Glutamide derivative as well as synthesis method and application thereof in reversing tumor drug resistance

Abstract

The invention belongs to the technical field of medicine, and discloses a glutarimide derivative and a synthesis method and application thereof in reversing tumor drug resistance, the structural general formula of the glutarimide derivative is shown as a formula I, a formula II or a formula III, and the glutarimide derivative specifically corresponds to 27 specific compounds (namely, compounds L1-L27). The glutarimide derivatives can be used for preparing anti-drug-resistant drugs for colon cancer drug-resistant cells, and especially have a drug resistance reversing effect aiming at the problem of drug resistance of paclitaxel to colon cancer drug-resistant cell strains SW620 / AD300. Moreover, the invention also provides a novel synthetic method for preparing the glutarimide derivative, and the preparation method has the characteristics of easily available initial reaction raw materials, simple steps, wide substrate range, high reaction speed, mild reaction conditions and the like.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, more particularly, to a glutarimide derivative, a synthetic method thereof and an application of the glutarimide derivative in reversing tumor drug resistance. When the glutarimide derivative is applied as a tumor drug resistance reversing agent, the glutarimide derivative can realize anti-tumor activity in a colon cancer drug resistant cell line SW620 / AD300, especially in combination with paclitaxel. BACKGROUND

[0002] Drug resistance refers to the resistance of tumor cells to a certain chemotherapeutic drug after long-term contact with the drug. It is the most important defense mechanism of tumor cells against chemotherapy drugs and one of the main reasons for chemotherapy failure. Drug resistance is one of the important reasons for the failure of cancer chemotherapy. Cancer patients can respond to drug treatment, but most patients develop resistance to targeted chemotherapy during treatment. There are many potential mechanisms of drug resistance, but the main mechanism is related to the high expression of ATP-binding cassette transporters in tumor drug-resistant cells. The most studied drug transporter is ABCB1 (also known as P-glycoprotein (P-gp) or MDR1), which is a member of the ABC family and is responsible for the efflux of chemotherapeutic drugs to reduce intracellular drug concentration. P-gp is widely expressed in many drug-resistant cancer cells such as colon cancer and prostate cancer cells, so it has become a promising therapeutic target for overcoming drug resistance. So far, there have been three generations of P-gp inhibitors for drug resistance research. The first generation includes verapamil and trifluorazine, but they have defects such as high toxicity and low clinical efficacy. The second generation includes diltiazem and biricodar, which have higher potency, specificity and lower toxicity than the first generation. However, these P-gp inhibitors can inhibit the activity of drug metabolizing enzyme CYP3A4, thereby interfering with drug metabolism kinetics, affecting normal cell metabolism, and causing unavoidable toxic side effects. The third generation, such as elacridar, can bind to P-gp with high affinity and has little effect on the pharmacokinetics of other drugs, but it still has problems such as high toxicity to normal cells. None of these drugs has been approved for clinical use. Therefore, developing a non-toxic and highly efficient P-gp inhibitor is one of the key ways to solve the problem of drug resistance.Paclitaxel (PTX) is clinically used for the treatment of prostate cancer, esophageal cancer, colon cancer and other cancers, but the inevitable problem of drug resistance also appears in the treatment of cancer, which limits its application in clinical cancer treatment. The main reason for drug resistance is that P-glycoprotein (P-gp) is overexpressed in drug-resistant tumor cells, which pumps the chemotherapeutic drugs such as PTX out of the tumor cells, reduces the concentration of the drugs in the tumor cells and thus reduces the therapeutic effect. (See: 1) S. Wang; S. Wang; Q. Teng; L. Yang; Z. Lei; X. Yuan; J. Huo; X. Chen, M. Wang; B. Yu; Z. Chen; H. Liu. J. Med. Chem. 2020, 63, 24, 15979-15996; 2) S. Yuan; B. Wang; Q. Dai; X. Zhang; J. Zhang; J. Zuo; H. Liu; Z. Chen; G. Li; S. Wang; H. Liu; B. Yu. J. Med. Chem. 2021, 64, 19, 14895-14911; 3) S. Wang; S. Wang; Q. Teng; Z. Lei; Z. Chen; X. Chen; H. Liu; B. Yu. J. Med. Chem. 2021, 64, 21, 16187-16204.) Therefore, it is necessary to find drug molecules that reverse drug resistance through chemical synthesis, natural compounds and other ways to combine with chemotherapeutic drugs for tumor treatment. It is of great significance to develop a new synthetic method to prepare a class of compounds that can reverse tumor drug resistance, and to find lead compounds to solve the problem of drug resistance of tumor chemotherapeutic drugs through activity screening and structure-activity relationship analysis.

[0003] Glutarimide derivatives are valuable nitrogen heterocyclic compounds, which exist in natural products such as glutarimide-containing polyketides and drug molecules, because they have various biological activities, including cytotoxic antitumor activity, antifungal activity, and inhibition of DNA / protein synthesis.

See: J. Blake Bartlett, Keith Dredge, Angus G. Dalgleish. Nat Rev Cancer 2004, 4, 314-322; b) Laura M. Luh, Ulrike Scheib, Katrin Juenemann, Lars Wortmann, Michael Brands, Philipp M. Cromm. Angew. Chem. Int. Ed. 2020, 59, 15448-15466; c) Hiromasa Kiyota, Yuko Shimizu, Takayuki Oritani. Tetrahedron Lett. 2000, 41, 5887; d) Jianhua Ju, Jeong-Woo Seo, Yeng Her, Si-Kyu Lim, and Ben Shen. Org. Lett. 2007, 9, 25, 5183-5186; e) Wei Lia, Gunda I. Georg. Chem. Commun., 2015, 51, 8634-8636

[0004] In view of the improvement needs for the drug resistance problem in existing tumor treatment, the purpose of the present application is to provide a glutarimide derivative and a synthetic method thereof and an application of reversing tumor drug resistance, the glutarimide derivative has a structural general formula as shown in formula I or formula II or formula III, and specifically corresponds to 27 specific compounds (i.e. compounds L1-L27), which can be used for preparing colon cancer drug-resistant cell anti-drug drugs, especially for reversing the drug resistance of paclitaxel to colon cancer drug-resistant cell strain SW620 / AD300. Moreover, the present application also provides a novel synthetic method for preparing the above glutarimide derivative, and the preparation method has the characteristics of easy to obtain starting materials, simple steps, wide substrate range, fast reaction speed and mild reaction conditions.

[0005] To achieve the above objectives, according to one aspect of the present invention, a glutarimide derivative is provided, characterized in that its general structural formula is shown as shown in Formula I, Formula II, or Formula III:

[0006]

[0007] In Equation I:

[0008] R1 is benzyl, R2 is H, R3 is H, R4 is trifluoromethyl, and R5 is H;

[0009] Alternatively, R1 is methyl, R2 is H, R3 is H, R4 is any one of phenyl, 4-methoxy-phenyl, 4-chloro-phenyl, and 4-fluoro-phenyl, and R5 is H;

[0010] Alternatively, R1 is methyl, R2 is H, R3 is ethoxy, R4 is ethyl, and R5 is H;

[0011] Alternatively, R1 = R2 = -(CH2CH2CH2)-, R3 is H, R4 is ethyl, and R5 is H;

[0012] Alternatively, R1 is methyl, R2 is H, R3 is H, R4 is trifluoromethyl, and R5 is 2-(2-sec-butyl(benzoic acid)ethyl)-piperidin-1-carboxylic acid tert-butyl ester;

[0013] In Formula II:

[0014] R1 is benzyl, R2 is methyl, and R3 is any one of H, 3,5-dimethyl, and 2-chloro-5-methoxy.

[0015] Alternatively, R1 is benzyl, R2 is benzyl, and R3 is H;

[0016] In Formula III:

[0017] R1 is benzyl, R2 is methyl, R3 is H, and R4 is any one of 4-methoxy-phenyl, 3,4-dimethoxy-phenyl, 3-methyl-4-methoxy-phenyl, methylenedioxyphenyl, 1,4-benzodioxane, 3,5-dimethyl-phenyl, phenylethynyl, 4-chloro-phenyl, 3,4,5-trimethoxy-phenyl, 3,5-dimethyl-4-methoxy-phenyl, 2-anthrayl, and benzofuranyl.

[0018] Alternatively, R1 is 3,4-dimethoxybenzyl, R2 is methyl, R3 is H, and R4 is 3,4-dimethoxy-phenyl;

[0019] Alternatively, R1 is benzyl, R2 = R3 = -(CH2CH2CH2CH2)-, and R4 is phenyl;

[0020] Alternatively, R1 is benzyl, R2 is H, R3 is methyl, and R4 is phenyl.

[0021] As a further preferred embodiment of the present application, the glutarimide derivative is specifically compound L24, whose structural formula is as follows:

[0022]

[0023] According to another aspect of the present application, the present application provides a preparation method of the above-mentioned glutarimide derivative, characterized in that, when the glutarimide derivative is any one of compounds L1-L7 and L20, the preparation method is to subject a compound of formula 1 to a cyclization reaction of 1,6-alkynyl under the promotion of an acid to generate the glutarimide derivative, and the synthetic route of the preparation method is as follows:

[0024]

[0025] Among them, compound L1 corresponds to R1 being Bn, R2 being H, R3 being H, R4 being CF3, and R5 being H;

[0026] Compound L2 corresponds to R1 being Me, R2 being H, R3 being H, R4 being Ph, and R5 being H;

[0027] Compound L3 corresponds to R1 being Me, R2 being H, R3 being H, R4 being 4-F-Ph, and R5 being H;

[0028] Compound L4 corresponds to R1 being Me, R2 being H, R3 being H, R4 being 4-Cl-Ph, and R5 being H;

[0029] Compound L5 corresponds to R1 being Me, R2 being H, R3 being H, R4 being 4-OMe-Ph, and R5 being H;

[0030] Compound L6 corresponds to R1=R2=-(CH2CHCH2)-, R3 being H, R4 being Et, and R5 being H;

[0031] Compound L7 corresponds to R1 being Me, R2 being H, R3 being OEt, R4 being Et, and R5 being H;

[0032] Compound L20 corresponds to R1 being Me, R2 being H, R3 being H, R4 being CF3, and R5 being

[0033] When the glutarimide derivative is any one of compounds L8-L11, the preparation method is to subject a compound of formula 2 to a cyclization reaction of 1,6-alkynyl under the promotion of an acid to generate the glutarimide derivative, and the synthetic route of the preparation method is as follows:

[0034] When the glutarimide derivative is any one of compounds L8-L11, the preparation method is to subject a compound of formula 2 to a cyclization reaction of 1,6-alkynyl under the promotion of an acid to generate the glutarimide derivative, and the synthetic route of the preparation method is as follows:

[0034]

[0035] In compound L8, R1 is Bn, R2 is Me, and R3 is H;

[0036] For compound L9, R1 is Bn, R2 is Bn, and R3 is H;

[0037] For compound L10, R1 is Bn, R2 is Me, and R3 is 3,5-dimethyl.

[0038] For compound L11, R1 is Bn, R2 is Me, and R3 is 2-chloro-4-methoxy.

[0039] When the glutarimide derivative is any one of compounds L12-L19, the preparation method is to prepare the compound of formula 3 in... The cyclization reaction of 1,6-enyne under acid-promoted conditions generates a glutarimide derivative. The synthetic route for its preparation is as follows:

[0040]

[0041] In compound L12, R1 is Bn, R2 is Me, R3 is H, and R is 4-methoxy.

[0042] For compound L13, R1 is Bn, R2 is Me, R3 is H, and R is 3,4-dimethoxy.

[0043] For compound L14, R1 is Bn, R2 is H, R3 is Me, and R is 3,4-dimethoxy.

[0044] For compound L15, R1 is Bn, R2 = R3 is -(CH2CH2CH2)-, and R is 3,4-dimethoxy.

[0045] In compound L16, R1 is 3,4-dimethoxybenzyl, R2 is Me, R3 is H, and R is 3,4-dimethoxy.

[0046] For compound L17, R1 is Bn, R2 is Me, R3 is H, and R is 3-methyl-4-methoxy.

[0047] For compound L18, R1 is Bn, R2 is Me, R3 is H, and R is 3-(-OCH2O-).

