Oxazolone matrine derivatives and preparation method and application thereof

Abstract

The invention discloses an oxazolone matrine derivative and a preparation method and application thereof, belonging to the technical field of chemical medicine. The oxazolone matrine derivative can be used to prepare anti-tumor drugs, with readily available raw materials, simple operation, simplified synthesis steps, mild reaction conditions, and high product yield. In vitro anti-tumor activity studies have shown that the compounds are promising anti-tumor drugs, and the compounds can be used in the treatment of clinical tumors.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical chemistry, and specifically relates to an oxazolone matrine derivative and a preparation method and application thereof. Background Art

[0002] The harm of cancer cannot be underestimated. All parts of the world are working hard to fight cancer and develop new anti-tumor drugs, and have made remarkable progress.

[0003] Matrine is a type of quinolizidine alkaloid isolated from plants such as Sophora flavescens and Sophora alopecuroides. Its chemical formula is C 15 H 24 N 2O , with a relative molecular mass of 248.58, and can be hydrolyzed and ring-opened to generate matrine under strong alkaline conditions. Matrine has multiple biological activities such as anti-tumor, anti-viral, neuroprotective, and insecticidal. Matrine has medicinal advantages, such as a special chemical skeleton, flexible structure, high solubility and good safety, making it a good precursor for structural modification. The current modifications include the modification of the C-13, C-14, and C-15 carbonyl groups of matrine, the opening of the D ring, the formation of ionic complexes, etc. The structure of matrine is as follows:

[0004]

[0005] In the field of medicinal chemistry, the family of heterocyclic compounds containing nitrogen, sulfur and oxygen plays an important role in five-membered and six-membered ring structures. Oxazolone is a five-membered heterocyclic compound containing oxygen and nitrogen. Substituted oxazole derivatives are associated with a variety of biological activities and important pharmacophores in drug development. Compounds containing the skeleton structure of oxazolone have broad-spectrum activities, such as anti-inflammatory, anticoagulant, and anti-tumor. In addition, studies have shown that the anti-tumor activity of matrine derivatives containing nitrogen, oxygen and sulfur elements will be improved to a certain extent. Summary of the invention

[0006] The purpose of the present invention is to overcome the deficiencies of the prior art and provide an oxazolone-type matrine derivative and a preparation method and application thereof, wherein the oxazolone-type matrine derivative has strong anti-tumor activity.

[0007] To achieve the above object, the present invention provides the following technical solutions:

[0008] One of the technical solutions of the present invention: an oxazolone-type matrine derivative having a structure as shown in general formula I, general formula II or general formula III:

[0009]

[0010] In the general formula Ⅰ, R 1=H, 9-fluorenylmethoxycarbonyl (Fmoc), p-tert-butylbenzenesulfonyl, tert-butyloxycarbonyl (Boc), R 2 =α-naphthyl ring, β-naphthyl ring, anthracene ring, 3-indolyl, N,N-dimethylphenyl, 3,5-dichlorophenyl, 2-chloroquinolyl;

[0011] In general formula II, R 1 =H, tert-butyloxycarbonyl, R 2 =N,N-dimethylphenyl, 3-indolyl;

[0012] In general formula III, R 1 =H, R 2 =α-naphthalene ring, anthracene ring.

[0013] The second technical solution of the present invention is a method for preparing the above-mentioned oxazolone matrine derivatives. When preparing the oxazolone matrine derivatives having the general formula I, II, and III structures, matrine is used as the starting material, the amide bond of the D ring of matrine is hydrolyzed to obtain potassium salt of matrine, and then reacted with MeOH / SOCl 2 The reaction yielded methyl matrine, which was then reacted with R 1 Cl, K 2 CO 3 After the reaction, the intermediate is acidified and then reacted with SOCl 2 The reaction obtains acyl chloride, which reacts with glycine in a 10% sodium hydroxide solution. The lone pair of electrons on the glycine amino group acts as a nucleophilic reagent to attack the carbonyl group of the acyl chloride to generate an alcohol acid intermediate. The intermediate group rearranges to generate N-acetylglycine when the chlorine is lost, and obtains oxazolone matrine derivatives having the general formula Ⅰ, Ⅱ, and Ⅲ. The chemical reaction formula is:

[0014]

[0015] The preparation method comprises the following steps:

[0016] 1) Mix matrine and potassium hydroxide solution, stir and reflux, cool to room temperature after the reaction, adjust to pH=7-8, filter and dry to obtain a white solid; dissolve the white solid in methanol, filter, concentrate the filtrate under reduced pressure and precipitate with acetone to obtain intermediate 1 potassium salt of matrine, which is directly used in the next reaction without purification; the structural formula of the intermediate 1 is as follows:

