Synthesis method of WRN inhibitor
WRN inhibitor compound A was prepared by Suzuki coupling, chlorosilanization, hydrolysis and condensation reactions, which solved the problem of lack of efficient synthesis methods in the existing technology and achieved high yield and feasibility for industrial production.
Patent Information
- Application Number
- CN202511082694.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-04
- Publication Date
- 2025-11-14
AI Technical Summary
The lack of efficient, stable, and scalable synthetic methods for WRN inhibitor compounds hinders their application in the treatment of MSI-H/dMMR tumors.
WRN inhibitor compound A was prepared by using Suzuki coupling, chlorosilanization, hydrolysis, oxidation, and condensation reactions, with specific solvents, catalysts, and bases selected. The specific steps included coupling of compounds I and II, chlorosilanization of compound III, hydrolysis of compound IV, and condensation of compound VI. The reaction conditions were optimized to improve the yield.
This invention provides a simple, reproducible, and high-yield synthetic method suitable for industrial production, filling the gap in the preparation of WRN inhibitor compounds and possessing significant market value and practical significance.
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Figure CN120943824A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to a method for synthesizing a WRN inhibitor, belonging to the field of pharmaceutical and chemical technology. Background Technology
[0002] Malignant tumors are a major threat to human life and health worldwide, and innovative treatment strategies have always been a core challenge in medical research. In recent years, precision medicine targeting specific molecular targets has brought breakthroughs in the field of oncology. Among them, Werner syndrome protein (WRN), as an ATP-dependent helicase, plays a crucial role in the DNA repair pathway. Studies have confirmed that WRN protein activity is essential for maintaining cancer cell survival in malignant tumor cells with microsatellite instability-high (MSI-H) or DNA mismatch repair system (dMMR) defects. This characteristic establishes a synthetic lethal effect between WRN and MSI malignant tumors, namely, inhibiting WRN can selectively kill such cancer cells while having minimal impact on normal cells. This mechanism makes WRN an important target for the treatment of MSI-H / dMMR tumors.
[0003] Based on the above findings, the development of small molecule inhibitors targeting WRN has become a new direction in anticancer drug development, and several WRN inhibitors have already been reported. Glaxosmithkline has developed a series of ATP-dependent helicase (WRN) inhibitors for Wilmer syndrome, with exemplary structures shown in WRN inhibitor 5:
[0004]
[0005] These compounds can effectively inhibit WRN helicase activity. Preclinical studies have shown that they have significant antitumor activity against MSI-H / dMMR type cancers (including colorectal cancer, endometrial cancer, etc.) and have important translational value.
[0006] However, despite the proven biological value of WRN inhibitors, no published literature or patents have disclosed complete synthetic routes for compounds such as WRN inhibitor 5. The lack of efficient, stable, and scalable preparation methods in current technology severely hinders subsequent pharmacological research, formulation development, and clinical application of these compounds. Therefore, developing a simple, high-yield, and purity-controllable synthetic process is not only a key technological requirement for advancing the drug development of these inhibitors but also a prerequisite for achieving industrial-scale production. Summary of the Invention
[0007] In view of the above technical background and to solve the problems in the prior art, the purpose of this invention is to provide a method for preparing WRN inhibitors (compound A) that is simple to operate, has good reproducibility, and a high overall yield, as shown in the following reaction:
[0008]
[0009] Where R1 is arbitrarily selected from C 1-6 Alkyl group; preferably, R1 is arbitrarily selected from methyl, ethyl, n-propyl, isopropyl, and tert-butyl;
[0010] R2 is selected from the following groups:
[0011]
[0012] R3 is selected from H and C. 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 The alkoxy group, -O-CH2-R6;
[0013] When R3 is not H, R4 and R5 are independently selected from H and C. 1-6 alkyl, C 1-6 The alkoxy group and halogen; when R3 is H, R4 and R5 are independently selected from H and C. 1-6 alkyl, C 1-6 Alkyl groups, halogens, or R4 and R5 together constitute C 5-7 The cycloalkyl group, wherein one or more carbon atoms of the cycloalkyl group, namely R4 and R5, can be replaced by oxygen;
[0014] R6 is selected from C 3-6 cycloalkyl and aryl groups;
[0015] X is selected from Cl, Br, and I.