[0048] For compound L19, R1 is Bn, R2 is Me, R3 is H, and R is 3-(-OCH2CH2O)-.

[0049] When the glutarimide derivative is any one of compounds L21-L27, the preparation method is to prepare the compound of formula 4 in... The cyclization reaction of 1,6-enyne under acid-promoted conditions generates an intermediate, which is then further coupled with a compound of formula 7 to generate a glutarimide derivative. The synthetic route for this preparation method is as follows:

[0050]

[0051]

[0052] In this compound, R4 corresponding to compound L21 is 3,5-dimethylphenyl, and R is...

[0053] The R4 corresponding to compound L22 is phenylacetylene, and R is H;

[0054] The R4 of compound L23 is 4-chlorophenyl, and R is B(OH)2;

[0055] The R4 corresponding to compound L24 is 3,4,5-trimethoxyphenyl, and R is B(OH)2;

[0056] The R4 corresponding to compound L25 is 3,5-dimethyl-4-methoxyphenyl, and the R is B(OH)2;

[0057] The R4 of compound L26 is 2-anthrayl, and R is B(OH)2;

[0058] The R4 corresponding to compound L27 is benzofuranyl, and R is B(OH)2.

[0059] As a further preferred embodiment of the present invention, when the glutarimide derivative is any one of compounds L1-L7 and L20, the preparation method is to dissolve the raw material with the structural formula shown in Formula 1 in an ultra-dry mixed solvent DCM / HFIP and add it to a drying reaction flask, and then add pre-treated dehydration solution. Molecular sieve, added The acid was stirred at a pre-set temperature for 30 minutes to 3 hours. After the reaction was complete, the molecular sieve was removed by filtration through a Buchner funnel, the organic layer was enriched, and the crude product was obtained by evaporation and concentration. The product was then separated by silica gel column chromatography. The silica gel column was pre-treated with petroleum ether solvent containing 0.1% triethylamine. The raw material with the structural formula shown in Formula 1 and the... The molar ratio of acid to feed is 1:1-1.2; the mixed solvent DCM / HFIP is obtained by mixing DCM and HFIP; the preset temperature is 20℃-35℃.

[0060] When the glutarimide derivative is any one of compounds L8-L11, the preparation method is to dissolve the raw material with the structural formula shown in Formula 2 in an ultra-dry mixed solvent DCM / HFIP and add it to a dry reaction flask, then add the dehydrated raw material. Molecular sieve, add TfOH, stir at room temperature for 1 hour, after the reaction is completed, remove the molecular sieve by filtering through a Buchner funnel, enrich the organic layer, evaporate and concentrate to obtain the crude product, and separate the product by silica gel column chromatography; wherein the molar ratio of the raw material with the structural formula as shown in formula 2 to the TfOH is 1:0.4-0.5, and the mixed solvent DCM / HFIP is obtained by mixing DCM and HFIP.

[0061] When the glutarimide derivative is any one of compounds L12-L19, the preparation method is to dissolve a raw material with a structural formula as shown in formula 3 in an ultradry solvent HFIP, add it to a dry reaction bottle, add a dehydrated catalyst, stir at a pre-set temperature under an argon environment for 3 hours-24 hours, after the reaction is completed, extract with ethyl acetate / water, enrich the organic layer, evaporate and concentrate to obtain the crude product, and separate the product by silica gel column chromatography. Molecular sieve, add a raw material with a structural formula as shown in formula 5, stir at room temperature for 1 hour, after the reaction is completed, remove the molecular sieve by filtering through a Buchner funnel, enrich the organic layer, evaporate and concentrate to obtain the crude product, and separate the product by silica gel column chromatography; the molar ratio of the raw material formula 3, the TfOH and the raw material formula 5 is 1:(1-1.2):(3-5).

[0062] When the glutarimide derivative is any one of compounds L21-L27, the preparation method is to dissolve a raw material with a structural formula as shown in formula 6 in a solvent, add it to a dry reaction bottle, add a base, a raw material with a structural formula as shown in formula 7 and a catalyst, stir at a pre-set temperature under an argon environment for 3 hours-24 hours, after the reaction is completed, extract with ethyl acetate / water, enrich the organic layer, evaporate and concentrate to obtain the crude product, and separate the product by silica gel column chromatography; wherein the molar ratio of the raw material with the structural formula as shown in formula 6 to the raw material with the structural formula as shown in formula 7 is 1:1.2-1.5, the catalyst is a palladium catalyst and cuprous iodide, the molar ratio of the raw material with the structural formula as shown in formula 6 to the cuprous iodide is 1:0.04-0.05, the molar ratio of the raw material with the structural formula as shown in formula 6 to the palladium catalyst is 1:0.05-0.1, the solvent is pure THF or a THF / H2O mixed solvent, the base is one of potassium phosphate trihydrate, triethylamine and sodium carbonate, the molar ratio of the raw material with the structural formula as shown in formula 6 to the base is 1:1.5-2.5, and the pre-set temperature is 40-75°C; the THF / H2O mixed solvent is obtained by mixing THF and H2O.

[0063] According to still another aspect of the present application, the present application provides use of the above glutarimide derivative in preparation of a tumor drug resistance reversing agent.

[0064] As a further preferred embodiment of the present application, the drug resistance reversing agent can exert an anti-tumor effect in tumor cells in combination with paclitaxel.

[0065] Preferably, the tumor drug resistance reversing agent can reverse the drug resistance of paclitaxel in colon cancer drug resistance cell line SW620 / AD300.

[0066] According to another aspect of the present application, the present application provides the use of the above glutarimide derivative in the preparation of a P-gp inhibitor.

[0067] According to the last aspect of the present application, the present application provides a drug resistance reversing agent, characterized in comprising the above glutarimide derivative.

[0068] Compared with the prior art, the present application provides new glutarimide derivatives, a synthesis method thereof and an anti-tumor drug resistance application, the glutarimide derivative has a structural general formula as shown in formula I or formula II or formula III, and specifically corresponds to 27 specific compounds (i.e., compounds L1-L27), the compounds have the effect of reversing drug resistance and play the role of anti-drug resistance, and are non-toxic.

[0069] The present application first reports the biological activity application of glutarimide derivatives as tumor drug resistance reversing agents. As exemplified in the following examples, the anti-tumor activity of the glutarimide derivatives obtained by the present application combined with paclitaxel in colon cancer drug resistance cell line SW620 / AD300 is evaluated, and the results show that the glutarimide derivatives can reverse drug resistance and can be used for preparing a drug resistance reversing agent. As detailed in the following examples, the anti-drug resistance activity of the compounds L1-L27 is evaluated, and the experiments prove that the compounds provided by the present application have the effect of reversing drug resistance and play the role of anti-drug resistance, for example, can be used for preparing anti-drug resistance drugs for colon cancer drug resistance cell line SW620 / AD300. Specifically, the anti-tumor drug resistance activity of the compounds L1-L27 is evaluated, and it is proved that the glutarimide derivatives provided by the present application have the effect of anti-tumor drug resistance. When paclitaxel and the glutarimide derivative compounds L1-L27 are added at the same time in colon cancer drug resistance cell line SW620 / AD300, compared with the control group with only paclitaxel, the obvious anti-tumor effect is shown, which shows that the compounds L1-L27 can be a good drug resistance reversing agent, and the best compound L24 for reversing drug resistance is explored as a safe P-gp inhibitor to play the role of reversing drug resistance, specifically by inhibiting the P-gp efflux function.

[0070] Moreover, the present application also provides a new preparation method for synthesizing glutarimide derivatives, the synthesis method is to use 1,6-alkyne with diversified structure as a substrate, the substrate generates a Michael addition reaction through generating a allene alcohol intermediate under the promotion of an acid to generate a glutarimide compound. ​The acid-promoted one-step construction of CO / CC and CC bonds in 1,6-enyne, leading to a cyclization reaction and the formation of a six-membered ring product containing a tetrasubstituted olefin, demonstrates the high efficiency of this synthetic method. The preparation of the starting material, 1,6-enyne, only requires the condensation reaction of an amine with a carboxylic acid to obtain an acetylide, followed by a nucleophilic substitution reaction of an acyl chloride. Therefore, the starting material is readily available and the reaction is easy to operate. Most of the obtained glutarimide compounds can be converted within 30 minutes to 1 hour, indicating the rapid reaction rate of this synthetic method. The synthesis of 27 glutarimide derivatives with different electronegativity and functional groups under these conditions demonstrates the broad substrate range of this synthetic method. Among the obtained compounds L1-L27, the highest yield was 86%, showcasing excellent reaction yields.

[0071] It is known in the prior art that the construction of tetrasubstituted olefins via allenol intermediates is mainly carried out under basic conditions [see: (a) HYKim, J.-Y.Li, K.Oh, Angew. Chem., Int. Ed. 2013, 52, 3736-3740; b) HYKim, E.O. Rooney, R.P. Meury, K.Oh, Angew. Chem., Int. Ed. 2013, 52, 8026-8030; c) TEReynolds, K.S. Cheidt, Angew. Chem., Int. Ed. 2007, 46, 7806-7809; d) R. Chinchilla, C. Nájera, Chem. Rev. 2000, 100, 1891-1928; e) HYKim, J.-Y.Li, K.Oh, J.Org. Chem. 201 2,77,11132-11145; f)P.Maity,SD Lepore,J.Org.Chem.2009,74,158-162; g)RV Kolakowski,M.Manpadi,Y.Zhang,TJ Emge,LJ Williams,J.Am.Chem.Soc.2009,131,12910-12911; h)TEReynolds,ARBharadwaj,KAScheidt,J.Am.Chem.Soc.2006,128,15382-15383; i)TEReynolds,MS Binkley,KAScheidt,Org.Lett.2008,10,2449-2452. Under acidic conditions, protonation is more likely to occur [see: Aleksander]. V. Vasilyev, Stéphane Walspurger, Stefan Chassaing, Patrick Pale, and Jean Sommer. Eur. J. Org. Chem. 2007, 5740–5748. Furthermore, the 1,6-enyne substrate yields cyclohexanone primarily through a cationic cyclization reaction under acidic conditions [see: Xiang Liu, Yuhan Wang, Jinlei Zhou, Yue Yu, Hua Cao. J. Org. Chem. 2020, 85, 2406-2414.].The synthetic method of this invention, synthesizing tetrasubstituted glutarimide derivatives containing allenol intermediates under acidic conditions, is challenging. Furthermore, the synthetic method of this invention has a broader substrate range than previous methods that produce single products. For example, it can construct compounds L1, L20, and L6 containing OTf groups, and compound L6 can be further derivatized to obtain compounds L21-L27 and all-carbon tetrasubstituted derivatives L8-L19.

[0072] During the research and development process, this invention mainly investigated different acids ( The reaction was carried out with different reaction parameters (using Lewis acids and solvents) and other acids, such as NH(Tf)2, HBr, AcOH, CF3CO2H, and Lewis acids, such as TMSOTf, BF3-Et2O, ZnBr2, and ZnCl2. TMSOTf, various sulfonic acids such as TfOH, arylsulfonic acids, methanesulfonic acids, and ethanesulfonic acids all yielded glutarimide derivatives. The reaction was carried out in various solvents, including CH3CN, DCM, HFIP, and DCM / HFIP mixtures. The preferred solvents were the mixed solvents DCM / HFIP or HFIP, which improved the conversion yield of the raw materials.

[0073] The present invention preferably uses raw material of formula 1 (1.0 equivalent) added to a mixed solvent DCM / HFIP (v / v = 20:1, 0.2M), and reacts with 1.0 equivalent of TfOH at room temperature or 35°C for 0.5-3 hours to prepare glutarimide derivatives L1-L7 and L20, which have the characteristics of high yield.

[0074] The present invention preferably uses raw material of formula 2 (1.0 equivalent) to add to a mixed solvent DCM / HFIP (v / v = 10:1, 0.2M), and reacts with 0.4 equivalent of TfOH at room temperature for 1 hour to prepare glutarimide derivatives L8-L11, which has the characteristics of high yield.

[0075] The present invention preferably uses raw material of formula 3 (1.0 equivalent) added to solvent HFIP (0.2M), 5.0 equivalent of electron-donating aromatic ring added, and 1.0 equivalent of TfOH reacted at room temperature for 1 hour to prepare glutarimide derivatives L12-L19, which has the characteristics of high yield.

