Terphenyl compounds, preparation methods and applications thereof

Abstract

The present invention relates to the field of pharmaceutical technology, specifically, to terphenyl compounds, their preparation methods, and their applications. Terphenyl compounds include the compound represented by Formula 1: #imgabs0#, wherein R1 is selected from H, R2 is selected from halogen and C1-C6 alkyl, and R3 is selected from a substituted secondary amine group. These compounds exhibit potent inhibitory effects on PD-1 / PD-1 and can be used as small molecule PD-1 / PD-1 inhibitors, thereby potentially treating tumors.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and in particular to terphenyl compounds, preparation methods and applications thereof. Background Art

[0002] Cancer immunotherapy has developed rapidly in recent years. Immune checkpoint inhibitors targeting programmed cell-death receptor 1 (PD-1) and programmed cell-death ligand 1 (PD-L1) have achieved remarkable success in clinical oncology. Within the tumor microenvironment, the binding of PD-1 in tumor-infiltrating lymphocytes to PD-L1 expressed on the surface of tumor cells enables tumor immune evasion and promotes tumor growth. Currently, 15 antibody drugs targeting PD-1 / PD-L1 have been approved for marketing by the US Food and Drug Administration (FDA) and Chinese drug regulatory authorities, including nine PD-1 monoclonal antibodies and six PD-L1 monoclonal antibodies. However, inherent limitations of antibody drugs, including poor tissue and tumor penetration, long half-life, poor oral bioavailability, high production costs, and immunogenicity, have severely hampered the effectiveness of immunotherapy. Small molecule inhibitors targeting PD-1 / PD-L1 are receiving increasing attention due to their better pharmacokinetic characteristics and oral availability, and their ability to be used in combination with antibody drugs or even directly replace antibody drugs.

[0003] However, the development of small molecule inhibitors of PD-1 / PD-L1 lags far behind that of monoclonal antibodies, with only a few entering clinical trials. Early on, the lack of structural information on the target hampered the rational design of small molecule inhibitors for PD-1 / PD-L1. Following the publication of the crystal structure of the hPD-1 / hPD-L1 complex in 2015, a growing number of patents for antibodies, cyclic peptides, and small molecule inhibitors targeting PD-1 and PD-L1 were published. Among these, the most notable is a class of 2-methyl-3-biphenyl derivatives disclosed by Bristol-Myers Squibb (BMS). Research by Hoalk's group discovered that these compounds induce polymerization of the PD-L1 protein, allowing two PD-L1 monomers to form a cylindrical hydrophobic pocket, providing a druggable target for the design of small molecule drugs. Consequently, small molecule inhibitors targeting PD-L1 have become a hot topic of research. However, to date, no small molecule inhibitors targeting PD-L1 have been approved for marketing, demonstrating that the development of novel small molecule inhibitors for PD-1 / PD-1 remains a challenging task and represents an unmet clinical need.

[0004] In view of this, the present invention is proposed. Summary of the Invention

[0005] The present invention aims to provide terphenyl compounds, preparation methods and applications thereof. The terphenyl compounds provided in the embodiments of the present invention have a good inhibitory effect on PD-1 / PD-1 and can be used as PD-1 / PD-1 small molecule inhibitors, thereby achieving the effect of treating tumors.

[0006] The present invention is achieved in that:

[0007] In a first aspect, the present invention provides a terphenyl compound, which includes a compound represented by Formula 1: Formula 1, wherein R1 is selected from H, R2 is selected from halogen and C1-C6 alkyl; and R3 is selected from substituted secondary amine groups.

[0008] In an optional embodiment, R2 is selected from any one of F, Cl, Br, I and C1-C3 alkyl.

[0009] In an optional embodiment, R2 is selected from any one of Cl, Br, methyl and ethyl.

[0010] In an alternative embodiment, the substituted secondary amine group is selected from , wherein n represents any integer from 1 to 5, and R4 is selected from hydroxyl, acetamide and piperazinone groups.