[0016] The specific steps for this route are as follows:
[0017] (1) Compound I and compound II were coupled via Suzuki coupling to obtain compound III;
[0018] (2) Compound III reacts with trimethylchlorosilane and sodium iodide to give compound IV;
[0019] (3) Compound IV was hydrolyzed in the presence of a base to give compound V;
[0020] (4) In the presence of an oxidant and a base, compound V reacts to give compound VI;
[0021] (5) Compound VI and compound VII react in the presence of a base and a condensing agent to obtain target compound A.
[0022] Further, in step (1), the reaction solvent is selected from any one of water, ether, amide solvents or a mixture thereof, preferably any one of water, dioxane, tetrahydrofuran, dimethylformamide or a mixture thereof, and more preferably a mixed solvent of water and dioxane.
[0023] Further, in step (1), the catalyst is selected from tetra(triphenylphosphine)palladium PdP4, PdCl2, Pd(dppf)Cl2, Pd(OAc)2, and preferably tetra(triphenylphosphine)palladium PdP4.
[0024] Furthermore, in step (1), the alkali is selected from cesium carbonate, potassium carbonate, and sodium carbonate, preferably potassium carbonate.
[0025] Further, in step (1), the molar ratio of compound I, compound II and base is 1:(0.9-1.1):(2-5).
[0026] Furthermore, in step (1), the reaction temperature is selected from 60 to 100°C, preferably 70 to 90°C.
[0027] Furthermore, in step (1), the reaction time is preferably 8 to 16 hours.
[0028] Further, in step (2), the reaction solvent is selected from one or any mixture of water, ether solvents, nitrile solvents, haloalkane solvents, amide solvents, and sulfone solvents; the ether solvent is, for example, tetrahydrofuran; the nitrile solvent is, for example, acetonitrile; the haloalkane solvent is, for example, dichloromethane or dichloroethane; the amide solvent is, for example, N,N-dimethylformamide; and the sulfone solvent is, for example, dimethyl sulfoxide or sulfolane. The reaction solvent is preferably a nitrile solvent, and more preferably acetonitrile.
[0029] Furthermore, the reaction solvent is preferably acetonitrile.
[0030] Furthermore, in step (2), the molar ratio of compound III, trimethylchlorosilane and sodium iodide is 1:(3-10):(3-10).
[0031] Furthermore, in step (2), the reaction temperature is preferably room temperature.
[0032] Furthermore, in step (2), the reaction time is preferably 1 to 5 hours.
[0033] Furthermore, in step (3), the alkali is selected from sodium hydroxide, potassium hydroxide, and lithium hydroxide; preferably lithium hydroxide.
[0034] Furthermore, in step (3), the molar ratio of compound IV to the base is selected from 1:(0.1~1).
[0035] Furthermore, in step (3), the reaction time is preferably 8 to 16 hours.
[0036] Furthermore, in step (3), the reaction temperature is selected from 30 to 80°C, preferably 40 to 60°C.
[0037] Further, in step (3), the reaction solvent is selected from one or any mixture of water, alcohol solvents, ether solvents, ester solvents, haloalkane solvents, and amide solvents; the alcohol solvent is, for example, methanol, ethanol, or isopropanol; the ether solvent is, for example, tetrahydrofuran or 1,4-dioxane; the ester solvent is, for example, ethyl acetate; the haloalkane solvent is, for example, dichloromethane or dichloroethane; and the amide solvent is, for example, N,N-dimethylformamide. The reaction solvent is preferably an ether solvent, and more preferably tetrahydrofuran.
[0038] Furthermore, in step (4), the oxidant is selected from any one of potassium persulfate, potassium peroxymonosulfate, ammonium persulfate, and hydrogen peroxide, preferably potassium persulfate.
[0039] Furthermore, in step (4), the molar ratio of compound V to oxidant is selected from 1:(1 to 10), preferably 1:(2 to 5).
[0040] Further, in step (4), the alkali is selected from one or more of sodium hydroxide, potassium hydroxide, and lithium hydroxide, preferably potassium hydroxide; more preferably, the potassium hydroxide is an aqueous solution of potassium hydroxide.
[0041] Furthermore, in step (4), the reaction temperature is preferably room temperature.
[0042] Furthermore, in step (4), the reaction time is preferably 8 to 16 hours.
