A phenylthiazolylbenzamide compound and its application

Abstract

The present invention provides a phenylthiazolyl benzamide compound and its application. The phenylthiazolyl benzamide compound has a structure as shown in the following formula I: wherein R1 is selected from R a , R b Independently selected from -Me, -CD3, or any one of; said R f Selected from the R g is selected from any one of hydrogen, -Me, -n-Pr, -n-Bu, or; R2 is selected from hydrogen or halogen; R3 is selected from any one of hydrogen, substituted or unsubstituted C6-C12 aryl; represents the connection site of the group. The phenylthiazolylbenzamide compounds provided by the present invention can effectively inhibit the activity of COUP-TFII and NRF2, and also have excellent inhibitory effect on the proliferation of prostate cancer cells.

Description

Technical Field

[0001] The invention belongs to the field of medicinal chemistry, and specifically relates to a phenylthiazolyl benzamide compound and an application thereof, and in particular relates to a phenylthiazolyl benzamide compound with high activity and an application thereof. Background Art

[0002] Nuclear receptors are a superfamily of ligand-dependent transcription factors that are homologous to steroid hormone receptors. They are involved in regulating the growth and development of the body and cell differentiation, thereby affecting a variety of physiological and metabolic processes in the body. Chicken ovalbumin upstream promoter transcription factor 2 (COUP-TFII) is one of the important members of the nuclear receptor family. The COUP-TF family includes two highly homologous subtypes, COUP-TFI and COUP-TFII, which are located on chromosomes 5 and 15, respectively, and are also called nuclear receptor 2 family 1 and 2 (NR2F1 and NR2F2). Since its specific binding ligand has not yet been identified, COUP-TF is currently considered to be an orphan nuclear receptor.

[0003] Current studies have shown that COUP-TFI plays an important role in neural development and COUP-TFII plays an important role in organ development. Other studies have shown that COUP-TFII plays an important role in all aspects of the body's physiological growth and development, can regulate multiple signaling pathways, and participate in controlling tumor growth, angiogenesis, and the regeneration of body tissues or cells.

[0004] It is worth mentioning that in embryonic development, tissue regeneration, and disease, COUP-TFII expression is abnormally upregulated; while in normal tissues or the mature stage of the body, COUP-TFII expression is maintained at a low level. Current research results also show that when the COUP-TFII gene is knocked out in adult mice, the mice do not show abnormal physiological phenomena such as phenotypic changes. This to a certain extent shows that inhibiting COUP-TFII does not produce toxic side effects. Therefore, the development of small molecule drugs that target and inhibit the regulation of COUP-TFII activity is feasible and has great potential.

[0005] During tumor growth, COUP-TFII is a necessary condition for angiogenesis, and its abnormal expression can lead to the occurrence and metastasis of cancer. Therefore, COUP-TFII is a potential ideal new target for cancer treatment.

[0006] NRF2 (nuclear factor E rythroid 2-related factor) was cloned by Moi et al. in 1994 as a factor that binds to the nuclear factor E2 repeat sequence in the β-globin gene promoter. It belongs to the CNC family of transcription factors and contains a leucine zipper structure.

[0007] The NRF2 / Keap1 (Kelch-like ECH-associated protein 1)-ARE (antioxidant response element) signaling pathway is an endogenous antioxidant signaling pathway discovered in recent years.

[0008] Activation of the NRF2 / Keap 1-ARE pathway is one of the important mechanisms of anti-tumor development. In the early stages of tumor development, activation of NRF2 enables cancer cells to adapt to adverse microenvironments and high endogenous ROS levels; in rapidly proliferating cells, NRF2 can support intermediary metabolism by promoting the biosynthesis of nucleotides and amino acids. Studies have shown that low-activity NRF2 promotes tumor development, while sustained high-activity NRF2 can accelerate cancer progression and tolerance to treatment.

[0009] Therefore, the development of compounds that can inhibit both COUP-TFII and NRF2 has very important practical value and has broad application prospects in tumor treatment. Summary of the invention

[0010] In view of the deficiencies of the prior art, the object of the present invention is to provide a phenylthiazolyl benzamide compound and its application, in particular to provide a highly active phenylthiazolyl benzamide compound and its application. The phenylthiazolyl benzamide compound provided by the present invention can effectively inhibit the activity of COUP-TFII and NRF2, and also has an excellent inhibitory effect on the proliferation of prostate cancer cells.

[0011] To achieve this object, the present invention adopts the following technical solutions:

[0012] In a first aspect, the present invention provides a phenylthiazolyl benzamide compound, wherein the phenylthiazolyl benzamide compound has a structure as shown in the following formula I:

[0013]

[0014] Wherein R1 is selected from

[0015] R a , R bIndependently selected from -Me, -CD3, Any one of .

[0016] The R f Selected from

[0017] The R g Selected from hydrogen, -Me, -n-Pr, -n-Bu, Any one of .

[0018] R2 is selected from hydrogen or halogen.

[0019] R3 is selected from any one of hydrogen, substituted or unsubstituted C6-C12 aryl.

[0020] Indicates the attachment site of a group.

[0021] The above-mentioned compounds with specific structures can effectively inhibit the activity of COUP-TFII and NRF2, and also have excellent inhibitory effects on the proliferation of prostate cancer cells.

[0022] Preferably, the substituent of the C6-C12 aryl group is selected from C1-C10 straight chain alkyl or branched chain alkyl, C1-C10 haloalkyl or -COOR c Any one of; said R c Selected from hydrogen or C1-C10 straight chain alkyl.

[0023] Preferably, said R2 is selected from hydrogen or -F.

[0024] Preferably, said R3 is selected from hydrogen or

[0025] The R i Any one selected from -CF3, -COOH or -COOEt.

