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CN114957248B - Pyrrolo pyrimidine compound and preparation method, pharmaceutical composition and application thereof - Google Patents

Pyrrolo pyrimidine compound and preparation method, pharmaceutical composition and application thereof Download PDF

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CN114957248B
CN114957248B CN202210498359.0A CN202210498359A CN114957248B CN 114957248 B CN114957248 B CN 114957248B CN 202210498359 A CN202210498359 A CN 202210498359A CN 114957248 B CN114957248 B CN 114957248B
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CN114957248A (en
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杨爽
欧洋
李建军
杨光
张坤
王璇
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Nankai University
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    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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Abstract

The invention discloses a compound shown as a formula (I) or pharmaceutically acceptable salt thereof, wherein R 1 Any one of H, alkyl, aryl, substituted aryl, alkylamide and arylamide structures; r is R 2 Any one of H, alkyl, halogen, aryl and substituted aryl structures; r is R 3 Any one of H, alkyl, aryl and substituted aryl structures; x is selected from CH 2 One of O, NH. The compound or the pharmaceutically acceptable salt thereof has the efficient and reversible effect of inhibiting the protease activity of the deubiquitinase USP 51.

Description

Pyrrolo pyrimidine compound and preparation method, pharmaceutical composition and application thereof
Technical Field
The invention belongs to the field of chemical drugs, and particularly relates to a pyrrolopyrimidine compound, a preparation method thereof, a pharmaceutical composition and application thereof in inhibition of deubiquitinase USP 51.
Background
Malignant tumor is one of the major diseases threatening the life safety of human beings. At present, the treatment means of malignant tumors mainly comprise: surgery, radiation therapy, chemotherapy, and the like. The traditional antitumor drug has the defect of large toxic and side effects and the like, so that the traditional antitumor drug is prevented from being further applied in clinic. A number of studies have shown that deubiquitinase exerts vital physiological functions by modulating protein interactions, localization and enzymatic activity, thereby affecting cellular processes including transcription, DNA damage signaling and DNA repair, cell cycle progression, oxidative stress, apoptosis, etc. It has a close relationship with the development of tumor. Deubiquitinase inhibitors have also been found to exert significant antitumor effects in various preclinical studies of tumors, and have become a hotspot in the development of new antitumor drug fields.
Epithelial-mesenchymal transition (EMT) and its reversal processes and mesenchymal-epithelial transition (MET) play a key role in tumor metastasis. During EMT, epithelial cells lose the adhesive and tight junctions that remain in close association with neighboring cells. Activation of EMT by multiple transcription factors can induce migration, invasion and metastasis of tumor cells. While ZEB1 is a transcription factor for controlling EMT induction, and can promote tumor invasion, metastasis and drug resistance treatment. In many cancer types, aberrant expression of ZEB1 is associated with invasive behavior, high tumor index, drug resistance, high metabolic plasticity and metastasis. For example, in breast cancer patients, ZEB1 overexpression was found to occur in tumor cells of triple negative/basal-like breast cancer.
USP51 is a deubiquitinase of ZEB1 (Zinc Finger E-Box Binding Homeobox 1), and targeting US P51 is also likely to be an alternative approach to targeting the pro-cancerous transcription factor ZEB 1. USP51 was found to be a ubiquitin enzyme that binds, de-ubiquitinates and stabilizes ZEB1 by screening a library of human de-ubiquitin enzymes. The absence of USP51 in stromal-like breast cancer cells results in down-regulation of ZEB1 protein and interstitial markers, up-regulation of cadherin-E, and inhibition of cell invasion. In contrast, USP51 overexpression in epithelial cells resulted in upregulation of ZEB1 and mesenchymal markers. Furthermore, USP51 was able to regulate the expression of ZEB1 target genes. Importantly, USP51 is overexpressed in breast cancer patients and is associated with poor survival.
The development of the USP family inhibitor is mainly aimed at targets such as USP7, USP1, USP9 and the like of hot spots. USP51 has been less frequently studied and reported as a newer deubiquitinase for its protease itself and its inhibitors. And the selective inhibitors of deubiquitinase developed at present still have a plurality of problems: (1) related studies on deubiquitinase have focused on USP family members, particularly USP7, but most inhibitors reported thereto exhibit weaker inhibitory activity or contain poor chemical characteristics or have poor selectivity for the currently known deubiquitinase family. (2) At present, the understanding of the structure and functional substrates of the deubiquitinase protein is insufficient and unbalanced, and the inherent mechanism related to the deubiquitinase and tumorigenesis still lacks exploratory knowledge. This also results in the disadvantages of poor specificity of the current small molecule inhibitors of most deubiquitinating enzymes. In addition, the original inhibitor screening method and technology have the defects of poor compatibility with the actual physiological environment, easy occurrence of false positive and the like. Therefore, the targeted USP51 small molecule inhibitor which has novel structure, high inhibition activity, high selectivity and high stability is researched and designed to be synthesized, and has important scientific significance.
Disclosure of Invention
In view of the above, the invention aims to find a novel-skeleton and better-activity and selectivity USP51 small-molecule inhibitor, and provides a pyrrolopyrimidine compound, a preparation method thereof, a pharmaceutical composition and application thereof in preparing medicines for preventing and treating cancers.
In order to achieve the above purpose, the technical scheme of the invention is realized as follows:
a compound of formula (I) or a pharmaceutically acceptable salt thereof:
wherein R is 1 Any one selected from aryl, substituted aryl, alkylamide and arylamide; r is R 2 Any one selected from H, alkyl, halogen, aryl and substituted aryl; r is R 3 Any one of H, alkyl, aryl and substituted aryl structures; x is selected from CH 2 One of O, NH.
Preferably, the structure of the compound is shown as formula II, formula III, formula IV, formula V, formula VI, formula VII or formula VIII:
wherein R is 4 The structure of (2) is one of the compounds 1-4:
；
R 5 the structure of (2) is one of the compounds 5-8:
；
R 6 is NH or O; when R is 6 In the case of O, X has the structure of compound 41:
；
when R is 6 When NH, the structure of X is one of the compounds 9-40 and 42-46:
；
R 7 the structure of (a) is one of the compounds 49-54:
；
R 8 the structure of (a) is one of the compounds 55-58:
；
R 9 the structure of (a) is one of the compounds 59-60:
；
R 10 the structure of (a) is one of compounds 61-62:
the invention also provides a preparation method of the compound or the pharmaceutically acceptable salt thereof, and the preparation route is a first route, a second route, a third route, a fourth route, a fifth route, a sixth route, a seventh route or an eighth route;
route one:
route two:
route three:
route four:
route five:
route six:
route seven:
route eight:
preferably, the cat (catalyst) in each route is taken from Pd 2 (dba) 3 、Pd(OAc) 2 、Pd(PPh 3 ) 4 、Pd(PPh 3 ) 2 Cl 2 、Pd(dppf)Cl 2 One of CuI, cuBr, cuCl; ligand is taken from brettphos, XPhos, SP hos, xantPhos, ruPhos, PPh 3 One of trans-Cyclohexane-1, 2-diamine; base (Base) is taken from K 3 PO 4 、K 2 CO 3 、Cs 2 CO 3 、KO t Bu、Na 2 CO 3 One of the following; sol (solvent) is taken from dioxane, H 2 O, THF, DMF, DMSO, DCM, meCN, toluene, one or both.
The above compound or a pharmaceutically acceptable salt thereof can be used for preparing medicines for inhibiting deubiquitinase USP51 and medicines for preventing and/or treating cancers related to deubiquitinase USP 51.