[0017] 2) The intermediate 1 obtained in step 1) and SOCl 2 Place in methanol, reflux under nitrogen protection, then remove the solvent under reduced pressure to obtain intermediate 2 methyl berberine; the structural formula of intermediate 2 is as follows:

[0018] 3) Intermediate 2 obtained in step 2) and K2 CO 3 Dissolve in anhydrous acetonitrile, add substituted acyl, sulfonyl or benzyl halide, and stir to react; filter after the reaction, concentrate the filtrate, extract, and dry the organic phase to obtain a residue; elute and purify the residue to obtain intermediate 3; the structural formula of intermediate 3 is as follows: Among them, R 1 =9-fluorenylmethoxycarbonyl, p-tert-butylbenzenesulfonyl or tert-butyloxycarbonyl;

[0019] 4) placing the intermediate 3 obtained in step 3) in an HCl solution, reflux, and cooling to room temperature after the reaction is completed, adjusting the pH to 5-6, and then extracting, drying the organic phase to obtain a residue; the residue is eluted and purified to obtain an intermediate 4; the structural formula of the intermediate 4 is as follows:

[0020] 5) The intermediate 4 obtained in step 4) is dissolved in dichloromethane, and dichlorothionyl and dimethylformamide are added. After the reaction is completed, the solvent is removed in vacuo to obtain the intermediate 5; the structural formula of the intermediate 5 is as follows:

[0021] 6) The intermediate 5 obtained in step 5) is added to a sodium hydroxide solution of glycine in portions, stirred, cooled, and acidified to obtain a white solid, which is washed, dried, and recrystallized to obtain an intermediate 6; the structural formula of the intermediate 6 is as follows:

[0022] 7) The intermediate 6 obtained in step 6) is mixed with aldehyde, anhydrous sodium acetate and acetic anhydride, refluxed, filtered, washed and recrystallized to obtain the target product 7, i.e., the oxazolone matrine derivative having the general formula I, II and III.

[0023] Furthermore, when n=1 in intermediate 6, the target product 7 is an oxazolone-type matrine derivative having the general structure of formula I; when n=2 in intermediate 6, the target product 7 is an oxazolone-type matrine derivative having the general structure of formula II; when n=3 in intermediate 6, the target product 7 is an oxazolone-type matrine derivative having the general structure of formula III.

[0024] Furthermore, in step 1), the mass fraction of the potassium hydroxide solution is 10%, the material-liquid ratio of matrine to potassium hydroxide solution is 10 mmol:40 mL; the stirring reflux temperature is 120° C., and the time is 8 h; and the pH is adjusted under ice bath conditions.

[0025] Further, in step 2), the intermediate 1, SOCl 2 The solid-liquid ratio of methanol is 10 mmol: 2.4 mL: 30 mL; the reflux time is 3 h.

[0026] Further, in step 3), the intermediate 2, K 2 CO 3 , anhydrous acetonitrile and acyl, sulfonyl or benzyl halide in a solid-liquid ratio of 10mmol:35mmol:30mL:10mmol; the extractant for the extraction is a mixture of ethyl acetate and water in a volume ratio of 2:1; the drying agent is Na 2 SO 4 ; The eluent for the elution is CH 2 Cl 2 and CH 3 OH are mixed in a volume ratio of 40:1; and the purification is performed by silica gel column chromatography.

[0027] Further, in step 4), the concentration of the HCl solution is 3 mol / L, the material-liquid ratio of the intermediate 3 and the HCl solution is 10 mmol: 20 mL; the reflux temperature is 70° C., and the time is 3 h; the extraction agent is a mixture of ethyl acetate and water in a volume ratio of 2: 1; the drying agent is Na 2 SO 4 ; The eluent for the elution is CH 2 Cl 2 and CH 3 OH are mixed in a volume ratio of 40:1; and the purification is performed by silica gel column chromatography.

[0028] Furthermore, in step 5), the solid-liquid ratio of the intermediate 4, dichloromethane, dichlorothionyl and dimethylformamide is 50 mmol: 50 mL: 75 mmol: 0.05 mmol; and the reaction time is 1 h.

[0029] Furthermore, in step 6), the preparation method of the sodium hydroxide solution of glycine is: dissolving 10 mmol of glycine in 30 mL of a 10% NaOH solution; the mass ratio of glycine to intermediate 5 is 1:2; the stirring time is 1 hour; and the recrystallization is recrystallization from ethanol.