[0076] The present invention preferably uses raw material of formula 4 (1.0 equivalent) added to a mixed solvent DCM / HFIP (v / v = 20:1, 0.2M), and reacts with 1.0 equivalent of TfOH at room temperature for 1 hour to prepare precursor glutarimide compound 6. Compounds L21-L27 are then converted in one step by palladium-catalyzed coupling reaction, which has the characteristics of high yield.

[0077] Specifically, the present invention can achieve the following beneficial effects:

[0078] (1) In response to the problem of drug resistance in tumor treatment by chemotherapy drugs, this invention has developed a new type of glutarimide derivative with the structure shown in Formula I, Formula II or Formula III. This type of compound has the function of reversing drug resistance and exhibits high inhibitory activity against the drug-resistant colon cancer cell line SW620 / AD300 when combined with paclitaxel (PTX).

[0079] (2) Based on the present invention, compounds L1-L27 have good activity against drug resistance in tumor drug-resistant cells. Among them, L24 combined with PTX has the best effect on the colorectal cancer drug-resistant cell line SW620 / AD300, with a half-maximal inhibitory concentration of 1.38 nM. Among the preferred compounds L1-L27, the most active compound L24 significantly increases the accumulation rate of rhodamine 123 in the drug-resistant cell line SW620 / AD300 at a concentration of 20 μM.

[0080] (3) Among the preferred compounds L1-L27, the most active compound L24 is provided by the present invention. The glutarimide derivative L24 has no significant inhibitory effect on the drug-metabolizing enzyme CYP3A4. The effect of compound L24 on CYP3A4 enzyme is significantly lower than that of the positive drug verapamil, indicating that L24 is a safe and effective P-gp inhibitor that reverses drug resistance.

[0081] (4) Furthermore, the method for synthesizing glutarimide derivatives in this invention is easy to prepare with readily available raw materials, simple in method, and free from transition metal catalysis, thus avoiding heavy metal pollution. Attached Figure Description

[0082] Figure 1 The tumor inhibition rate of compound L1-L27 in combination with paclitaxel in the drug-resistant tumor cell line SW620 / AD300.

[0083] Figure 2 The IC50 of the drug-resistant tumor cell line SW620 / AD300 with the addition of compound L24 and paclitaxel is... 50 picture.

[0084] Figure 3 The effect of paclitaxel combined with compound L24 on the proliferation of drug-resistant tumor cell line SW620 / AD300.

[0085] Figure 4 The effect of compound L24 on the accumulation of rhodamine 123 in the drug-resistant tumor cell line SW620 / AD300 is investigated.

[0086] Figure 5 The effect of compound L24 on the drug-metabolizing enzyme cytokinin P450 3A4 (CYP3A4 enzyme).

[0087] Figure 6 It is compound L24 1H NMR spectrum.

[0088] Figure 7 It is compound L24 13 C10 NMR spectrum.

[0089] Figure 8 It is compound L1 1 H NMR spectrum.

[0090] Figure 9 It is compound L1 13 C10 NMR spectrum.

[0091] Figure 10 It is compound L8 1 H NMR spectrum.

[0092] Figure 11 It is compound L8 13 C10 NMR spectrum. Detailed Implementation

[0093] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention. Furthermore, the technical features involved in the various embodiments of this invention described below can be combined with each other as long as they do not conflict with each other.

[0094] The glutarimide derivatives in this invention have the general structural formula shown in Formula I, Formula II, or Formula III:

[0095]

[0096] In Formula I:

[0097] R1 is benzyl, R2 is H, R3 is H, R4 is trifluoromethyl, and R5 is H;

[0098] Alternatively, R1 is methyl, R2 is H, R3 is H, R4 is any one of phenyl, 4-methoxy-phenyl, 4-chloro-phenyl, and 4-fluoro-phenyl, and R5 is H;

[0099] Alternatively, R1 is methyl, R2 is H, R3 is ethoxy, R4 is ethyl, and R5 is H;

[0100] Alternatively, R1 = R2 = -(CH2CH2CH2)-, R3 is H, R4 is ethyl, and R5 is H;

[0101] Alternatively, R1 is methyl, R2 is H, R3 is H, R4 is trifluoromethyl, and R5 is 2-(2-sec-butyl(benzoic acid)ethyl)-piperidine-1-carboxylic acid tert-butyl ester.

[0102] In Formula II:

[0103] R1 is benzyl, R2 is methyl, and R3 is any one of H, 3,5-dimethyl, and 2-chloro-5-methoxy.

[0104] Alternatively, R1 is benzyl, R2 is benzyl, and R3 is H.

[0105] In Formula III:

[0106] R1 is benzyl, R2 is methyl, R3 is H, and R4 is any one of 4-methoxy-phenyl, 3,4-dimethoxy-phenyl, 3-methyl-4-methoxy-phenyl, methylenedioxyphenyl, 1,4-benzodioxane, 3,5-dimethyl-phenyl, phenylethynyl, 4-chloro-phenyl, 3,4,5-trimethoxy-phenyl, 3,4,5-trimethyl-phenyl, 2-anthrayl, and benzofuranyl.

[0107] Alternatively, R1 is 3,4-dimethoxybenzyl, R2 is methyl, R3 is H, and R4 is 3,4-dimethoxy-phenyl;

[0108] Alternatively, R1 is benzyl, R2 = R3 = -(CH2CH2CH2CH2)-, and R4 is phenyl;

[0109] Alternatively, R1 is benzyl, R2 is H, R3 is methyl, and R4 is phenyl.

[0110] The following are the specific structural formulas of glutarimide derivatives L1-L27:

[0111]

[0112] The following are specific examples:

[0113] Example 1: Preparation and structural identification of glutarimide derivatives L1-L27

[0114] Synthetic route (I) of glutarimide derivatives L1-L27 in this invention: Synthetic steps of compound L1:

[0115]

[0116] The acetylide imide derivative of formula 1 (0.2 mmol, 1.0 equivalent) is soluble in a solution containing... In a mixed solvent of ultra-dry dichloromethane and hexafluoroisopropanol (v / v = 20:1, 1.0 mL), trifluoromethanesulfonic acid (0.2 mmol, 1.0 equivalent) was added and stirred at room temperature for 30 minutes. After filtration, the mixture was concentrated under vacuum using petroleum ether / ethyl acetate (v / v = 30:1) and then subjected to silica gel column chromatography (the silica gel powder could be pre-treated with petroleum ether containing 0.1% triethylamine by volume) to obtain the target compound L1. Its proton and carbon spectra are as follows (e.g.) Figure 8 , Figure 9 As shown):

[0117] Compound L1:82%yield(86.8mg), 1 H NMR (600MHz, CDCl3) δ7.46 (td, J=7.3,1.4

[0118] Hz,1H),7.37(t,J=7.8Hz,2H),7.34–7.22(m,10H),7.20–7.15(m,2H),4.98–4.89(m,2H),3.42(dd,J=13.9,4.6 Hz,1H),3.06(ddt,J=11.3,9.4,4.8Hz,1H),2.94(ddd,J=19.4,14.2,6.9Hz,2H),2.44(dd,J=14.5,11.1Hz,1H). 13 C NMR (150MHz, CDCl3) δ172.9,163.7,153.8,137.2,136.7,131.5,131.2,129.3,129.3,128.9,1 28.8,128.5,128.3,127.7,127.0,122.8,121.1,119.0,116.8,114.7,43.7,43.6,36.8,27.0. 19 F NMR(564MHz,CDCl3)δ-74.01.HRMS(ESI)m / z:[M+Na]+calcd for C 27 H 22 F3NO5SNa 552.1063; found:552.1053.

[0119] (II) Synthesis steps of compound L2:

[0120]

[0121] The acetylide imide derivative of formula 1 (0.2 mmol, 1.0 equivalent) is dissolved in a solution containing... In a mixed solvent of ultra-dry dichloromethane and hexafluoroisopropanol (v / v = 10:1, 1.0 mL), benzenesulfonic acid (0.2 mmol, 1.0 equivalent) was added, and the mixture was stirred at 35 °C for 3 hours. After filtration, the mixture was concentrated under vacuum and then subjected to silica gel column chromatography in petroleum ether / ethyl acetate (v / v = 10:1) (the silica gel powder could be pretreated with petroleum ether containing 0.1% triethylamine by volume) to obtain the target compound L2. The 1H and 1C spectra are as follows:

[0122] Compound L2:70%yield(64.6mg), 1 H NMR (400MHz, CDCl3) δ7.73–7.64(m,3H),7.64–7.35(m,8H),7.34–7.22(m,4H),5.23–5.03(m,2H),3.37(dd,J=1 4.2,5.1Hz,1H),3.06(dddd,J=12.5,8.9,7.0,3.5Hz,1H),2.73(dd,J=14.2,10.6Hz,1H),1.62(d,J=7.0Hz,3H). 13 C NMR (100MHz, CDCl3) δ175.0,165.2,154.8,137.3,136.9,134.0,132.2,130.2,129.7,129.0,1 28.9,128.5,128.0,127.9,127.6,121.8,43.4,37.7,30.1,16.7.HRMS(ESI)m / z:[M+Na]+calcd for C 26 H 23 NO5SNa 484.1189; found:484.1190.

[0123] (III) The synthetic procedures for compounds L3-L5 are similar to those for compound L2, and their synthetic routes are as follows:

[0124]

[0125] The only difference is:

[0126] The raw material used to synthesize L3 is 4-fluorobenzenesulfonic acid, specifically with an R5 group of 4-F.

[0127] The raw material used to synthesize L4 is 4-chlorobenzenesulfonic acid, specifically with an R5 group of 4-Cl.

[0128] The raw material used to synthesize L5 is 4-methoxybenzenesulfonic acid, specifically with an R5 group of 4-OMe.

[0129]

[0130] The sulfonic acid used in the synthesis of L6 is ethanesulfonic acid.

[0131]

[0132] The specific starting material compound of formula 1 used in the synthesis of L7 is an R3 group of 4-OEt and an ethanesulfonic acid.

[0133] The proton and carbon spectra of the synthesized compounds L3-L7 are as follows:

[0134] Compound L3:66%yield(63.3mg), 1 H NMR (400MHz, CDCl3) δ7.43 (ddd, J=9.0,

[0135] 5.0,2.5Hz,2H),7.34–7.18(m,6H),7.15–7.01(m,4H),6.96–6.83(m,2H),4.97–4.87(m,2H),3.19(dd,J=1 4.3, 5.1Hz, 1H), 2.90 (dqd, J=14.0, 7.0, 5.0Hz, 1H), 2.55 (dd, J=14.3, 10.6Hz, 1H), 1.44 (d, J=7.0Hz, 3H). 13 C NMR (100MHz, CDCl3) δ174.9,167.0,165.1,164.5,154.5,137.3,132.9,132.8,132.1,130.9,1 30.8,130.2,129.7,128.9,128.5,128.0,127.6,122.0,116.4,116.2,43.4,37.7,30.1,16.7. 19 F NMR(564MHz,CDCl3)δ-102.24.HRMS(ESI)m / z:[M+Na]+calcd for C 26 H 22 FNO5SNa 502.1095; found:502.1096.

[0136] Compound L4: 65% yield (64.5mg), 1H NMR(400MHz,CDCl3)δ7.41–7.26(m,9H),7.26–7.23(m,1H),7.19–7.05(m,4H),4.97(d,J=2.5Hz,2H),3.23(dd,J=14.2,5.1Hz,1H),2.94(tdd,J=12.0,8.7,6.0Hz,1H),2.60(dd,J=14.2,10.5Hz,1H),1.48(d,J=7.0Hz,3H). 13 C NMR(100MHz,CDCl3)δ174.9,165.0,154.5,140.8,137.3,135.3,132.1,130.3,129.7,129.4,129.3,129.0,128.5,128.0,127.6,122.0,43.4,37.7,30.1,16.8.HRMS(ESI)m / z:[M+Na]+calcd for C 26 H 22 ClNO5SNa 518.0799;found:518.0788.

