Diarylpyrimidine compounds, their preparation methods and uses

Abstract

This invention belongs to the fields of medicinal chemistry and pharmaceutical technology, and particularly relates to diarylpyrimidine compounds, their preparation methods, and uses. The diarylpyrimidine compounds of this invention underwent microtubule target activity testing, binding site confirmation, and in vitro and in vivo anti-cervical cancer pharmacological activity testing. Experimental results show that, in vitro, the diarylpyrimidine compounds inhibit microtubule polymerization by binding to the colchicine binding site of microtubules, thereby inhibiting the proliferation, cell cycle, apoptosis, cloning, and migration of cervical cancer cells. In vivo, the diarylpyrimidine compounds can be used in combination with cisplatin to further enhance the in vivo anti-cervical cancer efficacy and reduce the nephrotoxicity caused by high-dose cisplatin use, while also exhibiting good drug-like properties. The compounds and their salts of this invention can be used to prepare microtubule colchicine binding site inhibitors or in combination with cisplatin for the treatment of cervical cancer.

Description

Technical Field

[0001] This invention belongs to the fields of medicinal chemistry and pharmaceutical technology, and in particular relates to diarylpyrimidine compounds, their preparation methods and uses. Background Technology

[0002] According to the latest statistics released by the International Agency for Research on Cancer (IARC), a branch of the World Health Organization, in 2020 (CA: Acancer Journal for Clinicians, 2018, 68(6): 394-424.), there were approximately 19,292,789 new cases of malignant tumors and 9,958,133 new deaths worldwide in 2020 alone, indicating an increasingly serious situation regarding the incidence of malignant tumors. Among them, cervical cancer had 604,127 new cases and 341,831 new deaths, ranking fourth in both incidence and mortality among malignant tumors in women worldwide. Approximately 84% of cervical cancer cases and 88% of deaths worldwide occur in low-resource and underdeveloped countries, resulting in relatively poor awareness of cervical cancer and its precancerous lesions among women in these countries, as well as generally low vaccination rates and widespread screening rates (The Lancet Global Health, 2020, 8(2): e191-e203.). It is projected that over the next 10 years, the number of new cervical cancer cases worldwide will increase from 600,000 to 700,000 annually, while the number of deaths will increase from 341,000 to 400,000 annually (International Journal of Gynecological Cancer, 2020, 30(4):426-427.). Clearly, cervical cancer not only seriously threatens women's lives and health but has also become a global public health issue that cannot be ignored (Papillomavirus Research, 2019, 8:100170).

[0003] Currently, the main clinical treatments for cervical cancer include five regimens and methods: surgery, radiotherapy, chemotherapy, targeted therapy, and immunotherapy (Chinese Journal of Practical Gynecology and Obstetrics, 2018, 34(06):613-622). Among them, the first-line chemotherapy regimen for cervical cancer mainly uses cisplatin (DDP) alone or in combination with cisplatin (Gynecologic Oncology, 2014, 133(1):117-23). ​​However, this regimen has serious drug resistance and nephrotoxicity in clinical practice, thus hindering its widespread application (Cancer, 1992, (6):479-480). Furthermore, while the two currently available large-molecule biological drugs, bevacizumab and pembrolizumab, can improve patient survival to some extent in clinical treatment, they still have several drawbacks, including immunogenicity, limited treatment population, and high cost (Gynecologic Oncology, 2020, 159(1):142-149.; Expert Opinion on Drug Delivery, 2021, 18(2):187-203.). Therefore, discovering highly effective, low-toxicity, and drug-resistant small-molecule CBSIs for cervical cancer and realizing their clinical application in cervical cancer treatment or in combination with cisplatin is a promising and feasible approach.

[0004] Tubulin is a mature and classic target for anti-tumor drugs. Tubulin heterodimers mainly include seven sites: ① vincaline binding site, ② laulimalide binding site, ③ pirone binding site, ④ maytansine binding site, ⑤ paclitaxel binding site, ⑥ colchicine binding site, and ⑦ the seventh site. Among these, tubulin drugs, represented by paclitaxel (a paclitaxel binding site inhibitor) and vincaline (a vincaline binding site inhibitor), have been marketed, not only improving and prolonging the quality of life and lifespan of many cancer patients but also bringing much hope to their families. However, due to their inherent physicochemical properties and long-term clinical use, they have also revealed defects such as significant side effects and toxicity, and marked multidrug resistance (China Metallurgical Industry Medical Journal, 2011, 28(2):137-139; Cell Motility and the Cytoskeleton, 2003, 55(2):77-96). Compared with paclitaxel and vinblastine binding sites, the colchicine binding site (CBS) of tubulin has unique tumor angiogenesis disruption and anti-drug resistance advantages, and has become one of the hot topics in anti-tumor drug research (Trends in Cell Biology, 2018, 28(10):776-792.; Science advances, 2020, 7:eabg4168.). According to literature reports, during normal microtubule assembly, the tubulin heterodimer changes from its free state "bent" conformation to a "straight" conformation after obtaining energy from GTP, thus completing the normal assembly of the microtubule. During the transformation of tubulin from a curved to a straight conformation, the key difference between the colchicine site and the other six sites lies in the fact that during this transformation, the T7 ring is easily pushed towards the colchicine binding site (occupation site). In other words, the morphological change of the T7 ring prevents small molecule ligands from binding to the colchicine site. Thus, microtubule growth is a dynamic equilibrium process, exhibiting two reversible states of "occupation" and "release" as the T7 ring dynamically moves. Only when the T7 ring releases space and allows small molecule ligands (such as colchicine) to bind can the ligands occupy the colchicine binding site and bind effectively.Therefore, the mechanism of action of colchicine is that when tubulin assembles, small molecule ligands (such as colchicine) competitively bind to the binding site, which restricts the spatial movement of the T7 ring, thus preventing the conformational change from "bent" to "straight". As a result, tubulin cannot assemble into microtubules in a "straight" conformation, thereby disrupting the kinetics of microtubule growth and inhibiting microtubule assembly (J.Med.Chem.,2016,59(19):8685-8711.; PNAS,2009,106(33):13775-13779.). To date, no colchicine binding site inhibitors (CBSIs) have been approved for marketing.

[0005] Tubulin CBSIs have a similar mechanism to colchicine. Significant progress has been made in the research of CBSIs under development, especially in the structural modification of Combretastatin A. CA-4P, CA-1P, and Ombrabulin have entered Phase III clinical trials, and BNC105P has entered Phase II clinical trials for the treatment of various solid tumors. However, these drugs still have certain toxic side effects and are insufficient in prolonging patient survival. Furthermore, there are few clinical studies using cervical cancer as a development target and indication (Journal of Medicinal Chemistry, 2005, 48(12):4087-4099; British journal of haematology, 2020, 189(5):e211-e213; The Lancet Oncology, 2015, 16(5):531-540; Biochemical and Biophysical Research). Communications, 2020, 525(1):148-154.), To date, no small molecule anti-cervical cancer drugs with this structure have been approved for marketing.

[0006] The structure-activity relationship study based on stilbene compound CA-4 is shown in [link to study]. Figure 1 Various modifications to the two aromatic rings and double bond of CA-4 have been reported. Among them, the modification of the double bond of CA-4 is a hot topic of research. Cyclostructochemical modifications to the double bond of CA-4 mainly include three-membered rings (cyclopropane, three-membered epoxides and 2H-azacyclopropane, etc.), four-membered rings (azacyclobutane-2-one and cyclobutane, etc.), five-membered rings (azole five-membered rings, 2(5H)-furanone, cyclopentenone, oxazolone, furan and pyrrole, etc.), six-membered rings (pyrimidine, pyridine, pyrazine and unsaturated six-membered aliphatic rings), seven-membered rings and five-membered and six-membered heterocycles, etc. Summary of the Invention

[0007] Based on the shortcomings of existing colchicine binding site inhibitors, this invention provides diarylpyrimidine compounds, their preparation methods, and uses.

[0008] Specifically, the diarylpyrimidine compounds provided by this invention have been verified to have microtubule polymerization inhibitory activity and antitumor activity, and can be used as a novel microtubule colchicine binding site inhibitor. Based on this research, this invention provides diarylpyrimidine compounds and their preparation methods, as well as the application of compound drugs containing them as one of the components in the preparation of drugs for the prevention and / or treatment of tumor-related diseases. In particular, it provides a technical solution for the application of diarylpyrimidine compounds in combination with cisplatin in the preparation of drugs for the treatment of cervical cancer.

[0009] The objective of this invention can be achieved through the following technical solutions:

[0010] The present invention first provides diarylpyrimidine compounds as shown in formula (I) or formula (II) or pharmaceutically acceptable salts thereof (hereinafter referred to as pharmaceutical salts).

[0011]

[0012] In formula (I),

[0013] X is selected from either a carbon atom or a nitrogen atom;

[0014] R1 is selected from methoxy, ethoxy, methylthio, ethylthio, oxygen atom, nitrogen atom, halogen, formamido, N-methylformamido, N,N-dimethylformamido, or borate group;

[0015] R2 is selected from hydrogen atom, oxygen atom, nitrogen atom, halogen, trifluoromethyl, hydroxyl, amino, nitro, methyl carbonate or ethyl carbonate;

[0016] n is 0 or 1;

[0017] In formula (II),

[0018] R is selected from dimethoxy or trimethoxy groups with different substitution positions;

[0019] R1 is selected from methoxy, ethoxy, methylthio, or ethylthio;

[0020] R2 is selected from hydrogen atom, halogen, hydroxyl, amino or nitro group;

[0021] R3 is selected from methoxy groups.

[0022] In one embodiment of the present invention, the diarylpyrimidine compound represented by formula (I) is specifically selected from the following compounds:

[0023]

[0024]

[0025] In one embodiment of the present invention, the diarylpyrimidine compound represented by formula (II) is specifically selected from the following compounds:

[0026]

[0027]

[0028] In one embodiment of the present invention, the diarylpyrimidine compound or its pharmaceutically acceptable salt is specifically selected from the following compounds or their pharmaceutical salts: SV-6, SV-8, or SV-10, preferably SV-10.

[0029] In one embodiment of the present invention, the diarylpyrimidine compound or its pharmaceutically acceptable salt is specifically selected from the following compounds or their pharmaceutical salts: SZ-10.

[0030] In one embodiment of the invention, the diarylpyrimidine compound or its pharmaceutically acceptable salt refers to a compound whose salt, within a reliable pharmaceutical evaluation range, is suitable for contact with human or lower animal tissues without undue toxicity, irritation, or allergic reactions, has a reasonably reasonable benefit-risk ratio, is typically water- or oil-soluble or dispersible, and is effectively used for its intended purpose. This includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts, which are herein suitable for the intended use and chemically compatible with the compounds of Formula I and Formula II.

[0031] In one embodiment of the invention, the diarylpyrimidine compound or a pharmaceutically acceptable salt thereof, a list of suitable salts can be found in SM Birge et al., J. Pham. Sci., 1977, 66, 1-19.

[0032] The present invention also provides a method for preparing the diarylpyrimidine compound or a pharmaceutically acceptable salt thereof.

[0033] The preparation route for the diarylpyrimidine compounds represented by formula (I) is as follows:

[0034]

[0035] In one embodiment of the present invention, in step i, 4-chloro-5-bromopyrimidine (V1) is used as the initial raw material and reacted with 3,4,5-trimethoxyphenylboronic acid in a reaction solvent to prepare intermediate compound V2.

[0036] In one embodiment of the present invention, in step i, the reaction of 4-chloro-5-bromopyrimidine (V1) with 3,4,5-trimethoxyphenylboronic acid needs to be carried out in the presence of the metal catalysts tetratriphenylphosphine palladium and potassium carbonate.

[0037] In one embodiment of the present invention, in step i, the reaction solvent is selected as a mixed solvent with a volume ratio of toluene:ethanol:water = 3:2:1.

[0038] In one embodiment of the present invention, in step i, the reaction conditions are: reacting at 70°C under N2 protection until the raw materials are completely reacted.

[0039] In one embodiment of the present invention, in step i, after the reaction is completed, intermediate V2 is obtained by separation using a chromatographic column.

[0040] In one embodiment of the present invention, a specific method for synthesizing intermediate V2 in step i is provided: using commercially available 4-chloro-5-bromopyrimidine (V1) as the initial raw material, and using toluene:ethanol:water in a volume ratio of 3:2:1 as the reaction solvent, the intermediate V2 is obtained by reacting with 3,4,5-trimethoxyphenylboronic acid at 70°C under N2 protection conditions in the presence of the metal catalysts tetraphenylphosphine palladium and potassium carbonate until the reaction of the raw material is complete, and then separated by chromatographic column chromatography.

[0041] In one embodiment of the present invention, in step ii, the intermediate compound V2 is reacted with arylboronic acid in a reaction solvent to prepare the target compound of general formula (I).

[0042] In one embodiment of the present invention, in step ii, the reaction of intermediate compound V2 with arylboronic acid needs to be carried out in the presence of the metal catalysts tetraphenylphosphine palladium and potassium carbonate.

[0043] In one embodiment of the present invention, in step ii, the reaction solvent is selected as a mixed solvent with a volume ratio of toluene:ethanol:water = 3:2:1.

[0044] In one embodiment of the present invention, in step ii, the reaction conditions are: reacting at 70°C under N2 protection until the intermediate compound V2 is completely reacted.

[0045] In one embodiment of the present invention, in step ii, after the reaction is completed, the target compound of general formula (I) is obtained by separation using a chromatographic column.

