Artemisinin trimer derivative, its preparation method and application

Abstract

This invention relates to an artemisinin trimer derivative, its preparation method, and its application. This invention is the first to synthesize and isolate β,β,α trimer, β,α,α trimer, and α,α,α trimer. The isomers of these three trimers were successfully isolated by the new synthetic method and characterized and their activity was tested. The test results showed that they have high antitumor activity.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to an artemisinin trimer derivative, its preparation method, and its application. Background Technology

[0002] Artemisinin is a medicinal component extracted from the Artemisia annua plant, also known as artemisinin. It was first discovered in the late 1970s by Chinese scientist Tu Youyou and her team. Through research on the traditional Chinese herbal medicine Artemisia annua, Tu Youyou and others discovered that it contained an effective antimalarial component. After years of research and experimentation, they successfully extracted this component from Artemisia annua and named it artemisinin. It is an effective antimalarial and anti-inflammatory drug, particularly effective in treating malaria, and is widely used in clinical treatment.

[0003] Artemisinin not only has therapeutic effects on malaria but also possesses anti-tumor properties, primarily through inhibiting tumor cell proliferation: artemisinin can inhibit tumor cell proliferation, blocking tumor cell growth and division; inducing tumor cell apoptosis: artemisinin can induce tumor cell apoptosis, prompting tumor cells to die; blocking tumor blood supply: artemisinin can inhibit tumor angiogenesis, blocking tumor blood supply, thereby limiting tumor growth and spread; and enhancing immune function: artemisinin can enhance the body's immune function and improve the body's resistance to tumors.

[0004] In recent years, many scientists have modified and engineered artemisinin, leading to further research on its anti-tumor effects. Studies have shown that artemisinin dimers and trimers are significantly more effective than monomers, inhibiting tumor growth, and their activity is several orders of magnitude higher than that of the monomers.

[0005] Patent documents CN 109467565 A, CN 111372583A, and AU2007235813A1 report the synthesis of artemisinin trimers. These methods involve using dihydroartemisinin and a small molecule such as a polyol or haloalcohol (e.g., bromoethanol, ethylene glycol, glycerol, resorcinol, phloroglucinol, etc.) in the presence of a Lewis acid to form an intermediate with hydroxyl, phenolic, or bromine content. This intermediate is then reacted with a linker in the presence of a Lewis acid or base to form an ether, and a protecting group is grafted onto it. Finally, the protecting group is removed under acidic, alkaline, or neutral conditions to obtain the dihydroartemisinin trimer. However, the yields of dihydroartemisinin trimers prepared using these methods are relatively low, with the yield of a single configuration being less than 10%, and separation is difficult. Most of the trimeric compounds reported in the literature are mixtures of multiple configurations, and their antitumor activity and mechanisms of action remain unclear. While the synthesis and isolation of βββ-configuration trimers have been reported, the other three isomers have not been successfully isolated. Dihydroartemisinin possesses multiple chiral centers, and the synthesis of its derivatives mainly occurs at the hydroxyl group attached to carbon 10. However, the configuration of carbon 10 in the dimer and trimer derivatives reported in the existing literature is still unclear.

[0006] Therefore, a synthetic method for artemisinin trimers with more defined configurations and higher yields needs to be further developed. Summary of the Invention

[0007] The purpose of this invention is to address the aforementioned problems by providing an artemisinin trimer derivative, its preparation method, and its applications. This invention provides an artemisinin trimer with higher yield and more defined configuration, along with its synthesis method, offering a new approach for studying the configuration of artemisinin trimers.

[0008] The objective of this invention is achieved through the following technical solution:

[0009] In one aspect of this application, a compound of this application is provided.

[0010] The compound of this application is an artemisinin trimer derivative, which has the structure shown in the following formula:

[0011]

[0012] A method for preparing an artemisinin trimer derivative involves reacting compound SM1044 and compound 6 in an organic solvent under the action of an alkaline catalyst to generate the target product.

[0013] The chemical structural formula of the compound SM1044 is one of the following:

[0014]

[0015] The chemical structural formula of compound 6 is one of the following:

[0016]

[0017] As a preferred embodiment of the present invention, the organic solvent is solvent 1, which is a solvent capable of dissolving the compound SM1044 and the compound 6, including one or more of dimethylformamide, methanol, ethanol, isopropanol, acetone, dimethylacetamide, and acetonitrile. Preferably, solvent 1 is dimethylformamide.