[0026] The R j Selected from -F or -Cl;

[0027] Indicates the attachment site of a group.

[0028] Preferably, the phenylthiazolylbenzamide compound is selected from any one of the following formulas 1 to 20:

[0029]

[0030]

[0031] The compound of formula I can be prepared by the following method:

[0032]

[0033] Starting from p-aminoacetophenone, compound III is obtained by substitution, and then further substituted and cyclized to obtain intermediate M (R1 is selected from Starting from the intermediate M (R1 has the same limited range as above), compound 1-2 is obtained by substitution, and finally the bromine atom is replaced to obtain the compound shown in formula I.

[0034] In a second aspect, the present invention provides stereoisomers of the phenylthiazolylbenzamide compounds described above, pharmaceutically acceptable salts thereof or pharmaceutical compositions comprising the same.

[0035] Preferably, the pharmaceutical composition further comprises pharmaceutically acceptable excipients.

[0036] In a third aspect, the present invention also provides the use of the phenylthiazolyl benzamide compounds as described above or the stereoisomers of the phenylthiazolyl benzamide compounds as described above, their pharmaceutically acceptable salts, and pharmaceutical compositions comprising the same in the preparation of drugs for treating and / or preventing diseases associated with excessive COUP-TFII activity or overexpression of COUP-TFII.

[0037] Preferably, the disease associated with excessive COUP-TFII activity or overexpression of COUP-TFII includes prostate cancer.

[0038] In a fourth aspect, the present invention also provides the use of the phenylthiazolyl benzamide compounds as described above or the stereoisomers of the phenylthiazolyl benzamide compounds as described above, their pharmaceutically acceptable salts, and pharmaceutical compositions containing them in the preparation of drugs for treating and / or preventing diseases related to excessive activity of NRF2 (nuclear factor erythroid 2-related factor 2) or overexpression of NRF2.

[0039] Compared with the prior art, the present invention has the following beneficial effects:

[0040] The present invention provides a phenylthiazolylbenzamide compound with a specific structure, which can effectively inhibit the activity of COUP-TFII and NRF2, and also has an excellent inhibitory effect on the proliferation of prostate cancer cells, and is expected to become a new type of tumor prevention and treatment drug. DETAILED DESCRIPTION

[0041] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0042] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0043] The term "pharmaceutically acceptable salt" means that a compound can be converted into a corresponding salt by conventional methods, which is chemically or physically compatible with other ingredients constituting a pharmaceutical dosage form and physiologically compatible with receptors. The salt can be an acidic and / or basic salt formed by the compound and an inorganic and / or organic acid and / or an inorganic and / or organic base, and also includes zwitterionic salts (inner salts), and also includes quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly in the final separation and purification of the compound. It can also be obtained by appropriately mixing the compound of the present invention or its stereoisomer or solvate with a certain amount of acid or base. These salts may form a precipitate in the solution and be collected by filtration, or be recovered after the solvent is evaporated, or be obtained by cooling and drying after reaction in an aqueous medium. In particular, the salt is preferably a water-soluble, pharmaceutically acceptable, non-toxic acid addition salt, examples of which are salts formed between an amino group and an inorganic acid (such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid) or with an organic acid (such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid), or salts formed by using other conventional methods in the art (such as ion exchange methods).

[0044] The term "alkyl" refers to a saturated hydrocarbon containing a primary (normal) carbon atom, or a secondary carbon atom, or a tertiary carbon atom, or a quaternary carbon atom, or a combination thereof. Phrases containing this term, for example, "C1-C3 alkyl" refers to an alkyl group containing 1 to 3 carbon atoms, each occurrence of which can be independently C1 alkyl, C2 alkyl, or C3 alkyl. Suitable examples include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 2-propyl (i-Pr, i-propyl, -CH(CH3)2).

[0045] The term "heterocycloalkyl" refers to a non-aromatic cyclic group in which one or more atoms constituting the ring are heteroatoms and the rest are carbon atoms, and the heteroatoms include but are not limited to nitrogen atoms, oxygen atoms and sulfur atoms, etc. Preferred heterocycloalkyls are 3-10 membered saturated heterocycloalkyls. Unless otherwise specifically indicated in the specification, the heterocycloalkyl group may be a monocyclic ring ("monocyclic heterocycloalkyl"), or a bicyclic, tricyclic or higher heterocyclic ring system, which may include a paracyclic (fused), bridged (bridged) or spiro ring system (e.g., a bicyclic system ("bicyclic heterocycloalkyl"). The bicyclic ring system of the heterocycloalkyl group may include one or more heteroatoms in one or both rings; and is saturated. Heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetane, thietanyl, oxetanyl, tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydropyranyl, or isomers and stereoisomers thereof; in the scheme, "heterocycloalkyl" is a 4-6 membered heterocycloalkyl group, wherein the heteroatom is selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3.

[0046] The term "alkoxy" refers to a group having -O-alkyl, i.e. an alkyl group as defined above connected via an oxygen atom to a parent core structure. Phrases containing this term, for example, "C1-C3 alkoxy" means that the alkyl portion contains 1 to 3 carbon atoms.

[0047] The term "aryl" refers to an aromatic hydrocarbon group derived from an aromatic ring compound by removing a hydrogen atom, which can be a monocyclic aromatic group, a condensed aromatic group, or a polycyclic aromatic group. For polycyclic rings, at least one is an aromatic ring system. Preferred aryl groups are 6-10 membered aromatic groups, which can be selected from phenyl and naphthyl as examples.