[0046] In one embodiment of the present invention, a method for synthesizing the target compound of general formula (I) in specific step ii is provided: using intermediate compound V2 as a raw material, and in a volume ratio of toluene:ethanol:water = 3:2:

[0047] Using 1 as the reaction solvent, under the action of the metal catalysts tetraphenylphosphine palladium and potassium carbonate, the target compound of general formula (I) was obtained by reacting with different arylboronic acids at 70°C under N2 protection until the intermediate compound V2 was completely reacted and separated by chromatographic column.

[0048] The preparation route for the diarylpyrimidine compounds represented by formula (II) is as follows:

[0049]

[0050] In one embodiment of the present invention, in step iii, 2,4-dichloropyrimidine (V3) is used as the initial raw material and reacted with dimethoxy or trimethoxyphenylboronic acid with different substitution positions in a reaction solvent to prepare intermediate compound V4.

[0051] In one embodiment of the invention, in step iii, the reaction of 2,4-dichloropyrimidine (V3) with dimethoxy or trimethoxyphenylboronic acid with different substitution positions needs to be carried out in the presence of the metal catalysts tetratriphenylphosphine palladium and potassium carbonate.

[0052] In one embodiment of the present invention, in step iii, the reaction solvent is selected as a mixed solvent with a volume ratio of toluene:ethanol:water = 3:2:1.

[0053] In one embodiment of the present invention, in step iii, the reaction conditions are: reacting at 70°C under N2 protection until the raw materials are completely reacted.

[0054] In one embodiment of the present invention, in step iii, after the reaction is completed, intermediate compound V4 is obtained by separation using a chromatographic column.

[0055] In one embodiment of the present invention, a specific method for synthesizing intermediate compound V4 in step iii is provided: using commercially available 2,4-dichloropyrimidine (V3) as the initial raw material, and using toluene:ethanol:water in a volume ratio of 3:2:1 as the reaction solvent, under the action of the metal catalyst tetra-triphenylphosphine palladium and potassium carbonate, the intermediate compound V4 is obtained by reacting dimethoxy or trimethoxyphenylboronic acid with different substitution positions at 70°C under N2 protection conditions until the raw material reacts completely, and then separated by chromatographic column separation.

[0056] In one embodiment of the present invention, in step iv, intermediate compound V4 is reacted with dimethoxy or trimethoxyphenylboronic acid with different substitution positions in a reaction solvent to prepare the target compound of general formula (II).

[0057] In one embodiment of the invention, in step iv, the reaction of intermediate V4 compound with dimethoxy or trimethoxyphenylboronic acid with different substitution positions needs to be carried out in the presence of the metal catalysts tetratriphenylphosphine palladium and potassium carbonate.

[0058] In one embodiment of the present invention, in step iv, the reaction solvent is selected as a mixed solvent with a volume ratio of toluene:ethanol:water = 3:2:1.

[0059] In one embodiment of the present invention, in step iv, the reaction conditions are: reacting at 100°C under N2 protection until the intermediate compound V4 is completely reacted.

[0060] In one embodiment of the present invention, in step iv, after the reaction is completed, the target compound of general formula (II) is obtained by separation using a chromatographic column.

[0061] In one embodiment of the present invention, a method for synthesizing the target compound of general formula (II) in specific step ii is provided: using intermediate compound V4 as a raw material, and mixing it with toluene:ethanol:water in a volume ratio of 3:2:

[0062] Using 1 as the reaction solvent, under the action of the metal catalysts tetraphenylphosphine palladium and potassium carbonate, the compound reacts with different arylboronic acids at 100°C under N2 protection until the intermediate compound V4 is completely reacted. The target compound of general formula (II) is obtained by separation by chromatographic column.

[0063] The pharmaceutically acceptable salts of the aforementioned diarylpyrimidine compounds can be prepared using the aforementioned diarylpyrimidine compounds as raw materials and by means of technical means in this field.

[0064] The present invention also provides the use of the aforementioned diarylpyrimidine compounds or pharmaceutically acceptable salts thereof.

[0065] The use of the diarylpyrimidine compounds or their pharmaceutically acceptable salts in the preparation of antitumor drugs.

[0066] In relation to the use of the diarylpyrimidine compounds or their pharmaceutically acceptable salts in the preparation of antitumor drugs, the diarylpyrimidine compounds are compounds having the structure of the above general formula (I) or general formula (II), preferably including the various specific compounds defined above, and also including SZ-1 and SZ-2.

[0067]

[0068] In one embodiment of the present invention, the tumor is selected from cervical cancer.

[0069] In one embodiment of the present invention, the diarylpyrimidine compound or a pharmaceutically acceptable salt thereof is used in the preparation of a microtubule colchicine binding site inhibitor.

[0070] The mechanism by which the diarylpyrimidine compounds or their pharmaceutical salts inhibit tumor cell growth in this invention is through inhibition of microtubule polymerization by binding to the colchicine binding site. In vitro and in vivo anti-cervical cancer experiments have confirmed that these compounds have significant inhibitory effects on the proliferation of cervical cancer cells and their drug-resistant cells, and show good potential for drug development.

[0071] The present invention also provides an antitumor pharmaceutical composition comprising: a therapeutically effective amount of the diarylpyrimidine compound or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0072] In the antitumor drug composition, the diarylpyrimidine compound is a compound having the structure of the above general formula (I) or general formula (II), preferably including the various specific compounds defined above, and also including SZ-1 and SZ-2.

[0073]

[0074] In one embodiment of the present invention, the antitumor drug composition is a tablet, capsule, pill, injection, sustained-release formulation, spray, or nano-drug delivery system.

[0075] The present invention also provides an antitumor pharmaceutical composition comprising: a therapeutically effective amount of the diarylpyrimidine compound or its pharmaceutically acceptable salt and platinum-based drug as the active ingredient, and a pharmaceutically acceptable carrier.

[0076] In one embodiment of the present invention, the platinum-based drug is selected as cisplatin.

[0077] In one embodiment of the present invention, the antitumor drug composition is an anticervical cancer drug composition.

[0078] Based on the mechanism of action of small molecule ligands and colchicine binding sites described in the background art, the diarylpyrimidine compounds provided in this application are a novel type of microtubule protein CBSIs. They have the same mechanism as colchicine and, while avoiding the high toxicity of colchicine, can competitively bind to the colchicine binding site. This can solve the problems of drug resistance and nephrotoxicity that have occurred with cisplatin in current clinical practice, and provide new treatment options and medication regimens for the clinical treatment of cervical cancer.

[0079] This invention provides and demonstrates the mechanism by which diarylpyrimidine compounds or their pharmaceutical salts inhibit microtubule polymerization and thus suppress tumor cell growth by binding to the colchicine binding site. In vitro and in vivo experiments have confirmed that these compounds have significant inhibitory effects on cervical cancer and drug-resistant cells, as well as on in vivo cervical cancer xenograft models.

[0080] Furthermore, this invention demonstrated, using a human HeLa cell subcutaneous xenograft tumor model in female BALB / c nude mice, that the diarylpyrimidine compounds of this invention, or their pharmaceutical salts, can be used in combination with cisplatin and exhibit good in vivo anti-cervical cancer efficacy. Simultaneously, the nude mice in the combination therapy group did not show significant weight loss, and their serum renal function indicators were not significantly different from the control group. Combining these compounds with the drugs of this invention can reduce the nephrotoxicity caused by high-dose cisplatin use, thus mitigating cisplatin toxicity. The compounds or their pharmaceutical salts can be used in combination with platinum-based drugs for the prevention or treatment of cervical cancer and can reduce the nephrotoxicity caused by high-dose cisplatin use, potentially meeting urgent clinical needs.

[0081] The main innovation of this invention is that it targets tubulin CBS and uses CA-4 as a lead compound. Through a conformational restriction strategy, unlike the pyrimidine ring modification structures reported in previous articles (Bioorganic & Medicinal Chemistry, 2005, 13(6):2097-2107.; Bioorganic Chemistry, 2018, 78:130-140.), this invention replaces the CA-4 double bond with the classic parent ring pyrimidine ring of antitumor drugs (4,5-ortho-diaryl substitution and 2,4-meta-diaryl substitution) to improve the instability of its double bond structure. At the same time, while ensuring the dominant group of CA-4, and combining the characteristics of colchicine binding site and small molecule action mode, the invention focuses on the structural modification and optimization of aromatic heterocycles that affect the physicochemical properties and antitumor activity of small molecules. A new class of CBSIs with diarylpyrimidine structural skeletons with antitumor activity, especially anti-cervical cancer activity, was designed, synthesized and discovered.

[0082] Compared with the prior art, the beneficial effects of the present invention are reflected in:

[0083] This invention targets the colchicine binding site, which possesses unique advantages in tumor angiogenesis disruption and resistance to drug resistance. It addresses the critical clinical need for small-molecule therapies for cervical cancer and the serious drug resistance and nephrotoxicity associated with prolonged and high-dose use of cisplatin, a first-line drug. The invention discovers novel diarylpyrimidine CBSIs and their pharmaceutically acceptable salts for use in antitumor drugs. Existing compounds SZ-1 and SZ-2 have only been reported in terms of synthetic methodologies, with no reports on their activity. Furthermore, no other diarylpyrimidine CBSIs and their pharmaceutically acceptable salts have been reported by SciFinder. These compounds exert their antitumor effects by binding to the colchicine binding site, exhibiting superior anti-cervical cancer activity compared to the marketed drug cisplatin, with some compounds showing superior target activity against colchicine in inhibiting microtubule polymerization, and superior activity against drug-resistant cervical cancer cells compared to CA-4, cisplatin, and carboplatin. Meanwhile, it is superior to CA-4 in terms of solubility in buffer solutions at pH 2.0 and pH 7.4 and in vitro hepatic microsomal metabolic stability. It can also achieve better antitumor effects when used in combination with cisplatin and reduce nephrotoxicity caused by high doses of cisplatin. This provides the possibility for its clinical use in the treatment of cervical cancer or its combination with cisplatin to enrich the clinical treatment options for cervical cancer, and is expected to meet the urgent clinical needs. Attached Figure Description

[0084] Figure 1 Brief structure-activity relationship of Combretastatin A-4;

[0085] Figure 2 Compound SV-10 inhibits tubulin polymerization kinetics at the molecular level;

[0086] Figure 3 Compound SV-10 was confirmed at the molecular level to bind to the colchicine binding site;

[0087] Figure 4 Detection of the effect of compound SV-10 on microtubule morphology using immunofluorescence assay;

[0088] Figure 5: Effects of compound SV-10 on cervical cancer cell function at the cellular level;

[0089] Figure 5 includes Figure 5-A , Figure 5-B , Figure 5-C , Figure 5-D ;

[0090] Figure 6: Results of in vivo anti-cervical cancer efficacy study of compound SV-10;

[0091] Figure 6 includes Figure 6-A , Figure 6-B , Figure 6-C , Figure 6-D ;

[0092] Figure 7: Serum renal function evaluation results of compound SV-10 in nude mice;

[0093] Figure 7 includes Figure 7-A , Figure 7-B , Figure 7-C . Detailed Implementation

[0094] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments.

[0095] The specific synthetic routes involved in the diarylpyrimidine compounds of general formula (I) in the examples are as follows:

[0096] Synthesis of SV-1, SV4 to SV-21:

[0097]

[0098]

[0099]

[0100] Reagents and reaction conditions:

[0101] (i / ii)2,4-Dimethoxyphenylboronic acid, Pd[P(C6H5)3]4, K2CO3; Toluene:EtOH:H2O=3:2:1, 70℃;

[0102] (iii) Benzyl chloride, DMF, 50°C;

[0103] (iv) Pinaryl diboronate, AcOK, C 35 H 30 Cl4P2Pd, (1,4-Dioxane:EtOH:H2O=3:1:1), N2, 80℃;

[0104] (v)H2, Pd / C, CH3OH, rt; (vi)Et3N, ethyl chloroformate, anhydrous CH2Cl2, N2, 0℃~rt

[0105] Example 1: Preparation of 5-(4-methoxyphenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SV-1)

[0106]

[0107] Take a 250 mL inclined two-necked flask and dissolve the starting materials 4-chloro-5-bromo-pyrimidine (5.00 g, 25.8 mmol), (3,4,5-trimethoxyphenyl)boric acid (6.30 g, 14.9 mmol), and potassium carbonate (7.14 g, 51.6 mmol) in a 75 mL mixed solvent of Toluene:EtOH:H2O = 3:1:1. Then quickly add (beta-4)-platinum (3.00 g, 2.58 mmol), evacuate under N2 protection, and gradually heat to 70 °C and reflux for 5 h. After the reaction is complete as monitored by TLC, cool to room temperature, filter with diatomaceous earth, and evaporate the filtrate to dryness. Dissolve in EA (3 × 50 mL), extract successively with water (50 mL) and saturated NaCl solution (50 mL), combine the organic phases, and dry with anhydrous Na2SO4. The sample was filtered and mixed with silica gel, then purified by column chromatography at a ratio of PE:EA = 8:1 to obtain a white solid V2 (3.12 g, yield 36%). Mp 121.5-122.6℃; 1 H NMR (400MHz, DMSO-d6) δ9.16(s,1H),8.94(s,1H),7.14(s,2H),3.95(s,9H)ppm; 13 C NMR (100MHz, CDCl3) δ163.7,160.4,156.8,153.0,140.1,131.8,118.8,107.1,60.9,56.3ppm;

[0108] Take a 25 mL inclined two-necked flask and dissolve the raw material V5 (120 mg, 369 μmol), 4-methoxyphenylboronic acid (64 mg, 424 μmol), and potassium carbonate (100 mg, 738 μmol) sequentially in a Toluene:ETOH:H2O = 3:1:1 solution. Then, quickly add (beta-4)-platinum (42.7 mg, 36.9 μmol), evacuate under N2 protection, and gradually heat to 70 °C and reflux for 40 min. After the reaction is complete as monitored by TLC, cool to room temperature, filter with diatomaceous earth, and evaporate the filtrate to dryness. Dissolve in EA (3 × 15 mL), extract successively with water (15 mL) and saturated NaCl solution (20 mL), combine the organic phases, and dry with anhydrous Na2SO4. The sample was filtered and mixed with silica gel, then purified by column chromatography at a PE:EA ratio of 4:1 to obtain a white solid SV-1 (101 mg, yield 78%). MPa: 114.5-116.0℃; 1H NMR (400MHz, CDCl3) δ9.20(s,1H),8.69(s,1H),7.17(d,J=8.8Hz,2H),6.92(d,J=8.8Hz,2H),6.74(s,2H),3.85(s,3H),3.82(s,3H),3.65(s,6H)ppm; 13 C NMR (100MHz, CDCl3) δ162.8,159.8,158.4,157.3,153.0,139.5,132.9,132.5,130.6,128.9,114.6,107.5,61.1,56.1,55.6ppm; HRMS(ESI)calcd for C 20 H 21 N2O4 + [(M+H) + ]:353.1501,found:353.1495.

