater than 75%, T1 / 2 is reported as >60 min.

[0098] Example 12

[0099] In this example, the bioavailability of the preferred compound 2 was evaluated in mice. After the mice were fasted overnight, three mice were gavaged (50 mg / kg) and three mice were administered by tail vein (10 mg / kg). Then, 0.1 mL-0.2 mL of whole blood was collected at 5 min, 15 min, 30 min, 1h, 2 h, 4 h, 6 h, and 8 h, respectively, and placed in a pre-heparinized sodium tube. The blood and sodium heparin were fully mixed by flicking several times and then centrifuged (4 ℃, 3000 rpm, 12 min) to obtain plasma. 50 μL of plasma sample was taken, 50 μL of diluent (50% methanol / water) and 250 μL of methanol precipitant were added, vortexed and centrifuged (4 ℃, 12000rpm, 12 min), and the supernatant was sealed and sent to LC-MS / MS for detection. The parameters were analyzed by WinNonlin software, and the detection results are shown in Table 3 below:

[0100] Table 3

[0101] PK parameters IV (10 mg / Kg) PO (50 mg / Kg) Cmax (ng / mL) 10953 9433 AUC(0-t) (h*ng / mL) 32386 59198 Tmax (h) 0.083 0.5 T1 / 2 (h) 3.90 5.03 MRTinf (h) 4.25 6.37 F% NA 37.8%

[0102] Table 3 above shows that compound 2 has a higher C max and AUC, the oral half-life reached 5.03 h, and the oral bioavailability was 37.8%, which had high bioavailability.

[0103] Although the description of the present invention has been quite detailed and specifically describes several described embodiments, it is not intended to be limited to any of these details or embodiments or any particular embodiment, but should be regarded as providing a broad possible interpretation of these claims in view of the prior art by reference to the appended claims, thereby effectively covering the intended scope of the present invention. In addition, the above description of the present invention is based on the embodiments foreseeable by the inventor, and its purpose is to provide a useful description, and those non-substantial changes to the present invention that have not yet been foreseen may still represent equivalent changes to the present invention.

Claims

1. A substituted aminophenyl furanopyridine compound, characterized in that: Its structure is shown in Formula I: Formula I; Wherein, NR2R3 is selected from any one of a 3-9 membered saturated heterocyclic amino group and a dialkylamino group; the heterocyclic amino group contains at least one N atom and an O atom as ring atoms and is connected to the rest of the molecule through the amino part; the dialkylamino group is selected from any one of a dimethylamino group, a diethylamino group and a dipropylamino group; and n is an integer of 0-4.

2. A substituted aminophenyl furanopyridine compound, characterized in that: The substituted aminophenyl furanopyridine compound is one of Formula 1 to Formula 8, and the specific structural formula is as follows: 。 3. The substituted aminophenyl furanopyridine compound according to claim 1, characterized in that: Its structure is shown in Formula II: Formula II.

4. The method for preparing the substituted aminophenyl furanopyridine compound according to claim 2, characterized in that: Its preparation route is: ; wherein said Selected from any one of Formula 1 to Formula 8 of claim 2.

5. Use of the substituted aminophenyl furanopyridine compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 in the preparation of a deubiquitinating enzyme USP51 inhibitor.

6. The use according to claim 5, characterized in that: The deubiquitinase USP51 inhibitor can be used to prepare drugs for treating diseases related to abnormal deubiquitinase USP51.

7. The use according to claim 6, characterized in that: The related diseases are breast cancer, ovarian cancer, lymphoma, lung cancer, colorectal cancer, prostate cancer, bladder cancer, liver cancer, neuroblastoma, and leukemia.

8. A pharmaceutical composition, characterized in that The invention comprises a substituted aminophenyl furanopyridine compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3.

9. The pharmaceutical composition according to claim 8, characterized in that The pharmaceutical composition comprises an excipient.

10. The pharmaceutical composition according to claim 9, characterized in that The excipient is at least one of gum arabic, syrup, lanolin and starch.

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