[0048] The terms "halo", "halogen", "halo" refer to a halogen atom or substituted by a halogen atom, including a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

[0049] In various parts of the present invention, connecting substituents are described. When the structure clearly needs a linking group, the Markush variables listed for the group should be understood as a linking group. For example, if the structure needs a linking group and the Markush group definition for the variable lists "alkyl" or "aryl", it should be understood that the "alkyl" or "aryl" represents the alkylene group or arylene group connected respectively. In some specific structures, when the alkyl group is clearly expressed as a linking group, the alkyl group represents the alkylene group connected, for example, the alkyl in the group "-C1-C3 haloalkyl" should be understood as an alkylene group.

[0050] In addition, the term “including” is an open limitation rather than a closed one, that is, it includes the contents specified in the present invention but does not exclude other contents.

[0051] Unless otherwise specified, the present invention uses conventional methods such as mass spectrometry and nuclear magnetic resonance to identify compounds, and each step and condition can refer to conventional operating steps and conditions in the art.

[0052] Those skilled in the art will appreciate that, according to the conventions used in the art, the structural formulas used in this application to describe groups It means that the corresponding group is connected with other fragments and groups in the compound through this site. Without violating the common sense in the art, the above-mentioned preferred conditions can be combined arbitrarily to obtain the preferred embodiments of the present invention.

[0053] The reagents and raw materials used in the present invention are commercially available.

[0054] Intermediate Ma:

[0055]

[0056] Synthesis route:

[0057]

[0058] Compound Ma-1 (300 mg, 2.22 mmol) and triethylamine (449 mg, 4.44 mmol) were added to dichloromethane (10 mL), and compound Ma-2 (694 mg, 2.44 mmol) was slowly added under ice bath conditions, followed by stirring at 20°C for 2 hours. After the reaction was completed as detected by LCMS and TLC, water (100 mL) was added to quench the reaction. After extraction with dichloromethane, the organic phase was dried and concentrated, and then purified by column chromatography to obtain a yellow solid compound Ma-3 (530 mg, yield 62.3%).

[0059] Compound Ma-3 (530 mg, 1.38 mmol) and potassium carbonate (286 mg, 2.08 mmol) were added to N,N-dimethylformamide (10 mL), and compound Ma-4 (241 mg, 1.66 mmol) was slowly added under ice bath conditions, and then stirred at 20°C for 2 hours. After the reaction was completed by LCMS and TLC, water (100 mL) was added to quench the reaction, and the organic phase was dried and concentrated after extraction with ethyl acetate, and then purified by column chromatography to obtain yellow solid compound Ma-5 (520 mg, yield 94.2%). LCMS: [M+H] + =400.1.

[0060] Compound Ma-5 (520 mg, 1.30 mmol), compound Ma-6 (198 mg, 2.61 mmol) and elemental iodine (331 mg, 1.30 mmol) were added to ethanol (10 mL) and stirred at 90°C overnight. After the reaction was completed by LCMS and TLC, 1 M sodium hydroxide solution was added to adjust the pH of the solution to between 12 and 13. After extraction with ethyl acetate, the organic phase was dried and concentrated, and then purified by column chromatography to obtain a gray solid compound Ma-7 (300 mg, yield 64.9%). LCMS: [M+H] + =356.1.

[0061] Compound Ma-7 (300 mg, 0.85 mmol) and potassium carbonate (233 mg, 1.69 mmol) were added to N,N-dimethylformamide (10 mL), and di-tert-butyl dicarbonate (221 mg, 1.0 mmol) was slowly added under ice bath conditions, and then stirred at 20°C for 2 hours. After the reaction was completed by LCMS and TLC, water (50 mL) was added, and the organic phase was dried and concentrated after extraction with ethyl acetate, and then purified by column chromatography to obtain yellow solid compound Ma (270 mg, yield 70.1%). LCMS: [M+H] + =456.1.

[0062] Embodiment 1:

[0063]

[0064] Synthesis route:

[0065]

[0066] Referring to the synthesis of intermediate Ma, compound Ma-2 was replaced with an equal amount of N-tert-butoxycarbonyl-4-acetylchloropiperidine to obtain intermediate Mb.

[0067] Compound Mb (7.73 mmol), pyridine (6.1 g, 77.3 mmol) and 4-dimethylaminopyridine (189 mg, 1.55 mmol) were added to dichloromethane (40 mL), and a solution of p-bromobenzoyl chloride 1-1 (2.0 g, 9.28 mol) in dichloromethane (10 mL) was slowly added under ice bath conditions, and then stirred at 20°C overnight. After the reaction was completed, the mixture was concentrated and purified by column chromatography to obtain solid compound 1-2.

[0068] Compound 1-2 (5.3 mmol), compound 1-3 (6.4 mmol), [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (0.53 mmol) and sodium carbonate (10.6 mmol) were added to a mixed solution of 1,4-dioxane (40 mL) and water (10 mL), stirred at 100°C overnight, and after LCMS and TLC detection, the reaction was concentrated and purified by column chromatography to synthesize compound 1-4, which was then deprotected to obtain yellow solid compound 1 (17.9 mg, yield 29.2%). LCMS [M+H] + =568.1.

[0069] 1 H NMR (400MHz, DMSO-d6) δ12.88(s,1H),9.01(s,1H),8.80(s,1H),8.25(d,J=8.4Hz,2H),8.04-7.99(m,4H),7.93(d,J=8.0H z,2H),7.85(d,J=8.0Hz,2H),7.80(s,1H),7.41(d,J=7.2Hz,2H),3.17-3.11(m,1H),2.69-2.53(m,4H),1.86-1.68(m,4H).

[0070] Embodiment 2:

[0071]

[0072] Synthesis route:

[0073]

[0074] Referring to Example 1, Compound 1-3 was replaced with an equal amount of Compound 2-2 to synthesize yellow solid Compound 2 (67.8 mg, yield 72.4%). LCMS [M+H] + =572.1.