[0109] Example 2: Preparation of 2-methoxy-5-(4-(3,4,5-trimethoxyphenyl)pyrimidin-5-yl)phenol (SV-2)

[0110]

[0111] 2-(benzyloxy)-4-bromo-1-methoxybenzene (Sub-1)

[0112] In a 100 mL round-bottom flask, dissolve 2.00 g (9.85 mmol) of 5-bromo-2-methoxyphenol and 4.09 g (29.6 mmol) of potassium carbonate in 30 mL of DMF. Then add benzyl chloride (1.47 mL, 12.81 mmol). Heat to 50 °C and react for 2 h. Monitor the reaction by TLC until complete. Add 30 mL of water to the reaction mixture. A solid precipitates out; filter and dry to obtain a grayish-white solid, Sub-1 (1.98 g, yield 68%). Mp 106.8-107.2 °C; 1 H NMR (400MHz, CDCl3) δ7.47-7.33(m,5H),7.12-7.01(m,2H),6.79(d,J=8.3Hz,1H),5.14(s,2H),3.88(s,3H)ppm; 13 C NMR (100MHz, CDCl3) δ149.1,149.0,136.4,128.6,128.1,127.4,124.0,117.3,113.1,112.6,71.2,56.2ppm;

[0113] 2-(3-(benzyloxy-4-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (Sub-2)

[0114]

[0115] Take a 50 mL inclined double-necked flask and dissolve the raw materials Sub-1 (1.5 g, 5.11 mmol), pinacol diborate (1.56 g, 6.13 mmol), and potassium acetate (1.5 g, 15.33 mmol) sequentially in 25 mL of a mixed solvent of 1,4-dioxane, ethanol, and water (1,4-dioxane:ethanol:water = 3:1:1). Then, quickly add 1,1'-bis(diphenylphosphine)ferrocene palladium(II) dichloromethane complex (0.29 g, 0.36 mmol), evacuate under N2 protection, and gradually heat to 80 °C and reflux overnight. After the reaction is complete as monitored by TLC, cool to room temperature, filter with diatomaceous earth, and evaporate the filtrate to dryness. Dissolve in EA (3 × 20 mL), extract successively with water (20 mL) and saturated NaCl solution (30 mL), combine the organic phases, and dry with anhydrous Na2SO4. The sample was filtered and mixed with silica gel, then purified by column chromatography at a PE:EA ratio of 10:1 to obtain a white solid Sub-2 (0.77 g, yield 44%). MPa: 119.9-122.0℃; 1 H NMR (400MHz, CDCl3) δ7.65-7.19(m,1H),6.94(d,J=8.0Hz,1H),5.18(s,2H),3.92(s,3H),1.36(s,12H)ppm; 13 C NMR (100MHz, CDCl3) δ152.5,147.8,137.2,129.1,128.4,127.8,127.7,119.4,111.1,83.6,71.0,55.8ppm;

[0116] 5-(3-(benzyloxy)-4-methoxyphenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SV-2')

[0117]

[0118] Compound SV-2' was processed in the same manner as SV-1, using Sub-2 boric acid (144 mg, 424 μmol) to give a white solid SV-2' (108 mg, yield 64%). It was then directly added to the next step.

[0119] 2-Methoxy-5-(4-(3,4,5-trimethoxyphenyl)pyrimidin-5-yl)phenol (SV-2)

[0120]

[0121] Take a 50 mL inclined two-necked flask and dissolve the raw material SV-2' (250 mg, 545 μmol) and Pd / C (25 mg) in 10 mL of methanol. Vacuum the mixture, replace with hydrogen gas, and stir at room temperature for 6 h. After the reaction is complete as monitored by TLC, add diatomaceous earth for filtration to remove Pd / C, and directly add silica gel for mixing. Purify by column chromatography (PE:EA = 3:1) to obtain a white solid SV-2 (113 mg, yield 57%). Mp 142.2-142.8℃; 1 H NMR(400MHz,DMSO-d6)δ9.20(s,1H),9.17(s,1H),8.72(s,1H),7.01(d,J=8.2Hz,1H),6.80(s,2H ),6.73(dd,J=8.2,2.2Hz,1H),6.67(d,J=2.2Hz,1H),3.79(s,3H),3.69(s,3H),3.58(s,6H)ppm; 13 C NMR (100MHz, CDCl3) δ162.9,158.3,157.2,153.0,146.8,146.2,139.6,132.9,1 32.4,129.9,121.4,115.6,111.1,107.5,61.0,56.2,56.1ppm; HRMS(ESI)calcd for C 20 H 21 N2O5 + [(M+H) + ]:369.1450,found:369.1441.

[0122] Example 3: Preparation of ethyl (2-methoxy-5-(4-(3,4,5-trimethoxyphenyl)pyrimidin-5-yl)phenyl)carbonate (SV-3)

[0123]

[0124] A 25 mL inclined two-necked flask was used to stir a mixture of SV-2 (120 mg, 326 μmol), anhydrous CH2Cl2 (10 mL), and triethylamine (135 μL, 978 μmol) for 5 min under nitrogen protection at 0 °C. Then, ethyl chloroformate (53 mg, 489 μmol) was added dropwise. The system was reacted at 0 °C for 30 min, then slowly restored to room temperature and reacted for another 12 h. The reaction endpoint was monitored by TLC. Silica gel was added, and the mixture was purified by PE:EA = 5:1 column chromatography to obtain a pale yellow solid SV-3 (44 mg, yield 31%). Mp 107.7-108.1 °C. 1H NMR (400MHz, CDCl3) δ9.23 (s, 1H), 8.71 (s, 1H), 7.09 (d, J = 8.1Hz, 1H, 1H), 7.00 (d, J = 8.1Hz, 1H), 6. 76(s,2H),4.31(q,J=7.1Hz,2H),3.89(s,3H),3.87(s,3H),3.70(s,6H),1.39(t,J=7.1Hz,3H)ppm; 13 C NMR (100MHz, CDCl3) δ162.8,158.1,157.4,153.0,152.9,151.4,140.5,139.4,131.9,131 .8,129.0,128.0,123.4,112.9,107.2,65.1,60.9,56.2,55.9,14.2ppm; HRMS(ESI)calcd for C 23 H 25 N2O7 + [(M+H) + ]:441.1667,found:441.1662.

[0125] Example 4: Preparation of compounds SV-4 to SV-21

[0126] 5-(3-fluoro-4-methoxyphenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SV-4)

[0127]

[0128] Compound SV-4 was prepared in the same manner as SV-1, but the boric acid used was 3-fluoro-4-methoxyphenylboronic acid (72 mg, 424 μmol), yielding a white solid SV-4 (86 mg, yield 63%). Mp 128.8-129.4 °C; 1 H NMR (400MHz, CDCl3) δ9.24(s,1H),8.71(s,1H),7.04-6.99(m,3H),6.75(s,2H),3.93(s,3H),3.88(s,3H),3.70(s,6H)ppm; 13 CNMR(100MHz,CDCl3)δ162.9,158.1,157.4,153.0,152.4(d,J C,F =224Hz), 147.7(d,J) C,F =10Hz),139.5,132.0,131.8,129.2(d,J C,F =10Hz), 125.4(d,J) C,F=3Hz), 117.0(d,J C,F =19Hz),113.7,107.2,60.9,56.4,56.0ppm; HRMS(ESI)calcd for C 20 H 20 FN2O4 + [(M+H) + ]:371.1402,found:371.1409.

[0129] 5-(3-chloro-4-methoxyphenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SV-5)

[0130]

[0131] Compound SV-5 was prepared in the same manner as SV-1, but the boric acid used was 3-chloro-4-methoxyphenylboronic acid (78 mg, 424 μmol), yielding a white solid SV-5 (87 mg, yield 61%). Mp 126.3-127.0 °C; 1 H NMR (400MHz, CDCl3) δ9.26 (s, 1H), 8.74 (s, 1H), 7.36 (s, 1H), 7.09 (d, J = 7.6Hz, 1H ),6.95(d,J=7.6Hz,1H),6.76(s,2H),3.94(s,3H),3.88(s,3H),3.70(s,6H)ppm; 13 C NMR (100MHz, CDCl3) δ162.9,158.1,157.4,155.0,153.0,139.5,132.0,130.8,129.6,129.0,123.0,112.3,107.3,61.0,56.3,56.0ppm; HRMS(ESI)calcd for C 20 H 20 ClN2O4 + [(M+H) + ]:387.1106,found:387.1116.

[0132] 5-(4-(methylthio)phenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SV-6)

[0133]

[0134] Compound SV-6 was prepared in the same manner as SV-1, but the boric acid used was 4-methylthiophenylboronic acid (71 mg, 424 μmol), yielding a white solid SV-6 (83 mg, yield 61%). Mp 103.9-104.1 °C;1 H NMR (400MHz, CDCl3) δ9.24(s,1H),8.71(s,1H),7.28(d,J=7.9Hz,2H),7.19(d,J=7.9Hz,2H),6.75(s,2H),3.87(s,3H),3.66(s,6H),2.51(s,3H)ppm; 13 C NMR (100MHz, CDCl3) δ162.8,158.0,157.3,152.9,139.4,139.3,133.0,132.6,132.0,129.6,126.6,107.3,60.9,55.9,15.6ppm; HRMS(ESI)calcd forC 20 H 21 N2O3S + [(M+H) + ]:369.1267,found:369.1279.

[0135] 5-(4-(ethylthio)phenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SV-7)

[0136]

[0137] Compound SV-7 was prepared in the same manner as SV-1, but the boric acid used was 4-(ethylthio)phenylboronic acid (77 mg, 424 μmol), yielding a white solid SV-7 (91 mg, yield 65%). Mp 100.6-101.8 °C; 1 H NMR (400MHz, CDCl3) δ9.25(s,1H),8.72(s,1H),7.34(d,J=7.9Hz,2H),7.19(d,J=7.9Hz,2H ),6.75(s,2H),3.88(s,3H),3.67(s,6H),2.99(q,J=7.3Hz,2H),1.35(t,J=7.3Hz,3H)ppm; 13 C NMR (100MHz, CDCl3) δ162.8,158.0,157.3,152.9,139.4,137.6,133.6,132.6,132.0,129.7,128.6,107.3,60.9,55.9,27.1,14.2ppm; HRMS(ESI)calcd for C 21 H 23 N2O3S + [(M+H) + ]:383.1424,found:383.1436.

[0138] 5-(4-ethoxyphenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SV-8)

[0139]

[0140] Compound SV-8 was prepared in the same manner as SV-1, but the boric acid used was 4-ethoxyphenylboronic acid (70 mg, 424 μmol), yielding a white solid SV-8 (98 mg, yield 73%). Mp 86.4-87.2 °C; 1 H NMR (400MHz, CDCl3) δ9.22(s,1H),8.71(s,1H),7.17(d,J=8.4Hz,2H),6.92(d,J=8.4Hz,2H ), 6.76 (s, 2H), 4.06 (q, J = 7.0Hz, 2H), 3.87 (s, 3H), 3.67 (s, 6H), 1.45 (t, J = 7.0Hz, 3H) ppm; 13 C NMR (100MHz, CDCl3) δ162.7,159.0,158.2,157.0,152.9,139.2,132.8,132.4,130.4,128.5,115.0,107.2,63.6,60.9,55.9,14.7ppm; HRMS(ESI)calcd for C 21 H 23 N2O4 + [(M+H) + ]:367.1652,found:367.1664.

[0141] 5-(4-ethoxy-3-fluorophenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SV-9)

[0142]

[0143] Compound SV-9 was prepared in the same manner as SV-1, but the boric acid used was 3-fluoro-4-ethoxyphenylboronic acid (78 mg, 424 μmol), yielding a white solid SV-9 (92 mg, yield 65%). Mp 100.2-101.5 °C; 1 H NMR(400MHz, CDCl3)δ9.24(s,1H),8.70(s,1H),7.06-6.92(m,3H),6.75(s,2H) ), 4.15 (q, J = 7.0Hz, 2H), 3.88 (s, 3H), 3.70 (s, 6H), 1.49 (t, J = 7.0Hz, 3H) ppm; 13C NMR (100MHz, CDCl3) δ162.9,158.0,157.3,152.8(d,J C,F =246Hz), 151.4, 147.1 (d, J) C,F =10Hz),139.5,132.0,131.8,129.2(d,J C,F =6Hz), 125.3, 117.1 (d, J) C,F =19Hz),115.0,107.2,65.1,60.9,56.0,14.7ppm; HRMS(ESI)calcd forC 21 H 22 FN2O4 + [(M+H) + ]:385.1558,found:385.1572.