[0030] Furthermore, in step 7), the solid-liquid ratio of the intermediate 6, aldehyde, anhydrous sodium acetate and acetic anhydride is 10mmol:20mmol:10mmol:30mmol; the reflux reaction time is 2h, and the temperature is 95-100°C; and the recrystallization is recrystallization from ethanol.

[0031] The third technical solution of the present invention: the use of the above-mentioned oxazolone matrine derivatives in the preparation of drugs for liver cancer, lung cancer, cervical cancer and breast cancer.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] The preparation method of the oxazolone matrine derivative of the present invention has the advantages of easy-to-obtain reaction raw materials, simple operation, mild reaction conditions and high product yield. Experiments show that the oxazolone matrine derivative of the present invention has a strong inhibitory effect on the proliferation of various cancer cells, and the compound can be used for the treatment of tumors in clinic. DETAILED DESCRIPTION

[0034] Now, various exemplary embodiments of the present invention are described in detail, and this detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics and embodiments of the present invention. It should be understood that the terms described in the present invention are only for describing specific embodiments and are not used to limit the present invention.

[0035] In addition, for the numerical range in the present invention, it is understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0036] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0037] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are exemplary only.

[0038] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0039] In the following examples, the room temperature is 25±2°C.

[0040] In the following examples, the design of the oxazolone matrine derivatives is as follows:

[0041]

[0042] Example 1

[0043] Preparation of target compound G1:

[0044]

[0045] 1) Place 10 mmol of matrine in a 250 mL flask, add 40 mL of 10% KOH solution, stir and reflux at 120 °C for 8 h; after the reaction is completed, cool to room temperature, and add 20% H 2 SO 4 Adjust to pH=7, vacuum filter, and dry to obtain a white solid; dissolve the white solid in methanol, filter, and concentrate the filtrate under reduced pressure and precipitate with acetone to obtain intermediate 1 potassium salt of berberine acid, which is directly used in the next reaction without purification;

[0046] 2) 10 mmol of the intermediate 1 obtained in step 1) and 2.4 mL of SOCl 2 Place in 30 mL of methanol, reflux under nitrogen protection for 3 h, then remove the solvent under reduced pressure to obtain intermediate 2 methyl berberine;

[0047] 3) 10 mmol of the intermediate 2 obtained in step 2) and 35 mmol of K 2 CO 3 Dissolve in 30 mL of anhydrous acetonitrile, add 10 mmol of fluorenylmethoxycarbonyl chloride, and stir at room temperature overnight; filter after TLC monitoring of the reaction completion, concentrate the filtrate, extract with ethyl acetate-water (ethyl acetate: water = 2: 1 (v: v)), and then add Na 2 SO 4 The ethyl acetate phase was dried to obtain a residue; the residue was purified by CH 2 Cl 2 :CH 3 The mixture was eluted with an eluent of OH=40:1 (v:v) and purified by silica gel column chromatography to obtain intermediate 3;

[0048] 4) 10 mmol of the intermediate 3 obtained in step 3) was placed in 20 mL of 3 mol / L HCl solution and refluxed at 70°C for 3 h; after the reaction was completed, the mixture was cooled to room temperature, 3 mol / L KOH solution was added dropwise to adjust the pH to 5, and then extracted with ethyl acetate-water (ethyl acetate: water = 2: 1 (v: v)), Na 2 SO 4 The ethyl acetate phase was dried to obtain a residue; the residue was purified by CH 2 Cl 2 :CH 3 The mixture was eluted with an eluent of OH=40:1 (v:v) and purified by silica gel column chromatography to obtain intermediate 4;

[0049] 5) 50 mmol of the intermediate 4 obtained in step 4) was dissolved in 50 mL of anhydrous DCM (dichloromethane), and 75 mmol 1.5 eq of dichlorothionyl and 0.05 mmol of dimethylformamide were slowly added, and the reaction was stirred at room temperature for 1 h, and the solvent was removed in vacuo to obtain the intermediate 5;

[0050] 6) dissolving 10 mmol of glycine in 30 mL of a 10% by mass NaOH solution to obtain a sodium hydroxide solution of glycine, and then mixing 20 mmol of the intermediate 5 obtained in step 5) with the sodium hydroxide solution of glycine in portions, and stirring continuously for 1 hour; then adding crushed ice to cool the solution and adding 15 mL of a 30% by mass hydrochloric acid to acidify, filtering to obtain a white solid, washing the white solid with ice water, drying, and recrystallizing from ethanol to obtain an intermediate 6;

[0051] 7) 10 mmol of the intermediate 6 obtained in step 6) was mixed with 20 mmol of α-naphthaldehyde, 10 mmol of anhydrous sodium acetate and 30 mmol of acetic anhydride, and refluxed at 98° C. for 2 h. The mixture was filtered, washed and recrystallized from ethanol to obtain the target product 7. The yield of the target product 7 (G1) was calculated to be 78%.