[0075] 1H NMR (400MHz, DMSO-d6) δ12.90(s,1H),8.79(s,1H),8.56(s,1H),8.26(d,J=8.4Hz,2H),8.08-8.04(m,4H),7.94(d,J=8.0Hz,4H),7.81(s,1H), 7.43(d,J=6.8Hz,2H),4.34(q,J=7.2Hz,2H),3.36(s,1H),3.15(d,J=8.8Hz,2H),2.70-2.64(m,2H),1.81-1.75(m,4H),1.34(t,J=7.2Hz,3H).

[0076] Embodiment 3:

[0077]

[0078] Synthesis route:

[0079]

[0080] Referring to Example 1, compound Mb was replaced with an equal amount of Mf (ie, Ma), and compound 1-3 was replaced with an equal amount of 3-3 to synthesize yellow solid compound 3 (140.5 mg, yield 86.1%). LCMS [M+H] + =608.1.

[0081] 1 H NMR (400MHz, DMSO-d6) δ8.24(d,J=8.0Hz,2H),8.06(d,J=8.4Hz,2H),7.96-7.91(m,6H),7.86(d,J=8.0Hz,2H),7.71(s,1H),7.50(d,J=8.8 Hz, 2H), 4.33 (q, J = 7.2Hz, 2H), 3.07 (d, J = 4.4Hz, 3H), 2.60-2.54 (m, 2H), 1.93 (d, J = 12.0Hz, 2H), 1.63-1.54 (m, 2H), 1.33 (t, J = 7.2Hz, 3H).

[0082] Embodiment 4:

[0083]

[0084] Synthesis route:

[0085]

[0086] Compound 3 (50 mg, 0.08 mmol) was added to a mixed solution of tetrahydrofuran (1 mL) and ethanol (1 mL), and then an aqueous solution (1 mL) of sodium hydroxide (3.6 mg, 0.09 mmol) was added, and stirred at 45 °C overnight. After the reaction was completed by LCMS and TLC, the mixture was concentrated and purified by column chromatography to obtain a yellow solid compound 4 (12.8 mg, yield 27%). LCMS: [M+H] + =580.1.

[0087] 1 H NMR (400MHz, DMSO-d6) δ8.13(d,J=8.0Hz,2H),7.94(d,J=8.4Hz,2H),7.86-7.81(m,6H),7.73(d,J=8.0Hz,2H),7.59 (s,1H),7.37(d,J=8.8Hz,2H),3.02(d,J=4.4Hz,3H),2.49-2.43(m,2H),1.86(d,J=12.0Hz,2H),1.54-1.45(m,2H).

[0088] Embodiment 5:

[0089]

[0090] Synthesis route:

[0091]

[0092] Referring to Example 1, compound 1-2 was replaced with an equal amount of 3-2 to synthesize yellow solid compound 5 (2.21 g, yield 46.8%). LCMS [M+H] + =604.1.

[0093] 1 H NMR (400MHz, DMSO-d6) δ8.25(d,J=8.0Hz,2H),8.00(d,J=8.0Hz,2H),7.94(t,J=9.2Hz,4H),7.86(d,J=8.0Hz,2H),7.72(s,1H ), 7.50 (d, J = 8.4Hz, 2H), 3.05 (d, J = 12.8Hz, 2H), 2.59-2.52 (m, 2H), 2.43-2.38 (m, 1H), 1.94-1.88 (m, 2H), 1.61-1.52 (m, 2H).

[0094] Embodiment 6:

[0095]

[0096] Synthesis route:

[0097]

[0098] Compound 5 (100 mg, 0.166 mmol) and potassium carbonate (30 mg, 0.22 mmol) were added to N,N-dimethylformamide (5 mL), and then iodomethane (25.6 mg, 0.18 mmol) was added, and stirred at 20°C overnight. After the reaction was completed by LCMS and TLC, water (20 mL) was added, and the organic phase was dried and concentrated after extraction with ethyl acetate, and then purified by column chromatography to obtain white solid compound 6 (80.8 mg, yield 78.9%). LCMS: [M+H] + =618.1.

[0099] 1 H NMR (400MHz, DMSO-d6) δ12.88(s,1H),8.25(d,J=8.4Hz,2H),8.01-7.92(m,6H),7.86(d,J=8.4Hz,2H),7.73(s,1H),7 .51(d,J=8.8Hz,2H),2.65-2.62(m,1H),2.41-2.36(m,2H),2.33-2.29(m,3H),2.02-1.99(m,3H),1.75-1.67(m,3H).

[0100] Embodiment 7:

[0101]

[0102] Synthesis route:

[0103]

[0104] Referring to Example 6, methyl iodide was replaced with an equal amount of ethyl iodide to synthesize yellow solid compound 7 (84.2 mg, yield 78.6%). LCMS [M+H] + =646.1. 1 H NMR (400MHz, DMSO-d6) δ12.88(s,1H),8.27(d,J=7.6Hz,2H),8.04–7.84(m,8H),7.73(s,1H),7.51(d,J=8.0Hz,2H),3.29–3.21 (m,1H),2.95–2.85(m,2H),2.28–2.13(m,2H),2.01–1.84(m,4H),1.70–1.56(m,2H),1.47–1.34(m,2H),0.82(t,J=7.2Hz,3H).

[0105] Embodiment 8:

[0106]

[0107] Synthesis route:

[0108]

[0109] Referring to Example 6, methyl iodide was replaced with an equal amount of 1-bromopropane to synthesize yellow solid compound 8 (56.2 mg, yield 62.2%). LCMS [M+H] + =660.1.