[0018] And / or, the alkaline catalyst is alkaline catalyst 1, comprising one or more of potassium carbonate, sodium carbonate, cesium carbonate, triethylamine, N,N-diisopropylethylamine, and DBU, preferably, the alkaline catalyst 1 is potassium carbonate;

[0019] And / or, the molar ratio of compound SM1044, compound 6, and base catalyst 1 is 1:1 to 5.

[0020] And / or, the reaction temperature is 0-45℃, and the reaction time is 2-72 hours; preferably, the reaction temperature is 40℃, and the reaction time is 48 hours;

[0021] And / or, after the reaction is complete, separation and purification are performed, including the following steps: adding saturated NH4Cl, extracting multiple times with EA, washing the organic layer with brine and drying on anhydrous Na2SO4, concentrating the mixture under reduced pressure, and purifying the residue by chromatographic column to obtain the product.

[0022] As a preferred technical solution of the present invention, the compound SM1044 is obtained by deprotecting compound 4. In an organic solvent, compound 4 generates compound SM1044 in a free state under the action of an alkaline catalyst.

[0023] The chemical structural formula of compound 4 is as follows:

[0024]

[0025] As a preferred embodiment of the present invention, the organic solvent 2 is a solvent capable of dissolving the compound 4, including one or more of dimethylformamide, methanol, ethanol, isopropanol, acetone, dimethylacetamide, dichloromethane, ethyl acetate, and acetonitrile. Preferably, the solvent 2 is dimethylformamide.

[0026] And / or, the alkaline catalyst is alkaline catalyst 2, comprising one or more of piperidine, diethylamine, DBU, concentrated ammonia, morpholine, and cyclohexylamine, preferably, the alkaline catalyst 2 is piperidine;

[0027] And / or, the molar ratio of compound 4 to base catalyst 2 is 1:2-20;

[0028] And / or, the reaction temperature is 0℃ and the reaction time is 0.5-2 hours.

[0029] And / or, after the reaction is complete, separation and purification are performed, including the following steps: adding the mixture to saturated brine and filtering to obtain a crude product; washing the organic layer with brine and drying it on anhydrous Na2SO4; filtering the mixture and concentrating the filtrate under reduced pressure; and purifying the residue by chromatography to obtain the product.

[0030] As a preferred technical solution of the present invention, compound 4 is prepared by the following method: in a solvent, compound 1 is protected under alkaline conditions to obtain compound 2, and compound 2 and compound 3 are generated under the action of an acid catalyst;

[0031] As a preferred embodiment of the present invention, compound 6 is prepared by the following method: compound 3 and compound 5 are generated in an organic solvent under the action of an acid catalyst;

[0032] The chemical structural formula of compound 3 is as follows:

[0033]

[0034] The chemical structural formula of compound 5 is as follows:

[0035]

[0036] As a preferred embodiment of the present invention, the organic solvent is solvent 3, which is a solvent capable of dissolving compounds 3 and 5, including one or more of dichloromethane, methanol, ethanol, toluene, acetonitrile, ethyl acetate, tetrahydrofuran, and dimethylformamide, preferably dichloromethane;

[0037] And / or, the molar ratio of compound 3, compound 5 and acid catalyst is 1:0.05 to 1;

[0038] And / or, the acid catalyst includes BF3·Et2O, aluminum trichloride, ferric tribromide, and trifluoromethanesulfonic acid catalyst added at a solution temperature of 0°C;

[0039] And / or, the reaction temperature is 0-30℃, and the reaction time is 0.5-12 hours; preferably, the reaction temperature is 25℃, and the reaction time is 2 hours;

[0040] And / or, after the reaction is complete, separation and purification are performed, including the following steps: adding saturated NaHCO3, extracting multiple times with an organic solvent, washing the bound organic layer with brine, drying on anhydrous Na2SO4, concentrating the mixture under reduced pressure, and purifying the residue by chromatography to obtain the product.

[0041] Another aspect of the present invention provides a pharmaceutical composition comprising the aforementioned compound of the present application and its pharmaceutically acceptable salt, solvent compound or prodrug.

[0042] Another aspect of the present invention provides the use of the aforementioned compounds and pharmaceutical compositions in the preparation of a medicament for treating tumors.

[0043] In some embodiments, the tumor is pancreatic cancer, lung cancer, breast cancer, liver cancer, colorectal cancer, cervical cancer, esophageal cancer, stomach cancer, melanoma, pancreatic cancer, kidney cancer, leukemia, prostate cancer, bladder cancer, neuroblastoma, uterine cancer, ovarian cancer, or nasopharyngeal carcinoma.