[0011] In an optional embodiment, the terphenyl compound is selected from any one of the compounds represented by the following structural formulas:

[0012]

[0013] In a second aspect, the present invention provides a method for preparing the terphenyl compound described in the aforementioned embodiment, comprising: synthesizing according to the following synthesis route:

[0014]

[0015] In an optional embodiment, the process comprises: in step a, the molar ratio of 1-bromo-3-iodo-2-methylbenzene to phenylboric acid is 1:1-2, the catalyst used is a palladium catalyst, the temperature is 80-100° C., and the time is 8-12 hours;

[0016] The conditions of step b include: the catalyst used is a palladium catalyst, the temperature is 70-90°C, and the time is 10-16 hours;

[0017] The conditions of step c include: the catalyst used is a palladium catalyst, the temperature is 80-100° C., and the time is 8-12 hours;

[0018] The step d comprises: a temperature of 20-30° C. and a time of more than 12 hours.

[0019] In a third aspect, the present invention provides a PD-1 / PD-L1 inhibitor, which includes the terphenyl compound described in the aforementioned embodiment, or a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof.

[0020] In a fourth aspect, the present invention provides a use of the terphenyl compound described in the aforementioned embodiment, or a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof in the preparation of a PD-1 / PD-L1 inhibitor.

[0021] In a fifth aspect, the present invention provides a use of the terphenyl compound described in the aforementioned embodiment, or a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof in the preparation of an anti-tumor drug.

[0022] The present invention has the following beneficial effects: the terphenyl compounds provided in the embodiments of the present invention have a good inhibitory effect on PD-1 / PD-L1, can be used as PD-1 / PD-1 small molecule inhibitors, and then have the effect of treating tumors. DETAILED DESCRIPTION

[0023] To make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are described clearly and completely below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer are used. Where the manufacturer of the reagents or instruments is not specified, all are conventional products that can be purchased commercially.

[0024] The present invention provides a terphenyl compound, which includes a compound represented by Formula 1: Formula 1, wherein R1 is selected from H, R2 is selected from halogen and C1-C6 alkyl; and R3 is selected from substituted secondary amine groups.

[0025] Specifically, R2 is selected from any one of F, Cl, Br, I and C1-C3 alkyl. C1-C6 alkyl can be any one of methyl, ethyl, n-propyl, isopropyl, tert-butyl and the like.

[0026] The substituted secondary amine group is selected from wherein n represents any integer from 1 to 5, and R4 is selected from a hydroxyl group, an acetamide group, and a piperazinone group.

[0027] Specifically, the terphenyl compound is selected from any one of the compounds represented by the following structural formulas:

[0028]

[0029] Furthermore, the present invention provides a method for preparing the terphenyl compound described in the aforementioned embodiment, the preparation method comprising: using 1-bromo-3-iodo-2-methylbenzene and phenylboronic acid as starting materials, performing a three-step Suzuki-Miyaura coupling reaction to obtain a key intermediate 4; reacting intermediate 4 with the corresponding amine via a Schiff base, and finally reducing to obtain the target product.

[0030] The synthesis was carried out according to the following synthesis route:

[0031]

[0032] Reagents and conditions in the synthesis route: (a) phenylboronic acid, palladium catalysts such as Pd(PPh3)4, carbonates such as K2CO3, organic solvents such as toluene / water (v / v=2:1), 80-100°C under nitrogen, 8-12 h; (b) B2Pin2, palladium catalysts such as PdCl2(dtbpf), CH3COOK, 70-90°C under nitrogen, 10-16 h; (c) palladium catalysts such as Pd(PPh3)4, carbonates such as K2CO3, organic solvents such as toluene / water (V:V=2:1), 80-100°C under nitrogen, 8-12 h; (d) i: raw materials containing secondary amine groups, organic solvents such as CH2Cl2 / CH3OH (v / v=1:1), glacial acetic acid, room temperature, 2 h; ii: boron substances such as NaCNBH3, overnight at room temperature.

[0033] In a third aspect, the present invention provides a PD-1 / PD-L1 inhibitor, which includes the terphenyl compound described in the aforementioned embodiment, or a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof.

[0034] In a fourth aspect, the present invention provides a use of the terphenyl compound described in the aforementioned embodiment, or a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof in the preparation of a PD-1 / PD-L1 inhibitor.

[0035] In a fifth aspect, the present invention provides a use of the terphenyl compound described in the aforementioned embodiment, or a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof in the preparation of an anti-tumor drug.

[0036] The features and performance of the present invention are further described in detail below with reference to the embodiments.