[0137] Compound L5:61%yield(60.0mg), 1 H NMR(400MHz,CDCl3)δ7.43–7.34(m,2H),7.26(dddt,J=15.0,6.9,4.6,1.7Hz,6H),7.17–7.02(m,4H),6.75–6.62(m,2H),4.92(d,J=2.5Hz,2H),3.80(s,3H),3.17(dd,J=14.2,5.1Hz,1H),2.89–2.76(m,1H),2.51(dd,J=14.2,10.6Hz,1H),1.41(d,J=6.9Hz,3H). 13 C NMR(100MHz,CDCl3)δ175.0,165.3,164.0,154.8,137.4,132.5,130.3,130.1,129.7,128.9,128.5,128.1,127.9,127.6,121.6,114.2,55.9,43.3,37.8,30.1,16.7.HRMS(ESI)m / z:[M+Na]+calcd forC 27 H 25 NO6SNa514.1295;found:514.1305.

[0138] Compound L6:28%yield(24.6mg), 1 H NMR(600MHz,CDCl3)δ7.54–7.50(m,2H),7.47(td,J=5.0,2.4Hz,3H),7.40–7.36(m,2H),7.32–7.28(m,2H),7.26–7.22(m,1H),5.11–4.99(m,2H),3.42(dd,J=14.2,7.3Hz,1H),3.24(dq,J=14.7,7.4Hz,1H),2.92(td,J=7.5,2.0Hz,1H),2.89–2.82(m,1H),2.31–2.24(m,1H),2.01(t,J=7.6Hz,1H),1.93–1.86(m,1H),1.63–1.59(m,3H),1.40(t,J=7.4Hz,3H). 13 C NMR(150MHz,CDCl3)δ174.0,163.9,150.9,137.2,133.2,130.7,129.2,128.7,128.6,128.4,127.4,121.4,47.4,47.1,43.5,39.4,32.0,30.9,23.4,8.0.HRMS(ESI)m / z:[M+Na]+calcd for C 24 H 25 NO5SNa462.1346;found:462.1344.

[0139] Compound L7:75%yield(68.6mg), 1 H NMR(400MHz,CDCl3)δ7.30–7.17(m,7H),6.88–6.81(m,2H),4.90(d,J=1.8Hz,2H),4.03(q,J=7.0Hz,2H),3.12(dd,J=14.3,5.1Hz,1H),2.84(ddd,J=10.6,7.0,5.0Hz,1H),2.66(qd,J=7.3,4.0Hz,2H),2.49(dd,J=14.3,10.6Hz,1H),1.43–1.35(m,6H),1.25(t,J=7.4Hz,3H). 13C NMR (100MHz, CDCl3) δ174.7,164.9,160.5,154.0,137.1,130.6,128.6,128.2,127.2,124.8 ,121.3,114.1,63.4,47.8,43.0,37.3,29.9,16.3,14.6,7.9.HRMS(ESI)m / z:[M+Na]+calcd forC 24 H 27 NO6SNa480.1451; found:480.1452.

[0140] (iv) Synthetic steps of compounds L8-L11:

[0141] Taking compound L8 as an example, its synthetic route and steps are as follows:

[0142]

[0143] The acetylide imide derivative of formula 2 (0.2 mmol, 1.0 equivalent) was dissolved in a mixed solvent of ultra-dry dichloromethane and hexafluoroisopropanol (v / v = 10:1, 1.0 mL), and then added... Molecular sieves were used to slowly add trifluoromethanesulfonic acid (0.08 mmol, 0.4 equivalents), and the mixture was stirred at room temperature for 1 hour. After filtration, the crude product was concentrated under vacuum to obtain petroleum ether / ethyl acetate (v / v = 20:1). Purification by silica gel column chromatography yielded compound L8. Its 1H and 1C spectra are shown below (e.g., ...). Figure 10 , Figure 11 As shown):

[0144] Compound L8:84%yield(63.7mg), 1 H NMR (400MHz, CDCl3) δ7.80 (d, J=8.1Hz,

[0145] 1H),7.73(d,J=7.9Hz,1H),7.68–7.61(m,1H),7.58(dt,J=7.5,1.0Hz,1H),7.50–7.41(m,2H),7.39–7.21(m,6H),7.06(ddd,J=8.3, 7.4,1.3Hz,1H),5.14(s,2H),3.67(dd,J=13.4,5.2Hz,1H),3.07(dd,J=13.4,10.1Hz,1H),2.88–2.73(m,1H),1.35(d,J=7.0Hz,3H). 13C NMR (100MHz, CDCl3) δ174.7,167.5,145.5,142.5,141.3,138.1,137.5,136.9,130.3,130.1,129.0,128.7,1 28.4,128.0,127.7,127.5,126.9,126.8,120.2,119.5,43.7,38.5,35.3,17.0.HRMS(ESI)m / z:[M+Na]+calcd for C 26 H 21 NO2Na 402.1465; found:402.1458.

[0146] The synthetic procedures for compounds L9-L11 are similar to those for compound L8, and the synthetic route is as follows:

[0147]

[0148] The only difference is:

[0149] The starting material compound of formula 2 used in the synthesis of L9 has a benzyl group in R2 and an H group in R3;

[0150] The specific compound of formula 2 used in the synthesis of L10 has a methyl group at R2 and a 3,5-dimethylmethyl group at R3.

[0151] The specific compound of formula 2 used in the synthesis of L11 has a methyl group at R2 and a 2-chloro-4-methoxy group at R3.

[0152] The proton and carbon spectra of the synthesized compounds L9-L11 are as follows:

[0153] Compound L9:81%yield(73.8mg), 1 H NMR (400MHz, CDCl3) δ7.79 (d, J=8.0Hz,

[0154] 1H),7.66–7.56(m,2H),7.50–7.44(m,2H),7.38–7.28(m,6H),7.14(dd,J=4.7,1.9Hz,3H),7.07(tt,J=7.9,1.2Hz,2H),6.99(dd,J=6.6 ,2.8Hz,2H),5.19(d,J=4.3Hz,2H),3.41(d,J=8.3Hz,1H),3.31(dd,J=13.5,3.6Hz,1H),3.06(dd,J=8.0,2.5Hz,2H),2.85–2.76(m,1H). 13C NMR(100MHz,CDCl3)δ173.8,167.5,145.4,142.2,141.3,138.0,137.9,137.4,136.7,130.1,130.1,129.4,129.0,128.7,128.7,127.9,127.9,127.7,127.4,126.9,126.8,126.6,120.0,119.5,45.5,43.8,37.1,31.0.HRMS(ESI)m / z:[M+Na]+calcd for C 32 H 25 NO2Na 478.1778;found:478.1784.

[0155] Compound L10:80%yield(65.2mg),Z / E=1:0.96Mixture. 1 H NMR(400MHz,CDCl3)

[0156] δ7.93(d,J=8.0Hz,0.96H),7.73(d,J=7.9Hz,1H),7.60(dd,J=7.7,1.2Hz,1H),7.57–7.53(m,1H),7.44(ddt,J=10.0,5.9,1.6Hz,3.96H),7.34–7.27(m,6.88H),7.26–7.16(m,3.84H),7.04(td,J=7.7,1.3Hz,1H),6.91(s,0.96H),6.81(s,0.96H),5.14(s,2H),5.12–5.01(m,1.92H),3.58(dd,J=13.2,5.0Hz,0.96H),3.08(dd,J=13.2,10.0Hz,0.96H),2.99–2.94(m,0.96H),2.93–2.88(m,1H),2.76(dd,J=12.7,10.5Hz,1H),2.65–2.57(m,1H),2.40(s,2.88H),2.35(s,3H),2.32(s,3H),1.90(s,2.88H),1.47(d,J=6.9Hz,3H),1.19(d,J=6.9Hz,2.88H). 13C NMR(100MHz,CDCl3)δ175.1,174.9,167.4,166.6,149.4,148.0,142.7,142.2,142.1,140.7,140.4,140.2,139.8,137.9,137.6,135.5,135.4,133.7,132.2,131.7,129.8,129.8,129.3,128.9,128.6,128.5,127.8,127.6,127.6,127.6,127.3,126.5,125.3,125.3,120.1,119.6,118.5,118.2,43.9,43.6,38.9,38.8,37.9,35.2,23.3,22.2,21.7,21.6,16.9,16.5.HRMS(ESI)m / z:[M+Na]+calcd forC 28 H 25 NO2Na 430.1778;found:430.1771.

[0157] Compound L11:67%yield(59.5mg),Z / E=1:0.61Mixture. 1 H NMR(400MHz,CDCl3)

[0158] δ8.34–8.27(m,1H),8.27–8.22(m,0.61H),7.75(d,J=8.1Hz,0.61H),7.69(d,J=7.8Hz,1H),7.51(d,J=2.3Hz,1H),7.45(td,J=7.7,1.5Hz,3.22H),7.40–7.27(m,5.83H),7.26–7.20(m,2.22H),7.03(td,J=7.8,1.2Hz,0.61H),6.88(d,J=2.1Hz,0.61H),6.84(d,J=2.2Hz,1H),5.14(d,J=9.2Hz,3.22H),3.85(s,1.83H),3.69(dd,J=13.4,5.1Hz,1H),3.60(dd,J=13.4,5.2Hz,0.61H),3.57(s,3H),3.08(ddd,J=19.6,13.3,10.0Hz,1.61H),2.90–2.77(m,1.61H),1.36(t,J=6.9Hz,4.83H). 13C NMR (100MHz, CDCl3) δ175.0,174.8,167.8,167.7,159.8,159.4,145.9,145.2,141.9,141.7,140 .8,140.4,138.5,137.9,137.7,137.3,131.8,130.9,130.8,130.6,130.0,129.6,129.5,129.3,1 29.1,129.0,129.0,128.1,128.0,127.6,127.0,126.9,126.5,123.7,123.1,118.1,115.8,113.9 ,111.4,56.4,56.0,44.1,44.1,38.8,38.7,36.2,35.8,17.4,17.3.HRMS(ESI)m / z:[M+Na]+calcd for C 27 H 22 ClNO3Na466.1180; found:466.1178.

[0159] (V) Synthetic steps of compounds L12-L19:

[0160] The synthetic routes for compounds L12-L13 and L17-L19 are as follows:

[0161]

[0162] The compound acetylide derivative (Formula 3) (0.2 mmol, 1.0 equivalent) is dissolved in a solution containing... In an ultra-dry hexafluoroisopropanol (1.0 mL) solvent containing molecular sieves, aromatic compounds (1.0 mmol, 5.0 equivalents) and trifluoromethanesulfonic acid (0.2 mmol, 1.0 equivalents) were added sequentially. The mixture was stirred at room temperature for 1 hour and then concentrated under vacuum to obtain crude products. The crude products were purified by silica gel column chromatography using petroleum ether / ethyl acetate (v / v = any one of 20:1, 10:1, 8:1, 6:1, 3:1) to obtain the target compounds L12-L13 and L17-L19.

[0163] in:

[0164] The aromatic compound used to synthesize L12 is anisole (in this case, the R group is methoxy).

[0165] The aromatic compound used to synthesize L13 is 1,2-dimethoxybenzene (in this case, the R group is 1,2-dimethoxy).

[0166] The aromatic compound used in the synthesis of L17 is 2-methyl anisole (in this case, the R group is 1-methoxy-2-methyl);

[0167] The aromatic compound used to synthesize L18 is specifically 1,2-methylenedioxybenzene (in this case, the R group is methylenedioxy).

[0168] The aromatic compound used to synthesize L19 is 1,4-benzodioxane (in this case, the R group is bis(ethoxy)).

[0169] The synthetic route for compound L14 is as follows:

[0170]

[0171] The aromatic compound used to synthesize L14 is specifically 1,2-dimethoxybenzene.

[0172] The synthetic route for compound L15 is as follows:

[0173]

[0174] The aromatic compound used to synthesize L15 is specifically 1,2-dimethoxybenzene.

[0175] The synthetic route for compound L16 is as follows:

[0176]

[0177] The aromatic compound used to synthesize L16 is specifically 1,2-dimethoxybenzene.