[0144] 5-(3-chloro-4-ethoxyphenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SV-10)

[0145]

[0146] Compound SV-10 was processed in the same manner as SV-1, but the boric acid used was 3-chloro-4-ethoxyphenylboronic acid (85 mg, 424 μmol), yielding a white solid SV-10 (107 mg, yield 72%). Mp 131.9-132.4 °C; 1 H NMR (400MHz, CDCl3) δ9.24 (s, 1H), 8.70 (s, 1H), 7.35 (s, 1H), 7.06 (d, J = 8.5Hz, 1H), 6.92 (d, J = 8. 5Hz,1H),6.76(s,2H),4.14(q,J=7.0Hz,2H),3.88(s,3H),3.70(s,6H),1.50(t,J=7.0Hz,3H)ppm; 13 C NMR (100MHz, CDCl3) δ162.8,158.1,157.4,154.4,153.0,139.5,132.0,131.6,130 .7,129.4,128.8,123.4,113.4,107.3,64.9,60.9,56.0,14.6ppm; HRMS(ESI)calcd for C 21 H 22 ClN2O4 + [(M+H) + ]:401.1263,found:401.1274.

[0147] 5-(4-ethoxy-3-(trifluoromethyl)phenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SV-11)

[0148]

[0149] Compound SV-11 was prepared in the same manner as SV-1, but the boric acid used was 3-trifluoromethyl-4-ethoxyphenylboronic acid (99 mg, 424 μmol), yielding a white solid SV-11 (71 mg, yield 44%). Mp 104.7-106.0 °C; 1 H NMR (400MHz, CDCl3) δ9.30 (s, 1H), 8.81 (s, 1H), 7.54 (s, 1H), 7.31 (d, J = 7.9Hz, 1H), 7.00 (d, J = 7. 9Hz,1H),6.71(s,2H),4.16(q,J=6.9Hz,2H),3.87(s,3H),3.68(s,6H),1.48(t,J=6.9Hz,3H)ppm; 13 CNMR(100MHz, CDCl3)δ163.0,157.9,157.5,156.8,153.0,139.5,134.3,132.0,128.1,127.9,127.8,123.2(d,J C,F =271Hz), 119.7(d,J C,F =31Hz),113.4,107.2,64.8,61.0,55.9,14.5ppm; HRMS(ESI)calcd for C 22 H 22 F3N2O4 + [(M+H) + ]:435.1526,found:435.1534.

[0150] 5-(4-(2,2,2-trifluoroethoxy)phenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SV-12)

[0151]

[0152] Compound SV-12 was handled in the same manner as SV-1, but the boric acid used was 4-(2,2,2-trifluoroethoxy)phenylboronic acid (93 mg, 424 μmol), yielding a white solid SV-12 (56 mg, yield 36%). Mp 104.5-105.0 °C; 1H NMR (400MHz, CDCl3) δ9.22 (s, 1H), 8.69 (s, 1H), 7.22 (d, J = 8.6Hz, 2H), 6.98 (d, J=8.6Hz,2H),6.71(s,2H),4.38(q,J=8.1Hz,2H),3.86(s,3H),3.65(s,6H)ppm; 13 C NMR (100MHz, CDCl3) δ163.0,158.2,157.5,157.5,153.1,139.5,132.4,132.2,130.9,130.9,123.3(d,J C,F =277Hz),115.6,107.4,66.0(d,J C,F =35Hz),61.1,56.0ppm; HRMS(ESI)calcd for C 21 H 20 F3N2O4 + [(M+H) + ]:421.1375,found:421.1378.

[0153] 5-(4-Isopropoxyphenyl)-4-(3,4,5-Trimethoxyphenyl)pyrimidine (SV-13)

[0154]

[0155] Compound SV-13 was prepared in the same manner as SV-1, but the boric acid used was 4-isopropoxyphenylboronic acid (76 mg, 424 μmol), yielding a white solid SV-13 (102 mg, yield 73%). Mp 76.4-77.2 °C; 1 H NMR (400MHz, CDCl3) δ9.22 (s, 1H), 8.72 (s, 1H), 7.16 (d, J = 8.4Hz, 2H), 6.92 (d, J = 8.4Hz, 2H),6.77(s,2H),4.63-4.51(m,1H),3.87(s,3H),3.67(s,6H),1.36(d,J=6.0Hz,6H)ppm; 13 C NMR (100MHz, CDCl3) δ162.7,158.1,157.9,157.0,152.8,139.2,132.9,132.3,130.5,128.4,116.2,107.2,70.0,60.9,55.8,21.9ppm; HRMS(ESI)calcd for C 22 H 25 N2O4 + [(M+H)+ ]:381.1809,found:381.1819.

[0156] 5-(6-Methoxypyridin-3-yl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SV-14)

[0157]

[0158] Compound SV-14 was prepared in the same manner as SV-1, but the boric acid used was 2-methoxy-5-pyridineboronic acid (65 mg, 424 μmol), yielding a white solid SV-14 (95 mg, 73% yield). Mp 120.4-121.8℃; 1 H NMR (400MHz, CDCl3) δ9.26 (s, 1H), 8.72 (s, 1H), 8.14 (s, 1H), 7.40 (d, J = 8.6Hz, 1H ),6.76(d,J=8.6Hz,1H),6.72(s,2H),3.98(s,3H),3.88(s,3H),3.69(s,6H)ppm; 13 C NMR (100MHz, CDCl3) δ164.0,163.3,158.0,157.6,153.1,146.6,139.5,131.9,129.7,125.4,111.0,107.2,60.9,56.0,53.7ppm; HRMS(ESI)calcd for C 19 H 20 N3O4 + [(M+H) + ]:354.1448,found:354.1459.

[0159] N-Methyl-4-(4-(3,4,5-trimethoxyphenyl)pyrimidin-5-yl)benzamide (SV-16)

[0160]

[0161] Compound SV-16 was prepared in the same manner as SV-1, but the boric acid used was 4-(N-methylformamide)phenylboronic acid (76 mg, 424 μmol), yielding a white solid SV-16 (73 mg, yield 52%). Mp 179.4-180.4 °C; 1 H NMR (400MHz, CDCl3) δ9.31(s,1H),8.79(s,1H),7.83(s,2H),7.36(s,2H),6.70(s,2H),6.29(s,1H),3.86(s,3H),3.63(s,6H),3.05(s,3H)ppm;13 C NMR (100MHz, CDCl3) δ167.2,163.0,158.0,157.7,153.0,139.9,139.5,134.1,131.8,129.6,127.5,107.3,60.9,56.0,27.0ppm; HRMS(ESI)calcdfor C 21 H 22 N3O4 + [(M+H) + ]:380.1605,found:380.1615.

[0162] N,N-Dimethyl-4-(4-(3,4,5-trimethoxyphenyl)pyrimidin-5-yl)benzamide (SV-17)

[0163]

[0164] Compound SV-17 was prepared in the same manner as SV-1, but the boric acid used was 4-(N,N-dimethylcarbamoyl)phenylboronic acid (82 mg, 424 μmol), yielding a white solid SV-17 (90 mg, yield 62%). Mp 135.1-136.9 °C; 1 H NMR (400MHz, CDCl3) δ9.27(s,1H),8.73(s,1H),7.48(d,J=5.9Hz,2H),7.34(d,J= 5.9Hz,2H),6.74(s,2H),3.86(s,3H),3.66(s,6H),3.14(s,3H),3.02(s,3H)ppm; 13 C NMR (100MHz, CDCl3) δ170.6,162.9,158.1,157.7,153.0,139.6,138.0,136.2,132 .3,131.8,129.3,127.8,127.1,107.4,60.9,55.9,39.5,35.4ppm; HRMS(ESI)calcd for C 22 H 24 N3O4 + [(M+H) + ]:394.1761,found:394.1769.

[0165] 5-(4-Bromophenyl)-4-(3,4,5-Trimethoxyphenyl)pyrimidine (SV-18)

[0166]

[0167] Compound SV-18 was prepared in the same manner as SV-1, but the boric acid used was 4-bromophenylboronic acid (85 mg, 424 μmol), yielding a white solid SV-18 (100 mg, yield 68%). Mp 145.4-146.0 °C; 1 H NMR (400MHz, CDCl3) δ9.24(s,1H),8.68(s,1H),7.54(d,J=8.5Hz,2H),7.14(d,J=8.5Hz,2H),6.69(s,2H),3.86(s,3H),3.65(s,6H)ppm; 13 C NMR (100MHz, CDCl3) δ163.0,158.0,157.8,153.1,139.8,135.8,132.3,132.1,1 31.9,131.0,130.0,128.7,127.5,122.7,107.6,61.1,56.1ppm; HRMS(ESI)calcd forC 19 H 18 BrN2O3 + [(M+H) + ]:401.0495,403.0479,found:401.0501,403.0483.

[0168] (4-(4-(3,4,5-trimethoxyphenyl)pyrimidin-5-yl)phenyl)boronic acid (SV-19)

[0169]

[0170] Compound SV-19 was prepared in the same manner as SV-1, but the boric acid used was 1,4-phenylenediboric acid (70 mg, 424 μmol), yielding a white solid SV-19 (39 mg, yield 29%). Mp 184.9-186.6 °C; 1 H NMR(400MHz,DMSO-d6)δ9.26(s,1H),8.80(s,1H),8.17(s,2H),8.04(s,1H),7.83(d,J=7 .7Hz,2H),7.74(s,1H),7.28(d,J=7.7Hz,2H),6.72(s,2H),3.67(s,3H),3.51(s,6H)ppm; 13 C NMR (100MHz, DMSO-d6) δ162.2,158.6,157.7,152.7,139.0,138.5,134.9,133.5,133.2,132.4,128.7,107.8,60.6,55.9ppm; HRMS(ESI)calcd for C19 H 20 BN2O5 + [(M+H) + ]:367.1460,found:367.1471.

[0171] 5-(benzo[d][1,3]dioxol-5-yl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SV-20)

[0172]

[0173] Compound SV-20 was prepared in the same manner as SV-1, but the boric acid used was 3,4-(methylenedioxy)phenylboronic acid (70 mg, 424 μmol), yielding a white solid SV-20 (110 mg, yield 81%). Mp 105.8-107.0 °C; 1 H NMR (400MHz, CDCl3) δ9.23 (s, 1H), 8.70 (s, 1H), 6.88 (d, J = 7.7Hz, 1H), 6.89-6.79 (m, 3H), 6.68 (s, 1H), 6.00 (s, 2H), 3.88 (s, 3H), 3.71 (s, 6H) ppm; 13 C NMR (100MHz, CDCl3) δ162.8,158.2,157.2,152.9,148.1,147.7,139.4,132.8,1 32.2,130.3,122.9,109.7,108.9,107.2,101.4,61.0,56.0ppm; HRMS(ESI)calcd for C 20 H 19 N2O5 + [(M+H) + ]:367.1288,found:367.1300.

[0174] 5-(4-(3,4,5-trimethoxyphenyl)pyrimidin-5-yl)-1H-indole (SV-21)

[0175]

[0176] Compound SV-21 was processed in the same manner as SV-1, but the boric acid used was 5-indoleboric acid (68 mg, 424 μmol), yielding a white solid SV-21 (58 mg, yield 44%). Mp 167.8-169.1 °C; 1H NMR(400MHz,DMSO-d6)δ11.23(s,1H),9.22(s,1H),8.81(s,1H),7.56(s,1H),7.44-7.40 (m,2H),6.92-6.90(m,1H),6.77(s,2H),6.48-6.44(m,1H),3.64(s,3H),3.41(s,6H)ppm; 13 CNMR(100MHz,DMSO-d6)δ161.5,158.4,156.6,152.2,138.4,135.3,134.1,132.4,127 .9,126.8,126.4,122.4,120.7,111.6,107.3,101.4,60.0,55.3ppm; HRMS(ESI)calcd for C 21 H 20 N3O3 + [(M+H) + ]:362.1505,found:362.1507.

[0177] The synthetic routes involved in the diarylpyrimidine compounds of general formula (II) in the examples are as follows:

[0178]

[0179] Reagents and reaction conditions:

[0180] (i) 2,4-Dimethoxyphenylboronic acid, Pd[P(C6H5)3]4, K2CO3; Toluene:EtOH:H2O = 3:2:1, 70℃;

[0181] (ii)ArB(OH)2,Pd[P(C6H5)3]4,K2CO3; Toluene:EtOH:H2O=3:2:1,100℃

[0182] Example 5: Preparation of 2-(4-methoxyphenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SZ-1)

[0183] 2-(4-Chlorophenyl)-4-(3,4,5-Trimethoxyphenyl)pyrimidine (V4)

[0184]

[0185] Take a 250 mL inclined two-necked flask and dissolve the starting materials 2,4-dichloropyrimidine (5.00 g, 33.6 mmol), 3,4,5-trimethoxyphenylboronic acid (7.12 g, 33.6 mmol), and potassium carbonate (9.29 g, 67.2 mmol) in a Toluene:ETOH:H2O ratio of 3:1:1. Then, quickly add (beta-4)-platinum (3.88 g, 3.36 mmol), evacuate under N2 protection, and gradually heat to 70 °C and reflux for 5 h. After the reaction is complete as monitored by TLC, cool to room temperature, filter with diatomaceous earth, and evaporate the filtrate to dryness. Dissolve in EA (3 × 50 mL), extract successively with water (50 mL) and saturated NaCl solution (50 mL), combine the organic phases, and dry with anhydrous Na2SO4. The sample was filtered and mixed with silica gel, then purified by column chromatography at a ratio of PE:EA = 8:1 to obtain a white solid V4 (5.75 g, yield 61%). Mp 120.2-120.7℃; 1 H NMR (400MHz, CDCl3) δ8.60 (d, J = 5.3Hz, 1H), 7.59 (d, J = 5.3Hz, 1H), 7.33 (s, 2H), 3.97 (s, 6H), 3.93 (s, 3H) ppm; 13 C NMR(100MHz, CDCl3)δ166.8,161.9,159.8,153.9,141.9,130.4,115.0,105.0,61.1,56.6; HRMS(ESI)calcd for C 13 H 14 ClN2O3 + [(M+H) + ]:281.0687,found:281.0685.