[0037] Example 1

[0038] An embodiment of the present invention provides a terphenyl compound, the structural formula of which is shown below:

[0039] This embodiment also provides a method for synthesizing the terphenyl compound, comprising:

[0040] (1) Synthesis of 3-bromo-2-methyl-1,1'-biphenyl

[0041] 1-Bromo-3-iodo-2-methylbenzene (1.18 g, 4.0 mmol), phenylboronic acid (0.59 g, 4.8 mmol), and tetrakistriphenylphosphine palladium (0.23 g, 0.2 mmol) were dissolved in 30 ml of a mixture of toluene and water (volume ratio 2:1). Potassium carbonate (1.1 g, 8.0 mmol) was then added to the mixture. The mixture was reacted at 90°C under a nitrogen atmosphere for 9 h. TLC confirmed the reaction was complete. The reaction solution was extracted three times with ethyl acetate (30 ml). The combined organic phases were washed twice with 5 ml of water and then 5 ml of saturated brine, then dried over anhydrous Na₂SO₄. After filtration, the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography to obtain 0.82 g of a colorless liquid in an 83.2% yield.

[0042] The product characterization data are as follows: 1 H NMR (400MHz, DMSO) δ7.86-7.61 (m, 3H), 7.48-7.37 (m, 4H), 7.21 (t, J=8.4Hz, 1H), 2.31 (s, 3H).

[0043] (2) Synthesis of 4,4,5,5-tetramethyl-2-(2-methyl-[1,1'-biphenyl]-3-yl)-1,3,2-dioxaborane

[0044] 3-Bromo-2-methyl-1,1'-biphenyl (0.82 g, 3.33 mmol), pinacol diboron (1.0 g, 4.0 mmol), and 1,1'-bis(di-tert-butylphosphino)ferrocenedichloropalladium (0.1 g, 0.167 mmol) were dissolved in 15 ml of DMSO. Potassium acetate (0.962 g, 9.8 mmol) was then added. The mixture was allowed to react at 80°C under a nitrogen atmosphere for 12 hours. TLC confirmed the reaction was complete. The reaction solution was extracted three times with ethyl acetate (30 ml). The combined organic phases were washed sequentially with 5 ml of water and twice with 5 ml of saturated brine, then dried over anhydrous Na2SO4. After filtration, the solvent was removed under reduced pressure to yield 0.645 g of a colorless liquid, which was purified by silica gel column chromatography in a 66.5% yield.

[0045] The product characterization data are as follows: 1H NMR (400MHz, DMSO) δ7.92-7.60 (m, 4H), 7.49-7.45 (m, 3H), 7.15 (dd, J=7.4, 1.7Hz, 1H), 2.33 (s, 3H), 1.37 (d, J=8.0Hz, 12H).

[0046] (3) Synthesis of 3-chloro-2'-methyl-[1,1':3',1"-triphenyl]-4-carbaldehyde

[0047] 4,4,5,5-Tetramethyl-2-(2-methyl-[1,1'-biphenyl]-3-yl)-1,3,2-dioxaborolane (0.645 g, 2.19 mmol), 4-bromo-2-chlorobenzaldehyde (0.41 g, 1.9 mmol), and tetrakistriphenylphosphine palladium (0.11 g, 0.095 mmol) were dissolved in 30 ml of a mixture of toluene and water (volume ratio 2:1). Potassium carbonate (0.656 g, 4.75 mmol) was then added to the mixture. The mixture was reacted at 90°C under a nitrogen atmosphere for 12 hours. TLC confirmed the reaction was complete. The reaction solution was extracted three times with ethyl acetate (30 ml). The combined organic phases were washed twice with 5 ml of water and then 5 ml of saturated brine, then dried over anhydrous Na2SO4. After filtration, the solvent was removed under reduced pressure to yield 0.34 g of a white solid, which was purified by silica gel column chromatography in a yield of 58.5%.

[0048] The product characterization data are as follows: 1 H NMR (400MHz, DMSO) δ10.08 (s, 1H), 8.46 (d, J = 1.7Hz, 1H), 7.98-7.60 (m, 4H), 7.56-7.46 (m, 6H), 2.31 (s, 3H).