[0110] 1 H NMR (400MHz, DMSO-d6) δ12.88(s,1H),8.26(d,J=7.6Hz,2H),8.00(d,J=7.6Hz,2H),7.94(t,J=7.6Hz,4H),7.86(d,J=8.0Hz,2H ),7.72(s,1H),7.50(d,J=7.6Hz,2H),3.23-3.19(m,1H),2.88(d,J=10.8Hz,2H),2.21(t,J=7.2Hz,2H),1.93-1.84(m,4H),1.66 -1.57(m,2H),1.39 -1.32(m,2H),1.28 -1.19(m,2H),0.84(t,J=7.2Hz,3H).

[0111] Embodiment 9:

[0112]

[0113] Synthesis route:

[0114]

[0115] Referring to Example 6, methyl iodide was replaced with an equal amount of 1-fluoro-2-iodoethane to synthesize yellow solid compound 9 (82.3 mg, yield 50.8%). LCMS [M+H] + =647.1.

[0116] 1H NMR (400MHz, DMSO-d6) δ12.89(s,1H),8.26(d,J=8.4Hz,2H),8.00(d,J=8.0Hz,2H),7.94(t ,J=8.8Hz,4H),7.86(d,J=8.0Hz,2H),7.73(s,1H),7.51(d,J=8.0Hz,2H),4.54(t,J=4.8Hz, 1H),4.42(t,J=4.8Hz,1H),3.32-3.26(m,4H),2.93(d,J=11.6Hz,2H),2.62(t,J=4.8Hz,1H ), 2.55 (t, J = 4.8Hz, 1H), 2.04 (t, J = 12.0Hz, 2H), 1.93 (d, J = 12.0Hz, 2H), 1.67-1.59 (m, 2H).

[0117] Embodiment 10:

[0118]

[0119] Synthesis route:

[0120]

[0121] Referring to Example 6, methyl iodide was replaced with an equal amount of 1-fluoro-3-iodopropane to synthesize yellow solid compound 10 (45.3 mg, yield 53.2%). LCMS [M+H] + =664.1.

[0122] 1 H NMR (400MHz, DMSO-d6) δ12.88(s,1H),8.26(d,J=8.0Hz,2H),8.00(d,J=8.0Hz,2H),7.9 5(t,J=8.4Hz,4H),7.86(d,J=8.4Hz,2H),7.72(s,1H),7.51(d,J=8.0Hz,2H),4.50(t,J= 6.0Hz,1H),4.38(t,J=6.0Hz,1H),3.28–3.21(m,1H),2.90(d,J=11.2Hz,2H),2.33(t,J= 6.8Hz,2H),1.99–1.85(m,4H),1.82–1.75(m,1H),1.75–1.70(m,1H),1.68–1.55(m,2H).

[0123] Embodiment 11:

[0124]

[0125] Synthesis route:

[0126]

[0127] Referring to Example 6, methyl iodide was replaced with an equal amount of acetyl chloride to synthesize yellow solid compound 11 (13.9 mg, yield 16.1%). LCMS [M+H] + =646.1.

[0128] 1 H NMR(400MHz,DMSO-d6)δ12.89(s,1H),8.27(d,J=8.0Hz,2H),8.02(d,J=8.0Hz,2H), 7.96(t,J=9.2Hz,4H),7.87(d,J=8.4Hz,2H),7.75(s,1H),7.53(d,J=8.0Hz,2H),4.4 3(d,J=13.2Hz,1H),3.87(d,J=12.8Hz,1H),3.62(t,J=11.2Hz,1H),3.07(t,J=12.4 Hz,1H),2.63–2.55(m,1H),2.06–1.93(m,6H),1.71–1.51(m,1H),1.51–1.36(m,1H).

[0129] Embodiment 12:

[0130]

[0131] Synthesis route:

[0132]

[0133] Referring to Example 6, methyl iodide was replaced with an equal amount of acryloyl chloride to synthesize yellow solid compound 12 (22.7 mg, yield 19.8%). LCMS [M+H] + =658.1.

[0134] 1H NMR (400MHz, DMSO-d6) δ12.89(s,1H),8.27(d,J=8.4Hz,2H),8.03–7.92(m,6H),7.86(d,J=8.0H z,2H),7.74(s,1H),7.53(d,J=8.4Hz,2H),6.79(dd,J=16.8,10.4Hz,1H),6.09(dd,J=16.8,2.0 Hz,1H),5.66(dd,J=10.4,2.0Hz,1H),4.48(d,J=11.2Hz,1H),4.12(d,J=10.4Hz,1H),3.66(t,J =12.0Hz,1H),3.10(t,J=12.8Hz,1H),2.75–2.64(m,1H),2.09–1.98(m,2H),1.59–1.41(m,2H).

[0135] Embodiment 13:

[0136]

[0137] Synthesis route:

[0138]

[0139] Referring to Example 1, the compound Mb was replaced with an equal amount of Mg (ie, Ma) to obtain compound 13-5, and then referring to Example 6, methyl iodide was replaced with an equal amount of 1-fluoro-2-iodoethane to synthesize yellow solid compound 13 (82.3 mg, yield 50.8%). LCMS [M+H] + =647.1.

[0140] 1 H NMR (400MHz, DMSO-d6) δ12.89(s,1H),8.26(d,J=8.4Hz,2H),8.00(d,J=8.0Hz,2H),7.94(t ,J=8.8Hz,4H),7.86(d,J=8.0Hz,2H),7.73(s,1H),7.51(d,J=8.0Hz,2H),4.54(t,J=4.8Hz, 1H),4.42(t,J=4.8Hz,1H),3.32-3.26(m,4H),2.93(d,J=11.6Hz,2H),2.62(t,J=4.8Hz,1H ), 2.55 (t, J = 4.8Hz, 1H), 2.04 (t, J = 12.0Hz, 2H), 1.93 (d, J = 12.0Hz, 2H), 1.67-1.59 (m, 2H).