[0044] The general route of the artemisinin trimer derivative provided by this invention is as follows:

[0045]

[0046] Compound 1 was amino-protected with FmocCl under alkaline conditions. After the reaction was completed by TLC monitoring, compound 2 was obtained in a high yield by extraction, concentration, and drying. Compound 2 was etherified under boron trifluoride diethyl ether conditions, and purified to give compound 4. Compound 4 was deprotected to give compound SM1044 in its free state. Compound 3 was etherified under boron trifluoride diethyl ether conditions to give compound 6. Compound 6 and compound SM044 reacted to generate the target product 7 in a high yield.

[0047] Compared with the prior art, the present invention achieves the following technical effects:

[0048] (1) A well-defined artemisinin trimer with higher antitumor activity was designed and synthesized. This invention is the first to synthesize and isolate β,β,α trimer, β,α,α trimer and α,α,α trimer. The isomers of these three trimers were successfully isolated by the new synthesis method and characterized and activity tested. They have high antitumor activity and provide a new approach for the study of the configuration of artemisinin trimer.

[0049] (2) The preparation method is simple and the reaction conditions are mild: This invention starts from dihydroartemisinin, first synthesizes its dimer, and then further synthesizes the trimer. It is prepared by conventional reaction and commonly used reagent reaction. The reaction temperature is carried out at room temperature or low temperature, and the separation steps are simple.

[0050] It should also be understood that certain compounds of this application may exist in their free form for therapeutic purposes, or, where appropriate, in their pharmaceutically acceptable derivative forms. In this application, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, solvates, metabolites, or prodrugs, which, upon administration to a patient in need, can directly or indirectly provide the compound of this application or its metabolites or residues. Therefore, when referring to "compounds of this application" herein, it is also intended to cover the aforementioned various derivative forms of the compound.

[0051] Pharmaceutically acceptable salts of the compounds described in this application include their acid addition salts and base addition salts, and the types of salts are not particularly limited, as long as they are physiologically acceptable. Suitable examples of pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochlorides, hydrobroms, sulfates, nitrates, phosphates, acetates, trifluoroacetates, tartrates, fumarates, oxalates, maleates, citrates, succinates, methanesulfonates, benzenesulfonates, malates, aspartates, glucoheponicates, glucuronides, orotates, palmitates, and other similar salts. Suitable examples of pharmaceutically acceptable base addition salts include, but are not limited to, sodium salts, potassium salts, ammonium salts, calcium salts, magnesium salts, aluminum salts, iron salts, histidine salts, arginine salts, choline salts, and other similar salts.

[0052] The compounds of this application may exist in the form of solvates (preferably hydrates), wherein the compounds of this application contain a polar solvent, particularly such as water, methanol, or ethanol, as a structural element of the compound's crystal lattice. The amount of the polar solvent, particularly water, may be stoichiometric or non-stoichiometric.

[0053] The scope of this application also includes metabolites of the compounds of this application, i.e., substances formed in the body when the compounds of this application are administered. Such products can be generated by, for example, oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc., of the administered compound. Therefore, this application includes metabolites of the compounds of this application, including compounds prepared by methods that expose the compounds of this application to mammals for a time sufficient to produce their metabolites.

[0054] This application further includes, within its scope, prodrugs of the compounds of this application, which are certain derivatives of the compounds of this application that may have little or no pharmacological activity on their own, which, when administered to or onto the body, can be converted, for example, by hydrolysis and cleavage into the compounds of this application having the desired activity. Typically, such prodrugs are functional group derivatives of the compounds that are readily converted in vivo into the compounds with the desired therapeutic activity.

[0055] In this application, "pharmaceutically acceptable carrier" refers to a pharmacologically and pharmaceutically acceptable additive that is administered together with the active ingredient, and may include excipients, disintegrants, binders, lubricants, coating agents, dyes, diluents, bases, and isotropic agents, etc.

[0056] The dosage forms include, but are not limited to, tablets, capsules, lozenges, hard candies, powders, sprays, creams, ointments, drops, suppositories, gels, pastes, lotions, aqueous suspensions, injectable solutions, elixirs, and syrups.

[0057] Examples of dosage forms suitable for oral administration include tablets, capsules, powders, fine granules, granules, liquids, and syrups. Examples of dosage forms suitable for non-oral administration include injections, drops, and suppositories.