[0049] (4) Synthesis of N-(2-(((3-chloro-2'-methyl-[1,1':3',1"-triphenyl]-4-yl)methyl)amino)ethyl)acetamide

[0050] 3-Chloro-2'-methyl-[1,1':3',1"-triphenyl]-4-carbaldehyde (0.34 g, 1.1 mmol) and N-acetylethylenediamine (0.135 g, 1.32 mmol) were dissolved in 10 ml of a mixed solution of dichloromethane and methanol (volume ratio 1:1), and then 0.07 ml of glacial acetic acid was added. After reacting at room temperature for 2 h, sodium cyanoborohydride (0.242 g, 3.85 mmol) was added, and the reaction was stirred at room temperature overnight. TLC detection showed that the reaction was complete. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column to obtain 0.33 g of a white solid with a yield of 76.6%.

[0051] The product characterization data are as follows: 1H NMR (400MHz, DMSO) δ8.47 (d, J=1.8Hz, 1H), 7.71 (d, J=8.5Hz, 2H) 7.68-7.63 (m, 3H), 7.52 (t, J=8.5Hz, 1H), 7.49-7.4 5 (m, 3H), 7.38 (d, J=8.4Hz, 1H), 4.06 (s, 2H), 3.48 (t, J=7.1Hz, 2H), 2.82 (t, J=7.1Hz, 2H), 2.23 (s, 3H), 1.86 (s, 3H).

[0052] Examples 2-6

[0053] The corresponding terphenyl compounds were synthesized according to the synthesis method of Example 1. The structures and characterization data of the prepared terphenyl compounds are as follows:

[0054] Example 2: 1 H NMR (400MHz, DMSO) δ8.21 (d, J=1.9Hz, 1H), 7.72 (d, J=8.5Hz, 2H) 7.68-7.63 (m, 3H), 7.52 (t, J=8.5Hz, 1H), 7.48-7.45 (m, 3H), 7.36 (d, J = 8..3Hz, 1H), 4.02 (s, 2H), 3.48 (t, J = 7.1Hz, 2H), 2.82 (t, J = 7.1Hz, 2H), 2.23 (s, 3H), 1.86 (s, 3H).

[0055] Example 3: 1 H NMR (400MHz, DMSO) δ8.47 (d, J=1.8Hz, 1H), 7.71 (d, J=8.5Hz, 2H) 7.64-7.61 (m, 3H), 7.52 (t, J=8.5Hz, 1H), 7.4 8-7.45 (m, 3H), 7.38 (d, J=8..4Hz, 1H), 4.05 (s, 2H), 3.51 (t, J=6.7Hz, 2H), 2.79 (t, J=6.7Hz, 2H), 2.23 (s, 3H).

[0056] Example 4: 1H NMR (400MHz, DMSO) δ8.21 (d, J=1.9Hz, 1H), 7.72-7.70 (m, 2H) 7.65-7.61 (m, 3H), 7.52 (t, J=8.5Hz, 1H), 7.48 -7.45 (m, 3H), 7.36 (d, J = 8.3Hz, 1H), 4.00 (s, 2H), 3.51 (t, J = 6.7Hz, 2H), 2.78 (t, J = 6.7Hz, 2H), 2.23 (s, 3H).

[0057] Example 5: 1 H NMR (400MHz, DMSO) δ8.47 (d, J=1.8Hz, 1H), 7.72-7.70 (m, 2H) 7.65-7.61 (m, 3H), 7.52 (t, J=8.5Hz, 1H), 7.48-7.45 (m , 3H), 7.38 (d, J=8.4Hz, 1H), 4.06 (s, 2H), 3.55 (d, J=15.8Hz, 2H), 3.40-3.12 (m, 2H), 2.95-2.57 (m, 6H), 2.23 (s, 3H).

[0058] Example 6: 1 H NMR (400MHz, DMSO) δ8.21 (d, J=1.9Hz, 1H), 7.74-7.70 (m, 2H) 7.65-7.61 (m, 3H), 7.52 (t, J=8.5Hz, 1H), 7.48-7.46 (m , 3H), 7.36 (d, J=8.3Hz, 1H), 4.02 (s, 2H), 3.55 (d, J=15.8Hz, 2H), 3.40-3.13 (m, 2H), 2.95-2.57 (m, 6H), 2.23 (s, 3H).