[0052] G1-Chemical Formula: C 43 H 43 N 3 O 4

[0053] Light yellow powder, yield 65%, 1 H NMR (500 MHz, CDCl 3 )δ8.19(dd,1H),8.03(s,1H),7.97(tt,1H),7.94–7.86(m,5H),7.71(ddd,1H),7.64(dd, 2H),7.45(ddd,1H),7.34(ddd,2H),7.24(ddd,2H),5.00(d,2H),4.02(dd,1H),3.63(t,1H ),3.42(m,J=33.0Hz,2H),2.59(m,J=17.7Hz,2H),2.32(t,2H),2.18(m,J=17.2Hz,3H),2 .04(t,1H),1.82(m,J=5.9Hz,2H),1.67–1.52(m,6H),1.42(m,J=8.2Hz,4H),1.27(m,1H). 13 C NMR (126 MHz, CDCl 3)δ174.13(s),169.48(s),155.79(s),154.55(s),147.49(s),145.84(s),144.35(d,J=17.5Hz),143.32(s),141.53(d,J=15.0Hz) ,140.11(s),139.41(s),139.03(s),137.99(s),137.25(s),134.71(s),134.12(s),129.08(s),128.73(s),127.16–126.98(s),12 4.30(s),120.99(s),120.32(s),119.72(s),117.76(s),107.78(s),63.65(d,J=26.7Hz),57.20(d,J=8.6Hz),53.15(s),43.18(s) ),41.43(s),35.31(s),32.79(s),27.71(s),27.11(s),26.40(s),21.06(d,J=14.2Hz),20.70(s),18.93(s).m / z:665.3912(M+1).

[0054] Compounds G2 to G15 were prepared by referring to the preparation method of compound G1. The structural identification data of G2 to G15 are as follows:

[0055] G2-Chemical Formula: C 43 H 43 N 3 O 4

[0056] Reddish brown powder, yield 66%. 1 H NMR (500 MHz, CDCl 3)δ7.88(dd,2H),7.84–7.80(m,3H),7.76(m,J=10.0Hz,2H),7.67(dd,2H),7.65(s,1H),7.49(dd,1H),7.43(ddd,1H),7.32(ddd,2H),7.22(ddd,2H),4.79(d,2H),4.10(dd,1H),3.54(t,1H),3.31(dd,J=18.0Hz,2H),2.59(m,J=15.9Hz,2H),2.32(m,2H),2.27–2.09(m,3H),2.03(t,1H),1.83(m,J=23.5Hz,2H),1.68–1.57(m,5H),1.53(m,3H),1.41(m,J=11.0Hz,2H),1.31(m,1H). 13 C NMR(126MHz,CDCl 3 )δ176.15(s),174.30(s),167.71(s),154.82(s),132.27(s),130.91(s),128.81(s),65.56(s),55.21(s),43.30(s),42.92(s),35.34(s),30.54(s),29.63(d,J=9.5Hz),25.55(s),25.02(s),22.21(s),19.16(s),14.72(s),13.71(s).δ167.79(s),167.35(s),157.50(s),144.14(s),139.48(s),135.23(s),134.90(d,J=5.7Hz),131.00(s),129.77(d,J=12.3Hz),127.81(s),127.63(s),127.36(s),126.65(d,J=4.1Hz),126.28(s),125.95(s),125.14(s),121.89(s),120.96(s),67.50(s),64.79(s),55.58(s),53.09(s),48.11(s),46.35(s),40.68(s),36.68(s),30.86(s),28.37(d,J=6.9Hz),26.39(s),23.42(s),23.23(s),21.39(s).m / z:665.4532(M+1)。