[0186] 2-(4-Methoxyphenyl)-4-(3,4,5-Trimethoxyphenyl)pyrimidine (SZ-1)

[0187]

[0188] Take a 25 mL inclined two-necked flask and dissolve the raw material V4 (120 mg, 429 μmol), 4-methoxyphenylboronic acid (71.7 mg, 472 μmol), and potassium carbonate (119 mg, 858 μmol) sequentially in a Toluene:ETOH:H2O = 3:1:1 mixture. Then, quickly add (beta-4)-platinum (49.6 mg, 42.9 μmol), evacuate under N2 protection, and gradually heat to 100 °C and reflux for 5 h. After the reaction is complete as monitored by TLC, cool to room temperature, filter with diatomaceous earth, and evaporate the filtrate to dryness. Dissolve in EA (3 × 15 mL), extract successively with water (15 mL) and saturated NaCl solution (20 mL), combine the organic phases, and dry with anhydrous Na2SO4. The sample was filtered and mixed with silica gel, then purified by column chromatography at a PE:EA ratio of 4:1 to obtain a white solid SZ-1 (82 mg, yield 54%). MPa: 139.1-140.9℃. 1 H NMR (400MHz, CDCl3) δ8.75(d,J=5.3Hz,1H),8.51(d,J=8.9Hz,2H),7.46(d,J= 5.9Hz,3H),7.03(d,J=8.9Hz,2H),3.99(s,6H),3.94(s,3H),3.89(s,3H)ppm; 13 C NMR (100MHz, CDCl3) δ164.4,163.6,162.1,157.7,153.8,141.9,132.7,130.6,130.0,114.1,113.7,104.9,61.1,56.5,55.5ppm; HRMS(ESI)calcd for C 20 H 21 N2O4 + [(M+H) + ]:353.1496,found:353.1491.

[0189] Example 6: Preparation of SZ-2 to SZ-8

[0190] 2-(3-Fluoro-4-methoxyphenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SZ-2)

[0191]

[0192] Compound SZ-2 was prepared in the same manner as SZ-1, but the boric acid used was 3-fluoro-4-methoxyphenylboronic acid (80 mg, 472 μmol), yielding a white solid SZ-2 (96.8 mg, yield 61%). Mp 143.7-144.2 °C; 1H NMR (400MHz, CDCl3) δ8.74(d,J=5.3Hz,1H),8.35-8.23(m,2H),7.48(d,J=5.3Hz,1 H),7.43(s,2H),7.06(t,J=8.4Hz,1H),3.99(s,6H),3.96(s,3H),3.94(s,3H)ppm; 13 C NMR(100MHz,CDCl3)δ163.6,163.3(d,J C,F =3Hz), 157.7, 153.8, 152.5 (d, J) C,F =244Hz), 150.0(d,J C,F =11Hz),141.1,132.3,131.2(d,J C,F =6Hz), 124.7(d,J C,F =3Hz), 116.1(d,J C,F =20Hz),114.1,113.0,113.0,104.8,61.1,56.5,56.4ppm; HRMS(ESI)calcd for C 20 H 20 FN2O4 + [(M+H) + ]:371.1402,found:371.1398.

[0193] 2-(3-chloro-4-methoxyphenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SZ-3)

[0194]

[0195] Compound SZ-3 was prepared in the same manner as SZ-1, but the boric acid used was 3-chloro-4-methoxyphenylboronic acid (80 mg, 472 μmol), yielding a white solid SZ-3 (103 mg, yield 62%). Mp 188.4-190.2℃; 1 H NMR (400MHz, CDCl3) δ8.76(d,J=5.3Hz,1H),8.59(d,J=2.1Hz,1H),8.45(dd,J=8.7,2.1Hz,1H),7. 49(d,J=5.3Hz,1H),7.44(s,2H),7.05(d,J=8.7Hz,1H),4.00(s,6H),3.98(s,3H),3.94(s,3H)ppm; 13C NMR (100MHz, CDCl3) δ163.8,163.3,157.7,157.3,153.8,141.2,132.4,131.4,1 30.5,128.2,122.9,114.2,111.8,104.9,61.1,56.6,56.4ppm; HRMS(ESI)calcd for C 20 H 20 ClN2O4 + [(M+H) + ]:387.1106,found:387.1099.

[0196] 2-(4-ethoxyphenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SZ-4)

[0197]

[0198] Compound SZ-4 was prepared in the same manner as SZ-1, but the boric acid used was 4-ethoxyphenylboronic acid (78 mg, 472 μmol), yielding a white solid SZ-4 (145 mg, yield 84%). Mp 178.8-179.4 °C; 1 H NMR (400MHz, CDCl3) δ8.75(d,J=5.3Hz,1H),8.50(d,J=8.9Hz,2H),7.46-7.45(m,3H),7.01( d,J=8.9Hz,2H),4.13(q,J=7.0Hz,2H),3.99(s,6H),3.94(s,3H),1.46(t,J=7.0Hz,3H)ppm; 13 C NMR (100MHz, CDCl3) δ164.4,163.6,161.5,157.7,153.8,141.1,132.7,130.5,130.0,114.6,113.7,104.9,63.7,61.1,56.5,14.9ppm; HRMS(ESI)calcd for C 21 H 23 N2O4 + [(M+H) + ]:367.1652,found:367.1649.

[0199] 2-(4-ethoxy-3-fluorophenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SZ-5)

[0200]

[0201] Compound SZ-5 was prepared in the same manner as SZ-1, but the boric acid used was 4-ethoxy-3-fluorophenylboronic acid (87 mg, 472 μmol), yielding a white solid SZ-5 (80 mg, yield 49%). Mp 151.8-153.3 °C; 1 H NMR (400MHz, CDCl3) δ8.75(d,J=5.3Hz,1H),8.46-7.66(m,2H),7.49(d,J=5.3Hz,1H),7.44(s,2H) ,7.06(t,J=8.6Hz,1H),4.20(q,J=7.0Hz,2H),4.00(s,6H),3.94(s,3H),1.50(t,J=7.0Hz,3H)ppm; 13 C NMR(100MHz,CDCl3)δ163.7,163.4(d,J C,F =245Hz),157.8,153.8,152.7(d,J C,F =243Hz), 149.5(d,J C,F =11Hz),141.1,132.4,131.1(d,J C,F =7Hz), 124.7(d,J C,F =3Hz), 116.1(d,J C,F =20Hz),114.2,114.2,114.1,104.8,65.0,61.1,56.5,14.9ppm; HRMS(ESI)calcd forC 21 H 22 FN2O4 + [(M+H) + ]:385.1558,found:385.1553.

[0202] 2-(3-chloro-4-ethoxyphenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SZ-6)

[0203]

[0204] Compound SZ-6 was prepared in the same manner as SZ-1, but the boric acid used was 3-chloro-4-ethoxyphenylboronic acid (95 mg, 472 μmol), yielding a white solid SZ-6 (118 mg, yield 69%). Mp 193.2-194.3 °C; 1H NMR (400MHz, CDCl3) δ8.74(d,J=5.3Hz,1H),8.58(d,J=2.1Hz,1H),8.41(dd,J=8.7,2.1Hz,1H),7.48(d,J=5.3Hz,1 H),7.43(s,2H),7.02(d,J=8.7Hz,1H),4.19(q,J=7.0Hz,2H),3.99(s,6H),3.94(s,3H),1.51(t,J=7.0Hz,3H)ppm; 13 C NMR (100MHz, CDCl3) δ163.6,163.3,157.8,156.7,153.8,141.1,132.4,131.2,130 .4,128.0,123.2,114.1,112.8,104.8,64.9,61.1,56.5,14.8ppm; HRMS(ESI)calcd for C 21 H 22 ClN2O4 + [(M+H) + ]:401.1263,found:401.1260.

[0205] 2-(4-(methyl)phenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SZ-7)

[0206]

[0207] Compound SZ-7 was prepared in the same manner as SZ-1, but the boric acid used was 4-methylthiophenylboronic acid (79 mg, 472 μmol), yielding a white solid SZ-7 (129 mg, yield 82%). Mp 143.5-144.8 °C; 1 H NMR (400MHz, CDCl3) δ8.77(d,J=2.0Hz,1H),8.47(d,J=8.5Hz,2H),7.50(d,J=2.0Hz ,1H),7.45(s,2H),7.36(d,J=8.5Hz,2H),3.99(s,6H),3.94(s,3H),2.55(s,3H)ppm; 13 C NMR (100MHz, CDCl3) δ164.2,163.6,157.7,153.8,142.4,141.1,134.5,132.5,128.7,125.9,114.2,104.8,61.1,56.5,15.4ppm; HRMS(ESI)calcd for C 20 H 21 N2O3S +[(M+H) + ]:369.1267,found:369.1263.

[0208] 2-(4-(ethyl)phenyl)-4-(3,4,5-trimethoxyphenyl)pyrimidine (SZ-8)

[0209]

[0210] Compound SZ-8 was prepared in the same manner as SZ-1, but the boric acid used was 4-(ethylthio)phenylboronic acid (86 mg, 472 μmol), yielding a white solid SZ-8 (147 mg, 90% yield). Mp 149.9-150.6℃; 1 H NMR (400MHz, CDCl3) δ8.78(d,J=5.2Hz,1H),8.47(d,J=8.6Hz,2H),7.51(d,J=5.2Hz,1H),7.46(s,2H ),7.42(d,J=8.6Hz,2H),4.00(s,6H),3.94(s,3H),3.04(q,J=7.4Hz,2H),1.38(t,J=7.4Hz,3H)ppm; 13 C NMR (100MHz, CDCl3) δ164.2,163.7,157.7,153.8,141.2,140.9,135.0,132.5,128.8,127.8,114.2,104.9,61.1,56.5,27.0,14.3ppm; HRMS(ESI)calcd for C 21 H 23 N2O3S + [(M+H) + ]:383.1424,found:383.1421.

[0211] Example 7: Preparation of 4-(2,4-dimethoxyphenyl)-2-(4-methoxyphenyl)pyrimidine (SZ-9)

[0212] 2-Chloro-4-(2,4-dimethoxyphenyl)pyrimidine (V5)

[0213]

[0214] Take a 250 mL inclined two-necked flask and dissolve the starting materials 2,4-dichloropyrimidine (5.00 g, 33.6 mmol), 2,4-dimethoxyphenylboronic acid (6.12 g, 33.6 mmol), and potassium carbonate (9.29 g, 67.2 mmol) in a mixed solvent of Toluene:EtOH:H2O = 3:1:1. Then, quickly add (beta-4)-platinum (3.88 g, 3.36 mmol), evacuate, and under N2 protection, gradually raise the temperature to 70 °C and reflux for 5 h. After the reaction is complete as monitored by TLC, cool to room temperature, filter with diatomaceous earth, and evaporate the filtrate to dryness. Dissolve in EA (3 × 50 mL), extract successively with water (50 mL) and saturated NaCl solution (50 mL), combine the organic phases, and dry with anhydrous Na2SO4. The sample was filtered and mixed with silica gel, then purified by column chromatography at a ratio of PE:EA = 8:1 to obtain a white solid V5 (5.6 g, yield 67%). MPa: 128.9-129.2℃. 1 H NMR (400MHz, CDCl3) δ8.49(d,J=5.4Hz,1H),8.17(d,J=8.8Hz,1H),7.95(d,J=5.4Hz, 1H),6.63(dd,J=8.8,2.2Hz,1H),6.53(d,J=2.2Hz,1H),3.91(s,3H),3.88(s,3H)ppm; 13 C NMR(100MHz, CDCl3)δ165.3,163.9,161.1,160.1,158.9,132.9,119.2,117.3,105.9,98.9,55.7,55.7ppm; HRMS(ESI)calcd for C 12 H 12 ClN2O2 + [(M+H) + ]:251.0582,found:251.0582.