[0058] In this document, unless otherwise stated, all figures representing amounts of components, measurements, or reaction conditions as used herein should be understood to be modified in all cases by the term "about" to indicate possible measurement error. For example, when associated with a percentage, the term "about" may mean ±1%. Attached Figure Description

[0059] Figure 1 Compound 7(β,β,β) 1 H NMR spectrum;

[0060] Figure 2 Compound 7(β,β,β) 13 C NMR spectrum;

[0061] Figure 3 Compound 7(β,β,α) 1 H NMR spectrum;

[0062] Figure 4 Compound 7(β,β,α) 13 C NMR spectrum;

[0063] Figure 5 Compound 7(β,α,α) 1 H NMR spectrum;

[0064] Figure 6 Compound 7(β,α,α) 13 C NMR spectrum;

[0065] Figure 7 For compound 7(α,α,α) 1 H NMR spectrum;

[0066] Figure 8 For compound 7(α,α,α) 13 C10 NMR spectrum. Detailed Implementation

[0067] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0068] The purity of the products in the examples was tested using HPLC; the structure of the compounds was confirmed by 1H NMR spectroscopy.

[0069] Example 1

[0070] Synthesis of Compound 2

[0071] The reaction equation is as follows:

[0072]

[0073] Compound 1 (50.0 g, 476 mmol) and Et3N (83.0 mL, 951 mmol) were added separately to DCM (300 mL) solution, and FmocCl (123 g, 476 mmol) was added to DCM (100 mL) solution at 0 °C. The mixture was stirred at 25 °C under N2 for 2 hours. Thin-layer chromatography (PE:EA = 1:3) showed that compound FmocCl was completely consumed and formed a new spot. Saturated NH4Cl (20 mL) was added, and the mixture was extracted three times with DCM (500 mL). The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by chromatography to give compound 2 (110 g, yield 71%) as a white solid.

[0074] Example 2

[0075] Synthesis of Compound 4

[0076] The reaction equation is as follows:

[0077]

[0078] At 0 °C, BF3·Et2O (9.4 mL, 76.4 mmol) was added to a DCM (400 mL) solution of compound 2 (50.0 g, 153 mmol) and compound 3 (95.5 g, 336 mmol). The mixture was stirred at 25 °C for 2 hours under N2 conditions. After 1 hour, thin-layer chromatography (PE:EA = 1:1) showed that compound 2 was completely consumed and some new spots formed. Saturated NaHCO3 (100 mL) was added, and the mixture was extracted three times with DCM (200 mL). The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by chromatography to give compound 4 (95.0 g, 73% yield) as a white solid.

[0079] Example 3

[0080] Synthesis of intermediate SM1044

[0081] The structural formula of intermediate SM1044 is as follows:

[0082]

[0083] The reaction equation is:

[0084]

[0085] Piperidine (11.4 mL, 116 mmol) was added to a DMF (100 mL) solution of compound 4 (10.0 g, 11.6 mmol) at 0 °C. The mixture was stirred at 0 °C for 2 hours under N2 conditions. Thin-layer chromatography (DCM:MeOH = 15:1) showed that compound 4 was consumed and some new spots were formed. The mixture was added to saturated brine (50 mL) and filtered to obtain the crude product. The organic layer was washed with brine (20 mL) and dried over anhydrous Na2SO4. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by chromatography to obtain the desired intermediate products SM1044 (ββ, 3.2 g, 43% yield) as a white solid, SM1044 (βα, 2.7 g, 36% yield) as a white solid, and SM1044 (αα, 340 mg, 5% yield) as a white solid.

[0086] Example 4

[0087] Synthesis of Compound 6

[0088] The structural formula of compound 6 is as follows:

[0089]

[0090] The reaction equation is:

[0091]

[0092] At 0 °C, BF3·Et2O (744 mg, 5.28 mmol) was added to a DCM (80 mL) solution of compound 3 (3 g, 10.6 mmol) and compound 5 (1.32 g, 10.6 mmol). The mixture was stirred at 25 °C under N2 for 2 hours. Thin-layer chromatography (PE:EA = 5:1) showed that compound 3 was completely consumed and some new spots were formed. Saturated NaHCO3 (30 mL) was added, and the mixture was extracted three times with DCM (20 mL). The bound organic layer was washed with brine (50 mL) and dried on anhydrous Na2SO4. The mixture was concentrated under reduced pressure, and the residue was purified by chromatography to give the desired product compound 6 (β, 2.1 g, 51% yield) as a white solid and product compound 6 (α, 102 mg, 2% yield) as a colorless oil.