[0057] G3-Chemical Formula:C 47 H 45 N 3O 4

[0058] Pale yellow powder, yield 58%. 1 H NMR (500 MHz, CDCl 3 )δ8.64(t,1H),8.32(m,2H),8.20(s,1H),8.00(m,2H),7.90(dd,2H),7.47(m,J=15.0Hz,4H), 7.34(ddd,2H),7.24(ddd,2H),7.11(dd,2H),5.06(d,2H),3.84(dd,1H),3.70(q,1H),3.34–3 .13(m,2H),2.64(m,J=18.1Hz,2H),2.32(t,2H),2.28–2.11(m,3H),2.06(q,J=16.4Hz,2H),1 .85(m,J=57.1Hz,2H),1.62(m,2H),1.53(m,J=1.2Hz,4H),1.43(m,2H),1.37(m,J=9.8Hz,2H). 13 C NMR (126 MHz, CDCl 3 )δ176.15(s),174.30(s),167.71(s),154.82(s),132.27(s),130.91(s),128.8 1(s),65.56(s),55.21(s),43.30(s),42.92(s),35.34(s),30.54(s),29.63(d, J=9.5Hz),25.55(s),25.02(s),22.21(s),19.16(s),14.72(s),13.71(s).δ170 .10(s),167.35(s),157.50(s),144.14(s),139.48(s),132.79(d,J=14.7Hz),1 29.20(s),128.35(d,J=9.3Hz),127.63(s),126.86(s),126.28(s),125.90(s), 125.69(s),125.14(s),120.96(s),120.42(s),119.38(s),67.50(s),64.79(s) ,55.58(s),53.09(s),48.11(s),46.35(s),40.68(s),36.68(s),30.86(s),28. 37(d,J=6.9Hz),26.39(s),23.42(s),23.23(s),21.39(s).m / z:715.3341(M+1).

[0059] G4-Chemical Formula: C 39 H 39 Cl 2 N 3 O 4

[0060] Pale yellow powder, yield 70%. 1 H NMR (500 MHz, CDCl 3 )δ7.89(dd,2H),7.71(t,1H),7.45(d,2H),7.40(ddd,2H),7.34(dd,2H),7.24( ddd,2H),7.04(s,1H),5.10(d,2H),4.40(dd,1H),3.91(t,1H),3.29(dd,1H),2 .95(q,1H),2.64(m,J=2.2Hz,2H),2.39–2.16(m,5H),2.04(t,1H),1.89(m,J=4 5.5Hz,2H),1.76–1.59(m,5H),1.48(m,J=50.0Hz,4H),1.38(m,J=35.5Hz,2H). 13 C NMR (126 MHz, CDCl 3 )δ176.15(s),174.30(s),167.71(s),154.82(s),132.27(s),130.91(s), 128.81(s),65.56(s),55.21(s),43.30(s),42.92(s),35.34(s),30.54(s) ,29.63(d,J=9.5Hz),25.55(s),25.02(s),22.21(s),19.16(s),14.72(s) ,13.71(s).δ167.79(s),167.35(s),157.50(s),144.14(s),139.48(s),13 6.08(s),134.84(s),134.00(s),127.85(s),127.63(s),126.49(s),126. 28(s),125.14(s),122.00(s),120.96(s),67.50(s),64.79(s),55.58(s), 53.09(s),48.11(s),46.35(s),40.68(s),36.68(s),30.86(s),28.37(d, J=6.9Hz),26.39(s),23.42(s),23.23(s),21.39(s).m / z:683.4344(M+1).

[0061] G5-Chemical Formula: C 42 H 43 C1N 4 O 4

[0062] Pale yellow powder, yield 73%. 1 H NMR(500MHz,DMSO)δ7.88(dd,2H),7.56(ddd,2H),7.32(ddt,J=3.5Hz,3H),7.26–7.21(m,4H), 7.19(m,1H),7.04(m,1H),5.25(d,2H),4.43(dd,1H),4.05(q,1H),3.80(t,1H),3.37(dd,1H), 2.91(q,1H),2.63(m,J=7.0Hz,2H),2.46(dd,1H),2.36–2.18(m,5H),2.08(m,J=48.6Hz,2H),1 .89(m,J=30.7Hz,2H),1.66(m,J=4.4Hz,4H),1.53(m,2H),1.43(m,J=3.0Hz,3H),1.33(m,2H). 13 C NMR(125MHz,DMSO)δ173.10(s),166.62(s),157.50(s),144.14(s),140.01(s),139.48(s),136.87(s),12 9.04(s),128.64(s),127.63(s),126.89(s),126.28(s),125.70(s),125.14(s),123.14(s),121.69(s),1 20.96(s),67.50(s),64.79(s),55.58(s),53.09(s),48.11(s),46.35(s),40.74(d,J=16.0Hz),36.68(s) ,36.02(s),30.86(s),28.37(d,J=6.9Hz),26.39(s),23.42(s),23.23(s),21.39(s).m / z:702.8647(M+1).