[0178] The proton and carbon spectra of the synthesized compounds L12-L19 are as follows:

[0179] Compound L12:76%yield(62.5mg),Z / E=0.73:1Mixture. 1 H NMR (400MHz, CDCl3)

[0180] δ7.39–7.27(m,10.84H),7.26–7.22(m,3.46H),7.13–7.09(m,1.46H),7.06–6.99(m ,4H),6.92–6.84(m,3.46H),6.76–6.72(m,1H),5.02–4.90(m,3.46H),3.82(s,3H),3 .79(s,2.19H),2.89(dd,J=13.5,4.9Hz,1H),2.80(dd,J=13.4,5.0Hz,0.73H),2.75 –2.67(m,1.73H),2.52(ddd,J=26.9,13.5,9.4Hz,1.73H),1.25(t,J=6.8Hz,5.19H). 13C NMR(100MHz,CDCl3)δ175.2,175.1,167.8,167.7,160.1,159.8,153.6,153.3,142.3,141.2,137.8,134.1,133.2,131.7,130.9,130.1,129.4,129.2,129.1,128.7,128.5,128.5,128.3,128.2,127.4,127.4,125.1,125.0,113.8,113.7,55.5,55.3,43.2,43.1,38.7,38.6,34.6,34.5,16.6,16.6.HRMS(ESI)m / z:[M+Na]+calcd for C 27 H 25 NO3Na 434.1727;found:434.1730.

[0181] Compound L13:82%yield(72.4mg),Z / E=0.73:1Mixture. 1 H NMR(400MHz,CDCl3)

[0182] δ7.40–7.32(m,6.46H),7.32–7.26(m,4.19H),7.26–7.21(m,3.19H),7.12(dd,J=6.7,3.0Hz,1.46H),7.05–6.98(m,2H),6.84(d,J=8.3Hz,1H),6.71–6.65(m,1.73H),6.58(d,J=2.0Hz,1H),6.54(dd,J=8.2,2.0Hz,0.73H),6.48(d,J=2.1Hz,0.73H),5.02–4.90(m,3.46H),3.90(s,3H),3.87(s,2.19H),3.77(s,3H),3.60(s,2.19H),2.89(dd,J=13.4,5.0Hz,1H),2.82(dd,J=13.4,4.9Hz,0.73H),2.76–2.68(m,1.73H),2.58–2.48(m,1.73H),1.26(dd,J=7.0,5.8Hz,5.19H). 13C NMR(100MHz,CDCl3)δ175.3,175.1,167.7,167.6,153.7,153.6,149.6,149.4,148.7,148.6,142.0,141.0,137.8,137.7,134.4,133.4,130.1,129.3,129.1,129.1,128.8,128.5,128.5,128.4,128.2,127.4,127.4,125.3,125.0,123.2,122.3,113.2,112.7,110.9,110.6,56.1,56.1,56.0,55.9,43.2,43.1,38.6,38.6,34.7,34.6,16.6,16.6.HRMS(ESI)m / z:[M+Na]+calcd for C 28 H 27 NO4Na 464.1832;found:464.1825.

[0183] Compound L14:79%yield(69.8mg), 1 H NMR(400MHz,CDCl3)δ7.43–7.32(m,6H),7.29(d,J=8.5Hz,4.1H),7.26–7.18(m,3.5H),7.16–7.09(m,1.4H),7.02–6.94(m,2H),6.83(d,J=8.3Hz,1H),6.71–6.63(m,1.7H),6.60(d,J=2.0Hz,1H),6.51(dd,J=8.2,2.0Hz,0.7H),6.45(d,J=2.1Hz,0.7H),5.04–4.94(m,3.4H),3.88(s,3H),3.85(s,2.1H),3.77(s,3H),3.60(s,2.1H),3.10(ddd,J=7.2,5.3,2.0Hz,1H),2.97(dtd,J=7.7,5.5,2.9Hz,0.7H),2.85(dd,J=5.3,3.4Hz,0.7H),2.81(dd,J=5.3,3.3Hz,1H),2.63(dd,J=5.1,2.0Hz,1H),2.59(dd,J=5.1,2.0Hz,0.7H),1.23(dd,J=9.2,7.0Hz,5.1H). 13C NMR(100MHz,CDCl3)δ171.9,171.7,167.8,167.7,151.7,151.6,149.8,149.6,149.1,149.0,142.1,141.4,137.9,137.8,134.5,133.9,131.8,131.5,129.8,129.8,129.6,129.5,129.0,128.9,128.8,128.8,128.6,128.5,127.9,122.5,112.8,111.3,111.0,56.4,56.3,56.3,56.2,43.3,43.2,41.4,41.3,32.3,32.2,19.9,19.9.HRMS(ESI)m / z:[M+Na]+calcdfor C 28 H 27 NO4Na 464.1832;found:464.1823.

[0184] Compound L15:72%yield(69.3mg),Z / E=0.8:1Mixture. 1 H NMR(400MHz,CDCl3)

[0185] δ7.42–7.34(m,7.4H),7.29(d,J=8.6 Hz,3.6H),7.25–7.18(m,3.4H),7.16–7.10(m,1.6H),7.01–6.96(m,2H),6.84(d,J=8.3 Hz,1H),6.70–6.64(m,1.8H),6.62(d,J=2.0Hz,1H),6.51(dd,J=8.2,2.0 Hz,0.8H),6.45(d,J=2.0 Hz,0.8H),5.06–4.93(m,3.6H),3.90(s,3H),3.86(s,2.4H),3.77(s,3H),3.60(s,2.4H),2.86(s,1H),2.81(d,J=3.3 Hz,1.8H),2.74(dt,J=12.5,4.2 Hz,0.8H),2.49(dt,J=13.7,3.4 Hz,1.8H),1.89–1.80(m,2H),1.75–1.64(m,2.8H),1.33(d,J=13.2 Hz,4H),1.15(dd,J=11.4,8.2 Hz,3.8H). 13C NMR(100 MHz,CDCl3)δ173.6,173.4,167.8,167.7,151.4,151.2,149.4,149.2,148.7,148.7,142.1,141.2,137.8,137.7,134.6,133.6,132.7,132.1,131.9,129.5,129.5,129.3,129.2,129.0,128.7,128.7,128.5,128.4,128.4,128.2,128.2,127.5,122.3,122.2,112.5,112.5,111.0,110.6,56.1,56.0,55.9,55.9,43.7,43.6,43.2,43.1,40.9,40.8,29.2,29.1,26.6,24.9,24.9,22.0,21.9.HRMS(ESI)m / z:[M+Na]+calcd forC 31 H 31 NO4Na 504.2145;found:504.2147.

[0186] Compound L16:65%yield(65.2 mg),Z / E=0.75:1 Mixture. 1 H NMR(400 MHz,CDCl3)

[0187] δ7.37–7.21(m,5.50H),7.14–7.09(m,1.50H),7.03–6.93(m,5.50H),6.84(d,J=8.3 Hz,1H),6.78(d,J=8.2 Hz,1.75H),6.72–6.69(m,0.75H),6.69–6.66(m,0.75H),6.57(d,J=2.0 Hz,1H),6.53(dd,J=8.2,2.0 Hz,0.75H),6.47(d,J=2.0 Hz,0.75H),4.96–4.81(m,3.50H),3.89(s,3H),3.88–3.85(m,7.50H),3.83(d,J=2.2 Hz,5.25H),3.77(s,3H),3.63(s,2.25H),2.88(dd,J=13.4,5.0 Hz,1H),2.81(dd,J=13.4,5.0 Hz,0.75H),2.70(qt,J=7.3,5.7 Hz,1.75H),2.51(ddd,J=18.3,13.5,9.4 Hz,1.75H),1.25(dd,J=7.0,5.6 Hz,5.25H). 13 C NMR(100 MHz,CDCl3)δ175.3,175.2,167.7,153.6,153.3,149.6,149.3,148.8,148.8,148.7,148.5,148.5,142.1,141.0,134.4,133.5,130.5,130.0,129.3,128.8,128.5,128.3,128.2,125.3,125.2,123.2,122.3,122.0,113.2,113.0,113.0,112.7,111.1,111.0,110.9,110.6,56.1,56.1,56.1,55.9,55.9,43.0,42.9,38.6,34.7,34.6,16.6.HRMS(ESI)m / z:[M+Na]+calcd for C 30 H 31 NO6Na 524.2044;found:524.2037.

[0188] Compound L17:87%yield(74.0 mg),Z / E=0.93:1 Mixture. 1 H NMR(400 MHz,CDCl3)

[0189] δ7.35(dtd,J=7.7,6.3,5.5,1.7 Hz,6.65H),7.32–7.26(m,5H),7.23(dt,J=6.6,1.5 Hz,3.72H),7.11(dd,J=6.6,3.0 Hz,2H),7.05–6.99(m,2H),6.92(dd,J=8.4,2.3 Hz,1H),6.85(d,J=2.2 Hz,1H),6.82–6.75(m,2.79H),6.68–6.62(m,0.93H),4.97(qd,J=13.7,10.5 Hz,3.86H),3.84(s,3H),3.81(s,2.79H),2.90(dd,J=13.5,4.9 Hz,1H),2.79(dd,J=13.3,5.0 Hz,0.93H),2.75–2.66(m,1.93H),2.52(ddd,J=26.7,13.5,9.5 Hz,1.93H),2.17(s,3H),2.09(s,2.79H),1.25(t,J=6.6 Hz,5.79H). 13 C NMR(100MHz,CDCl3)δ175.3,175.2,167.9,167.8,158.3,158.1,154.0,153.8,142.4,141.3,137.9,137.8,133.7,132.7,132.4,131.8,130.1,129.3,129.1,129.1,128.7,128.5,128.4,128.4,128.2,128.2,128.2,127.4,127.4,126.7,126.4,124.9,124.7,109.5,109.4,55.5,55.3,43.2,43.1,38.7,38.6,34.6,34.6,16.6,16.5.HRMS(ESI)m / z:[M+Na]+calcd forC 28 H 27 NO3Na 4448.1883;found:448.1884.

[0190] Compound L18:86%yield(77.8 mg),Z / E=0.63:1 Mixture. 1 H NMR(400 MHz,CDCl3)

[0191] δ7.33(dddd,J=18.9,10.9,5.7,2.6 Hz,10.04H),7.26–7.20(m,3H),7.14–7.08(m,1.26H),7.03–6.98(m,2H),6.79(d,J=8.0 Hz,1H),6.68–6.58(m,1.63H),6.54(d,J=1.7Hz,1H),6.50–6.44(m,1.26H),5.98(s,2H),5.96(s,1.26H),5.05–4.88(m,3.26H),2.88(dd,J=13.5,5.0 Hz,1H),2.79(dd,J=13.5,5.0 Hz,0.63H),2.75–2.66(m,1.63H),2.58–2.50(m,1H),2.48(d,J=4.2 Hz,0.63H),1.27(d,J=7.0 Hz,3H),1.23(d,J=7.0 Hz,1.89H). 13 C NMR(100 MHz,CDCl3)δ175.1,175.1,167.7,167.6,153.3,153.1,148.1,147.9,147.8,147.7,142.0,140.8,137.7,135.7,134.7,130.0,129.2,129.1,129.0,128.8,128.5,128.5,128.4,128.3,127.5,127.4,125.6,125.6,124.2,123.3,110.4,109.9,108.3,108.2,101.6,101.4,43.2,43.2,38.6,34.6,34.5,16.6,16.6.HRMS(ESI)m / z:[M+Na]+calcd for C 27 H 23 NO4Na 448.1519;found:448.1512.

[0192] Compound L19:82%yield(72.0mg),Z / E=0.68:1Mixture. 1 H NMR(400MHz,CDCl3)

[0193] δ7.40–7.26(m,9.76H),7.26–7.21(m,4H),7.14–7.04(m,1.68H),7.03–6.97(m,1.68H),6.83(d,J=8.3 Hz,1H),6.70(d,J=8.3Hz,0.68H),6.63–6.53(m,2.36H),6.48(dd,J=8.3,2.1Hz,0.68H),5.04–4.87(m, 3.36H), 4.33–4.14 (m, 6.72H), 2.90 (dd, J=13.5, 5.0Hz, 1H), 2.78 (dd, J=13.4, 5.0Hz, 0.68H), 2.74–2. 63(m,1.68H),2.51(ddd,J=31.0,13.5,9.3Hz,1.68H),1.26(d,J=6.9Hz,3H),1.22(d,J=7.0Hz,2.04H). 13 C NMR (100MHz, CDCl3) δ175.2,175.2,167.8,167.6,153.3,153.1,144.2,143.9,143.4,143.3 ,142.1,141.0,137.8,135.1,134.1,130.0,129.2,129.0,129.0,128.7,128.5,128.5,128.4 ,128.2,128.2,127.4,125.4,125.2,123.5,122.8,119.1,118.5,117.2,117.1,64.6,64.5, 64.5,64.4,43.2,43.1,38.6,38.6,34.5,34.5,16.6,16.6.HRMS(ESI)m / z:[M+Na]+calcdfor C 28 H 25 NO4Na 462.1676,found:462.1666.