[0093] Compound 6(β): 1 H NMR (400MHz, CDCl3) δ5.49 (s, 1H), 4.84 (d, J = 3.6Hz, 1H), 4.15-4.09 (m, 1H), 3. 823.76(m,1H),3.53-3.50(m,2H),2.69-2.61(m,1H),2.41-2.33(m,1H),2.06-2 .01(m,1H),1.92-1.84(m,2H),1.78-1.73(m,1H),1.68-1.62(m,1H),1.53-1.4 5(m,2H),1.43(s,3H),1.37-1.32(m,1H),1.28-1.22(m,1H),0.96-0.90(m,7H).

[0094] Compound 6(α): 1 H NMR (400MHz, CDCl3) δ5.49 (s, 1H), 5.00 (d, J = 4.4Hz, 1H), 4.17-4.11 (m, 1H), 3.88-3.82 (m, 1H), 3.54-3.51 (m, 2H), 2.35-2.27 (m, 1H), 2 .05-1.99(m,1H),1.91-1.78(m,2H),1.64-1.57(m,4H),1.47-1.42(m,5H),1.33-1.26(m,2H),1.24-1.21(m,3H),0.94(d,J=6.0Hz,3H).

[0095] Example 5

[0096] Synthesis of Compound 7 (β,β,β)

[0097]

[0098] The reaction equation is as follows:

[0099]

[0100] Compound SM1044 (ββ300 mg, 0.471 mmol) and compound 6 (β219 mg, 0.565 mmol) were added to DMF (4 mL) solution with K2CO3 (195 mg, 1.41 mmol) at 25 °C. The mixture was stirred at 40 °C under N2 for 2 days. Thin-layer chromatography (PE:EA = 1:1) showed that compound SM1044 was completely consumed, forming a new spot. Saturated NH4Cl (10 mL) was added, and the mixture was extracted three times with EA (30 mL). The bound organic layer was washed with brine (20 mL) and dried over anhydrous Na2SO4. The mixture was concentrated under reduced pressure, and the residue was purified by column chromatography to give the desired product compound 7 (β,β,β) (360 mg, 80% yield).

[0101] Figure 1 Compound 7(β,β,β) 1 H NMR spectrum; Figure 2 Compound 7(β,β,β) 13 C10 NMR spectrum.

[0102] 1 H NMR (600MHz, CDCl3) δ5.39 (s, 3H), 4.77 (d, J = 2.0Hz, 3H), 3.90-3.87 (m, 3H), 3.46-3.42 (m, 3H), 2.81-2.78(m,3H),2.74-2.70(m,3H),2.63-2.61(m,3H),2.39-2.34(m,3H),2.03-2.01(m,3H), 1.90-1.86(m,3H),1..83-1.77(m,3H),1.73-1.70(m,4H),1.65-1.62(m,3H),1.53-1.45(m,6H) ,1.43(s,9H),1.35-1.32(m,3H),1.27-1.24(m,3H),0.95(d,J=6.4Hz,9H),0.92-0.86(m,12H).

[0103] 13C NMR (101MHz, CDCl3) δ117.39,104.33,104.15,102.49,100.44,91.24,88.02,81.09,80.36,66.44,65.68,52.59,52.29,51.63,51.61,45. 30,44.37,37.38,37.31,36.43,36.29,34.62,34.19,32.57,30.80,2 6.15,26.02,24.69,24.66,24.41,22.18,20.38,20.26,13.03,12.62.

[0104] Example 6

[0105] Synthesis of Compound 7 (β,β,α)

[0106]

[0107] The synthesis route is the same as in Example 5.

[0108] Compound SM1044 (β, α 100 mg, 0.157 mmol) and compound 6 (β 73.6 mg, 0.188 mmol) were added to DMF (4 mL) solution with K2CO3 (65.0 mg, 0.47 mmol) at 25 °C. The mixture was stirred at 40 °C under N2 for 2 days. Thin-layer chromatography (PE:EA = 1:1) showed that compound SM1044 was completely consumed, forming a new spot. Saturated NH4Cl (10 mL) was added, and the mixture was extracted three times with EA (30 mL). The bound organic layer was washed with brine (20 mL) and dried over anhydrous Na2SO4. The mixture was concentrated under reduced pressure, and the residue was purified by column chromatography to give the desired product, compound 7 (β, β, α) (91 mg, 61% yield).