[0141] Embodiment 14:

[0142]

[0143] Synthesis route:

[0144]

[0145] Referring to Example 9, Compound 5 was replaced with an equal amount of Compound 3 to synthesize yellow solid Compound 14 (82.3 mg, yield 50.8%). LCMS [M+H] + =647.1.

[0146] 1 H NMR (400MHz, DMSO-d6) δ12.89(s,1H),8.26(d,J=8.0Hz,2H),8.07(d,J=8.0Hz,2H) ,7.95(t,J=8.8Hz,6H),7.73(s,1H),7.51(d,J=8.8Hz,2H),4.54(t,J=4.8Hz,1H),4 .42(t,J=4.8Hz,1H),4.34(q,J=7.2Hz,2H),3.28–3.21(m,1H),2.93(d,J=11.2Hz,2 H),2.75–2.49(m,2H),2.16–1.87(m,4H),1.74–1.55(m,2H),1.34(t,J=7.2Hz,3H).

[0147] Embodiment 15:

[0148]

[0149] Synthesis route:

[0150]

[0151] Referring to Example 3, the compound Ma-4 was replaced with an equal amount of methyl iodide, the compound 3-2 was replaced with an equal amount of 15-1, and the compound 3-3 was replaced with an equal amount of 15-2 to synthesize 15-4. Then, referring to Example 6, methyl iodide was replaced with an equal amount of 1-fluoro-2-iodoethane to synthesize the yellow solid compound 15 (14.9 mg, yield 18.6%). LCMS [M+H] + =665.1.

[0152] 1H NMR(400MHz,DMSO-d6)δ12.90(s,1H),8.26(d,J=8.0Hz,2H),8.15(s,1H),8.00-7.94(m,6H),7.73(s,1H),7.51(d,J=8.4Hz,2H),4 .54(t,J=4.8Hz,1H),4.42(t,J=4.8Hz,1H),3.29-3.21(m,4H),2.93(d,J=10.8Hz,2H),2.75-2.49(m,2H),2.12-1.87(m,4H),1.67 -1.59(m,2H).

[0153] Embodiment 16:

[0154]

[0155] Synthesis route:

[0156]

[0157] Referring to Example 15, compound 15-2 was replaced with an equal amount of 16-2 to synthesize yellow solid compound 16 (5.0 mg, yield 6%). LCMS [M+H] + =665.1.

[0158] 1 H NMR(400MHz,DMSO-d6)δ12.88(s,1H),8.24-8.22(m,2H),8.02(s,1H),7.96-7.94(m,2H),7.84-7.82(m,2H),7.72-7.71(m,2H),7.67-7.65(m ,2H),7.51-7.49(m,2H),4.56(s,1H),4.44(s,1H),3.33(s,3H),2.95- 2.87(m,1H),2.33-2.31(m,1H),1.96-1.93(m,2H),1.67-1.64(m,4H).

[0159] Embodiment 17:

[0160]

[0161] Synthesis route:

[0162]

[0163] Referring to Example 15, compound 15-2 was replaced with an equal amount of 17-2 to synthesize yellow solid compound 17 (14.7 mg, yield 17.9%). LCMS [M+H]+ =681.1.

[0164] 1 H NMR(400MHz,DMSO-d6)δ12.90(s,1H),8.26(d,J=7.6Hz,2H),8.00-7.94(m,5H),7.92-7.83(m,2H),7.73(s,1H),7.51(d,J=8.0Hz,2H),4.54 (t,J=4.8Hz,1H),4.42(t,J=4.8Hz,1H),3.31-3.24(m,4H),2.93(d,J=10.8Hz,2H),2.72-2.49(m,2H),2.12-1.87(m,4H),1.68-1.58(m,2H).

[0165] Embodiment 18:

[0166]

[0167] Synthesis route:

[0168]

[0169] Referring to Example 3, compound 3-1 was replaced with an equal amount of 18-1 to obtain compound 18-2, which was then deprotected to synthesize yellow solid compound 18 (25 mg, yield 94.3%). LCMS [M+H] + =478.1.

[0170] 1 H NMR(400MHz,DMSO-d6)δ7.97-7.93(m,4H),7.65(s,1H),7.61-7.55(m,1H),7.49-7.44(m,3H),3. 15(s,1H),3.00(d,J=12.0Hz,2H),2.47-2.45(m,2H),1.88(d,J=11.6Hz,2H),1.56-1.46(m,2H).

[0171] Embodiment 19:

[0172]

[0173] Synthesis route:

[0174]

[0175] Referring to Example 9, Compound 5 was replaced with an equal amount of Compound 18 to synthesize yellow solid Compound 19 (11.8 mg, yield 15.4%). LCMS [M+H]+ =524.1.

[0176] 1 H NMR (400MHz, DMSO-d6) δ12.89(s,1H),7.98-7.93(m,4H),7.74(s,1H),7.63-7.58(m,1H),7.51-7.48(m,3H),4.54(t,J=4.4Hz, 1H), 4.42 (t, J = 4.4Hz, 1H), 3.27-3.26 (m, 1H), 2.93 (d, J = 11.2Hz, 2H), 2.71-2.49 (m, 2H), 2.12-1.84 (m, 4H), 1.67-1.58 (m, 2H).

[0177] Embodiment 20:

[0178]

[0179] Synthesis route:

[0180]

[0181] Referring to Example 12, Compound 5 was replaced with an equal amount of Compound 18 to synthesize yellow solid Compound 20 (11.6 mg, yield 20.8%). LCMS [M+H] + =532.1.