[0194] (vi) The synthetic procedure for compound L20 is similar to that for compound L1, and the synthetic route is as follows:

[0195]

[0196] L20 preparation method: The raw material of formula 1 (0.2 mmol, 1.0 equivalent) is dissolved in a solution containing... The target compound L20 was obtained by adding trifluoromethanesulfonic acid (0.2 mmol, 1.0 equivalent) to a mixed solvent of ultra-dry dichloromethane and hexafluoroisopropanol (v / v = 20:1, 1.0 mL), stirring at room temperature for 1 h, filtering, vacuum concentration, and silica gel column chromatography in petroleum ether / ethyl acetate (v / v = 10:1) (the silica gel powder could be pretreated with petroleum ether containing 0.1% triethylamine by volume).

[0197] The proton and carbon spectra of the synthesized compound L20 are as follows:

[0198] Compound L20: 68% yield (111.9mg), 1 H NMR (600MHz, CDCl3) δ7.95 (d, J=8.1

[0199] Hz,2H),7.52–7.46(m,1H),7.41(dd,J=8.6,7.0Hz,2H),7.38–7.30(m,4H),5.01–4.89(m,2H),4 .73(qd,J=6.3,1.3Hz,1H),4.51(s,1H),4.35–4.26(m,2H),4.08(s,1H),3.09(dd,J=14.5,5.1Hz ,1H),2.95–2.79(m,2H),2.61(dd,J=14.5,10.2Hz,1H),2.25–2.14(m,1H),1.87(dt,J=12.8,6. 4Hz,1H),1.71–1.47(m,8H),1.43(d,J=7.0Hz,3H),1.16(d,J=6.1Hz,3H),0.86(t,J=7.4Hz,3H). 13 C NMR (150MHz, CDCl3) δ174.0,166.2,163.7,155.5,155.5,154.4,141.7,131.4,131.3,129.8,129.6,129.3,128.6,128.4,122.6,121.2,1 19.1,117.0,114.8,73.0,73.0,62.7,62.7,48.0,43.2,39.0,38.9,37.2,29.8,29.1,28.9,28.6,25.5,25.5,19.8,19.1,16.5,9.7,9.7. 19 F NMR(565MHz,CDCl3)δ-73.82.HRMS(ESI)m / z:[M+Na]+calcd for C 34 H 39F3N2O9SNa 731.2221; found:731.2212.

[0200] (vii) The synthesis of compounds L21-L27 is based on compound 6. The synthetic steps of compound 6 are similar to those of compound L1. The synthetic route is as follows:

[0201]

[0202] The synthetic route for obtaining compound L21 from compound 6 is as follows:

[0203]

[0204] Using compound 6 as a substrate, compound 6 (0.1 mmol, 1.0 equivalent) was dissolved in a THF / H₂O (v / v = 10:1) mixed solvent. Potassium phosphate trihydrate (0.25 mmol, 2.5 equivalent), 1,1-bis(diphenylphosphine)dimerferropalladium dichloride (0.01 mmol, 0.1 equivalent), and 3,5-dimethylphenylboronic acid pinacol ester (0.12 mmol, 1.2 equivalent) were added sequentially. The reaction was carried out under an inert argon atmosphere at 75°C for 24 hours. Extraction was performed with ethyl acetate and water, and the crude product was concentrated under vacuum to obtain a petroleum ether / ethyl acetate (v / v = 30:1) crude product. Purification by silica gel column chromatography yielded compound L21. The 1H and 1C spectra are as follows:

[0205] Compound L21: 45% yield (18.4mg), 1 H NMR (400MHz, CDCl3) δ7.30–7.22(m,5H),7.22–7.10(m,3H),7.08–6.94(m,2H),6.83(s,1H),6.63–6.45(m,2H),4.96–4.76 (m,2H),2.72(dd,J=13.5,5.0Hz,1H),2.66–2.55(m,1H),2.40(dd,J=13.4,9.4Hz,1H),2.11(s,6H),1.15(d,J=6.9Hz,3H). 13 C NMR (100MHz, CDCl3) δ175.2,167.6,154.2,141.9,141.0,137.8,137.6,130.2,129.8,128.9,128. 6,128.5,128.5,127.4,126.9,125.5,43.2,38.6,34.4,21.5,16.6.HRMS(ESI)m / z:[M+Na]+calcd for C 28 H 27NO2Na 432.1934; found:432.1927.

[0206] The synthetic route for obtaining compound L22 from compound 6 is as follows:

[0207]

[0208] Compound 6 (0.1 mmol, 1.0 equivalent), bis(triphenylphosphine)palladium dichloride (0.005 mmol, 0.05 equivalent), and cuprous iodide (0.005 mmol, 0.05 equivalent) were added to a reaction flask, followed by the sequential addition of triethylamine (0.15 mmol, 1.5 equivalent) and phenylacetylene (0.13 mmol, 1.3 equivalent). The mixture was heated at 60 °C for 4 hours under an inert argon atmosphere with 1.0 mL of anhydrous THF. The crude product was concentrated under vacuum and purified by silica gel column chromatography with petroleum ether / ethyl acetate (v / v = 30:1) to obtain compound L22.

[0209] Compound L22:87%yield(35.3mg),Z / E=0.36:1Mixture. 1 H NMR (600MHz, CDCl3)

[0210] δ7.45–7.41(m,0.72H),7.39(ddd,J=6.5,3.5,1.6Hz,2.72H),7.36–7.29(m,2.08H),7.29–7.13 (m,14.88H),5.02(d,J=1.9Hz,0.72H),4.91–4.81(m,2H),3.24(dd,J=14.0,5.1Hz,1H),2.82(d ddd,J=13.9,9.7,6.9,5.0Hz,1H),2.72(dt,J=14.0,9.6Hz,1.36H),2.54(ddd,J=10.3,7.1,5.0 Hz, 0.36H), 2.38 (dd, J=14.5, 10.4Hz, 0.36H), 1.35 (d, J=6.9Hz, 3H), 1.13 (d, J=6.9Hz, 1.08H). 13C NMR (150MHz, CDCl3) δ173.9,173.5,164.4,164.3,137.2,136.8,136.5,136.3,13 3.2,132.4,131.0,130.7,130.7,130.7,128.4,128.0,127.9,127.8,127.7,127. 6,127.6,127.5,127.4,127.2,127.1,126.3,121.9,121.2,100.9,100.4,89.0,8 7.8,42.3,42.0,36.9,36.3,33.5,31.5,15.6,15.1.HRMS(ESI)m / z:[M+Na]+calcd for C 28 H 23 NO2Na 428.1621; found:428.1620.

[0211] The synthetic route for compounds L23-L27 is as follows:

[0212]

[0213] Compound 6 (0.1 mmol, 1.0 equivalent), arylphenylboronic acid (0.15 mmol, 1.5 equivalent), bis(triphenylphosphine)palladium dichloride (0.005 mmol, 0.05 equivalent), and 2M sodium carbonate solution (1.0 mL) were reacted in THF (4.0 mL) under an inert argon atmosphere at 40 °C for 3 hours. The mixture was extracted with ethyl acetate and water, and the crude product was concentrated under vacuum to obtain the crude product. The crude product was purified by silica gel column chromatography with petroleum ether / ethyl acetate (v / v = any one of 25:1, 3:1, or 20:1) to obtain compounds L23-L27.

[0214] in:

[0215] The arylphenylboronic acid used in the synthesis of L23 is specifically 4-chlorophenylboronic acid (in this case, the R group is 4-chloro group);

[0216] The arylphenylboronic acid used to synthesize L24 is specifically 3,4,5-trimethoxyphenylboronic acid (in this case, the R group is 3,4,5-trimethoxy);

[0217] The arylphenylboronic acid used to synthesize L25 is specifically 3,5-dimethyl-4-methoxy-phenylboronic acid (in this case, the R group is 3,5-dimethyl-4-methoxy).

[0218] The arylphenylboronic acid used in the synthesis of L26 is specifically 2-anthraylphenylboronic acid (in this case, the R group is 2-anthrayl);

[0219] The arylphenylboronic acid used to synthesize L27 is specifically benzofuran-5-boronic acid (in this case, the R group is 3-furanyl);

[0220] The proton and carbon spectra of the synthesized compounds L23-L27 are as follows:

[0221] Compound L23: 88% yield (36.6mg), Z / E=1:1Mixture. 1 H NMR (400MHz, CDCl3)δ

[0222] 7.28–7.06(m,20H),7.02–6.92(m,4H),6.91–6.77(m,4H),4.93–4.77(m,4H),2.72(dt,J=13 .4,5.1Hz,2H),2.66–2.56(m,2H),2.40(dd,J=13.4,9.3Hz,2H),1.15(dd,J=7.0,4.9Hz,6H). 13 C NMR (100MHz, CDCl3) δ175.0,174.9,167.4,167.3,152.1,152.1,141.4,140.4, 140.3,139.2,137.6,137.6,134.9,134.3,131.3,130.5,129.9,129.2,129.1, 129.0,128.9,128.7,128.6,128.5,128.5,128.5,128.4,127.6,127.5,126.5, 126.4,43.2,43.2,38.6,38.5,34.3,34.3,16.6.HRMS(ESI)m / z:[M+Na]+calcd forC 26 H 22 ClNO2Na438.1231; found:438.1227.

[0223] Compound L24: 89% yield (42.0mg), Z / E=0.84:1Mixture. 1 H NMR (400MHz, CDCl3)

[0224] δ7.33(ddt,J=5.6,3.9,1.6Hz,6.84H),7.30–7.24(m,4.2H),7.24–7.18(m,3.68H),7.13–7.08(m,1.68H),7.02–6.97(m,2H),6.28(s,2H),6.17(s,1.68H),5.00–4.88(m,3.68H),3.85(s,3H),3.84(s,2.52H),3.74(s,6H),3.57(s,5.04H),2.82(ddd,J=13.5,11.0,5.0Hz,1.84H),2.75–2.67(m,1.84H),2.54–2.46(m,1.84H),1.27(d,J=7.0Hz,3H),1.24(d,J=6.9Hz,2.52H). 13 C NMR(100MHz,CDCl3)δ175.2,175.1,167.6,167.3,153.8,153.6,153.2,153.0,141.4,140.7,138.4,138.3,137.8,137.7,137.3,136.3,129.9,129.2,129.1,129.0,128.9,128.5,128.5,128.5,128.2,127.5,127.5,125.9,125.5,107.2,106.7,61.1,56.4,56.2,43.3,43.2,38.7,38.5,34.7,34.6,16.7,16.5.HRMS(ESI)m / z:[M+Na]+calcd for C 29 H 29 NO5Na 494.1938;found:494.1934.

[0225] Compound L25:83%yield(36.5mg),Z / E=0.5:1Mixture. 1 H NMR(400MHz,CDCl3)

[0226] δ7.42–7.27(m,9H),7.23(d,J=6.7Hz,3H),7.14–7.07(m,1H),7.04–6.96(m,2H),6.74(s,2H),6.64(s,1H),5.05–4.87(m,3H),3.74(s,3H),3.72(s,1.5H),2.89–2.76(m,1.5H),2.70(dddd,J=9.4,7.3,4.9,2.5Hz,1.5H),2.50(ddd,J=13.4,11.9,9.4Hz,1.5H),2.24(s,6H),2.16(s,3H),1.25(dd,J=10.1,6.9Hz,4.5H). 13 C NMR(100MHz,CDCl3)δ175.3,175.3,167.7,167.6,157.4,157.2,153.7,153.7,142.1,141.2,137.8,137.8,137.2,136.2,131.0,130.6,130.4,129.9,129.8,129.1,129.1,128.9,128.6,128.5,128.5,128.2,127.4,125.4,125.1,59.9,43.2,43.1,38.6,34.6,34.4,16.6,16.6,16.4,16.3.HRMS(ESI)m / z:[M+Na]+calcd for C 29 H 29 NO3Na 462.2040;found:462.2046.