[0043] Further, in step (4), the post-reaction treatment includes: filtration, adjusting the pH of the filtrate to acidic with 20% sulfuric acid, adding 20% sulfuric acid again to adjust the pH to acidic after the solid precipitates, heating and stirring, cooling to precipitate the solid, filtration, and washing.
[0044] Furthermore, in step (5), the condensing agent is any one of the onium salt condensing agents, such as HATU, HBTU, HCTU, TBTU; preferably HATU.
[0045] Further, in step (5), the base is selected from one or more of triethylamine, diisopropylethylamine, pyridine, tri-n-butylamine, tert-butylamine or N-methylmorpholine, preferably diisopropylethylamine.
[0046] Furthermore, in step (5), the molar ratio of compound VI to compound VII is 1:(1-5), preferably 1:(1-3).
[0047] Further, in step (5), the molar ratio of compound VI to condensing agent and base is 1:(2-5):(0.1-0.5).
[0048] Furthermore, in step (5), the reaction temperature is preferably room temperature.
[0049] Furthermore, in step (5), the reaction time is preferably 8 to 16 hours.
[0050] Further, in step (5), the reaction solvent is selected from one or any mixture of sulfone solvents and amide solvents, such as dimethyl sulfoxide and sulfolane; the amide solvent is such as N,N-dimethylformamide. Further, the reaction solvent is preferably any one or a mixture of dimethyl sulfoxide and N,N-dimethylformamide.
[0051] Furthermore, compound VII can optionally be obtained using any conventional chiral resolution method via the following route:
[0052]
[0053] A second aspect of the present invention provides a series of intermediate compounds III-1, IV-1, V-1, or VI-1, with the following structural formulas:
[0054]
[0055] R1 is defined as above.
[0056] A third aspect of the present invention provides a method for preparing WRN inhibitor compound A, wherein WRN inhibitor compound A is further prepared by compound III-1, compound IV-1, compound V-1 or compound VI-1.
[0057] Beneficial technical effects of the present invention:
[0058] 1. Through creative labor, this invention synthesizes a novel intermediate for WRN inhibitor compound A and proposes a feasible method for preparing WRN inhibitor compound A. This is the first route for this compound and fills a gap in its preparation.
[0059] 2. Through creative labor, the present invention designs a novel route suitable for preparing compound A; using the novel intermediate provided by the present invention and by confirming key process parameters, the present invention obtains WRN inhibitor compound A in unexpectedly high yield, and the operation is simple, reproducible, and suitable for industrial-scale production.
[0060] 3. The route proposed in this invention is simple to operate, does not require harsh reaction conditions, and uses a mild reaction to obtain the final product. The synthetic route has a low risk factor and is conducive to industrial-scale production. Therefore, it has good market value and far-reaching practical significance. Attached Figure Description
[0061] Figure 1 For compound III-1 1 HNMR spectrum.
[0062] Figure 2 For compound V-1 1 HNMR spectrum.
[0063] Figure 3 For compound VI-1 1 HNMR spectrum. Detailed Implementation
[0064] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that the following detailed description of the technical solutions of the present invention using embodiments will help to further understand the advantages and effects of the technical solutions of the present invention. The embodiments do not limit the scope of protection of the present invention, which is determined by the claims.
[0065] Unless otherwise specified, the experimental methods described in the following examples are generally performed under standard conditions or as recommended by the manufacturer.
[0066] Unless otherwise specified, all raw materials and reagents used in the examples are commercially available. Unless otherwise indicated, the reagents are used directly without purification. All solvents are purchased from commercial suppliers and are ready for use without treatment.
[0067] Example 1
[0068]
[0069] Under argon protection, 8 g of compound I-1 and 9 g of compound II-1 were dissolved in dioxane / water (8 / 1200 mL), and 16 g of potassium carbonate and 2 g of tetrakis(triphenylphosphine)palladium were added. The mixture was incubated overnight at 80 °C. After the reaction was complete, the mixture was extracted with water and dichloromethane, evaporated to dryness, and purified by column chromatography (PE:DCM = 10:1) to give 13 g of a colorless oil, with a yield of 94.28%. Compound III-1... 1 HNMR spectra as follows Figure 1 As shown. 1H NMR (400MHz, DMSO-d6) δ8.13(d,J=7.9Hz,1H),7.94(dd,J=7.7,1.8Hz,1H),7.70(d,J=7.9Hz,1H),747-737(m ,5H),7.32(t,J=7.2Hz),7.27(d,J=7.8Hz,1H),7.11(t,J=7.9Hz,1H),5.24(s,2H),3.98(s,3H),3.81(s,3H).