[0059] Test Example 1

[0060] The following compounds were subjected to Autodock Vina molecular docking experiments to detect the binding energy between the following compounds and the PD-L1 protein. The results are shown in Table 1.

[0061]

[0062] Table 1

[0063]

[0064]

[0065] As shown in Table 1, by changing the groups provided by the compounds of the present invention, their binding ability is reduced, and they cannot bind well, and thus cannot effectively inhibit PD-1 / PD-L1 binding.

[0066] Test Example 2

[0067] 1. Test of compound's inhibitory activity against PD-1 / PD-L1

[0068] The in vivo anti-tumor activity of some of the compounds was tested using homogeneous time-resolved fluorescence (HTRF). Using the HTRF PD-1 / PD-L1 binding assay kit developed by Cisbio, the terphenyl derivatives provided in the examples of the present invention were tested for their inhibitory effects on PD-1 / PD-L1 according to the manufacturer's instructions.

[0069] Experimental procedure: In a 96-well plate, add 2 μl of diluent or target compound diluted with diluent to each well, then add 4 μl of PD-1 protein and 4 μl of PD-L1 protein to each well, incubate at room temperature for 15 minutes, and then add 10 μl of Lanti-Tag1-Eu to each well. 3+ After incubation with anti-Tag2-XL665 at room temperature for 1 hour, the fluorescence signals at 665 nm and 620 nm were detected. HRTR ratio = (665 nm / 620 nm) × 10 4 Each compound was tested at 8 concentrations, with 3 replicates per concentration. The inhibition rate and IC were then calculated. 50 , experimental data are shown in Table 2.

[0070] Table 2 Inhibitory activity of compounds against PD-1 / PD-L1

[0071] Compound <![CDATA[IC 50 (nM)]]> Example 1 7.3 Example 2 6.8 Example 3 15.4 Example 4 13.1 Example 5 4.5 Example 6 2.5 BMS202 24.3

[0072] As shown in Table 2, the inhibitory effects of the terphenyl derivatives provided in Examples of the present invention on PD-1 / PD-L1 were determined using HTRF technology according to standard operating procedures, with compound BMS202 used as a reference. The results show that the terphenyl derivatives provided in Examples 1-6 of the present invention all had significant inhibitory effects on PD-1 / PD-L1. The compounds provided in Examples of the present invention can be used to treat immune diseases such as hepatitis, HIV, and tumors.

[0073] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the present invention.

Claims

1. A terphenyl compound, characterized in that: It includes the compound shown in formula 1: Wherein, R1 is selected from H, R2 is selected from halogen and C1-C6 alkyl; R3 is selected from wherein n represents any integer from 1 to 5, R4 is selected from a hydroxyl group, an acetamide group, and a piperazinone group, and the N located at the meta position of the carbonyl group in the piperazinone group is connected to the alkylene represented by n.

2. The terphenyl compound according to claim 1, characterized in that R2 is selected from any one of F, Cl, Br, I and C1-C3 alkyl.

3. The terphenyl compound according to claim 2, characterized in that R2 is selected from any one of Cl, Br, methyl and ethyl.

4. The terphenyl compound according to claim 1, characterized in that The terphenyl compound is selected from any one of the compounds represented by the following structural formulas:

5. A method for preparing the terphenyl compound according to claim 1, characterized in that: include: The synthesis was carried out according to the following synthesis route:

6. The preparation method according to claim 5, characterized in that include: In step a, the molar ratio of 1-bromo-3-iodo-2-methylbenzene to phenylboric acid is 1:1-2, the catalyst used is a palladium catalyst, the temperature is 80-100° C., and the time is 8-12 hours; The conditions of step b include: the catalyst used is a palladium catalyst, the temperature is 70-90°C, and the time is 10-16 hours; The conditions of step c include: the catalyst used is a palladium catalyst, the temperature is 80-100° C., and the time is 8-12 hours; The step d comprises: a temperature of 20-30° C. and a time of more than 12 hours.

7. A PD-1 / PD-L1 inhibitor, characterized in that The invention comprises the terphenyl compound according to claim 1 and a pharmaceutically acceptable salt thereof.

8. Use of the terphenyl compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a PD-1 / PD-L1 inhibitor.

9. Use of the terphenyl compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of an anti-tumor drug.

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