[0227] Compound L26:84%yield(40.4mg),Z / E=0.51:1Mixture. 1 H NMR(400MHz,CDCl3)

[0228] δ8.44–8.33(m,2.51H),8.18(s,0.51H),8.02–7.91(m,4.02H),7.85(d,J=8.8Hz,0.51H),7.79(d,J=1.6Hz,1H),7.58(d,J=1.6Hz,0.51H),7.51–7.43(m,3.06H),7.26(s,11.06H),7.24(s,1H),7.19(dd,J=6.7,2.9Hz,1H),7.15–7.04(m,3.51H),5.07–4.89(m,3.02H),2.93(ddd,J=22.9,13.4,4.9Hz,1.51H),2.86–2.67(m,1.51H),2.61(ddd,J=20.4,13.4,9.4Hz,1.51H),1.30(d,J=6.9Hz,1.53H),1.23(d,J=6.9Hz,3H). 13 C NMR(100MHz,CDCl3)δ175.2,175.1,167.6,167.6,153.6,153.5,141.7,140.7,138.9,137.8,137.7,137.6,132.5,132.3,132.0,131.3,131.2,131.1,131.0,130.2,129.8,129.4,129.2,129.2,128.9,128.7,128.6,128.6,128.6,128.5,128.5,128.4,128.4,128.3,128.0,127.5,127.2,127.1,126.9,126.4,126.4,126.3,126.2,126.0,125.7,125.7,43.2,38.7,34.6,34.5,16.7,16.6.HRMS(ESI)m / z:[M+Na]+calcd for C 34 H 27 NO2Na 504.1934;found:504.1927.

[0229] Compound L27:97%yield(40.9mg),Z / E=0.69:1Mixture. 1 H NMR(400MHz,CDCl3)

[0230] δ7.65(d,J=2.2Hz,1H),7.59(d,J=2.3Hz,0.69H),7.47(d,J=8.5Hz,1H),7.34(td,J=4.5,3.9,1.4Hz,7H),7.32–7.26(m,5.45H),7.26–7.20(m,3.45H),7.12(dd,J=6.6,3.0Hz,1.38H),7.03(ddd,J=8.3,6.3,1.7Hz,3H),6.93(dd,J=8.5,1.8Hz,0.69H),6.76–6.72(m,1H),6.65–6.61(m,0.69H),4.96(qd,J=13.7,4.9Hz,3.38H),2.86(ddd,J=13.5,5.0,3.8Hz,1.69H),2.72(dddd,J=11.8,9.4,7.0,4.9Hz,1.69H),2.54(ddd,J=13.1,9.3,3.6Hz,1.69H),1.25(dd,J=12.6,7.0Hz,5.07H). 13 C NMR(150MHz,CDCl3)δ175.0,167.6,167.5,154.7,153.8,153.6,145.9,145.4,142.2,141.1,137.6,136.5,135.5,129.9,129.1,128.9,128.6,128.4,128.3,128.1,128.1,127.5,127.4,127.3,127.3,126.3,125.8,125.6,125.6,122.8,121.9,43.0,38.5,38.5,34.3,34.3,16.5,16.4.HRMS(ESI)m / z:[M+Na]+calcd for C 28 H 23 NO3Na 444.1570;found:444.1565.

[0231] It is known in the prior art that the construction of tetrasubstituted olefins via allenol intermediates is mainly carried out under basic conditions [see: (a) HYKim, J.-Y.Li, K.Oh, Angew. Chem., Int. Ed. 2013, 52, 3736-3740; b) HYKim, E.O. Rooney, R.P. Meury, K.Oh, Angew. Chem., Int. Ed. 2013, 52, 8026-8030; c) TEReynolds, K.S. Cheidt, Angew. Chem., Int. Ed. 2007, 46, 7806-7809; d) R. Chinchilla, C. Nájera, Chem. Rev. 2000, 100, 1891-1928; e) HYKim, J.-Y.Li, K.Oh, J.Org. Chem. 201 2,77,11132-11145; f)P.Maity,SD Lepore,J.Org.Chem.2009,74,158-162; g)RV Kolakowski,M.Manpadi,Y.Zhang,TJ Emge,LJ Williams,J.Am.Chem.Soc.2009,131,12910-12911; h)TEReynolds,ARBharadwaj,KAScheidt,J.Am.Chem.Soc.2006,128,15382-15383; i)TEReynolds,MS Binkley,KAScheidt,Org.Lett.2008,10,2449-2452. Under acidic conditions, protonation is more likely to occur [see: Aleksander]. V. Vasilyev, Stéphane Walspurger, Stefan Chassaing, Patrick Pale, and Jean Sommer. Eur. J. Org. Chem. 2007, 5740–5748. Furthermore, the 1,6-enyne substrate, under acidic conditions, primarily yields the cyclohexanone product via a cationic cyclization reaction [see: Xiang Liu, Yuhan Wang, Jinlei Zhou, Yue Yu, Hua Cao. J. Org. Chem. 2020, 85, 2406-2414.]. This method... Synthesizing tetrasubstituted glutarimide derivatives from acid-promoted 1,6-enyne substrates via allenol intermediates is challenging. This method offers a broader substrate range than previous single-product approaches, allowing the construction of compounds L1, L20, and L6 containing OTf groups. Furthermore, compound L6 can be further derivatized to yield compounds L21-L27 and all-carbon tetrasubstituted derivatives L8-L19.

[0232] Example 2: Evaluation of antitumor drug resistance activity of compounds L1-L27. This section will primarily focus on the evaluation of antitumor drug resistance activity and the application of compound L24 as a P-gp inhibitor to reverse drug resistance.

[0233] First, the activity of compounds L1-L27 in combination with PTX in the drug-resistant tumor cell line SW620 / AD300 (provided by Professor Liu Hongmin's group, School of Pharmacy, Zhengzhou University) was evaluated. The compounds were combined at a concentration of 20 μM with different concentrations (the concentrations are shown in Table 1 under "IC50"). 50 The drug-resistant tumor cell line SW620 / AD300 was treated with PTX (as shown in "of PTX(nM)"). After 72 hours, the cell survival rate was determined by CCK8 assay. The results showed that the combination of multiple compounds of the present invention with paclitaxel had good reversal activity against the drug-resistant tumor cell line SW620 / AD300.

[0234] Table 1 and Figure 2 IC shown 50 CCK-8 assay: SW620 / AD300 cells were seeded in 96-well microplates (5000 cells / well) and cultured overnight. After incubation with different concentrations of the compound for 24 hours, the culture medium was discarded, and 100 μL of fresh culture medium mixed with CCK-8 (100 μL / mL) was added. The 96-well microplates were then incubated in the dark for 2 hours. A450 nm was measured using a Varioskan LUX microplate spectrophotometer (Thermo Fisher, Waltham, MA). IC50 was calculated. 50 The data was computed using GraphPad Prism 8.0.2 software. Figure 2 The graph was created using the software GraphPad Prism 8.0.2.

[0235] Figure 1The results were obtained through a CCK-8 assay. Specifically, SW620 / AD300 cells were seeded in 96-well microplates (5000 cells / well) and cultured overnight. After incubation with compound L1-L27 at a concentration of 20 μmol / L for 24 h, the culture medium was discarded, and 100 μL of fresh culture medium mixed with CCK-8 (100 μL / mL) was added. The 96-well microplates were then incubated in the dark for 2 h. A450 nm was measured using a Varioskan LUX microplate spectrophotometer (Thermo Fisher, Waltham, MA). Graphs were plotted using GraphPad Prism 8.0.2 software.

[0236] Figure 3 The cells were obtained through a plate colony-forming assay. Specifically, SW620 / AD300 cells were seeded in 6-well plates (2000 cells / well) and cultured overnight. Cells were treated for 7 days with different concentrations (5, 10, 20 μmol / L) of compound L24, PTX alone (1 μmol / L), or PTX (1 μmol / L) combined with compound L24 (20 μmol / L). The culture medium was aspirated, and the cells were rinsed with PBS. Cells were then fixed with 4% paraformaldehyde (Biossci, Wuhan, China) for 30 minutes and stained with 0.1% (w / v) crystal violet staining solution (Biosharp, Anhui, China) for 30 minutes. Cells were washed with ultrapure water and air-dried.

[0237] Figure 4 The accumulation of rhodamine was determined using a specific assay: SW620 / AD300 cells were seeded in 6-well plates and cultured overnight. They were then treated with different concentrations (5, 10, 20 μmol / L) of compound L24 for 24 h, incubated with 5 μg / mL Rho 123 at 37°C for 30 min, and then washed twice with pre-chilled PBS. The cells were then resuspended in PBS and analyzed by flow cytometry.

[0238] Figure 5 The effect of compound L24 on human cytochrome P450 3A4 activity was investigated using an enzyme activity inhibition assay kit (Abcam, Shanghai, China). This assay utilized a non-fluorescent CYP3A4 substrate to convert it into a highly fluorescent metabolite detectable in the visible range (Ex / Em = 535 / 578 nm). Suitable reaction wells were prepared, containing the substrate, compound, and the potent inhibitor ketoconazole. The substrate mixture was then added to the wells, and fluorescence was measured at Ex / Em = 535 / 578 nm in kinetic mode for 40 min.

[0239] Figure 1The inhibition rate of glutarimide compound L1-L27 in combination with paclitaxel against drug-resistant tumor cell line SW620 / AD300; an inhibition rate greater than 50% indicates that the compound has the activity to reverse drug resistance.

[0240] Figure 2 To optimize the survival rate of drug-resistant tumor cell line SW620 / AD300 by combining compound L24 with PTX, the effects of compound L24 alone or in combination with PTX on the proliferation of SW620 / AD300 cell line were investigated using plate colony assay and cell cycle analysis.

[0241] Figure 4 To optimize the Rhodamine 123 accumulation assay of compound L24 with optimal activity, positive control verapamil and different concentrations (5, 10, 20 μM) of compound L24 were used. The higher the concentration of compound L24, the higher the proportion of Rhodamine 123 in SW620 / AD300 cells. This indicates that compound L24 inhibits the efflux of P-gp, and the inhibitory effect on P-gp efflux is significant at a concentration of 20 μM.

[0242] Figure 5 To select the compound with the best activity, L24 was evaluated for its effect on the CYP3A4 enzyme, a drug-metabolizing enzyme. Many reported P-gp inhibitors have been shown to affect the activity of the CYP3A4 enzyme, thus interfering with drug metabolism. Figure 5 Using verapamil and the widely used specific CYP3A4 inhibitor ketoconazole as positive controls, the results showed that compound L24 had a smaller effect on the CYP3A4 enzyme than verapamil, indicating that compound L24 is a safe P-gp inhibitor.

[0243] Based on this, the IC50 of the compound combined with PTX was calculated using the software GraphPad Prism 8.0.2 against drug-resistant colon cancer cells. 50 The results are shown in Table 1. The combination of compound L24 and PTX significantly enhanced the half-maximal inhibitory concentration (IC50) of drug-resistant colon cancer cells compared to the PTX-only group. For a more direct comparison, this application compared the IC50 values ​​of compound L24 combined with PTX and the PTX-only group. 50 Compare and plot as follows Figure 1 As shown. “IC” 50 "Inhibitor concentration" refers to the concentration of the inhibitor when "cell viability" is inhibited by half. This quantitative indicator measures the ability of a specific drug or other substance (inhibitor) to inhibit a specific biological or biochemical process. All experimental data are mean ± standard error, n = 6, with the solvent control group as the reference group (100% survival rate). RF (Reversal Fold) is a compound that can reverse the resistance fold of PTX in SW620 / Ad300 cells.