[0063] G6-Chemical Formula: C 41 H 46 N 4 O 4

[0064] Reddish brown oil, yield 74%. 1H NMR(500MHz,CDCl 3 )δ7.91(m,J=10.0Hz,4H),7.57(dd,2H),7.34(ddd,2H),7.27(s,1H),7.24dd,2H),6.99(tt,2H),5.43(d,2H),4.00(t,1H),3.83(m,1H),3.51(dd,1H),3.20(m,1H),3.02(s,6H),2.62(m,J=3.9Hz,2H),2.37–2.14(m,4H),2.06(m,J=24.5Hz,2H),1.85(m,J=47.1Hz,2H),1.63(m,J=12.7Hz,5H),1.48(m,J=50.0Hz,4H),1.35(m,J=28.4Hz,2H). 13 C NMR(126MHz,CDCl 3 )δ176.15(s),174.30(s),167.71(s),154.82(s),132.27(s),130.91(s),128.81(s),65.56(s),55.21(s),43.30(s),42.92(s),35.34(s),30.54(s),29.63(d,J=9.5Hz),25.55(s),25.02(s),22.21(s),19.16(s),14.72(s),13.71(s).δ167.79(s),167.35(s),157.50(s),152.56(s),144.14(s),139.48(s),134.16(s),129.62(s),127.63(s),126.28(s),125.14(s),122.86(d,J=4.8Hz),120.96(s),113.39(s),67.50(s),64.79(s),55.58(s),53.09(s),48.11(s),46.35(s),41.92(s),40.68(s),36.68(s),30.86(s),28.37(d,J=6.9Hz),26.39(s),23.42(s),23.23(s),21.39(s).m / z:658.2937(M+1)。

[0065] G7-Chemical Formula:C 33 H 41 N 3 O 4

[0066] White powder, yield 66%. 1 H NMR (500 MHz, CDCl 3 ) δ 8.18 (dd, 1H), 7.96 (tt, 1H), 7.90 (m, J = 6.0, 4.0 Hz, 4H), 7.70 (ddd, 1H), 7.44 (ddd, 1H),, 4.02 (dd, 1H), 3.32 (m, J = 4.8 Hz, 2H), 2.62 (m, J = 3.6 Hz, 2H), 2.32 (q, 2H), 2.23 (m, J = 0.7 Hz, 2H), 2.07 (m, J = 34.4 Hz, 2H), 1.85 (m, J = 33.1 Hz, 2H), 1.66 (m, 1H), 1.61 (m, J = 7.0 Hz, 4H), 1.53 (t, 2H), 1.50–1.44 (m, 2H), 1.42 (s, 9H), 1.36 (m, J = 32.7 Hz, 2H). 13 C NMR (126 MHz, CDCl 3 ) δ 176.15 (s), 174.30 (s), 167.71 (s), 154.82 (s), 132.27 (s), 130.91 (s), 128.81 (s), 65.56 (s), 55.21 (s), 43.30 (s), 42.92 (s), 35.34 (s), 30.54 (s), 29.63 (d, J = 9.5 Hz), 25.55 (s), 25.02 (s), 22.21 (s), 19.16 (s), 14.72 (s), 13.71 (s). δ 167.79 (s), 167.35 (s), 157.27 (s), 135.12 (d, J = 16.3 Hz), 133.47 (s), 130.39 (s), 128.84 (s), 128.44 (s), 127.83 (s), 126.77 (d, J = 19.9 Hz), 126.35 (s), 126.09 (s), 122.46 (s), 81.20 (s), 64.79 (s), 55.58 (s), 53.09 (s), 46.35 (s), 40.68 (s), 36.68 (s), 30.86 (s), 28.36 (t, J = 4.0 Hz), 26.39 (s), 23.42 (s), 23.23 (s), 21.39 (s). m / z: 543.2743 (M+1).