The cancers include: gynecological cancers, for example: ovarian cancer, cervical cancer, vaginal cancer, pudendum cancer, uterine/endometrial cancer, gestational trophoblastic tumor, fallopian tube cancer, uterine sarcoma; endocrine cancers, such as: adrenal cortex cancer, pituitary cancer, pancreatic cancer, thyroid cancer, parathyroid cancer, thymus cancer, and multiple endocrine tumors; bone cancers, for example: osteosarcoma, ewing's sarcoma, chondrosarcoma, etc.; lung cancer, for example: small cell lung cancer, non-small cell lung cancer; brain and CNS tumors, for example: neuroblastoma, acoustic neuroma, neuroglioma and other brain tumors, spinal cord tumors, breast cancer, colorectal cancer, advanced colorectal adenocarcinoma; gastrointestinal cancers, such as: liver cancer, extrahepatic cholangiocarcinoma, gastrointestinal carcinoid tumor, gallbladder cancer, gastric cancer, esophageal cancer, and small intestine cancer; genitourinary cancers, such as: penile cancer, delphinium cancer, prostate cancer; head and neck neoplasms, for example: nasal cancer, sinus cancer, nasopharyngeal cancer, oral cancer, lip cancer, salivary gland cancer, laryngeal cancer, hypopharyngeal cancer, and orthopharyngeal cancer; blood cancers, for example: acute myelogenous leukemia, acute lymphoblastic leukemia, childhood leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, acute promyelocytic leukemia, plasma cell leukemia; bone marrow cancer hematological disorders, such as: myelodysplastic syndrome, myeloproliferative disorders, fan Heni anemia, aplastic anemia, idiopathic macroglobulinemia; lymphomas, for example: hodgkin's disease, non-hodgkin's lymphoma, peripheral T-cell linezolid, cutaneous T-cell lymphoma, AIDS-related lymphoma; eye cancers, comprising: retinoblastoma, uveal melanoma; skin cancers, such as: melanoma, non-melanoma skin cancer, merkel cell carcinoma; soft tissue sarcomas, for example: kaposi's sarcoma, childhood soft tissue sarcoma, adult soft tissue sarcoma, and urinary system cancer, for example: kidney cancer wilms' tumor, skin cancer, urinary tract cancer and metastatic cell cancer. Preferably for the treatment of breast cancer, colorectal cancer, ovarian cancer, prostate cancer, lung cancer.
The invention also provides a pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof, which may further comprise one or more pharmaceutically acceptable excipients. The dosage form of the pharmaceutical composition is any pharmaceutically acceptable dosage form.
The beneficial effects of the invention are as follows: compared with the prior art, the compound or the pharmaceutically acceptable salt thereof has the effect of efficiently inhibiting the deubiquitinase USP51, and provides a new field of view and thought for widening the cognition of USP families, the research on related mechanisms of the deubiquitinase USP51 in the protein deubiquitination process and the development of novel antitumor small molecule drugs.
Detailed Description
The conception and technical effects of the present invention will be clearly and completely described in conjunction with examples below to fully understand the objects, aspects and effects of the present invention. Unless defined otherwise, technical terms used in the following examples have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention pertains. The test reagents used in the following examples, unless otherwise specified, are all conventional biochemical reagents; the experimental methods are conventional methods unless otherwise specified.
Example 1: preparation of Compound 1
The structures of the compounds S1, S2, S3, S5-1, S6-1 and 1 are shown below:
1) Preparation of compound S2:
compound S1 (3 g,19.53 mmol) was dissolved in acetonitrile (100 mL) under ice-bath and argon atmosphere, N-iodosuccinimide (5.28 g) was added, and after 5 minutes the ice-bath was removed and stirred at room temperature. After 2 hours, the liquid in the bottle was concentrated by rotary evaporator, the reaction solution was washed with saturated NaCl solution, extracted with ethyl acetate (3×150 mL), the organic phase was dried with anhydrous sodium sulfate, concentrated, and the obtained crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate=5:1 to 3:1) to obtain compound S2 (white solid, 4.50g, yield: 72%).
2) Preparation of compound S3:
compound S2 (4 g,15.21 mmol) was dissolved in tetrahydrofuran (130 mL) under ice-bath and argon, di-tert-butyl dicarbonate (3.40 g,18.56 mmol), triethylamine (1.42 g,16.42 mmol) and N, N-dimethylpiperidine (135 mg,1.08 mmol) were added. After 5 minutes the ice bath was removed and stirred at room temperature. After 3 hours, 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 obtained crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate=6:1 to 4:1) to obtain compound S3 (white solid, 5.87g, yield: 75%).
3) Preparation of Compound S5-1:
compound S3 (600 mg,1.6 mmol) was dissolved in (dioxane: water=2:1) (12 mL), and 3,4 dimethoxy phenylboronic acid (298 mg,1.74 mmol) and [1,1' -bis (diphenylphosphine) ferrocene ] palladium dichloride dichloromethane complex (118 mg, catalytic amount), potassium carbonate (4474 mg,3.20 mmol) were added under ice-bath. After 5 minutes, the ice bath was removed, and the temperature was raised to 80℃by transferring to an oil bath. After 3 hours, the liquid in the bottle was concentrated using a rotary evaporator. The reaction solution was washed with saturated NaCl solution, extracted with ethyl acetate (3×150 mL), the organic phase was dried over anhydrous sodium sulfate, concentrated, and the resulting crude product was purified by silica gel column chromatography (dichloromethane: methanol=60:1 to 40:1) to give compound S4-1. Compound S4-1 was redissolved in dichloromethane (5 mL) under ice-bath and argon protection, trifluoroacetic acid (5 mL) was added, the ice-bath was removed after 5 minutes, after stirring at room temperature for 1 hour, the liquid in the bottle was concentrated by rotary evaporator, the reaction solution was washed with saturated sodium bicarbonate solution, extracted with ethyl acetate (3X 50 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated to give deprotected product S5-1 (white solid, 356mg, yield: 67%).
4) Preparation of Compound S6-1:
compound S5-1 (300 mg,1.03 mmol) was dissolved in 1, 4-dioxane (5 mL), 4-iodoanisole (254 mg,0.49 mmol) was added, cuprous iodide solid (18 mg,0.08mmol, catalytic amount) was added, trans-1, 2-cyclohexanediamine (120 mg,0.70 mmol) and potassium phosphate (500 mg,3.01 mmol) were added. After being put into an oil bath and heated to 110 ℃ and refluxed for 3 hours, the mixture was filtered with celite, and the liquid in the bottle was concentrated by a rotary evaporator to purify the obtained crude product by silica gel column chromatography (petroleum ether: ethyl acetate=5:1 to 3:1) to obtain compound S6-1 (white solid, 310mg, yield: 70%).
5) Preparation of Compound 1:
compound S6-1 (100 mg,0.26 mmol) was dissolved in 1, 4-dioxane (2 mL), 3, 4-difluoroaniline (45 mg,0.34 mmol) was added, palladium acetate (3 mg,0.01 mmol) was added, 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (15 mg,0.02 mmol) and cesium carbonate (130 mg,0.35 mmol) were added. After being put into an oil bath and heated to 110 ℃ and refluxed for 8 hours, the mixture was filtered with celite, and the liquid in the bottle was concentrated by a rotary evaporator to purify the obtained crude product by silica gel column chromatography (petroleum ether: ethyl acetate=4:1 to 1:1) to obtain compound 1 (white solid, 109mg, yield: 65%). Compound 1 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.97(s,1H),8.09(s,1H),7.68(d,J＝8.9Hz,2H),7.35(s,1H),7.22(d,J＝8.5Hz,2H),7.15(s,1H),7.09(d,J＝9.0Hz,2H),7.05(d,J＝6.5Hz,2H),7.01(s,1H),3.95(s,6H),3.90(s,3H). 13 C NMR(100MHz,CDCl 3 )δ214.0,213.8,158.5,155.9,154.8,154.0,152.0,150.6,149.5,148.4,130.4,128.9,128.5,126.4,125.1,125.1,122.3,119.4,116.8,114.6,114.6,111.9,110.3,107.7,56.1,56.1,55.6.HRMS(ESI)calculated for C 27 H 23 F2N 4 O 3 + [M+H] + :489.1733,found:489.1750.