[0215] 4-(2,4-Dimethoxyphenyl)-2-(4-methoxyphenyl)pyrimidine (SZ-9)

[0216]

[0217] Take a 25 mL inclined two-necked flask and dissolve the raw material V5 (120 mg, 479 μmol), 4-methoxyphenylboronic acid (80.1 mg, 527 μmol), and potassium carbonate (132 mg, 958 μmol) sequentially in a mixed solvent of Toluene:EtOH:H2O = 3:1:1. Then, quickly add (beta-4)-platinum (55.4 mg, 47.9 μmol), evacuate under N2 protection, and gradually heat to 100 °C and reflux for 5 h. After the reaction is complete as monitored by TLC, cool to room temperature, filter with diatomaceous earth, and evaporate the filtrate to dryness. Dissolve in EA (3 × 15 mL), extract successively with water (15 mL) and saturated NaCl solution (20 mL), combine the organic phases, and dry with anhydrous Na2SO4. The sample was filtered and mixed with silica gel, then purified by column chromatography at a PE:EA ratio of 4:1 to obtain a white solid SZ-9 (88 mg, yield 52%). MPa 85.4-85.8℃; 1 HNMR (400MHz, CDCl3) δ8.68(d,J=5.3Hz,1H),8.49(d,J=9.0Hz,2H),8.30(d,J=8.7Hz,1H),7.80(d,J=5.4Hz,1H),7 .00(d,J=9.0Hz,2H),6.68(dd,J=8.7,2.3Hz,1H),6.56(d,J=2.3Hz,1H),3.91(s,3H),3.88(s,3H),3.88(s,3H)ppm; 13 C NMR (100MHz, CDCl3) δ164.0,163.0,162.1,161.8,159.9,156.8,132.5,131.2, 129.8,119.5,118.3,113.9,105.7,99.0,55.7,55.7,55.5ppm; HRMS(ESI)calcd for C 19 H 19 N2O3 + [(M+H) + ]:323.1390,found:323.1392.

[0218] Example 8: Preparation of SZ-10 to SZ-18

[0219] 5-(4-(2,4-dimethoxyphenyl)pyrimidin-2-yl)-2-methoxyphenol (SZ-10)

[0220]

[0221] Take a 50 mL inclined two-necked flask and dissolve 250 mg (603 μmol) of the hydroxyl-protected benzyl-protected raw material and 25 mg of Pd / C in 10 mL of methanol. Vacuum the mixture, replace with hydrogen gas, and stir at room temperature for 6 h. After the reaction is complete as monitored by TLC, add diatomaceous earth for filtration to remove palladium on carbon, and directly add silica gel for mixing. Purify by column chromatography (PE:EA = 3:1) to obtain a white solid SZ-10 (79 mg, yield 39%). Mp 85.1-86.8℃; 1 H NMR (400MHz, CDCl3) δ8.67(d,J=5.3Hz,1H),8.32(d,J=8.7Hz,1H),8.15(s,1H),8.09(d,J=8.5Hz,1H),7.81(d,J=5.3Hz,1H),6. 95(d,J=8.5Hz,1H),6.67(dd,J=8.7,1.5Hz,1H),6.55(d,J=1.5Hz,1H),5.79(s,1H),3.95(s,3H),3.90(s,3H),3.88(s,3H)ppm; 13 C NMR (100MHz, CDCl3) δ163.7,163.1,162.2,160.0,156.6,148.9,145.8,132.6,132.0, 120.8,119.4,118.4,114.6,110.5,105.7,99.0,56.1,55.7,55.6ppm; HRMS(ESI)calcd for C 19 H 19 N2O4 + [(M+H) + ]:339.1339,found:339.1339.

[0222] 4-(2,4-Dimethoxyphenyl)-2-(3-fluoro-4-methoxyphenyl)pyrimidine (SZ-11)

[0223]

[0224] Compound SZ-11 was processed in the same manner as SZ-9, but the boric acid used was 3-fluoro-4-methoxyphenylboronic acid (90 mg, 527 μmol), yielding a white solid SZ-11 (88 mg, yield 49%). Mp 125.2-126.8℃; 1H NMR (400MHz, CDCl3) δ8.67(d,J=5.4Hz,1H),8.33-8.25(m,3H),7.83(d,J=5.4Hz,1H),7.05(t,J=8.5H z,1H),6.68(dd,J=8.7,2.4Hz,1H),6.56(d,J=2.4Hz,1H),3.96(s,3H),3.91(s,3H),3.89(s,3H)ppm; 13 C NMR(100MHz,CDCl3)δ163.1,162.9(d,J C,F =3Hz),162.2,160.0,156.8,152.6(d,J C,F =243Hz), 149.8(d,J C,F =11Hz), 132.5, 131.8 (d, J) C,F =7Hz), 124.5(d,J C,F =3Hz), 119.2, 118.7, 116.0 (d, J) C,F =20Hz),113.0,105.7,99.0,56.4,55.7,55.7ppm; HRMS(ESI)calcd forC 19 H 18 FN2O3 + [(M+H) + ]:341.1296,found:341.1295.

[0225] 2-(3-chloro-4-methoxyphenyl)-4-(2,4-dimethoxyphenyl)pyrimidine (SZ-12)

[0226]

[0227] Compound SZ-12 was processed in the same manner as SZ-9, but the boric acid used was 3-chloro-4-methoxyphenylboronic acid (98 mg, 527 μmol), yielding a white solid SZ-12 (124 mg, yield 66%). Mp 140.2-142.2℃; 1H NMR (400MHz, CDCl3) δ8.67(d,J=5.3Hz,1H),8.58(d,J=1.7Hz,1H),8.43(dd,J=8.5,1.7Hz,1H),8.29(d,J=8.5Hz,1H),7.83(d, J=5.3Hz,1H),7.02(d,J=8.6Hz,1H),6.69(dd,J=8.6,1.9Hz,1H),6.56(d,J=1.9Hz,1H),3.97(s,3H),3.91(s,3H),3.89(s,3H). 13 C NMR (100MHz, CDCl3) δ163.2,162.8,162.4,160.0,157.0,156.7,132.6,132.0,130.3, 128.0,122.8,119.2,118.7,111.8,105.7,99.0,56.4,55.7,55.7ppm; HRMS(ESI)calcd for C 19 H 18 ClN2O3 + [(M+H) + ]:357.1000,found:357.0997.

[0228] 2,4-Bis(2,4-dimethoxyphenyl)pyrimidine (SZ-13)

[0229]

[0230] Compound SZ-13 was processed in the same manner as SZ-9, but the boric acid used was 2,4-dimethoxyphenylboronic acid (96 mg, 527 μmol), yielding a white solid SZ-13 (119 mg, yield 64%). Mp 116.6-117.7 °C; 1 H NMR (400MHz, CDCl3) δ8.74(d,J=5.4Hz,1H),8.21(d,J=8.7Hz,1H),7.85(d,J=8.4Hz,1H),7.80(d ,J=5.4Hz,1H),6.65-6.58(m,3H),6.55(d,J=2.3Hz,1H),3.90(s,3H),3.89(s,3H),3.86(s,6H). 13C NMR (100MHz, CDCl3) δ165.2,162.9,162.2,162.14,159.8,159.5,156.4,133.2,132.7,122.3,11 9.6,118.1,105.7,105.1,99.7,98.9,77.5,77.2,76.8,56.3,55.7,55.6,55.6.HRMS(ESI)calcd for C 20 H 21 N2O4 + [(M+H) + ]:353.1496,found:353.1491.

[0231] 4-(2,4-Dimethoxyphenyl)-2-(4-ethoxyphenyl)pyrimidine (SZ-14)

[0232]

[0233] Compound SZ-14 was processed in the same manner as SZ-9, but the boric acid used was 4-ethoxyphenylboronic acid (87 mg, 527 μmol), yielding a white solid SZ-14 (148 mg, yield 84%). Mp 116.6-117.7℃; 1 H NMR (400MHz, CDCl3) δ8.67(d,J=5.4Hz,1H),8.48(d,J=8.9Hz,2H),8.30(d,J=8.7Hz,1H),7.79(d,J=5.4Hz,1H),6.99(d,J=8.9Hz, 2H),6.68(dd,J=8.7,2.3Hz,1H),6.56(d,J=2.3Hz,1H),4.12(q,J=7.0Hz,2H),3.91(s,3H),3.88(s,3H),1.45(t,J=7.0Hz,3H)ppm; 13 C NMR (100MHz, CDCl3) δ164.0,163.0,162.2,161.2,160.0,156.7,132.6,131.0,129 .9,119.6,118.3,114.5,105.7,99.0,63.7,55.7,55.7,15.0ppm; HRMS(ESI)calcd for C 20 H 21 N2O3 + [(M+H) + ]:337.1547,found:337.1545.

[0234] 4-(2,4-Dimethoxyphenyl)-2-(4-ethoxy-3-fluorophenyl)pyrimidine (SZ-15)

[0235]

[0236] Compound SZ-15 was prepared in the same manner as SZ-9, but the boric acid used was 4-ethoxy-3-fluorophenylboronic acid (97 mg, 527 μmol), yielding a white solid SZ-15 (95 mg, 51% yield). Mp 139.2-140.9℃; 1 H NMR (400MHz, CDCl3) δ8.67(d,J=5.4Hz,1H),8.29(d,J=8.8Hz,3H),7.82(d,J=5.4Hz,1H),7.04(t,J=8.6Hz,1H),6.68(d d,J=8.6,2.3Hz,1H),6.56(d,J=2.3Hz,1H),4.19(q,J=7.0Hz,2H),3.91(s,3H),3.88(s,3H),1.49(t,J=7.0Hz,3H)ppm; 13 C NMR(100MHz,CDCl3)δ163.1,163.0(d,J C,F =3Hz),162.2,160.0,158.9,156.8,152.8(d,J C,F =243Hz), 149.1(d,J C,F =11Hz), 132.5, 131.8 (d, J) C,F =7Hz), 124.4(d,J C,F =3Hz), 119.3, 118.6, 116.1 (d, J C,F =10Hz),114.2,105.9,105.7,99.0,65.0,55.7,55.7,14.9ppm; HRMS(ESI)calcd for C 20 H 20 FN2O3 + [(M+H) + ]:355.1452,found:355.1452.

[0237] 2-(3-chloro-4-ethoxyphenyl)-4-(2,4-dimethoxyphenyl)pyrimidine (SZ-16)

[0238]

[0239] Compound SZ-16 was prepared in the same manner as SZ-9, but the boric acid used was 4-ethoxy-3-chlorophenylboronic acid (106 mg, 527 μmol), yielding a white solid SZ-16 (119 mg, yield 61%). Mp 127.8-128.4 °C; 1 H NMR (400MHz, CDCl3) δ8.67(d,J=5.4Hz,1H),8.57(d,J=2.1Hz,1H),8.40(dd,J=8.6,2.1Hz,1H),8.29(d,J=8.6Hz,1H),7.82(d,J=5.4Hz,1H),7.0 0(d,J=8.7Hz,1H),6.69(dd,J=8.7,2.3Hz,1H),6.56(d,J=2.3Hz,1H),4.19(q,J=7.0Hz,2H),3.91(s,3H),3.89(s,3H),1.50(t,J=7.0Hz,3H)ppm; 13 C NMR (100MHz, CDCl3) δ163.1,162.9,162.3,160.0,156.8,156.4,132.6,131.9,130.3,127 .9,123.1,119.3,118.7,112.9,105.7,99.0,65.0,55.7,55.7,14.8ppm; HRMS(ESI)calcd for C 20 H 20 ClN2O3 + [(M+H) + ]:371.1157,found:371.1154

[0240] 4-(2,4-Dimethoxyphenyl)-2-(4-(methyl)phenyl)pyrimidine (SZ-17)

[0241]

[0242] Compound SZ-17 was prepared in the same manner as SZ-9, but the boric acid used was 4-methylthiophenylboronic acid (88 mg, 527 μmol), yielding a white solid SZ-17 (139 mg, 78% yield). Mp 113.9-115.6℃; 1H NMR (400MHz, CDCl3) δ8.69(d,J=5.4Hz,1H),8.46(d,J=8.6Hz,2H),8.30(d,J=8.7Hz,1H),7.83(d,J=5.4Hz,1H),7. 35(d,J=8.6Hz,2H),6.68(dd,J=8.7,2.3Hz,1H),6.56(d,J=2.3Hz,1H),3.91(s,3H),3.88(s,3H),2.54(s,3H)ppm; 13 C NMR (100MHz, CDCl3) δ163.8,163.1,162.3,160.0,156.8,141.7,135.2,132.6, 128.6,126.0,119.4,118.8,105.7,99.0,55.7,55.7,15.5ppm; HRMS(ESI)calcd for C 19 H 19 N2O2S + [(M+H) + ]:339.1162,found:339.1159.

[0243] 4-(2,4-Dimethoxyphenyl)-2-(4-(Ethyl)phenyl)pyrimidine (SZ-18)

[0244]

[0245] Compound SZ-18 was prepared in the same manner as SZ-9, but the boric acid used was 4-(ethylthio)phenylboronic acid (96 mg, 527 μmol), yielding a white solid SZ-18 (152 mg, yield 82%). Mp 102.2-103.6 °C; 1 H NMR (400MHz, CDCl3) δ8.69(d,J=5.4Hz,1H),8.45(d,J=8.5Hz,2H),8.30(d,J=8.7Hz,1H),7.84(d,J=5.4Hz,1H),7.40(d,J=8.5Hz, 2H),6.68(dd,J=8.7,2.3Hz,1H),6.56(d,J=2.3Hz,1H),3.91(s,3H),3.88(s,3H),3.02(q,J=7.4Hz,2H),1.36(t,J=7.4Hz,3H)ppm; 13C NMR (100MHz, CDCl3) δ163.7,163.1,162.3,160.0,156.8,140.1,135.7,132.6,128 .7,128.0,119.4,118.8,105.7,99.0,55.7,55.6,27.1,14.4ppm; HRMS(ESI)calcd for C 20 H 21 N2O2S + [(M+H) + ]:353.1315,found:353.1318.

[0246] Example 9: In vitro cervical cancer cell proliferation inhibition experiment

[0247] Cells and culture conditions:

[0248] Human cervical cancer cell lines Hela, Siha, and MS751 were ordered from the Cell Bank of the Chinese Academy of Sciences. All Hela, Siha, and MS751 cells were cultured in DMEM medium (purchased from Meilun Biotechnology) containing 10% fetal bovine serum. The human cisplatin-resistant cervical cancer cell line Hela / DDP was ordered from Shanghai Aolu Biotechnology Co., Ltd.; the human cisplatin-resistant cervical cancer cell line Siha / DDP was ordered from Hunan Fenghui Biotechnology Co., Ltd. Cell culture of the drug-resistant lines was performed according to the company's operating instructions. All cells were cultured at 37°C, saturated humidity, 5% CO2, and 95% atmospheric atmosphere.