[0070] Example 3
[0071]
[0072] Under argon protection, 13 g of compound III-1 was dissolved in 200 mL of acetonitrile, and 28 g of sodium iodide and 20 g of trimethylchlorosilane were added. The mixture was reacted at room temperature for 2 h. After the reaction was complete, it was extracted with water and dichloromethane, and evaporated to dryness to obtain 13 g of crude solid product, which was used directly in the next step. [M+H] + =151.1.
[0073] Example 4
[0074]
[0075] Under argon protection, 13 g of compound IV-1 and 77 mL of 2M lithium hydroxide solution were added to 150 mL of tetrahydrofuran. The mixture was stirred overnight at 50 °C. After the reaction was complete, the mixture was washed with water and dichloromethane. The pH of the aqueous phase was adjusted to acidic with 1M HCl, and the solution was filtered to give 11 g of a white solid. The yield was 90%. (The last sentence appears to be incomplete and possibly refers to the reaction of compound V-1.) 1 HNMR spectra as follows Figure 2 As shown. 1 H NMR(400MHz,DMSO-d6)δ14.81(s,1H),13.34(s,1H),8.41(s,1H),7.60-7.47(m,2H),7.44 (d,J=70Hz,2H),741-726(m,4H),7.13(d,J=15.6Hz,1H),6.83(d,J=76Hz,1H),5.19(s,2H)
[0076] Example 5
[0077]
[0078] Under argon protection, 11 g of compound V-1 was added to 200 mL of 10% potassium hydroxide aqueous solution, followed by the addition of 19 g of potassium persulfate at 5°C. The reaction was allowed to proceed overnight at room temperature, and the mixture was filtered directly after the reaction was complete. The filtrate was adjusted to acidic pH with 20% sulfuric acid, precipitating a solid. The mixture was stirred for 2 hours, and then adjusted to acidic pH again with 20% sulfuric acid. The mixture was stirred overnight at 100°C. After the reaction was complete, the temperature was lowered to 0°C, precipitating a solid. The solid was filtered, washed with water and ether, yielding 10 g of a brownish-yellow solid. The yield was 90%. (The text abruptly ends here, likely due to an incomplete translation or a missing section.) 1 HNMR spectra as follows Figure 3 As shown. 1 HNMR(400MHz,DMSO-d6)δ15.64(s,1H),13.22(s,1H),9.61(s,1H),7.47-7.39(m, 4H),7.37-7.28(m,4H,7.22(d,J=8.3Hz,1H),7.07(t,J=7.4Hz,1H),5.15(s,2H).
[0079] Example 6
[0080]
[0081] Under argon protection, 10 g of compound VI-1 and 6 g of compound VI were dissolved in 200 mL of dimethyl sulfoxide, followed by the addition of 13 g of diisopropylethylamine and 28 mL of propyl phosphate cyclic anhydride. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was extracted with water and dichloromethane, evaporated to dryness, and purified by column chromatography (DCM:MEOH = 50:1, DCM:MEOH = 10:1) to give 10 g of a yellow solid. Yield: 75%.
[0082] The above are merely preferred embodiments of the present invention and are not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.
Claims
1. A method for preparing compound A: in, R1 is selected from C 1-6 Alkyl groups; R2 is selected from the following groups: R3 is selected from H and C. 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 The alkoxy group, -O-CH2-R6; When R3 is H, R4 and R5 are independently selected from H and C. 1-6 alkyl, C 1-6 alkoxy, halogen or R4 and R5 Together they constitute C 5-7 The cycloalkyl group, wherein one or more carbon atoms of the cycloalkyl group, namely R4 and R5, can be replaced by oxygen; R6 is selected from C 3-6 cycloalkyl and aryl groups; X is selected from Cl, Br, and I; Includes the following steps: (1) Compound I and compound II were coupled via Suzuki coupling to obtain compound III; (2) Compound III reacts with trimethylchlorosilane and sodium iodide to give compound IV; (3) Compound IV was hydrolyzed in the presence of a base to give compound V; (4) In the presence of an oxidant and a base, compound V reacts to give compound VI; (5) Compound VI and compound VII react under the conditions of alkali and condensing agent to obtain compound A.