example 2: preparation of Compound 2
The structure of compound 2 is shown below:
compound 2 (white solid, 242mg, yield: 61%) was obtained according to the synthetic procedure of compound 1 using 4-methoxyphenylboronic acid. Compound 2 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.97(s,1H),8.09(s,1H),7.68(d,J＝8.9Hz,2H),7.35(s,1H),7.22(d,J＝8.5Hz,2H),7.15(s,1H),7.09(d,J＝9.0Hz,2H),7.05(d,J＝6.5Hz,2H),7.01(s,1H),3.95(s,6H),3.90(s,3H). 13 C NMR(100MHz,CDCl 3 )δ214.0,213.8,158.5,155.9,154.8,154.0,152.0,150.6,149.5,148.4,130.4,128.9,128.5,126.4,125.1,125.1,122.3,119.4,116.8,114.6,114.6,111.9,110.3,107.7,56.1,56.1,55.6.HRMS(ESI)calculated for C 27 H 23 F2N 4 O 3 + [M+H] + :489.1733,found:489.1750.
example 3: preparation of Compound 3
The structures of compounds S5-2 and 3 are shown below:
1) Preparation of Compound S5-3:
compound S3 (500 mg,1.30 mmol) was dissolved in (dioxane: water=2:1) (10 mL), and 3, 5-dimethoxyphenylacetylene (260 mg,1.50 mmol) and [1,1' -bis (diphenylphosphine) ferrocene ] palladium dichloride dichloromethane complex (103 mg, catalytic amount) and potassium carbonate (350 mg,2.70 mmol) were added under ice bath. After 5 minutes, the ice bath was removed, and the temperature was raised to 80℃by transferring to an oil bath. After 3 hours, the liquid in the bottle was concentrated using a rotary evaporator. The reaction solution was washed with saturated NaCl solution, extracted with ethyl acetate (3×150 mL), the organic phase was dried over anhydrous sodium sulfate, concentrated, and the resulting crude product was purified by silica gel column chromatography (dichloromethane: methanol=60:1 to 40:1) to give compound S5-2 (white solid, 580mg, yield: 63%).
2) Preparation of compound 3:
compound S6-3 was obtained according to the synthetic procedure of compound S6-1 using compound S5-2, and compound 3 (white solid, 242mg, yield: 61%) was obtained according to the synthetic procedure of compound 1 described in example 1 using compound S6-3. Compound 3 was tested as follows: 1 H NMR(400MHz,DMSO-d6)δ9.76(s,1H),9.56(s,1H),8.85(s,1H),8.17(s,2H),7.74(d,J＝8.1Hz,2H),7.64(d,J＝3.3Hz,3H),7.40(d,J＝3.3Hz,1H),7.30(d,J＝9.8Hz,1H),6.68(d,J＝3.7Hz,1H),3.33(s,3H),1.51(d,J＝9.8Hz,7H). 13 C NMR(100MHz,DMSO-d6)δ156.0,153.3,151.7,151.2,151.2,150.6,138.5,138.2,134.4,133.6,131.9,131.9,131.5,127.1,124.2,124.2,123.1,119.0,119.0,118.9,113.6,113.6,107.2,101.9,101.9,79.7,31.6,28.6,28.6.HRMS(ESI)calculated for C 29 H 23 F 2 N 4 O 3 + [M+H] + :513.1733,found:513.1745.
example 4: preparation of Compound 4
The structures of compounds S6-4 and 4 are shown below:
1) Preparation of Compound S6-4:
compound S6-4 was obtained according to the synthetic procedure of compound 6-1 using compound S3.
2) Preparation of Compound 4:
compound 4 (white solid, 142mg, yield: 57%) was obtained according to the synthetic procedure of compound 1 using compound S6-4.
Example 5: preparation of Compound 5
The structures of compounds S7-5 and 5 are shown below:
1) Preparation of Compound S7-5:
compound S1 (150 mg,0.97 mmol) was dissolved in tetrahydrofuran (9 mL) under ice-bath and argon atmosphere, and 4-methoxybenzyl alcohol (203 mg,1.47 mmol), diisopropyl azodicarboxylate (237 mg,1.47 mmol) and triphenylphosphine (284 mg,1.47 mmol) were added. After 5 minutes the ice bath was removed and stirred at room temperature. After 5 hours, 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 obtained crude product was purified by silica gel column chromatography (dichloromethane: methanol=50:1 to 30:1) to obtain compound S7-5 (white solid, 160mg, yield: 75%).
2) Preparation of compound 5:
the preparation method comprises the following steps: compound S7-5 (130 mg,0.41 mmol) was dissolved in 1, 4-dioxane (4 mL), 3, 4-difluoroaniline (80 mg,0.62 mmol) was added, palladium acetate (6 mg,0.02 mmol) was added, and 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (35 mg,0.02 mmol) and cesium carbonate (260 mg,0.73 mmol) were added. After being put into an oil bath and heated to 110 ℃ and refluxed for 8 hours, the mixture was filtered with celite, and the liquid in the bottle was concentrated by a rotary evaporator to purify the obtained crude product by silica gel column chromatography (petroleum ether: ethyl acetate=4:1 to 1:1) to obtain compound 5 (white solid, 40mg, yield: 31%). Compound 5 was tested as follows: 1 H NMR(400MHz,Methanol-d 4 )δ8.18(s,1H),7.57(s,1H),6.90(d,J＝8.3Hz,1H),6.82(d,J＝8.3Hz,3H),6.71(q,J＝5.7,4.3Hz,1H),6.49–6.39(m,3H),6.01(s,1H),4.51(s,3H),4.10(s,1H),2.90(s,1H). 13 C NMR(100MHz,Methanol-d 4 )δ159.32,159.11,155.44,151.50,149.92,133.26,129.49,129.21,128.57,128.23,126.78,116.21,113.81,113.44,106.83,99.69,63.52,54.24.HRMS(ESI)calculated for C 20 H 17 F 2 N 4 O + [M+H] + :367.1365,found:367.1389.
example 6: preparation of Compounds 6-7
The structures of compounds 6-7 are shown below:
(1) Preparation of Compound 6: compound 6 (white solid, 60mg, yield: 40%) was obtained according to the synthetic procedure of compound 5 using p-methoxyphenylethanol. Compound 6 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.62(s,1H),8.13(s,1H),7.12(d,J＝33.4Hz,4H),6.83(s,3H),6.37(s,1H),5.30(s,1H),4.35(d,J＝14.7Hz,2H),3.77(s,3H),3.09(d,J＝14.7Hz,2H). 13 C NMR(100MHz,CDCl 3 )δ158.5,151.6,150.0,150.0,130.2,129.8,129.8,126.8,116.8,114.1,114.1,114.1,113.8,113.6,113.2,107.7,107.5,99.8,55.3,46.6,35.7.HRMS(ESI)calculated for C 21 H 19 F 2 N 4 O + [M+H] + :281.1521,found:381.1537.
(2) Preparation of compound 7: compound 7 (white solid, 57mg, yield: 47%) was obtained according to the synthetic procedure of compound 5 using tetrahydro-2H-pyran-4-ol. Compound 7 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.63(s,1H),8.04(s,1H),7.43(s,1H),7.15-7.03(m,3H),6.44(s,1H),4.83(s,1H),4.18(d,J＝14.0Hz,2H),3.64(d,J＝14.0Hz,2H),2.22-1.96(m,4H). 13 C NMR(100MHz,CDCl 3 )δ155.2,151.3,150.6,137.3,137.2,123.1,117.0,116.9,113.6,107.7,107.5,100.3,67.3,67.3,50.8,32.9,32.9.HRMS(ESI)calculated for C 17 H 16 F 2 N 3 O + [M+H] + :316.1256,found:316.1267.
example 7: preparation of Compound 8
The structures of compounds S8, S9 and 8 are shown below:
1) Preparation of compound S8:
compound S8 was obtained according to the synthetic procedure of compound S6-1 using tert-butyl (4-iodophenyl) carbamate.
2) Preparation of compound S9:
compound S9 was obtained according to the synthetic procedure of compound 1 using compound S8.