[0182] 1 H NMR(400MHz,DMSO-d6)δ7.98-7.93(m,4H),7.72(s,1H),7.63-7.57(m,1H),7.52-7.46(m,3H),6.81-6.75(m,1H),6.17-5.5 8(m,2H),4.56–4.03(m,2H),3.68-3.62(m,1H),3.12-3.06(m,1H),2.70-2.66(m,1H),2.06-1.99(m,2H),1.51-1.44(m,2H).

[0183] Effects of COUP inhibitor on the proliferation activity of prostate cancer cells

[0184] 1. Methods

[0185] Prostate cancer cells PC-3 and DU-145 were cultured at 1.5×10 4The cells were inoculated in a 96-well plate at a density of 1 / well and cultured overnight at 5% CO2 and 37°C. The next day, the compound was diluted 3-fold with a maximum concentration of 100 μM and added to the cell culture medium for a total of 10 concentrations for a further 24 h, divided into 2 replicate wells.

[0186] After the cells were treated with the compounds for 24 h, the The cell viability was determined according to the instructions of the AQueous One Solution Cell Proliferation Assay Kit (Promega, G3580). The method was briefly described as follows: 20 μL of MTS reagent was added to each well, mixed and incubated in an incubator at 37°C for 1 hour, and the absorbance was determined using an MD i3x multi-function microplate reader at a wavelength of 490 nM.

[0187] The effect of the compound on cell proliferation was evaluated by cell viability, and the IC50 value was calculated using GraphPad Prism 8.0 software with three-parameter fitting of the dose-response curve. Cell viability = (signal value of the sample - signal value of the medium control) / (signal value of the DMSO control - signal value of the medium control) × 100%. Inhibition ratio = (1-cell viability) × 100%.

[0188] 2. Results

[0189]

[0190]

[0191] *: IC50>10μM; **: 10μM≥IC50>1μM; ***: 1μM≥IC50>0.1μM; ****: 0.1μM≥IC50

[0192] 30≥Max; 50≥Max>30; 70≥Max>50; Max>70

[0193] Summary: The compounds provided by the present invention generally have good inhibitory activity on the proliferation of prostate cancer cells.

[0194] Detection of the inhibitory activity of compounds against COUP-TF II based on reporter gene activity assay

[0195] 1. Methods

[0196] 1.1 Plasmid co-transfection into HEK293T cells and compound treatment

[0197] One day before plasmid transfection, HEK293T cells were cultured at 1×10 4 The density of 100 μg / well was inoculated into a 96-well plate. HD (Promega, #E2311) instructions for cell transfection. The main steps are: taking a single well as an example, add 20 ng and 5 ng of plasmid pCR3.1-COUP-TFII and pXP2-NGFIA-Luc to 10 μL of Opti-MEM TM I culture medium (Gibco, #11058021) and mix well; then add 0.2 μL of HD, mix well and let stand at 20℃ for 5min; then add this 10μL mixture to the cell well containing 100μL culture medium. 6h after cell co-transfection, the compound was diluted with half-log dilution multiples at 100μM as the highest concentration, and added to the cell culture medium for treatment for 24h, divided into 2 duplicate wells.

[0198] 1.2One-Glo Luciferase Assay

[0199] After the cells were treated with the compounds for 24 h, the ONE- Luciferase Assay System (Promega, #E6120) instructions were used for detection. The main steps were: 50 μL of culture medium was discarded from each well, and 50 μL of ONE- Luciferase reagent, shake at 20°C for 10 min; take 80 μL of the cleavage reaction solution to a white opaque optiPlate-96 well plate, and use the MD i3x multifunctional microplate reader to detect the luminescent signal value (Firefly-Luc) of firefly luciferase. The Firefly-Luc signal value was used as the inhibitory activity of the compound on COUP-TF II, and was normalized with the ratio of the solvent DMSO group. The GraphPad Prism8.0 software was used to fit the dose-response curve with three parameters to calculate the EC50 value. The inhibition rate was calculated using the following formula: inhibition rate = (1-Luc of the compound-treated sample / Luc of the DMSO group) × 100%. VCT-8 was used as a positive control compound (WO2019 / 222134).

[0200]

[0201] 2. Results

[0202] The experimental data are shown in the table below.

[0203]

[0204]

[0205] *: 20μM≥EC50>5μM; **: 5μM≥EC50>1μM; ***: 1μM≥EC50>0.1μM;

[0206] ****:0.1μM≥EC50

[0207] 20≥Max; 40≥Max>20; 70≥Max>40; Max>70

[0208] Summary: The compounds provided by the present invention generally have good inhibitory activity against COUP-TFII.

[0209] Detection of the inhibitory activity of compounds on NRF2 based on reporter gene activity assay

[0210] 1. Methods

[0211] 1.1 Plasmid co-transfection into HEK293T cells and compound treatment

[0212] pGL4.37[luc2P / ARE] (Promega, #E3641) and pRL-TK (Promega, #E2241) plasmids were purchased from Promega; the plasmids were transformed into DH5α Escherichia coli using the CaCl2 method, and the corresponding plasmid DNA was purified using a plasmid extraction kit (TIANGEN, #D107) after further culture and amplification. The day before plasmid transfection, HEK293T cells were cultured at 1×10 4 The density of 100 μg / well was inoculated into a 96-well plate. HD (Promega, #E2311) instructions for cell transfection. The main steps are: Taking a single well as an example, add plasmids pGL4.37[luc2P / ARE] and pRL-TK at a ratio of 80ng and 8ng to 10μL Opti-MEM TM I culture medium (Gibco, #11058021) and mix well; then add 0.2 μL of HD, mix well and let stand at 20℃ for 5min; then add this 10μL mixture to the cell well containing 100μL culture medium. 6h after cell co-transfection, the compound was diluted with half-log dilution multiples at 100μM as the highest concentration, and added to the cell culture medium for treatment for 24h, divided into 2 duplicate wells.