[0249] Cell proliferation inhibition experiment:

[0250] Before the cell proliferation experiment, the solutions for each concentration of drug-treated experimental group, negative control group, and blank group were prepared according to the above method. Then, following the cell passage procedure, the density of the cell suspension was measured. Based on the plating requirements (2000–3000 cells / 100 μL per well), 100 μL of cell suspension was added to each well of a sterile NEST-96 plate and incubated for 24 hours (ensuring complete cell adhesion). The supernatant was then carefully discarded. For the compound treatment group, 100 μL of cell culture medium containing the specified concentration of the target compound was added to each well. For the blank group (no cells inoculated) and the negative control group, equal volumes of cell culture medium and cell culture medium containing 0.1% DMSO were added, respectively. Each group was configured with three replicates. After incubating statically for 48 hours, thaw the CCK-8 solution, remove the cells to be tested, add 10 μL of CCK-8 solution to each well, and continue incubating statically for 1.5 hours. After shaking the culture plate for about 1 minute, measure the absorbance (OD) at 450 nm using a microplate reader. 450 The equation for calculating the inhibition rate of the target compound is: Inhibition rate = 1 - [(OD of the compound treatment group)] 450 Value - Mean OD of blank group450 (value) / (negative control group OD) 450 Value – Mean OD of blank group 450 [Value], IC 50 The values ​​can be calculated using GraphPad Prism 5 software. The above experiment was repeated three times, and the IC values ​​were calculated three times. 50 Mean and standard deviation.

[0251]

[0252] Table 1. Screening results of the anti-cervical cancer proliferative activity of SV series CBSIs *

[0253]

[0254] *The target compound was reacted for 48 hours, and the experimental results are expressed as IC50. 50 Mean ± standard deviation (μM) were determined by three independent repeated experiments.

[0255]

[0256] Table 2. Evaluation results of the anti-cervical cancer proliferation activity of the target compounds in the SZ series. *

[0257]

[0258]

[0259] *The target compound was reacted for 48 hours, and the experimental results were determined by three independent replicate experiments.

[0260] The experimental results (Tables 1 and 2) show that diarylpyrimidine CBSIs were generally most sensitive to HeLa cells, followed by MS751 cells, while Siha cells were relatively less sensitive. In both general formulas, compounds with 4,5-diaryl ortho-substituted positions showed better overall activity than those with 2,4-diaryl meta-substituted positions. All 38 compounds exhibited micromolar activity against the three cervical cancer cell lines, with IC50 values ​​of [missing information]. 50 The activity of 31 target compounds was generally superior to cisplatin, ranging from 0.091 to 33.91 μM. The five most active representative compounds (SV-6, SV-8–10, and SZ-10) exhibited the best inhibitory activity against the proliferation of three tumor cell lines (IC50). 50 The values ​​all reached the submicromolar level, which is 8 to 52 times that of cisplatin. Among them, SV-10 was the most active compound, with an IC50 value for inhibiting the proliferation of HeLa, Siha, and MS751. 50 The value is 0.091–0.15 μM, which is 47–52 times that of cisplatin.

[0261] Example 10: Kinetics of microtubule polymerization inhibition by representative diarylpyrimidine compounds

[0262] Table 3. Experimental results on the kinetics of microtubule polymerization inhibition by some diarylpyrimidine compounds*

[0263]

[0264]

[0265] *The microtubule polymerization kinetics experiment was conducted by Jinan Huawi Pharmaceutical Technology Co., Ltd.

[0266] Eleven representative compounds from the SV and SZ series were selected for experiments on their inhibitory activity against tubulin polymerization. The results showed that all eleven compounds exhibited inhibitory activity against tubulin polymerization. Based on the target activity test results, when R2 in general formula (I) is a hydrogen atom, the activity is best when R1 is an ethoxy group, followed by a methylthio group, and then a methoxy group. However, when R1 is either methoxy or ethoxy, the substituent relationship of R2 is unclear. When R1 in compounds of general formula (I) is an ethoxy group and R2 is a hydrogen or chlorine atom, i.e., compounds SV-8 and SV-10, the target activity against tubulin polymerization is optimal, with an IC50 value of [missing value]. 50 The values ​​were 3.7 μM and 4.6 μM, respectively, which were 1.8 to 1.5 times that of the positive control drug colchicine. Meanwhile, SV-6 and SV-8–SV-10, which exhibited the best anti-cervical cancer cell proliferation activity, also showed the best target inhibitory activity, indicating an intrinsic correlation and consistency between anti-tumor cell activity and anti-microtubule target activity. In general formula (II), compound SZ-10, which exhibited the best anti-cervical cancer cell activity, also showed good target activity, with an IC50 value of 3.7 μM and 4.6 μM, respectively. 50 The concentration was 5.2 μM, which is 1.3 times that of colchicine. Based on these results, it can be concluded that the SV and SZ series compounds can specifically inhibit tubulin polymerization kinetics and can exert antitumor effects as classic tubulin polymerization inhibitors.

[0267] Example 11: In vitro experiment on the inhibition of cisplatin-resistant cervical cancer cell proliferation by SV-10

[0268] Table 4. Inhibitory activity of SV-10 on the proliferation of cisplatin-resistant cervical cancer cell lines.

[0269]

[0270]

[0271] * Rf = [Hela(Siha) / DDP IC]50 ] / [Hela(Siha)IC 50 ]

[0272] Experimental results showed that SV-10 exhibited good anti-proliferative activity against both cervical cancer cell lines, HeLa / DDP and Siha / DDP, with an IC50 concentration of 100 mg / kg / dt. 50 The values ​​were 7.21 and 6.28 μM, respectively, with resistance indices of 79.23 and 41.87, respectively. For the two cisplatin-resistant cervical cancer strains, Hela / DDP and Siha / DDP, SV-10 showed superior inhibitory activity compared to CA-4, cisplatin, and carboplatin, and was a CA-4 IC50 inhibitor. 50 The activity of SV-10 was 1.4 and 2.2 times that of cisplatin, 2.7 and 4.2 times that of cisplatin, and 34 and 35 times that of carboplatin. Analysis of the resistance index showed that SV-10 was significantly more active than cisplatin, being 9.5 and 18.6 times more potent. Based on the above experimental data, it can be concluded that compound SV-10 has stronger inhibitory activity than CA-4 against cisplatin-resistant cervical cancer cells, warranting further research.

[0273] Example 12: In vitro microtubule polymerization inhibition experiment

[0274] (1) Tubulin purification: Porcine brain tubulin was obtained by three temperature-dependent assembly / disassembly cycles under the conditions of 100 mM PIPES (pH 6.5), 1 mM MgSO4, 2 mM EGTA, 1 mM GTP, and 1 mM 2-mercaptoethanol. In the first polymerization cycle, 4 M glycerol and 0.2 mM benzyl sulfonyl fluoride were added, respectively. Homogeneous tubulin was prepared from the tubulin using cellulose phosphate (P11) chromatography, and the purified protein was then aliquoted and stored at -70°C for later use.

[0275] (2) In vitro microtubule polymerization inhibition experiment: Under the conditions of PEM buffer (100 mM MPIPES, 1 mM MgCl2 and 1 mM EGTA) containing 1 mM GTP and 5% glycerol, different concentrations of compounds were mixed with microtubules, and the microtubule polymerization was monitored at 37°C. The absorbance value was calculated by light scattering at 340 nm using a SPECTRA MAX190 (Molecular Device) spectrophotometer.

[0276] Experimental results show that SV-10 can inhibit tubulin polymerization in a concentration-dependent manner, such as... Figure 2As shown, compared with the control DMSO group, SV-10 at dosage concentrations of 3, 6, and 12.5 μM resulted in 39%, 54%, and 77% microtubule polymerization, respectively. Simultaneously, SV-10 at dosage concentrations of 3 μM and 12.5 μM exhibited comparable inhibitory activity against microtubule polymerization to 6 μM colchicine and 6 μM CA-4, respectively. The in vitro IC50 value for SV-10 inhibiting microtubule polymerization was 4.6 ± 0.2 μM, indicating that SV-10 inhibits tumor cell growth by suppressing microtubule polymerization.

[0277] Example 13: SV-10 competitively binds to colchicine binding sites

[0278] Following the cell passage procedure, the density of the cell suspension was measured, and 5 × 10⁵ cells were added to each well of a 6-well plate. 5Cells were cultured overnight. The next day, target and control compounds of various concentrations were added to each well and incubated for 2 hours. Then, 100 μM EBI was added and incubated for 1.5 hours. After this time, the culture medium was aspirated, the cells were digested, and the cell suspension was collected. The cells were centrifuged at 1500 rpm for 5 minutes, and the supernatant was aspirated. The cell pellet was rinsed twice with PBS, centrifuged (1500 rpm for 5 minutes), and the supernatant was aspirated. A specified volume of RIAP protein lysis buffer (Radio Immunoprecipitation Assay Lysis Buffer) was added to the collected cells, and the cells were placed in crushed ice. To ensure complete cell lysis, the cells were vortexed every 10 minutes (5-10 seconds). After 30 minutes, the cells were centrifuged at 4°C / 12000 rpm for 15 minutes, and the supernatant was collected. The total protein concentration was determined by the BCA (bicinchoninic acid) method. 5× loading buffer was added to each sample, mixed, and boiled in a water bath for 10 minutes. After boiling, the mixture was cooled for later use or stored at -20°C. Prepare the mini gel according to the gel preparation procedure. Adjust the loading volume according to the concentration of each sample and control the final loading amount of total protein to 50 μg. Reserve pre-stained marker lanes as needed for the experiment. After loading the samples, set the electrophoresis voltage to 80V / 40min initially, then adjust it to 120V / 70min. The electrophoresis time can be appropriately extended or shortened according to experimental requirements. After electrophoresis, cut the methanol-soaked and activated PVDF membrane. Prepare the transfer system according to the sequence of [foam pad / 5 layers of filter paper / gel / PVDF membrane / 5 layers of filter paper / foam pad "sandwich"]. The transfer voltage is constant at 110V, and the transfer time is 90min. After the transfer, block with 5% skim milk at room temperature, then wash with 1X PBST solution for 10min. Place the PVDF membrane in Anti-β-Tubulin Mouse Monoclonal Antibody solution and incubate overnight at 4°C. The next day, the primary antibody solution was recovered, and the membrane was washed 6 times with 1X PBST for 5 minutes each time. Then, the secondary antibody dilution solution of the corresponding species was added, and the membrane was incubated at room temperature for 1 hour. After incubation, the membrane was washed 6 times with 1X PBST for 5 minutes each time. After washing, the ECL luminescent solution was mixed and gently dropped onto the PVDF membrane, which was then quickly placed into the imager for detection. The development results were statistically analyzed using ImageJ software.

[0279] Experimental results are as follows Figure 3As shown, compared with DMSO and EBI alone, SV-10 inhibited the formation of EBI-β-tubulin adducts in a concentration-dependent manner from 1 μM to 100 μM, resulting in a reduction in the adduct band, which is structurally similar to the positive control colchicine from 2 μM to 5 μM. However, vincristine, as a vincristine site inhibitor, did not show inhibition of adduct band formation with increasing concentration. The EBI competition assay revealed the mechanism by which SV-10 inhibits tubulin polymerization by binding to the colchicine binding site.

[0280] Example 14: SV-10 disrupts microtubule networks at the cellular level

[0281] Following the cell passage procedure, the density of the cell suspension was measured, and 5 × 10⁵ cells were added to each well of a 24-well plate containing cell spread sheets. 4 Cells were cultured overnight. The next day, different concentrations of SV-10 were added, with DMSO as a control, and cultured for another 24 hours. The culture medium was aspirated, and the cells were gently washed three times with PBS. 300 μL of a CH3OH:AcOH mixture of 3:1 was added to each well for fixation at room temperature for 10 min, followed by three gentle washes with PBS. 0.3% Triton X100 was added to PBS for permeation for 10 min, followed by three washes with PBS. Immediately afterward, PBS containing 5% BSA was added for blocking for 0.5 h. α-Tubulin antibody was added, and the cells were placed in a foam box containing well-moistened gauze at 4°C overnight. The next day, the cells were gently washed three times with PBS, then Alexa Fluor 488 was added, and the cells were incubated at 37°C in the dark for 2 h. After three gentle washes with PBS, anti-fluorescence quenching agent was dropped onto a glass slide, and the slide was flipped over to cover the slide. The experimental results were observed and photographed using a confocal microscope.

[0282] Experimental results are as follows Figure 4 As shown, immunofluorescence analysis of HeLa cells after drug administration using confocal microscopy also revealed disruption of microtubule dynamics in the drug-treated group. Figure 4 As shown, after DMSO treatment of HeLa cells for 24 hours, the microtubule network in the DMSO group was normally arranged and well-distributed. In contrast, SV-10 caused significant abnormalities and disruptions in tubulin in a concentration-dependent manner. These results indicate that SV-10 can inhibit tubulin polymerization and interfere with the formation of normal microtubule networks in a manner similar to CA-4 and colchicine.