3) Preparation of Compound 8:
compound S9 (250 mg) was dissolved in methylene chloride (5 mL) under an ice bath and argon atmosphere, and an appropriate amount of trifluoroacetic acid (5 mL) was added. After 5 minutes the ice bath was removed and stirred at room temperature. After 1 hour, the liquid in the bottle was concentrated by rotary evaporator, the reaction solution was washed with saturated sodium bicarbonate solution, extracted with ethyl acetate (3×50 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated to give a crude deprotected compound. The crude product was dissolved in DMF (3 mL), potassium carbonate (170 mg,0.86 mmol), KI (10 mg,0.07mmol, catalytic amount) were added, after stirring for 3 hours at 60 ℃, water (10 mL) was added to quench the reaction, extraction was performed three times with ethyl acetate (20 mL), the organic phases were combined, and the column layer was purified (petroleum ether: ethyl acetate=3: to 1:1) to give compound 8 (white solid, 158mg, yield: 57%).
Example 8: preparation of Compound 9
The structures of compounds S11 and 9 are shown below:
1) Preparation of compound S11:
compound S11 was obtained according to the synthetic procedure of compound S6-1 using 4-iodoanisole (294 mg,0.49 mmol).
2) Preparation of Compound 9:
compound S11 (150 mg,0.58 mmol) was dissolved in 1, 4-dioxane (6 mL), 4-methylaniline (93 mg,0.87 mmol) was added, palladium acetate (6.5 mg,0.03 mmol) was added, and 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (36 mg,0.06 mmol) and cesium carbonate (282 mg,0.87 mmol) were added. After being put into an oil bath and heated to 110 ℃ and refluxed for 8 hours, the liquid in the bottle was filtered with celite, and the obtained crude product was purified by column chromatography on silica gel (petroleum ether: ethyl acetate=4:1 to 2:1) to obtain compound 9 (white solid, 162mg, yield: 63%). Compound 9 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.69(s,1H),7.67(d,J＝14.3Hz,2H),7.61-7.54(m,2H),7.43(s,1H),7.16(s,1H),7.10(d,J＝11.5Hz,2H),7.01(s,2H),6.52(s,1H),3.85(d,J＝14.3Hz,3H),2.31(s,3H). 13 C NMR(100MHz,CDCl 3 )δ158.1,156.4,151.8,151.0,137.9,131.0,130.9,129.3,129.3,125.8,124.9,124.9,118.6,118.6,114.4,114.4,113.2,101.3,55.6,20.8.HRMS(ESI)calculated for C 20 H 19 N 4 O + [M+H] + :331.1553,found:331.1552.
example 9: preparation of Compounds 10-46
The structures of compounds 10-46 are shown below:
(1) Preparation of compound 10: compound 10 was obtained following the synthetic procedure of compound 9 using 4-dimethylaminoaniline. Compound 10 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.58(s,1H),7.60(d,J＝9.0Hz,2H),7.46(d,J＝9.0Hz,2H),7.19(s,1H),7.08(s,1H),6.96(d,J＝9.0Hz,2H),6.69(d,J＝6.9Hz,2H),6.44(s,1H),3.90-3.66(m,3H),2.84(s,6H). 13 C NMR(100MHz,CDCl 3 )δ156.9,155.9,151.0,149.9,145.7,130.0,129.8,124.5,123.8,123.6,120.0,119.6,113.4,113.3,112.8,112.8,111.9,100.2,54.6,40.4,28.7.HRMS(ESI)calculated for C 21 H 22 N 5 O + [M+H] + :360.1819,found:360.1820.
(2) Preparation of Compound 11: compound 11 was obtained following the synthetic procedure of compound 9 using 3, 4-methylenedioxyaniline. Compound 11 was tested as follows: 1 H NMR(400MHz,DMSO-d 6 )δ8.80(s,1H),7.77(s,2H),7.63(s,1H),7.37(s,1H),7.31(s,1H),7.15(d,J＝10.2Hz,2H),7.01(s,1H),6.86(s,1H),6.67(s,1H),6.01(s,2H),4.04(s,3H). 13 C NMR(100MHz,DMSO-d 6 )δ153.4,151.6,146.2,143.0,137.6,130.2,126.1,121.2,120.2,119.3,110.2,109.7,108.5,106.7,103.7,97.3,97.0,96.4,50.9,25.0.HRMS(ESI)calculated for C 20 H 17 N 4 O 3 + [M+H] + :361.1295,found:361.1294.
(3) Preparation of Compound 12: compound 12 was obtained following the synthetic procedure of compound 9 using aniline. Compound 12 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.73(s,1H),7.69(s,2H),7.68–7.63(m,2H),7.46(s,1H),7.32(s,2H),7.20(s,1H),7.05-7.01(m,2H),6.98(s,1H),6.53(s,1H),3.87(s,3H). 13 C NMR(100MHz,CDCl 3 )δ158.1,156.2,151.7,151.0,140.4,130.9,128.8,128.8,126.0,124.9,124.9,121.5,118.4,118.4,114.4,114.4,113.4,101.3,55.6.HRMS(ESI)calculated for C 19 H 17 N 4 O + [M+H] + :317.1397,found:317.1398.
(4) Preparation of Compound 13: compound 13 was obtained following the synthetic procedure of compound 9 using quinolin-8-amine. Compound 13 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ9.88(s,1H),8.95(s,1H),8.82(s,2H),8.14(s,1H),7.73(d,J＝3.3Hz,2H),7.51(s,1H),7.41(s,1H),7.34(s,1H),7.24(s,1H),7.07(s,2H),6.61(s,1H),3.89(d,J＝34.5Hz,3H). 13 C NMR(100MHz,CDCl 3 )δ158.2,156.0,151.7,151.0,147.6,138.6,136.8,136.1,131.0,128.2,127.5,126.2,125.1,125.1,121.4,118.3,114.4,114.4,113.4,113.4,101.3,55.6.HRMS(ESI)calculated for C 22 H 18 N 5 O + [M+H] + :368.1506,found:368.1530.
(5) Preparation of compound 14: compound 14 was obtained following the synthetic procedure of compound 9 using (Z) -4- (phenyldiazenyl) aniline. Compound 14 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.76(s,1H),7.97-7.82(m,6H),7.67(d,J＝2.7Hz,2H),7.54(s,2H),7.45(d,J＝24.5Hz,2H),7.25(d,J＝1.1Hz,2H),7.09(s,2H),6.59(s,1H),3.91(s,3H). 13 C NMR(100MHz,CDCl 3 )δ158.4,155.5,152.9,151.4,151.0,147.1,143.3,130.7,130.2,129.0,129.0,126.7,125.1,125.1,124.3,124.3,122.5,122.5,117.8,117.8,114.5,114.5,113.9,101.4,55.7.HRMS(ESI)calculated for C 25 H 21 N 6 O + [M+H] + :421.1771,found:421.1769.
(6) Preparation of compound 15: following the synthetic procedure of compound 9 using 3, 5-bistrifluoromethylanilineCompound 15 was obtained. Compound 15 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.77(s,1H),8.21(d,J＝12.3Hz,3H),7.59(d,J＝9.0Hz,2H),7.45(s,1H),7.27(s,1H),7.06(d,J＝8.9Hz,2H),6.60(s,1H),3.89(s,3H). 13 C NMR(100MHz,Chloroform-d)δ158.45,155.14,151.14,151.08,141.71,132.13,131.80,130.32,127.22,127.22,125.02,125.02,124.82,122.12,117.42,114.76,114.76,114.32,114.16,101.35,55.58.HRMS(ESI)calculated for C 21 H 15 F 6 N 4 O + [M+H] + :453.1145,found:453.1151.
(7) Preparation of Compound 16: compound 16 was obtained following the synthetic procedure of compound 9 using 9-ethyl-9H-carbazol-4-amine. Compound 16 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.72(s,1H),8.63(s,1H),7.88(s,1H),7.67(d,J＝8.9Hz,2H),7.37(s,1H),7.35(s,1H),7.32(s,1H),7.24(s,1H),7.18(s,1H),7.12(d,J＝5.1Hz,2H),6.99(d,J＝8.9Hz,2H),6.48(s,1H),4.29(d,J＝7.1Hz,2H),3.80(s,3H),1.35(t,J＝7.2Hz,3H). 13 C NMR(100MHz,CDCl 3 )δ171.2,158.1,156.9,151.1,140.4,135.9,132.5,130.9,128.8,125.6,125.5,125.0,124.4,123.1,120.7,118.6,118.2,114.9,114.5,112.9,110.9,109.0,108.4,101.3,60.4,59.6,55.6.HRMS(ESI)calculated for C 27 H 24 N 5 O + [M+H] + :434.1975,found:434.1971.