[0067] G8 - Chemical Formula: C 26 H 32 N 4 O 2

[0068] Reddish brown oil, yield 69%. 1 H NMR (500 MHz, CDCl 3 )δ8.10(s,1H),7.78(s,1H),7.65(dd,1H),7.62(s,1H),7.19(m,1H),7.11 (d,1H),7.08(q,1H),3.05(dd,1H),2.60(m,J=24.1Hz,3H),2.32(t,2H),2 .23(m,J=10.0Hz,2H),2.09(m,1H),2.04(d,1H),1.92–1.73(m,3H),1.72– 1.50(m,6H),1.49(s,1H),1.46(m,J=25.4Hz,3H),1.37(m,J=12.8Hz,2H). 13 CNMR (126MHz, CDCl 3 )δ176.15(s),174.30(s),167.71(s),154.82(s),132.27(s),130.91(s),128.81(s),65.56(s),55.21(s),43.30(s),42.92(s),35.3 4(s),30.54(s),29.63(d,J=9.5Hz),25.55(s),25.02(s),22.21(s),19.16(s),14.72(s),13.71(s).δ173.50(s),167.84(s),135.68( s),132.78(s),129.22(s),127.16(s),122.80(s),120.35(d,J=9.1Hz),119.96(s),112.56(s),107.91(s),65.58(s),57.82(s),53.0 9(s),48.20(s),41.73(s),35.91(s),32.21(s),28.41(d,J=16.6Hz),26.47(s),23.42(s),23.23(s),20.44(s).m / z:432.2810(M+1).

[0069] G9-Chemical Formula: C 28 H 33 N 3 O 2

[0070] Pale yellow oil, yield 79%. 1 H NMR (500 MHz, CDCl 3)δ8.19(dd,1H),7.97(tt,1H),7.94(s,1H),7.92(m,3H),7.90(ddd,J=5.0Hz,1H),7.71(ddd,1H),2.84(dd,1H),2.58(m,J=10.5Hz,2H),2.39(t ,1H),2.32(q,2H),2.27–2.12(m,3H),2.04(t,1H),1.72(m,J=16.8Hz,3 H),1.51(m,J=12.3Hz,6H),1.45–1.32(m,4H),1.31(s,1H),1.18(m,1H). 13 C NMR (126 MHz, CDCl 3 )δ176.15(s),174.30(s),167.71(s),154.82(s),132.27(s),130.91 (s),128.81(s),65.56(s),55.21(s),43.30(s),42.92(s),35.34(s), 30.54(s),29.63(d,J=9.5Hz),25.55(s),25.02(s),22.21(s),19.16( s),14.72(s),13.71(s).δ170.10(s),167.35(s),135.12(d,J=16.3Hz ),133.47(s),130.39(s),128.84(s),128.44(s),127.83(s),126.77( d,J=19.9Hz),126.35(s),126.09(s),122.46(s),65.58(s),57.82(s) ,53.09(s),48.20(s),41.73(s),35.91(s),32.21(s),28.41(d,J=16. 6Hz),26.47(s),23.42(s),23.23(s),20.44(s).m / z:443.2460(M+1).

[0071] G10-Chemical Formula: C 38 H 45 N 3 O 4 S

[0072] Pale yellow oil, yield 72%. 1 H NMR (500 MHz, CDCl 3)δ8.19(dd,1H),7.99(s,1H),7.97(t,1H),7.94–7.87(m,3H),7.71(dd,1H), 7.63(m,J=15.0Hz,4H),7.45(ddd,1H),3.29(dd,1H),2.91(dd,1H),2.52(m, J=14.6Hz,3H),2.37(m,J=43.8Hz,4H),1.82(m,J=40.9Hz,2H),1.69(m,2H), 1.63–1.52(m,4H),1.52–1.42(m,5H),1.35(m,1H),1.33(s,9H),1.30(m,1H). 13 C NMR (126 MHz, CDCl 3 )δ176.15(s),174.30(s),167.71(s),154.82(s),132.27(s),130.91(s),12 8.81(s),65.56(s),55.21(s),43.30(s),42.92(s),35.34(s),30.54(s),29 .63(d,J=9.5Hz),25.55(s),25.02(s),22.21(s),19.16(s),14.72(s),13.7 1(s).δ167.79(s),167.35(s),154.65(s),135.37–134.95(m),133.47(s),1 30.39(s),128.85(d,J=2.5Hz),128.41(d,J=6.3Hz),127.83(s),126.77(d, J=19.9Hz),126.35(s),126.09(s),122.46(s),64.79(s),60.40(s),53.09( s),49.16(s),40.89(s),35.99(s),34.94(s),31.36(s),29.04(s),28.37(d ,J=6.9Hz),26.39(s),23.42(s),23.23(s),21.39(s).m / z:639.4935(M+1).