[0283] Example 15: In vitro cell cycle inhibition experiment

[0284] Following the cell passage procedure, the density of the cell suspension was measured, and 2.5 × 10⁻⁶ cells were added to each well of a 6-well plate. 5Cells were collected and transferred to an incubator for overnight culture. The next day, the supernatant was aspirated. The drug-treated group received 2 mL of complete culture medium containing the target compounds at predetermined concentrations, while the control group received an equal volume of cell culture medium containing 0.1% DMSO and continued culture for 24 h. The supernatant was then discarded. Cells were carefully washed and digested with PBS and trypsin, respectively, to prepare single-cell suspensions. After preparation, the cells were centrifuged (1500 rpm / 5 min) and the supernatant was aspirated. The cell pellet was rinsed twice with pre-chilled PBS, centrifuged again (1500 rpm / 5 min) and the supernatant was aspirated. The cell pellet was resuspended in pre-chilled 70% ethanol and gently mixed, then fixed overnight at 4°C. The next day, the cells were centrifuged (1500 rpm / 5 min) and the supernatant was aspirated. The cell pellet was rinsed once with pre-chilled PBS, centrifuged again (1500 rpm / 5 min) and the supernatant was aspirated to obtain the cell pellet. Add 0.5 mL of staining buffer, 25 μL of 20×PI staining solution, and 10 μL of 50×RNase A solution to each cell pellet sample. After preparation, mix well and add to each cell sample. Then incubate at 37°C in the dark for 0.5 h. After incubation, perform the cell analysis using a Beckman flow cytometer according to the operating procedure. Figure 5-A The results showed that compound SV-10 could significantly arrest the G2 / M phase.

[0285] Example 16: In vitro apoptosis induction experiment

[0286] Following the cell passage procedure, the density of the cell suspension was measured, and 2.5 × 10⁻⁶ cells were added to each well of a 6-well plate. 5 Cells were collected and transferred to an incubator for overnight culture. The next day, the supernatant was aspirated. The drug-treated experimental group received 2 mL of complete culture medium containing the target compounds at predetermined concentrations, while the control group received an equal volume of cell culture medium containing 0.1% DMSO and continued culture for 24 h. The supernatant was then discarded, cells were enriched, and centrifuged (1500 rpm / 5 min) to obtain a cell pellet. The supernatant was aspirated, and the cell pellet was washed twice with PBS, centrifuged (1500 rpm / 5 min), and the supernatant was aspirated. The cell pellet was gently resuspended in Annexin V binding buffer. 5 μL of Annexin V-PE and 7-AAD staining solution were added to each cell sample, gently mixed, and incubated at room temperature in the dark for 15 min. Samples were then analyzed using a Beckmann flow cytometer according to the prescribed procedures. Figure 5-B The results showed that compound SV-10 could induce apoptosis in cells.

[0287] Example 17: Inhibition of cell colony formation in vitro

[0288] Following the cell passage procedure, cell suspension density was measured, and 500 cells were added to each well of a 6-well plate and cultured overnight. Both the control and drug-treated groups had three replicates. After cell attachment, drug administration was initiated based on cell count to prevent premature cell death. After drug administration began, the medium was changed and the drug was re-added every two days. The experiment was terminated when individual clones were visible to the naked eye, with the entire experimental period lasting approximately 10–14 days. At this point, the culture medium was aspirated, cells were gently washed with PBS, fixed with 4% paraformaldehyde for 0.5 h, the fixative was aspirated, cells were stained with 0.2% crystal violet for 30 min, gently washed with PBS, and the liquid was drained before photographic analysis. Figure 5-C The results showed that compound SV-10 could inhibit cell clone formation.

[0289] Example 18: Inhibition of cell migration in vitro

[0290] ① Mark 5 parallel straight lines evenly on the bottom of the six-hole plate; ② Add 3×10 to each hole 5 Cells were cultured overnight. ③ Using a white sterile pipette tip, evenly scratched the cells along the vertical marking line; ④ Immediately wash away floating cells with PBS. The drug-treated experimental group was given 2 mL of serum-free culture medium containing the target compound at the set concentration, while the control group was given an equal volume of serum-free culture medium containing 0.1% DMSO; ⑤ After culturing for 24 h, the cells were photographed under a microscope, and the scratch distance was analyzed and calculated using ImageJ software. Figure 5-D The results showed that compound SV-10 could inhibit cell migration and prevent scratch healing.

[0291] Example 19: Water solubility and LogP test

[0292] Water solubility is one of the key issues that need to be addressed in drug development. This experiment, based on the solubility test methods specified in the Chinese Pharmacopoeia, estimated the solubility of compounds, prepared standard solutions of the compounds, and established standard curves for the preferred compound SV-10 and the control compound CA-4. The solubility of the preferred compound SV-10 and the control compound CA-4 in different buffer solutions was calculated based on the established standard curve equations. Detailed data are summarized in Table 5.

[0293] Table 5. Solubility test results of SV-10 and CA-4

[0294]

[0295]

[0296] The solubility of SV-10 in water (pH=7.0) is 8.12 μg / mL, comparable to CA-4 (9.85 μg / mL). The solubility of SV-10 in weakly alkaline conditions (simulating human body fluids, pH=7.4) is 17.15 μg / mL, four times that of CA-4 (4.34 μg / mL). The solubility of SV-10 in strongly acidic conditions (simulating human gastric juice, pH=2.0) is 31.66 μg / mL, ten times that of CA-4 (3.23 μg / mL). The LogP of SV-10 is 2.36, comparable to CA-4 (2.04). Therefore, SV-10 exhibits good water solubility and has potential for further drug development.

[0297] Example 20: In vitro study of liver microsomal metabolic stability

[0298] The liver is the primary organ for drug metabolism in the human body, with most drug metabolism occurring primarily through hepatic drug-metabolizing enzyme systems. Therefore, studying the in vitro stability of drugs in hepatic microsomal metabolites is of significant importance for predicting preliminary drug-likeness. In this experiment, the test compound, hepatic microsomal protein (0.5 mg / mL), 1 mM NADPH, and 0.1 M phosphate buffer were mixed and incubated for different times. The compound concentration at different times was detected by LC-MS-MS, and its half-life (T0) was calculated. 1 / 2 ), Liver microparticle in vivo clearance rate CL int(mic) Intrahepatic clearance rate CL int(liver) The percentage of compounds remaining after 60 min was determined. Propafenone, diclofenac, and testosterone, which have moderate metabolic stability in liver microsomes, were selected as control drugs. The experimental results are shown in Table 6.

[0299] Table 6 Results of in vitro liver microsomal metabolic stability assay for SV-10

[0300]

[0301] As the experimental results show: the metabolic half-life T of SV-10 in human liver microsomes is... 1 / 2 The time to clearance was 29.9 min, and the intrinsic clearance rate was 46.4 μL / min / mg. This result is superior to the reported stability of CA-4 metabolism in human liver microsomes in vitro (T0) in the literature (Oncology Reports, 2013, 29(6):2451-2458.). 1 / 2 The half-life was 8.83 min, which is 3.4 times its metabolic half-life. The preferred compound SV-10 has a mouse liver microsomal half-life comparable to propafenone, T... 1 / 2The detection time was 1.8 minutes, and the drug was undetectable at 60 minutes. These experimental results indicate that SV-10 exhibits good metabolic stability in human liver microsomes and possesses good potential for drug development.

[0302] Example 21: In vivo pharmacodynamic study of SV-10 against cervical cancer

[0303] Following the cell passage procedure, the density of the HeLa cell suspension was measured at 6 × 10⁻⁶. 6 100 μL / cell per female Nude Balb / C nude mouse was injected into the right back of the mouse. The long axis (L) and short axis (W) of the tumor were observed and measured, and the tumor volume was calculated. The tumor volume was calculated when it reached 90 mm². 3 Nude mice were then randomly divided into 5 groups: a blank solvent group, a 30 mg / kg SV-10 administration group, a 60 mg / kg SV-10 group, a 6 mg / kg cisplatin administration group, and a 3 mg / kg cisplatin + 30 mg / kg SV-10 combination administration group. Administration began on day 1. SV-10 and DDP were administered intraperitoneally. SV-10 was administered once daily, 6 mg / kg DDP every 4 days, and 3 mg / kg DDP every 2 days. The weight of the nude mice and the long and short diameters of the tumors were measured every 3 days. After 21 days of continuous administration, the experiment ended. The nude mice were sacrificed, tumor tissue was obtained, photographed, and weighed to calculate the total glycated tumor (TGI). Three nude mice from each administration group were randomly selected, blood was collected from their eyeballs, and serum was separated for subsequent evaluation of renal function indicators. Subsequently, the heart, liver, spleen, lungs, and kidneys of the selected nude mice were fixed in 4% paraformaldehyde for 24 hours, then embedded, and 4 μm paraffin sections were prepared. Tumor volume = 0.5 × L × W 2 Where W and L represent the short diameter and long diameter, respectively. The formula for calculating the tumor inhibition rate (TGI) is: TGI = (1 - average tumor weight in the drug-treated group / average tumor weight in the solvent-treated group) × 100%. The tumor inhibition rates of each group are summarized in Table 7.

[0304] Table 7. Statistical table of the inhibition rate of anti-cervical cancer proliferation in each treatment group.

[0305]

[0306] The experimental results confirmed that SV-10 alone is effective and safe for inhibiting cervical cancer tumor growth in vivo. Furthermore, SV-10 can be used in combination with DDP, which not only improves the therapeutic effect but also reduces the dosage of cisplatin and mitigates the toxic side effects of high-dose DDP. This aligns with the initial research objectives and suggests that SV-10, as a microtubule colchicine site inhibitor, has potential anti-cervical cancer therapeutic effects and can be combined with cisplatin to enrich clinical treatment regimens for cervical cancer. Detailed experimental results are shown in Figure 6.

[0307] Example 22: Effects of SV-10 on renal function in nude mice in vivo

[0308] Blood was collected from the eyeballs and centrifuged at 4000 rpm for 5 min. The serum was then separated, transferred, and stored at -20℃ for later use. The analytical procedure was performed according to the Redu / Changchun Huili reagent kit instructions, with the appropriate parameters set and samples loaded onto the fully automated biochemical analyzer.

[0309] Figure 7-A Comparison of creatinine (CREA, μM) levels in blood samples from five groups of nude mice: Creatinine is an important indicator for assessing kidney function. The experimental results show that the average creatinine level (14.79 μM) in the cisplatin-only group was 67% higher than that in the control group, suggesting that cisplatin administration may cause kidney damage in mice. The creatinine levels in the other three groups were 31%, 23%, and 40% higher than those in the control group, respectively. There was no significant difference between the combined administration group and the control group.

[0310] Figure 7-B Comparison of uric acid (UA, μM) levels in blood samples from five groups of nude mice: The cisplatin-only group (149 μM) showed a 95% increase in uric acid levels compared to the control group (75 μM), suggesting that cisplatin administration may cause kidney damage or nephropathy in mice; the other three groups showed a 21% increase, a 36% decrease, and a 16% decrease in creatinine levels compared to the control group, respectively; there was no significant difference between the combined administration group and the control group.

[0311] Figure 7-C Comparison of blood urea nitrogen (UREA, mg / dL) levels in five groups of nude mice: The urea nitrogen levels in the four treatment groups were not significantly different from the control group; the increase or decrease remained within the range of 1.3%–23%. This indicates that cisplatin or SV-10 had no significant effect on urea nitrogen levels.

[0312] By combining in vivo pharmacological activity experiments against cervical cancer, it was found that cisplatin alone at 6 mg / kg exhibited slightly better antitumor activity than SV-10 at 60 mg / kg, but its toxicity was significantly stronger. Therefore, combining SV-10 with cisplatin can improve activity while significantly reducing the nephrotoxicity caused by high-dose cisplatin, providing a basis for subsequent research on combination therapy with cisplatin.

[0313] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. However, the present invention is not limited to the embodiments described. Various changes and modifications can be made to the present invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed.

[0314] The above description of the embodiments is provided to enable those skilled in the art to understand and use the invention. It will be apparent to those skilled in the art that various modifications can be made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present invention is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the invention should be within the protection scope of the present invention.

Claims

1. A diarylpyrimidine compound or a pharmaceutically acceptable salt thereof, characterized in that: The compounds selected are shown below:

2. The method for preparing the diarylpyrimidine compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The preparation route for the diarylpyrimidine compounds represented by formula (I) is as follows: In step i, 4-chloro-5-bromopyrimidine is used as the initial raw material and reacted with 3,4,5-trimethoxyphenylboronic acid in a reaction solvent to prepare intermediate compound V2. In step ii, intermediate compound V2 is reacted with arylboronic acid in a reaction solvent to prepare the target compound of general formula (I); The target compound of general formula (I) is selected from the following compounds:

3. The use of the diarylpyrimidine compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The use of the diarylpyrimidine compounds or their pharmaceutically acceptable salts in the preparation of antitumor drugs.

4. The application according to claim 3, characterized in that: The tumor was selected as cervical cancer.

5. The application according to claim 3, characterized in that: The use of the diarylpyrimidine compounds or their pharmaceutically acceptable salts in the preparation of microtubule colchicine binding site inhibitors.

6. An antitumor drug composition, characterized in that: It includes: A therapeutically effective amount of the diarylpyrimidine compound of claim 1 or a pharmaceutically acceptable salt thereof. And pharmaceutically acceptable carriers.

7. The antitumor drug composition according to claim 6, characterized in that: It includes: A therapeutically effective amount of the diarylpyrimidine compound of claim 1 or a pharmaceutically acceptable salt thereof, together with a platinum-based drug, serves as the active ingredient. And pharmaceutically acceptable carriers.

8. The antitumor drug composition according to claim 7, characterized in that: The platinum-based drug selected was cisplatin.

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