[0109] Figure 3 Compound 7(β,β,α) 1 H NMR spectrum Figure 4 Compound 7(β,β,α) 13 C10 NMR spectrum.

[0110] 1H NMR (400MHz, CDCl3) δ5.38 (s, 2H), 5.30 (s, 1H), 4.75 (d, J = 2.8Hz, 2H), 4.41 (d, J = 9. 2Hz,1H),4.00-3.94(m,1H),3.89-3.83(m,2H),3.51-3.41(m,3H),2.84-2.72(m,6H) ,2.61-2.57(m,2H),2.38-2.31(m,4H),2.02-1.99(m,3H),1.87-1.81(m,5H),1.77- 1.60(m,8H),1.53-1.47(m,4H),1.41(s,9H),1.31-1.19(m,8H),0.94-0.84(m,21H).

[0111] 13 C NMR (101MHz, CDCl3) δ104.23,104.02,102.17,100.25,91.19,87.88,81.15,80.34,67.78,67.35,54.65,54.29,52.60,51.66,45. 35,44.47,37.43,37.3836.47,36.34,34.65,34.25,32.60,30.88,26.20,26.04,24.70,24.45,22.21,20.38,20.28,13.11,12.71.

[0112] Example 7

[0113] Synthesis of Compound 7 (β,α,α)

[0114]

[0115] The synthesis route is the same as in Example 5.

[0116] Compound SM1044 (αα 100 mg, 0.157 mmol) and compound 6 (β 73.6 mg, 0.188 mmol) were added to DMF (4 mL) solution with K2CO3 (65.0 mg, 0.47 mmol) at 25 °C. The mixture was stirred at 40 °C under N2 for 2 days. Thin-layer chromatography (PE:EA = 1:1) showed that compound SM1044 was completely consumed, forming a new spot. Saturated NH4Cl (10 mL) was added, and the mixture was extracted three times with EA (30 mL). The bound organic layer was washed with brine (20 mL) and dried over anhydrous Na2SO4. The mixture was concentrated under reduced pressure, and the residue was purified by column chromatography to give the desired product, compound 7 (β, α, α) (90 mg, 61% yield).

[0117] Figure 5 Compound 7(β,α,α) 1 H NMR spectrum; Figure 6 Compound 7(β,α,α) 13 C10 NMR spectrum.

[0118] 1 H NMR (400MHz, CDCl3) δ5.37 (s, 1H), 5.30 (s, 2H), 4.74 (d, J = 3.2Hz, 1H), 4.41 (d, J = 9. 2Hz,2H),3.98-3.93(m,2H),3.88-3.82(m,1H),3.53-3.42(m,3H),2.85-2.74(m,6H ),2.60-2.56(m,1H),2.38-2.30(m,5H),2.06-1.8(m,5H),1.87-1.81(m,3H),1.75- 1.60(m,7H),1.53-1.47(m,4H),1.41(s,9H),1.31-1.21(m,8H),0.98-0.83(m,21H).

[0119] 13 C NMR (101MHz, CDCl3) δ102.31,102.10,100.30,98.34,89.28,86.01,79.29,78.47,65.87,65.42,52.96,52.47,50.73,49.78,43.49,42. 60,35.51,35.47,34.60,64.34.47,32.77,32.38,30.75,29.01,24.32,34.16,22.83,22.80,22.56,20.31,18.50,18.40,11.23,10.83.

[0120] Example 8

[0121] Synthesis of Compound 7 (α,α,α)

[0122] The synthesis route is the same as in Example 5.

[0123]

[0124] Compound SM1044 (αα166 mg, 0.260 mmol) and compound 6 (α102 mg, 0.260 mmol) were added to DMF (4 mL) solution with K2CO3 (108 mg, 0.782 mmol) at 25 °C. The mixture was stirred at 40 °C under N2 for 4 days. Thin-layer chromatography (PE:EA = 1:1) showed that compound SM1044 was completely consumed, forming a new spot. Saturated NH4Cl (10 mL) was added, and the mixture was extracted three times with EA (30 mL). The bound organic layer was washed with brine (20 mL) and dried over anhydrous Na2SO4. The mixture was concentrated under reduced pressure, and the residue was purified by column chromatography to give the desired product, compound 7 (α,α,α) (78 mg, 52% yield).