[0244] Table 1: 72-hour half-maximal inhibitory concentration (IC50) of compounds L1-L27 (20 μM), verapamil (VPM, 5 μM), and different concentrations of PTX against drug-resistant colon cancer cell line SW620 / AD300. 50 (nM)

[0245]

[0246] As shown in Table 1, “IC 50 The smaller the value of "of PTX(nM)", the better the anti-tumor effect against the drug-resistant colon cancer cell line SW620 / AD300; PTX alone (1μM) at the half-maximal inhibitory concentration (IC50) of the drug-resistant tumor cell line SW620 / AD300. 50 =4301±153nM, indicating that only PTX (1μM) had poor antitumor effect. The prepared compound L1-L27 (20μM) combined with different concentrations of paclitaxel showed improved antitumor effects, as listed in the table. The compound L1 (20μM) combined with PTX showed the following IC50: 50 =20.29±1.08 nM, which is 212 times the resistance reversal fold (RF) compared to PTX alone; similarly, in compounds L2-L27, the half-maximal inhibitory concentration (IC50) of compound L6 is 20.29±1.08 nM. 50 =161.40±7.06 nM, the resistance reversal fold was 26.6, indicating that compound L6 has weak resistance reversal activity. The half-maximal inhibitory concentration (IC50) of compounds L2, L4, L7-L9, L11, L21-L23, and L27 was 10 nM. <IC 50 <100 nM, the reversal fold of resistance of 52.9 to 276.2 times indicates that these compounds have moderate to good reversal activity against drug resistance. Compounds L3, L5, L12, L14, L15, L18, and L25 have a half-maximal inhibitory concentration (IC50) of >10 nM. 50 Between 5 nM, the reversal folds of drug resistance ranged from 533 to 851.7 times, indicating that these compounds possess excellent drug resistance reversal activity. The half-maximal inhibitory concentrations (IC50) for compounds L10, L13, L16, L17, L19, L20, L24, and L26 were also high. 50 The compounds have a resistance reversal fold of <5 nM and a resistance reversal fold of >1000 times, indicating that these compounds have good resistance reversal activity. Among them, compound L24 combined with PTX has the best anti-tumor effect, with a half-maximal inhibitory concentration of 1.38 nM and a resistance reversal fold of 3116.7 times.

[0247] Those skilled in the art will readily understand that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A glutarimide derivative, characterized in that, Its general structural formula is shown in Formula I, Formula II, or Formula III: In Equation I: R1 is benzyl, R2 is H, R3 is H, R4 is trifluoromethyl, and R5 is H; Alternatively, R1 is methyl, R2 is H, R3 is H, R4 is any one of phenyl, 4-methoxy-phenyl, 4-chloro-phenyl, and 4-fluoro-phenyl, and R5 is H; Alternatively, R1 is methyl, R2 is H, R3 is ethoxy, R4 is ethyl, and R5 is H; Alternatively, R1 = R2 = -(CH2CH2CH2)-, R3 is H, R4 is ethyl, and R5 is H; Alternatively, R1 is methyl, R2 is H, R3 is H, R4 is trifluoromethyl, and R5 is 2-(2-sec-butyl(benzoic acid)ethyl)-piperidin-1-carboxylic acid tert-butyl ester; In Formula II: R1 is benzyl, R2 is methyl, and R3 is any one of H, 3,5-dimethyl, and 2-chloro-5-methoxy. Alternatively, R1 is benzyl, R2 is benzyl, and R3 is H; In Formula III: R1 is benzyl, R2 is methyl, R3 is H, and R4 is any one of 4-methoxy-phenyl, 3,4-dimethoxy-phenyl, 3-methyl-4-methoxy-phenyl, methylenedioxyphenyl, 1,4-benzodioxane, 3,5-dimethyl-phenyl, phenylethynyl, 4-chloro-phenyl, 3,4,5-trimethoxy-phenyl, 3,5-dimethyl-4-methoxy-phenyl, 2-anthrayl, and benzofuranyl. Alternatively, R1 is 3,4-dimethoxybenzyl, R2 is methyl, R3 is H, and R4 is 3,4-dimethoxy-phenyl; Alternatively, R1 is benzyl, R2 = R3 = -(CH2CH2CH2CH2)-, and R4 is phenyl; Alternatively, R1 is benzyl, R2 is H, R3 is methyl, and R4 is phenyl.

2. The glutarimide derivative as described in claim 1, characterized in that, The glutarimide derivative is specifically compound L24, whose structural formula is as follows:

3. The method for preparing the glutarimide derivative as described in claim 1, characterized in that, When the glutarimide derivative is any one of compounds L1-L7 and L20, the preparation method is to prepare the compound of formula 1 in... The cyclization reaction of 1,6-enyne under acid-promoted conditions generates a glutarimide derivative. The synthetic route for its preparation is as follows: In this case, compound L1 corresponds to Bn for R1, H for R2, H for R3, CF3 for R4, and H for R5; For compound L2, R1 is Me, R2 is H, R3 is H, R4 is Ph, and R5 is H; For compound L3, R1 is Me, R2 is H, R3 is H, R4 is 4-F-Ph, and R5 is H; For compound L4, R1 is Me, R2 is H, R3 is H, R4 is 4-Cl-Ph, and R5 is H; For compound L5, R1 is Me, R2 is H, R3 is H, R4 is 4-OMe-Ph, and R5 is H; For compound L6, R1 = R2 = -(CH2CHCH2)-, R3 is H, R4 is Et, and R5 is H; For compound L7, R1 is Me, R2 is H, R3 is OEt, R4 is Et, and R5 is H; For compound L20, R1 is Me, R2 is H, R3 is H, R4 is CF3, and R5 is... When the glutarimide derivative is any one of compounds L8-L11, the preparation method is to prepare the compound of formula 2 in... The cyclization reaction of 1,6-enyne under acid-promoted conditions generates a glutarimide derivative. The synthetic route for its preparation is as follows: In compound L8, R1 is Bn, R2 is Me, and R3 is H; For compound L9, R1 is Bn, R2 is Bn, and R3 is H; For compound L10, R1 is Bn, R2 is Me, and R3 is 3,5-dimethyl. For compound L11, R1 is Bn, R2 is Me, and R3 is 2-chloro-4-methoxy. When the glutarimide derivative is any one of compounds L12-L19, the preparation method is to prepare the compound of formula 3 in... The cyclization reaction of 1,6-enyne under acid-promoted conditions generates a glutarimide derivative. The synthetic route for its preparation is as follows: In compound L12, R1 is Bn, R2 is Me, R3 is H, and R is 4-methoxy. For compound L13, R1 is Bn, R2 is Me, R3 is H, and R is 3,4-dimethoxy. For compound L14, R1 is Bn, R2 is H, R3 is Me, and R is 3,4-dimethoxy. For compound L15, R1 is Bn, R2 = R3 is -(CH2CH2CH2)-, and R is 3,4-dimethoxy. In compound L16, R1 is 3,4-dimethoxybenzyl, R2 is Me, R3 is H, and R is 3,4-dimethoxy. For compound L17, R1 is Bn, R2 is Me, R3 is H, and R is 3-methyl-4-methoxy. For compound L18, R1 is Bn, R2 is Me, R3 is H, and R is 3-(-OCH2O-). For compound L19, R1 is Bn, R2 is Me, R3 is H, and R is 3-(-OCH2CH2O)-. When the glutarimide derivative is any one of compounds L21-L27, the preparation method is to prepare the compound of formula 4 in... The cyclization reaction of 1,6-enyne under acid-promoted conditions generates an intermediate, which is then further coupled with a compound of formula 7 to generate a glutarimide derivative. The synthetic route for this preparation method is as follows: In this compound, R4 corresponding to compound L21 is 3,5-dimethylphenyl, and R is... The R4 corresponding to compound L22 is phenylacetylene, and R is H; The R4 of compound L23 is 4-chlorophenyl, and R is B(OH)2; The R4 corresponding to compound L24 is 3,4,5-trimethoxyphenyl, and R is B(OH)2; The R4 corresponding to compound L25 is 3,5-dimethyl-4-methoxyphenyl, and the R is B(OH)2; The R4 of compound L26 is 2-anthrayl, and R is B(OH)2; The R4 corresponding to compound L27 is benzofuranyl, and R is B(OH)2.

4. The method for preparing the glutarimide derivative as described in claim 1, characterized in that, When the glutarimide derivative is any one of compounds L1-L7 and L20, the preparation method is to dissolve the raw material with the structural formula shown in Formula 1 in an ultra-dry mixed solvent DCM / HFIP and add it to a dry reaction flask, then add the dehydrated raw material. Molecular sieve, added The acid was stirred at a pre-set temperature for 30 minutes to 3 hours. After the reaction was complete, the molecular sieve was removed by filtration through a Buchner funnel, the organic layer was enriched, and the crude product was obtained by evaporation and concentration. The product was then separated by silica gel column chromatography. The silica gel column was pre-treated with petroleum ether solvent containing 0.1% triethylamine. The raw material with the structural formula shown in Formula 1 and the... The molar ratio of acid to feed is 1:1-1.2; the mixed solvent DCM / HFIP is obtained by mixing DCM and HFIP; the preset temperature is 20℃-35℃. When the glutarimide derivative is any one of compounds L8-L11, the preparation method is to dissolve the raw material with the structural formula shown in Formula 2 in an ultra-dry mixed solvent DCM / HFIP and add it to a dry reaction flask, then add the dehydrated raw material. Molecular sieves were added to TfOH, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the molecular sieves were removed by filtration through a Buchner funnel, the organic layer was enriched, and the crude product was obtained by evaporation and concentration. The product was then separated by silica gel column chromatography. The molar ratio of the raw material with the structural formula shown in Formula 2 to the TfOH was 1:0.4-0.5, and the mixed solvent DCM / HFIP was obtained by mixing DCM and HFIP. When the glutarimide derivative is any one of compounds L12-L19, the preparation method is to dissolve the raw material with the structural formula shown in Formula 3 in the ultra-dry solvent HFIP and add it to a dry reaction flask, then add the dehydrated material. Molecular sieves were added to the raw material with the structural formula shown in Formula 5 and stirred at room temperature for 1 hour. After the reaction was completed, the molecular sieves were removed by filtration through a Buchner funnel, the organic layer was enriched, the crude product was obtained by evaporation and concentration, and the product was obtained by silica gel column chromatography. The molar ratio of the raw material of Formula 3, the TfOH and the raw material of Formula 5 was 1:(1-1.2):(3-5). When the glutarimide derivative is any one of compounds L21-L27, the preparation method is as follows: the raw material with the structural formula shown in Formula 6 is dissolved in a solvent and added to a dry reaction flask. A base, the raw material with the structural formula shown in Formula 7, and a catalyst are added. The mixture is stirred for 3-24 hours under argon atmosphere and at a pre-set temperature. After the reaction is complete, the mixture is extracted with ethyl acetate / water to enrich the organic layer. The crude product is obtained by evaporation and concentration, and the product is separated by silica gel column chromatography. The molar ratio of the raw material with the structural formula shown in Formula 6 to the raw material with the structural formula shown in Formula 7 is 1:1.2-1.

5. The catalyst is a palladium catalyst. The raw material with the structural formula shown in Formula 6 has a molar ratio of 1:0.04-0.05 to the cuprous iodide, and the raw material with the structural formula shown in Formula 6 has a molar ratio of 1:0.05-0.1 to the palladium catalyst. The solvent is pure THF or a THF / H2O mixed solvent. The base is one of potassium phosphate trihydrate, triethylamine, and sodium carbonate. The raw material with the structural formula shown in Formula 6 has a molar ratio of 1:1.5-2.5 to the base. The preset temperature is 40-75℃. The THF / H2O mixed solvent is obtained by mixing THF and H2O.

5. The use of the glutarimide derivative as described in claim 1 or 2 in the preparation of tumor drug resistance reversal agents.

6. The application as described in claim 5, characterized in that, The drug resistance reversal agent can exert an anti-tumor effect in tumor cells in combination with paclitaxel; Preferably, the tumor resistance reversal agent can reverse the resistance to paclitaxel induced in the colon cancer cell line SW620 / AD300.

7. The use of the glutarimide derivative as described in claim 2 in the preparation of P-gp inhibitors.

8. A drug resistance reversal agent, characterized in that, Including the glutarimide derivatives as described in claim 1 or 2.

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