(8) Preparation of compound 17: compound 17 was obtained following the synthetic procedure of compound 9 using 3, 4-difluoroaniline. Compound 17 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.79(s,1H),8.41(s,1H),7.72(d,J＝8.8Hz,2H),7.37(s,2H),7.33(s,2H),7.17(d,J＝9.8Hz,2H),6.65(s,1H),3.96(s,3H). 13 C NMR(100MHz,CDCl 3 )δ158.3,155.2,151.1,149.1,139.8,132.6,130.5,130.1,129.8,126.6,124.9,124.9,119.5,117.5,114.7,114.7,113.5,101.4,55.6.HRMS(ESI)calculated for C 19 H 15 Cl 2 N 4 O + [M+H] + :385.0617,found:385.0625.
(9) Preparation of compound 18: use of 3, 4-dimethoxy aniline as followsCompound 18 was obtained following the synthetic procedure of compound 9. Compound 18 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.73(s,1H),7.61(s,1H),7.59(s,2H),7.16(s,1H),7.00(d,J＝8.8Hz,2H),6.96(s,2H),6.53(s,1H),6.11(s,1H),3.85(s,3H),3.68(s,6H). 13 C NMR(100MHz,CDCl 3 )δ161.0,158.3,156.2,151.7,150.9,142.2,142.1,130.8,126.5,125.5,125.4,114.4,114.4,113.3,101.2,96.5,96.4,94.2,94.2,55.6,55.3,55.3.HRMS(ESI)calculated for C 21 H 21 N 4 O 3 + [M+H] + :377.1608,found:377.1623.
(10) Preparation of compound 19: using dibenzo [ b, d]Furan-3-amine compound 19 was obtained according to the synthetic procedure of compound 9. Compound 19 was tested as follows: 1 H NMR(400MHz,DMSO-d 6 )δ9.91(s,1H),8.85(s,1H),8.52(s,1H),7.97(d,J＝28.5Hz,2H),7.82(d,J＝8.5Hz,2H),7.68(s,1H),7.62(s,2H),7.38(s,1H),7.32(s,1H),7.17(s,2H),6.68(s,1H),3.87(d,J＝20.0Hz,3H). 13 C NMR(100MHz,DMSO)δ158.2,157.0,156.3,155.9,153.1,151.7,151.4,141.8,130.9,128.9,127.2,126.3,125.4,124.6,123.4,121.1,120.5,114.9,114.8,113.4,111.7,101.8,100.9,100.5,56.0.HRMS(ESI)calculated for C 25 H 19 N 4 O 2 + [M+H] + :407.1503,found:407.1516.
(11) Preparation of compound 20: using benzo [ d ]]The thiazol-5-amine was synthesized according to the procedure of compound 9 to give compound 20. Compound 20 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.77(s,1H),8.21(s,3H),7.59(d,J＝15.7Hz,2H),7.42(s,1H),7.28(s,1H),7.07(s,2H),6.60(s,1H),4.07-3.78(m,3H). 13 C NMR(100MHz,CDCl 3 )δ158.2,156.1,154.5,154.4,151.7,151.0,139.2,130.7,126.3,124.9,124.9,121.5,118.2,114.6,114.6,113.8,112.6,101.3,55.6.HRMS(ESI)calculated for C 20 H 16 N 5 OS + [M+H] + :374.1070,found:374.1070.
(12) Preparation of compound 21: compound 21 was obtained following the synthetic procedure of compound 9 using 3, 4-dichlorophenylamine. To compoundingObject 21 was detected as follows: 1 H NMR(400MHz,Chloroform-d)δ8.72(d,J＝1.2Hz,1H),8.26-7.84(m,1H),7.64(dd,J＝9.4,2.8Hz,2H),7.31(s,1H),7.24(dd,J＝3.8,1.2Hz,1H),7.09-6.99(m,4H),6.57(dd,J＝3.6,1.2Hz,1H),3.89(d,J＝1.1Hz,3H). 13 C NMR(100MHz,CDCl 3 )δ158.4,155.6,151.4,151.0,130.6,126.4,125.0,125.0,116.9,116.9,116.8,114.5,114.5,113.7,113.5,107.8,107.5,101.3,55.6.HRMS(ESI)calculated for C 19 H 15 Cl 2 N 4 O + [M+H] + :385.0617,found:385.0655.
(13) Preparation of compound 22: compound 22 was obtained following the synthetic procedure of compound 9 using 4-alkynylaniline. Compound 22 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.66(s,1H),7.70-7.54(m,4H),7.35(d,J＝13.3Hz,2H),7.18(d,J＝10.8Hz,2H),7.01(d,J＝15.7Hz,2H),6.50(s,1H),3.83(s,3H),2.96(s,1H). 13 C NMR(100MHz,CDCl 3 )δ158.3,155.6,151.5,151.0,140.9,133.1,132.9,130.7,126.5,125.3,125.0,117.8,117.7,114.6,114.4,114.3,113.7,101.3,84.2,75.8,55.6.HRMS(ESI)calculated for C 21 H 17 N 4 O + [M+H + :441.1397,found:441.1390.
(14) Preparation of compound 23: compound 23 was obtained following the synthetic procedure of compound 9 using 3,4, 5-trimethoxyaniline. Compound 23 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.71(s,1H),7.56(d,J＝21.6Hz,2H),7.15(d,J＝15.4Hz,2H),6.98(s,4H),6.55(s,1H),3.88(s,3H),3.76(d,J＝34.0Hz,10H). 13 C NMR(100MHz,CDCl 3 )δ161.0,158.3,156.2,151.7,150.9,142.2,142.1,130.8,126.5,125.5,125.4,114.4,114.4,113.3,101.2,96.5,96.4,94.2,94.2,55.6,55.3,55.3.HRMS(ESI)calculated for C 22 H 23 N 4 O 4 + [M+H] + :407.1714,found:407.1721.
(15) Preparation of compound 24: compound 24 was obtained following the synthetic procedure of compound 9 using 3-phenoxyaniline. Compound 24 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.71(s,1H),7.75-7.60(m,4H),7.35-7.28(m,2H),7.24(s,1H),7.20(d,J＝3.7Hz,1H),7.08-6.95(m,7H),6.55(d,J＝3.7Hz,1H),3.86(s,3H). 13 C NMR(100MHz,CDCl 3 )δ158.4,158.2,156.2,151.7,151.0,136.4,136.4,130.8,129.6,129.6,126.1,125.0,125.0,122.5,120.2,119.9,119.9,117.8,117.8,114.4,114.4,113.4,101.3,55.6.HRMS(ESI)calculated for C 25 H 21 N 4 O 2 + [M+H] + :409.1659,found:409.1665.
(16) Preparation of compound 25: compound 25 was obtained following the synthetic procedure of compound 9 using pentafluoroaniline. Compound 25 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.68(s,1H),7.54(d,J＝23.6Hz,2H),7.23(d,J＝18.0Hz,2H),6.98(d,J＝13.9Hz,2H),6.60(s,1H),3.96-3.78(m,3H). 13 C NMR(100MHz,CDCl 3 )δ158.2,155.5,151.6,151.6,150.9,150.9,130.3,130.3,126.9,126.9,124.8,124.8,114.5,114.3,114.3,114.3,101.3,101.3,55.6.HRM S(ESI)calculated for C 19 H 11 F 5 N 4 O + [M+H] + :406.0853,found:406.0866.
(17) Preparation of compound 26: compound 26 was obtained following the synthetic procedure of compound 9 using 2,3, 4-trifluoroaniline. Compound 26 was tested as follows: 1 H NMR(400MHz,DMSO-d 6 )δ9.17(s,1H),8.77(s,1H),7.70(d,J＝8.9Hz,2H),7.60(d,J＝14.4Hz,2H),7.27(s,1H),7.06(d,J＝8.9Hz,2H),6.65(s,1H),3.81(s,3H). 13 C NMR(100MHz,DMSO)δ158.0,156.5,151.6,151.5,130.8,127.2,127.0,126.9,125.0,125.0,114.7,114.7,113.9,111.8,111.8,111.6,111.6,101.7,55.9.HRMS(ESI)calculated for C 19 H 14 F 3 N 4 O + [M+H] + :371.1114,found:371.1119.