[0125] Figure 7 For compound 7(α,α,α) 1 H NMR spectrum; Figure 8 For compound 7(α,α,α) 13 C10 NMR spectrum.

[0126] 1 H NMR (400MHz, CDCl3) δ5.44(s,1H),5.32(s,2H),4.94(d,J=4.4Hz,1H),4.44(d,J=9.2Hz,2H),4 .04-3.97(m,2H),3.92-3.84(m,1H),3.57-3.54(m,3H),2.88-2.73(m,5H),2.40-2.27(m,5H), 2.04-1.98(m,3H),1.90-1.85(m,3H),1.77-1.63(m,8H),1.56-1.47(m,6H),1.42(s,7H),1.41 (s,3H),1.33-1.24(m,9H),1.18(d,J=7.2Hz,3H),1.00-0.931(m,12H),0.87(d,J=7.2Hz,6H).

[0127] 13 C NMR (101MHz, CDCl3) δ104.35,103.18,100.36,91.33,89.12,81.75,80.52,67.80,54.73,54.55,52.10,51.82,46.6 2,45.54,39.81,37.49,37.89,36.70,34.61,34.41,32.79,26.20,26.13,24.85,22.34,20.42,20.23,19.74,12.88.

[0128] Example 8

[0129] Culture of tumor cells:

[0130] MCF-7 is a human breast cancer tumor cell. MCF-7 cells were cultured in DMEM medium. The medium contained 10% fetal bovine serum (FBS) and 1% penicillin-drug antibodies. The medium was 1640, and the DMEM was manufactured by Corning. The FBS was manufactured by Life Technologies Company (Life Technologies, Karlsruhe, Germany). Other reagents were manufactured by Sigma Company.

[0131] Preparation of cell suspension:

[0132] Thaw cells stored in liquid nitrogen in a 37°C water bath. Transfer the cell suspension to a 15mL centrifuge tube, add 2mL of culture medium, and centrifuge at 1000rpm for 3min. After centrifugation, aspirate and discard the supernatant, resuspend the cells in 1mL of culture medium, and then transfer the suspension to a 90mm culture dish. Add 7mL of culture medium and 4μL of mycoplasma prevention agent, and incubate at 37°C in a 5% CO2 incubator.

[0133] After the cells adhered and multiplied to 80%-90%, the culture medium was aspirated, and the cells were gently washed several times with PBS (without calcium and magnesium ions). Then, the cells were digested with 0.25% trypsin solution. The resulting cell suspension was added to 2 mL of culture medium and transferred to a 15 mL centrifuge tube. The cells were centrifuged at 1000 rpm for 3 min. After centrifugation, the supernatant was discarded, and the cells were resuspended in 1 mL of culture medium to obtain the cell suspension required for the experiment.

[0134] To evaluate the antiproliferative activity of the aforementioned artemisinin trimer derivatives, the prepared MCF-7 cell suspension was subjected to a 6×10⁻⁶ ppm concentration. 3 Cells were injected evenly into 96-well plates at a rate of 100 μL / well. After cell adhesion (24 hours), different concentrations of the test compound were added. The 96-well plates were then incubated at 37°C, 95% air, and 5% CO2 for 48 hours. The antiproliferative activity of the test compound was evaluated using the MTT assay. The final quantitative analysis data were analyzed using Graphpad Prism 9 to obtain the IC50 values. 50 The above experiments all require three replicates.

[0135] The MTT assay is used to determine cell proliferation and viability in studies of cytotoxicity and antiproliferative substances. In metabolically active cells, the yellow tetrazolium salt (MTT) is reduced to formazan by mitochondrial dehydrating enzymes. Water-insoluble formazan crystals accumulate in the cells, and their dissolution requires organic detergents that disrupt the membrane and simultaneously dissolve the dye; therefore, a triple solution at pH 7.4 is used to dissolve the formazan crystals. Cell proliferation is quantitatively evaluated at 570 nm using a 96-well microplate reader.

[0136] The MTT assay procedure is as follows: 5 mg / mL MTT in PBS solution is added to a 96-well plate that has been incubated for 48 hours after the addition of the test sample, at a dose of 20 μL / well. The plate is incubated at 37°C for 1-4 hours. Then, 100 μL / well of triplet solution is added, and the plate is incubated overnight at 37°C. After 12 hours, the absorbance at 570 nm is measured using a microplate reader to obtain the final data. The IC50 of each compound against breast cancer MCF-7 is then determined. 50 As shown in Table 1.