[0213] 1.2 Dual-Glo Luciferase Assay

[0214] After the cells were treated with the compounds for 24 h, the Luciferase Assay System (Promega, #E2940) instructions were used for detection. The main steps were: 50 μL of culture medium was discarded from each well, and 50 μL of Dual- Luciferase reagent, shake at 20℃ for 10min; take 80μL of cleavage reaction solution to a white opaque optiPlate-96 well plate, and use MD i3x multifunctional microplate reader to detect the luminescent signal value (Firefly-Luc) of firefly luciferase; then add 40μL Dual- Stop& Reagent, shake at 20℃ for 10min; then use MD i3x multifunctional microplate reader to detect the luminescent signal value (Renilla-Luc) of Renilla luciferase. The ratio of Firefly-Luc / Renilla-Luc was used as the inhibitory activity of the compound on NRF2, and the ratio of the solvent DMSO group was normalized. The EC50 value was calculated using GraphPad Prism8.0 software with three-parameter fitting of the dose-response curve. The inhibition rate was calculated using the following formula: inhibition rate = (1-the ratio of Firefly-Luc / Renilla-Luc of the compound-treated sample / the ratio of Firefly-Luc / Renilla-Luc of the DMSO group) × 100%.

[0215] 2. Results

[0216] The experimental data are shown in the table below.

[0217]

[0218] *: EC50>20μM; **: 20μM≥EC50>10μM; ***: 10μM≥EC50>1μM;

[0219] 20≥Max; 40≥Max>20; 70≥Max>40; Max>70

[0220] Summary: The compounds provided by the present invention also have good inhibitory activity against NRF2.

[0221] Pharmacokinetic evaluation

[0222] In mice, the bioavailability and pharmacokinetic behavior of the compound were evaluated. Six male ICR mice of similar weight were selected, of which three mice were given a single oral gavage of 10 mg / kg, and three mice were given a single dose of 1 or 5 mg / kg intravenously. Blood samples were collected at 5 minutes (intravenous administration), 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours and 7 hours after administration, and the concentration of plasma samples was analyzed by LC-MS / MS, and the pharmacokinetic parameters of the compound were analyzed using the PKSolver free tool and non-compartmental model (NCA) software.

[0223] Experimental protocol:

[0224] Experimental animals: Each compound test group included 6 healthy male ICR mice, 18-25 g, purchased from Charles River, and randomly divided into 2 groups, 3 mice in each group.

[0225] Preparation: Weigh a certain amount of compound and add it to 2% DMSO + 15% Solutol + 83% saline to prepare a clear solution.

[0226] Dosage: ICR mice were fasted overnight and given the compound at a 10 mg / kg oral dose or 1 or 5 mg / kg intravenous dose. The oral and intravenous administration volumes were 10 mL / kg and 1 or 5 mL / kg, respectively. Food was given uniformly 2 hours after administration.

[0227] Sample collection: About 30 μL of blood was collected from the great saphenous vein at 5 minutes (intravenous administration), 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours and 7 hours after administration. The blood was placed in a commercial test tube containing K2-EDTA, and then the blood samples were centrifuged at 4°C, 4600 rpm for 5 minutes to obtain plasma samples, and then all plasma samples were quickly frozen on dry ice and kept at -70°C until LC-MS / MS analysis.

[0228] Sample preparation: 10 μL of plasma sample was aspirated and precipitated with a methanol solution containing 50 nmol / L α-naphthoflavone (internal standard). The mixture was thoroughly mixed and centrifuged at 14000 rpm for 5 min at 4°C. Then 75 μL of the supernatant was mixed with 75 μL of methanol for LC-MS / MS analysis.

[0229] The pharmacokinetic parameters are shown in Table 1.

[0230] Table 1

[0231]

[0232] Conclusion: This class of compounds has good absorption in mice, slow elimination, high exposure, and high bioavailability, which warrants further study.

[0233] The applicant declares that the present invention illustrates the phenylthiazolyl benzamide compounds and their applications through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A phenylthiazolylbenzamide compound, characterized in that: The phenylthiazolyl benzamide compound has a structure shown in the following formula I: Wherein R1 is selected from R a , R b Independently selected from -Me, -CD3, Any of the following: The R f Selected from The R g Selected from hydrogen, -Me, -n-Pr, -n-Bu, Any of the following: R2 is selected from hydrogen or halogen; R3 is selected from hydrogen or The R i Any one selected from -CF3, -COOH or -COOEt; The R j Selected from -F or -Cl; Indicates the attachment site of a group.

2. The phenylthiazolylbenzamide compound according to claim 1, characterized in that: The R2 is selected from hydrogen or -F.

3. The phenylthiazolylbenzamide compound according to claim 1, characterized in that: The phenylthiazolylbenzamide compound is selected from any one of the following formulas 1 to 20:

4. A stereoisomer of the phenylthiazolylbenzamide compound according to any one of claims 1 to 3, a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the same.

5. The stereoisomer of the phenylthiazolylbenzamide compound, its pharmaceutically acceptable salt or a pharmaceutical composition comprising the same according to claim 4, characterized in that: The pharmaceutical composition also includes pharmaceutically acceptable excipients.

6. Use of the phenylthiazolyl benzamide compound according to any one of claims 1 to 3 or the stereoisomer of the phenylthiazolyl benzamide compound according to claim 4 or 5, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same in the preparation of a medicament for treating and / or preventing prostate cancer associated with excessive COUP-TFII activity or overexpression of COUP-TFII.

7. Use of a phenylthiazolyl benzamide compound according to any one of claims 1 to 3 or a stereoisomer of a phenylthiazolyl benzamide compound according to claim 4 or 5, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same in the preparation of a medicament for treating and / or preventing prostate cancer associated with excessive NRF2 activity or overexpression.

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