(18) Preparation of compound 27: compound 27 was obtained following the synthetic procedure of compound 9 using 4-trifluoromethylaniline. Compound 27 was tested as follows: 1 H NMR(400MHz,DMSO-d 6 )δ9.98(s,1H),8.88(s,1H),8.06(s,2H),7.75(d,J＝12.0Hz,2H),7.60(d,J＝24.0Hz,3H),7.17(d,J＝3.5Hz,2H),6.72(s,1H),3.85(s,3H). 13 C NMR(100MHz,DMSO)δ158.3,155.9,151.6,151.2,145.2,130.7,127.7,126.6,126.2,126.2,126.2,125.4,123.9,120.8,120.5,117.9,115.0,113.9,101.7,56.0.HRMS(ESI)calculated for C 20 H 16 F 3 N 4 O + [M+H] + :385.1271,found:385.1283.
(19) Preparation of Compound 28: compound 28 was obtained following the synthetic procedure of compound 9 using 3-chloromethylaniline. Compound 28 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.84(s,1H),8.60(s,1H),8.49(s,1H),7.70(s,1H),7.59(s,1H),7.54(s,2H),7.08(s,2H),6.96(s,2H),6.44(s,1H),3.75(s,3H). 13 C NMR(100MHz,CDCl3)δ158.2,155.6,151.4,151.0,141.5,134.6,130.6,129.6,126.4,124.8,124.8,121.3,118.1,116.2,114.6,114.6,113.7,101.4,55.6.HRMS(ESI)calculated for C 19 H 16 ClN 4 O + [M+H] + :351.1107,found:351.1112.
(20) Preparation of compound 29: compound 29 was obtained following the synthetic procedure of compound 9 using 3,4, 5-trifluoroaniline. Compound 29 was tested as follows: 1 H NMR(400MHz,DMSO-d 6 )δ9.93(s,1H),8.86(s,1H),7.80(d,J＝6.5Hz,1H),7.78-7.69(m,2H),7.63(s,1H),7.13(d,J＝4.6Hz,2H),6.69(s,1H),3.84(s,3H). 13 C NMR(100MHz,DMSO)δ171.5,167.7,158.3,158.0,155.7,151.8,151.1,130.6,127.9,127.8,125.6,124.4,115.7,114.9,113.8,113.3,102.2,102.0,101.8,55.9.HRMS(ESI)calculated for C 19 H 14 F 3 N 4 O + [M+H] + :371.1114,found:371.1122.
(21) Preparation of compound 30: compound 30 was obtained following the synthetic procedure of compound 9 using 3-methoxyaniline. Compound 30 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.72(s,1H),7.7-7.57(m,3H),7.34(s,1H),7.17(d,J＝4.8Hz,2H),7.04(d,J＝5.9Hz,3H),6.53(d,J＝14.4Hz,2H),3.90(s,3H),3.71(d,J＝19.1Hz,3H). 13 C NMR(100MHz,CDCl 3 )δ160.2,158.2,156.1,151.7,150.9,141.7,130.8,129.4,126.3,125.3,125.3,114.4,114.4,113.3,110.7,107.3,103.9,101.0,54.9,29.9.HRMS(ESI)calculated for C 20 H 19 N 4 O 2 + [M+H] + :347.1503,found:347.1521.
(22) Preparation of Compound 31: compound 31 was obtained following the synthetic procedure of compound 9 using 3-methyl-4-fluoroaniline. Compound 31 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.70(s,1H),8.33(s,1H),7.59(s,2H),7.36(s,1H),7.25(s,1H),7.21(s,1H),7.05-6.95(m,3H),6.55(s,1H),3.90(d,J＝9.2Hz,3H),2.41(d,J＝1.5Hz,3H).13C NMR(100MHz,CDCl 3 )δ158.2,156.4,151.7,151.0,139.7,130.7,126.7,126.4,126.0,125.9,124.8,124.6,123.2,120.2,120.1,120.1,114.4,113.8,101.3,55.6,50.7.HRMS(ESI)calculated for C 21 H 18 F 3 N 4 O + [M+H] + :399.1427,found:399.1438.
(23) Preparation of compound 32: compound 32 was obtained following the synthetic procedure of compound 9 using 4-fluoroaniline. Compound 32 was tested as follows: 1 H NMR(400MHz,Chloroform-d)δ8.70(s,1H),7.7-7.57(m,4H),7.21(d,J＝6.2Hz,2H),7.09-6.94(m,4H),6.55(s,1H),3.89(d,J＝37.8Hz,3H). 13 C NMR(100MHz,CDCl 3 )δ159.1,158.2,156.7,156.2,151.7,151.0,136.4,130.8,126.1,125.0,125.0,120.0,115.4,115.2,114.4,114.4,113.5,101.3,55.6.HRMS(ESI)c alculated for C 19 H 16 FN 4 O + [M+H] + :345.1303,found:345.1324.
(24) Preparation of compound 33: compound 33 was obtained following the synthetic procedure of compound 9 using 3-fluoroaniline. Compound 33 was tested as follows: 1 H NMR(400MHz,DMSO-d 6 )δ9.81(s,1H),8.86(s,1H),8.00(s,1H),7.77(d,J＝8.9Hz,2H),7.62(s,1H),7.48(s,1H),7.26(s,1H),7.12(s,2H),6.70(s,2H),3.85(s,3H). 13 C NMR(100MHz,CDCl 3 )δ160.1,155.6,151.0,141.5,134.6,130.6,126.4,125.1,124.9,124.8,121.3,121.3,118.1,116.3,116.2,114.6,114.6,101.4,55.6.HRMS(ESI)calculated for C 19 H 16 FN 4 O + [M+H] + :355.1303,found:355.1326.
(25) Preparation of Compounds 34-40: compound 34 was obtained following the synthetic procedure of compound 9 using 4- (1, 2-triphenylvinyl) aniline. Compound 35 was obtained following the synthetic procedure of compound 9 using 4-nitriloaniline. Compound 36 was obtained following the synthetic procedure of compound 9 using 9H-fluoren-2-amine. Compound 37 was obtained following the synthetic procedure of compound 9 using 4-vinylaniline. Compound 38 was obtained following the synthetic procedure of compound 9 using 1- (4-aminophenyl) ethan-1-one. Compound 39 was obtained following the synthetic procedure of compound 9 using (E) -4-styrylaniline. Compound 40 was obtained following the synthetic procedure of compound 9 using ethyl 4-aminobenzoate.
(26) Preparation of Compound 41: pterostilbene (90.0 mg) was dissolved in DMF, potassium carbonate (68.0 mg,0.470 mmol) was added to the reaction system followed by stirring at room temperature for 2 hours, the reaction was quenched with water (10 mL), extracted three times with ethyl acetate (20 mL), the organic phases were combined, and the column layer was purified (petroleum ether: ethyl acetate=3:1 to 1:1) to give compound 41 (white solid, 158mg, yield: 53%).
(27) Preparation of Compounds 42-46: compound 42 was obtained following the synthetic procedure of compound 9 using pyrene-2-amine. Compound 43 was obtained following the synthetic procedure of compound 9 using anthracene-2-amine. Compound 44 was obtained following the synthetic procedure of compound 9 using naphthalene-2-amine. Compound 45 was obtained following the synthetic procedure of compound 9 using benzofuran-6-amine. Compound 46 was obtained following the synthetic procedure of compound 9 using 3-aminobenzofuran-2-carboxylic acid methyl ester.