[0137] Table 1. Toxicity of artemisinin derivatives in tumor cell lines (IC50) 50 / μM)

[0138] Compound types MCF-7 Compound 7(α,α,α) 0.06±0.01 Compound 7(β,β,α) 0.003±0.05 Compound 7(β,α,α) 0.005±0.02 Etoposide 0.09±0.05

[0139] 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. An artemisinin trimer derivative, or a pharmaceutically acceptable salt, solvate or prodrug thereof, characterized in that, The artemisinin trimer derivative is a compound with the structure shown in the following formula:

2. The method for preparing an artemisinin trimer derivative as described in claim 1, characterized in that, In an organic solvent, compound SM1044 and compound 6 are reacted in the presence of a base catalyst; The chemical structural formula of the compound SM1044 is one of the following: The chemical structural formula of compound 6 is one of the following:

3. The method for preparing an artemisinin trimer derivative as described in claim 2, characterized in that, The organic solvent is solvent 1, which is a solvent capable of dissolving the compound SM1044 and the compound 6, including one or more of dimethylformamide, methanol, ethanol, isopropanol, acetone, dimethylacetamide, and acetonitrile. Preferably, solvent 1 is dimethylformamide. And / or, the alkaline catalyst is alkaline catalyst 1, comprising one or more of potassium carbonate, sodium carbonate, cesium carbonate, triethylamine, N,N-diisopropylethylamine, and DBU, preferably, the alkaline catalyst 1 is potassium carbonate; And / or, the molar ratio of compound SM1044, compound 6, and base catalyst 1 is 1:1 to 5. And / or, the reaction temperature is 0-45℃, and the reaction time is 2-72 hours; preferably, the reaction temperature is 40℃, and the reaction time is 48 hours; And / or, after the reaction is complete, separation and purification are performed, including the following steps: adding saturated NH4Cl, extracting multiple times with EA, washing the organic layer with brine and drying on anhydrous Na2SO4, concentrating the mixture under reduced pressure, and purifying the residue by chromatographic column to obtain the product.

4. The method for preparing an artemisinin trimer derivative as described in claim 2, characterized in that, The compound SM1044 is obtained by deprotecting compound 4. In organic solvent 2, compound 4 generates compound SM1044 under the action of an alkaline catalyst. The chemical structural formula of compound 4 is as follows:

5. The method for preparing an artemisinin trimer derivative as described in claim 4, characterized in that, The organic solvent is solvent 2, which is a solvent capable of dissolving compound 4, including one or more of dimethylformamide, methanol, ethanol, isopropanol, acetone, dimethylacetamide, dichloromethane, ethyl acetate, and acetonitrile. Preferably, solvent 2 is dimethylformamide. And / or, the base catalyst is base catalyst 2, including one or more of piperidine, diethylamine, DBU, concentrated ammonia, morpholine, cyclohexylamine, preferably, the base catalyst 2 is piperidine; And / or, the molar ratio of compound 4 to base catalyst 2 is 1:2-20; And / or, the reaction temperature is 0℃ and the reaction time is 0.5-12 hours. And / or, after the reaction is complete, separation and purification processes are performed. The process includes the following steps: adding the mixture to saturated brine and filtering to obtain a crude product; washing the organic layer with brine and drying it on anhydrous Na2SO4; filtering the mixture and concentrating the filtrate under reduced pressure; and purifying the residue by chromatography to obtain the product.

6. The method for preparing an artemisinin trimer derivative as described in claim 4, characterized in that, Compound 4 was prepared by the following method: Compound 1 was protected with a protecting group in an organic solvent under alkaline conditions to obtain Compound 2, and Compound 2 and Compound 3 were generated under the action of an acid catalyst.

7. A pharmaceutical composition, characterized in that, Contains artemisinin derivatives as claimed in claim 1, pharmaceutically acceptable salts, solvent compounds, or prodrugs thereof.

8. Use of the artemisinin derivative of claim 1 or the pharmaceutical composition of claim 7 in the preparation of a medicament for treating tumors.

9. The use as described in claim 8, characterized in that... The tumors mentioned are pancreatic cancer, lung cancer, breast cancer, liver cancer, colorectal cancer, cervical cancer, esophageal cancer, stomach cancer, melanoma, pancreatic cancer, kidney cancer, leukemia, prostate cancer, bladder cancer, neuroblastoma, uterine cancer, ovarian cancer, or nasopharyngeal carcinoma.

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