2. The method as described in claim 1, characterized in that, In step (1), one or more of the following conditions must be met: The reaction solvent is selected from any one of water, ethers, amides, or a mixture thereof, preferably from any one of water, dioxane, tetrahydrofuran, dimethylformamide, or a mixture thereof, and more preferably from a mixed solvent of water and dioxane; and / or, the catalyst is selected from tetra(triphenylphosphine)palladium PdP4, PdCl2, Pd(dppf)Cl2, Pd(OAc)2, preferably tetra(triphenylphosphine)palladium PdP4; And / or, the base is selected from cesium carbonate, potassium carbonate, sodium carbonate, preferably potassium carbonate; And / or, the molar ratio of compound I, compound II and base is 1:(0.9–1.1):(2–5); And / or, the reaction temperature is selected from 60 to 100°C, preferably 70 to 90°C; And / or, the reaction time is preferably 8 to 16 hours.
3. The method as described in claim 1, characterized in that, In step (2), one or more of the following conditions must be met: The reaction solvent is selected from one or any mixture of water, ether solvents, nitrile solvents, haloalkane solvents, amide solvents, and sulfone solvents, preferably nitrile solvents, and more preferably acetonitrile; And / or, the molar ratio of compound III, trimethylchlorosilane and sodium iodide is 1:(3-10):(3-10); And / or, the reaction time is preferably 1 to 5 hours.
4. The method as described in claim 1, characterized in that, In step (3), one or more of the following conditions must be met: The molar ratio of compound IV to the base is 1:(0.1~1); And / or, in step (3), the alkali is selected from sodium hydroxide, potassium hydroxide, and lithium hydroxide, preferably lithium hydroxide; And / or, the reaction time is preferably 8 to 16 hours; And / or, the reaction temperature is selected from 30 to 80°C, preferably 40 to 60°C; And / or, the reaction solvent is selected from one or any mixture of water, alcohol solvents, ether solvents, ester solvents, haloalkane solvents, and amide solvents, preferably ether solvents, and more preferably tetrahydrofuran.
5. The method according to any one of claims 1 to 4, characterized in that, The oxidant in step (4) is selected from any one of potassium persulfate, potassium peroxymonosulfate, ammonium persulfate, and hydrogen peroxide; potassium persulfate is preferred.
6. The method as described in claim 5, characterized in that, In step (4), the molar ratio of compound V to oxidant is 1:(1-10), preferably 1:(2-5).
7. The method as described in claim 5, characterized in that, In step (4), the alkali is selected from one or more of sodium hydroxide, potassium hydroxide, and lithium hydroxide; preferably, it is potassium hydroxide. And / or, preferably, the reaction time is 8 to 16 hours; And / or, preferably, the post-reaction treatment includes: filtration, adjusting the pH of the filtrate to acidic with 20% sulfuric acid, adjusting the pH to acidic again with 20% sulfuric acid after the solid precipitates, heating and stirring, cooling to precipitate the solid, filtration, and washing.
8. The method according to claims 1 to 4, characterized in that, In step (5), one or more of the following conditions must be met: The condensing agent is any one of the onium salt condensing agents; And / or, preferably, the condensing agent is selected from one or more of HATU, HBTU, HCTU, and TBTU; And / or, preferably, the condensing agent is HATU; And / or, the base is selected from one or more of triethylamine, diisopropylethylamine, pyridine, tri-n-butylamine, tert-butylamine or N-methylmorpholine; preferably diisopropylethylamine; And / or, the molar ratio of compound VI to compound VII is 1:(1-5), preferably 1:(1-3); And / or, the molar ratio of compound VI, condensing agent and base is 1:(2-5):(0.1-0.5); And / or, the reaction time is 8–16 hours; And / or, the reaction solvent is selected from one of sulfone solvents, amide solvents, or any mixture thereof; preferably, dimethyl sulfoxide, N,N-dimethylformamide, or a mixture thereof.
9. The structural formulas of compounds III-1, IV-1, V-1, and VI-1 are as follows: in, R1 is selected from C 1-6 Alkyl groups.
10. A method for preparing WRN inhibitor compound A, comprising further preparing WRN inhibitor compound A by means of compound III-1, compound IV-1, compound V-1 or compound VI-1 as described in claim 9.