Example 10: preparation of Compound 49
The structures of compounds S12-49 and 49 are shown below:
the preparation method comprises the following steps:
1) Preparation of Compounds S13-49:
at N 2 Under protection, p-alkynylaniline (300 mg,2.56 mmol) was dissolved in THF (16 mL), followed by addition of triethylamine @ to the reaction system1.53mL,5.12 mmol), cuprous iodide (48.8 mg,0.256 mmol), bis triphenylphosphine palladium dichloride (180 mg,0.256 mmol) and iodobenzene (891 mg,3.84 mmol), followed by overnight reaction at room temperature, after completion of the reaction, the solvent was distilled off by rotary evaporation, then extracted with water (10 mL) and ethyl acetate (20 mL), the organic phases were combined and purified by subsequent column chromatography (petroleum ether: ethyl acetate = 3: 1) Compound S13-49 was obtained.
2) Preparation of Compound 49:
compound S11 (150 mg,0.580 mmol) was dissolved in 1, 4-dioxane (6 mL), S13-49 (122 mg, 0.553mmol) was added, palladium acetate (6.50 mg,0.0290 mmol) was added, 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (36.0 mg,0.0580 mmol) and cesium carbonate (282 mg,0.870 mmol) were added. After being put into an oil bath and heated to 110 ℃ and refluxed for 8 hours, the mixture was filtered with celite, and the liquid in the bottle was concentrated by a rotary evaporator to purify the obtained crude product by silica gel column chromatography (petroleum ether: ethyl acetate=4:1 to 2:1) to obtain compound 49 (white solid, 192mg, yield: 71%).
Example 11: preparation of Compounds 50-54
The structures of compounds 50-54 are shown below:
preparation of Compounds 50-54: compound 50 was obtained following the synthetic procedure of compound 49 using 4-nitrilodibenzene. Compound 51 was obtained following the synthetic procedure of compound 49 using 4-iodobenzaldehyde. Compound 52 was obtained following the synthetic procedure of compound 49 using 3-iodo-2-methoxypyridine. Compound 53 was obtained following the synthetic procedure of compound 49 using 2-acetyl-6-iodopyridine. Compound 54 was obtained following the synthetic procedure of compound 49 using 4-hydroxy iodobenzene.
Example 12: preparation of Compound 55
The structures of compound 55 and S15-55 are shown below:
the preparation method comprises the following steps:
1) Preparation of Compounds S15-55:
at N 2 Under protection, 4-acetylaniline (300 mg,2.22 mmol) was dissolved in EtOH (6 mL), followed by addition of sodium hydroxide (266 mg,6.66 mmol) to the reaction system, followed by addition of benzaldehyde (349 mg,2.66 mmol) after 10 minutes, followed by reaction overnight at room temperature, after completion of the reaction, the solvent was distilled off by rotary evaporation, followed by extraction with water (10 mL) and ethyl acetate (20 mL), and the combined organic phases were purified by column chromatography (petroleum ether: ethyl acetate=3:1) to give compound S15-55.
2) Preparation of compound 55:
compound S11 (150 mg,0.580 mmol) was dissolved in 1, 4-dioxane (6 mL), S15-55 (144 mg, 0.553mmol) was added, palladium acetate (6.50 mg,0.0290 mmol) was added, 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (36.0 mg,0.0580 mmol) and cesium carbonate (282 mg,0.870 mmol) were added. After being put into an oil bath and heated to 110 ℃ and refluxed for 8 hours, the liquid in the bottle was filtered with celite, and the obtained crude product was purified by column chromatography on silica gel (petroleum ether: ethyl acetate=4:1 to 2:1) to obtain compound 55 (white solid, 135mg, yield: 52%).
Example 13: preparation of Compounds 56-58
The structures of compounds 56-58 are shown below:
preparation of Compounds 56-58: compound 56 was obtained following the synthetic procedure of compound 55 using 4-nitrilobenzaldehyde. Compound 57 was obtained following the synthetic procedure of compound 55 using cinnamaldehyde. Compound 58 was obtained following the synthetic procedure of compound 55 using 2-naphthaldehyde.
Example 14: preparation of Compound 59
The structures of compounds 59 and S17 are shown below:
the preparation method comprises the following steps:
1) Preparation of compound S17:
at N 2 Under protection, compound S16 (7H-pyrrolo [2, 3-d)]Pyrimidine-2-amine) (250 mg,1.86 mmol) was dissolved in dioxane (6 mL), followed by addition of cuprous iodide (36.0 mg,0.186 mmol), trans-1, 2-cyclohexanediamine (213 mg,1.86 mmol), potassium phosphate (792 mg,3.73 mmol) and 4-methoxyiodobenzene (460 mg,1.96 mmol) to the reaction system, followed by reflux overnight, filtration over celite after completion of the reaction and cooling to room temperature, rotary evaporation of the solvent, followed by column chromatography purification (dichloromethane: methanol=40: 1) Compound S17 was obtained.
2) Preparation of compound 59:
compound S17 (50.0 mg,0.208 mmol) was dissolved in dichloromethane (2 mL), triethylamine (45 mg, 0.264 mmol), cinnamoyl chloride (29.0 mg,0.229 mmol) were added, reacted overnight at room temperature, then extracted with water (10 mL) and dichloromethane (20 mL), combined and the organic phases were purified by column chromatography (petroleum ether: ethyl acetate=3:1) to give compound 59 (white solid, 49.0mg, yield: 72%).
Example 15: preparation of Compound 60
The structure of compound 58 is shown below:
the preparation method comprises the following steps: compound 60 was obtained following the synthetic procedure of compound 59 using (E) -2-cyano-3-phenylpropionyl chloride.
Example 16: preparation of Compound 61
The structures of compound 61 and S19-61 are shown below:
the preparation method comprises the following steps:
1) Preparation of Compound S19-61:
at N 2 Under protection, the compound ethyl benzoate (1.00 g,6.66 mmol) was dissolved in methyl acetate (60 mL), then hydrazine hydrate (541 mg,8.66 mmol) was added to the reaction system, followed by reflux reaction overnight, after the reaction was completed and cooled to room temperature, the solvent was distilled off by rotary evaporation, and then purified by column chromatography (dichloromethane: methanol=50:1) to obtain compound S19-61.
2) Preparation of Compound 61:
compound 38 (263 mg, 0.284 mmol) was dissolved in absolute ethanol (7 mL), two drops of glacial acetic acid and compound S19-61 (100 mg, 0.284 mmol) were added, the reaction was refluxed overnight, after completion of the reaction and cooling to room temperature, the solvent was distilled off by rotary evaporation, then extracted with water (10 mL) and ethyl acetate (20 mL), and the combined and organic phases were purified by column chromatography (dichloromethane: methanol=100:1) to give compound 61 (white solid, 260mg, yield: 74%).
Example 17: preparation of Compound 62
The structure of compound 62 is shown below:
the preparation method comprises the following steps: compound 62 was obtained following the synthetic procedure of compound 61 using ethyl 2-furancarboxylate.
Example 18: USP51 enzymatic research
Ub-AMC (C-terminal-7-amido mark ubiquitin-4-methyl coumarin mark) method is used for carrying out USP in vitro kinase activity test, and the direct action capability of the drug small molecules and protein kinase and the small molecule activity are detected. The fluorescent substrate A MC is combined with ubiquitin chain to emit no fluorescence, when the fluorescent substrate AMC is cut off from ubiquitin chain, the AMC emits fluorescence, and the inhibition degree of the USP is analyzed by detecting fluorescence intensity. Meanwhile, compounds dihydromyricetin (Dihydromyric etin, DIH) and Epigallocatechin (EPI) were selected as positive controls, and specific test results are shown in Table 1. IC (integrated circuit) 50 Between 0.01 and 0.1. Mu.M, labeled A, IC 50 At 0.1-1. Mu.M, labeled B, live IC 50 At 1-10. Mu.M, labeled C, IC 50 Labeled D at 10-30. Mu.M.
TABLE 1
From the results of table 1, it is clear that compounds 17, 19, 34, 39, 40, 41, 42 and 50 all showed excellent activity against USP51 superior to positive controls DHI and EPI, and that compounds 34 and 42 were far more active than the other analogues, and were expected to be developed as potential USP51 small molecule inhibitors.
The present invention is not limited to the above embodiments, but is merely preferred embodiments of the present invention, and the present invention should be construed as being limited to the above embodiments as long as the technical effects of the present invention are achieved by the same means. Various modifications and variations are possible in the technical solution and/or in the embodiments within the scope of the invention.

Claims (5)

1. A compound of formula II, formula III, formula IV, formula V, formula VI, formula VII or formula VIII, or a pharmaceutically acceptable salt thereof:
formula II, & lt>Formula III>A formula IV;a formula V;Formula VI;Formula VII;formula VIII;
wherein R is 4 The structure of (2) is one of the compounds 1-4:
；R 5 the structure of (2) is one of the compounds 5-8:
the method comprises the steps of carrying out a first treatment on the surface of the X is NH or O; when X is O, R 6 The structure of (a) is compound 41:
；
x when NH, R 6 The structure of (a) is one of compounds 9-40 and 42-46:
；
R 7 the structure of (a) is one of the compounds 49-54:
；
R 8 the structure of (a) is one of the compounds 55-58:
；
R 9 the structure of (a) is one of the compounds 59-60:
；
R 10 the structure of (a) is one of compounds 61-62:
。
2. a process for the preparation of a compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the preparation is in route one, route two, route three, route four, route five, route six, route seven or route eight;
route one:
；
route two:
；
route three:
；
route four:
；
route five:
；
route six:
；
route seven:
；
route eight:
。
3. use of a compound of claim 1, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prevention and/or treatment of cancer associated with deubiquitinase USP 51; the cancer is breast cancer, ovarian cancer, lung cancer, gastric cancer and endometrial cancer.
4. A pharmaceutical composition comprising a compound of claim 1 or a pharmaceutically acceptable salt thereof.
5. The pharmaceutical composition of claim 4, further comprising one or more pharmaceutically acceptable excipients.
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CN118660872A (en) * 2022-04-29 2024-09-17 江苏亚虹医药科技股份有限公司 Pyrimidine compound, preparation method and medical application thereof
CN115246832B (en) * 2022-06-15 2024-05-24 深圳湾实验室 Deubiquitinase USP25 and USP28 targeted inhibitors, and preparation and application thereof
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CN116768902B (en) * 2023-05-19 2024-09-24 南开大学 Substituted phenol hydroxyl phenyl pyrrolopyrimidine compound and preparation method and application thereof

Citations (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005107760A1 (en) * 2004-04-30 2005-11-17 Irm Llc Compounds and compositions as inducers of keratinocyte differentiation
CN1918158A (en) * 2004-02-14 2007-02-21 Irm责任有限公司 Compounds and compositions as protein kinase inhibitors
CN101981036A (en) * 2008-02-06 2011-02-23 诺瓦提斯公司 Pyrrolo [2, 3-D] pyridines and use thereof as tyrosine kinase inhibitors
CN102482277A (en) * 2009-05-05 2012-05-30 达纳-法伯癌症研究所有限公司 Egfr inhibitors and methods of treating disorders
CN102869358A (en) * 2010-04-13 2013-01-09 诺华有限公司 Combination comprising a cyclin dependent kinase 4 or cyclin dependent kinase (cdk4/6) inhibitor and an mtor inhibitor for treating cancer
CN103059030A (en) * 2012-12-28 2013-04-24 北京师范大学 Pyrimidine compound with effect of adhesion kinase inhibition and preparation method and application thereof
CN105705504A (en) * 2013-10-10 2016-06-22 密歇根大学董事会 Deubiquitinase inhibitors and methods for use of the same
CN107266453A (en) * 2012-08-06 2017-10-20 美国艾森生物科学公司 It is used as the new pyrrole and pyrimidine compound of kinases inhibitor
CN107406451A (en) * 2014-12-30 2017-11-28 福马治疗股份有限公司 Pyrrolopyrimidine and pyrazolopyrimidine as the inhibitor of ubiquitin-specific protease 7
WO2017211245A1 (en) * 2016-06-06 2017-12-14 深圳市塔吉瑞生物医药有限公司 Substituted pyrrolopyrimidine compound and application thereof
CN107613769A (en) * 2015-02-17 2018-01-19 润新生物公司 Some chemical entities, composition and method
WO2018082444A1 (en) * 2016-11-02 2018-05-11 叶宝欢 Pyrazolopyrimidine compound as pi3k inhibitor and use thereof
CN110467615A (en) * 2018-05-10 2019-11-19 四川科伦博泰生物医药股份有限公司 Azolopyrimidines include its pharmaceutical composition and its preparation method and application
CN110691782A (en) * 2016-12-01 2020-01-14 艾普托斯生物科学公司 Fused pyrimidine compounds as dual inhibitors of BRD4 and JAK2 and methods of use thereof
CN112552294A (en) * 2019-09-10 2021-03-26 上海翰森生物医药科技有限公司 Piperazine heterocyclic derivative inhibitor, preparation method and application thereof

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007038387A2 (en) * 2005-09-23 2007-04-05 Yale University Compounds and methods for the treatment of viruses and cancer
US20130225527A1 (en) * 2008-05-21 2013-08-29 Ariad Pharmaceuticals, Inc. Phosphorus Derivatives as Kinase Inhibitors
US20160222014A1 (en) * 2013-09-10 2016-08-04 Asana Biosciences, Llc Compounds for regulating fak and/or src pathways
US10570141B2 (en) * 2016-03-25 2020-02-25 Chia Tai Tianqing Pharmaceutical Group Co., Ltd. Substituted pyrrolopyrimidine CDK inhibitor, pharmaceutical composition containing same and use thereof

Patent Citations (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1918158A (en) * 2004-02-14 2007-02-21 Irm责任有限公司 Compounds and compositions as protein kinase inhibitors
WO2005107760A1 (en) * 2004-04-30 2005-11-17 Irm Llc Compounds and compositions as inducers of keratinocyte differentiation
CN101981036A (en) * 2008-02-06 2011-02-23 诺瓦提斯公司 Pyrrolo [2, 3-D] pyridines and use thereof as tyrosine kinase inhibitors
CN102482277A (en) * 2009-05-05 2012-05-30 达纳-法伯癌症研究所有限公司 Egfr inhibitors and methods of treating disorders
CN102869358A (en) * 2010-04-13 2013-01-09 诺华有限公司 Combination comprising a cyclin dependent kinase 4 or cyclin dependent kinase (cdk4/6) inhibitor and an mtor inhibitor for treating cancer
CN107266453A (en) * 2012-08-06 2017-10-20 美国艾森生物科学公司 It is used as the new pyrrole and pyrimidine compound of kinases inhibitor
CN103059030A (en) * 2012-12-28 2013-04-24 北京师范大学 Pyrimidine compound with effect of adhesion kinase inhibition and preparation method and application thereof
CN105705504A (en) * 2013-10-10 2016-06-22 密歇根大学董事会 Deubiquitinase inhibitors and methods for use of the same
CN107406451A (en) * 2014-12-30 2017-11-28 福马治疗股份有限公司 Pyrrolopyrimidine and pyrazolopyrimidine as the inhibitor of ubiquitin-specific protease 7
CN107613769A (en) * 2015-02-17 2018-01-19 润新生物公司 Some chemical entities, composition and method
WO2017211245A1 (en) * 2016-06-06 2017-12-14 深圳市塔吉瑞生物医药有限公司 Substituted pyrrolopyrimidine compound and application thereof
WO2018082444A1 (en) * 2016-11-02 2018-05-11 叶宝欢 Pyrazolopyrimidine compound as pi3k inhibitor and use thereof
CN110691782A (en) * 2016-12-01 2020-01-14 艾普托斯生物科学公司 Fused pyrimidine compounds as dual inhibitors of BRD4 and JAK2 and methods of use thereof
CN110467615A (en) * 2018-05-10 2019-11-19 四川科伦博泰生物医药股份有限公司 Azolopyrimidines include its pharmaceutical composition and its preparation method and application
CN112552294A (en) * 2019-09-10 2021-03-26 上海翰森生物医药科技有限公司 Piperazine heterocyclic derivative inhibitor, preparation method and application thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
刘晓飞 ; 王庭芳 ; 鞠曹云 ; 张灿 ; .4-苯基-吡咯并[2,3-d]嘧啶类JAK2激酶抑制剂的设计、合成和抗肿瘤活性.中国药科大学学报.2017,(02),150-156. *

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