[0073] G11-Chemical Formula: C 36 H 44 N 4 O 4 S

[0074] Pale yellow oil, yield 69%. 1 H NMR (500 MHz, CDCl 3)δ8.25(s,1H),7.89(s,1H),7.78(s,1H),7.64(m,J=15.0Hz,5H),7.19(ddd,1H),7.10(ddd,J=15.0Hz,2H),3.89(dd,1H),3.33(dd,1H),3.08(q,1H),2.55(m,J=1.2Hz,2H),2.32(t,2H),2.17(m,J=2.7Hz,2H),1.80(m,J=6.4Hz,2H),1.67(t,J=7.0Hz,1H),1.57(m,2H),1.51(m,J=19.3Hz,4H),1.43(m,3H),1.38(m,2H),1.33(d,J=2.5Hz,9H),1.32–1.25(m,1H). 13 C NMR(126MHz,CDCl 3 )δ176.15(s),174.30(s),167.71(s),154.82(s),132.27(s),130.91(s),128.81(s),65.56(s),55.21(s),43.30(s),42.92(s),35.34(s),30.54(s),29.63(d,J=9.5Hz),25.55(s),25.02(s),22.21(s),19.16(s),14.72(s),13.71(s).δ173.10(s),167.84(s),154.65(s),135.68(s),135.26(s),132.78(s),129.22(s),128.86(s),128.39(s),127.16(s),122.80(s),120.35(d,J=9.1Hz),119.96(s),112.56(s),107.91(s),64.79(s),60.40(s),53.09(s),49.16(s),40.89(s),35.99(s),34.94(s),31.36(s),29.04(s),28.37(d,J=6.9Hz),26.39(s),23.42(s),23.23(s),21.39(s).m / z:628.2868(M+1).

[0075] G12-Chemical Formula:C 32 H 46 N 4 O 4

[0076] White powder, yield 77%.1 H NMR (500 MHz, CDCl 3 )δ7.63(d,2H),6.78(d,2H),5.64(t,1H),4.27(dd,1H),3.20(tt,J=14.9Hz,3H),3.02(s,6H),2.77(q,1H),2.62(m,J=1.8Hz,2H),2.34 –2.20(m,4H),2.09(m,J=50.7Hz,2H),1.87(m,J=52.5Hz,2H),1.64(m,J=10.4Hz,5H),1.53(m,4H),1.42(s,9H),1.35(m,J=38.0Hz,2H). 13 C NMR (126 MHz, CDCl 3 )δ176.15(s),174.30(s),167.71(s),154.82(s),132.27(s),130.91(s),128.81(s),65.56(s),55.21(s),43.30(s),42.92(s),35.34( s),30.54(s),29.63(d,J=9.5Hz),25.55(s),25.02(s),22.21(s),19.16(s),14.72(s),13.71(s).δ168.80(s),164.68(s),157.27(s),1 50.44(s),137.80(s),130.00(s),123.11(s),117.40(s),112.85(s),81.20(s),64.79(s),55.58(s),53.09(s),46.35(s),41.92(s),4 0.68(s),36.68(s),35.49(s),34.17(s),30.86(s),28.37(d,J=7.9Hz),26.39(s),23.42(s),23.23(s),21.39(s).m / z:550.3976(M+1).

[0077] G13-Chemical Formula: C 27 H 34 N 4 O 2

[0078] Pale yellow oil, yield 65%. 1 H NMR (500 MHz, CDCl 3)δ8.39(s,1H),7.97(tt,1H),7.59(dd,1H),7.09(ddd,1H),6.97(ddd,1H ),6.46(d,1H),6.21(t,1H),3.53(s,2H),3.01(dd,1H),2.67–2.39(m,3H) ,2.33–2.14(m,5H),2.04(t,1H),1.92–1.67(m,4H),1.66(s,J=0.6Hz,1H ),1.54(m,J=12.6Hz,3H),1.45(m,J=25.5Hz,5H),1.31(m,J=53.7Hz,2H). 13 CNMR (126MHz, CDCl 3 )δ176.15(s),174.30(s),167.71(s),154.82(s),132.27(s),130.91(s),128.81(s),65.56(s),55.21(s),43.30(s),42.92(s),35.34(s) ),30.54(s),29.63(d,J=9.5Hz),25.55(s),25.02(s),22.21(s),19.16(s),14.72(s),13.71(s).δ168.80(s),162.49(s),138.07(s),13 6.55(s),132.45(s),127.64(s),121.59(d,J=1.5Hz),120.36(s),111.82(s),110.22(s),105.60(s),65.58(s),57.82(s),53.09(s),48 .20(s),41.73(s),35.91(s),34.72(s),34.17(s),32.21(s),28.48(s),26.47(s),23.42(s),23.23(s),20.44(s).m / z:446.2595(M+1).

[0079] G14-Chemical Formula: C 34 H 39 N 3 O 2

[0080] Pale yellow oil, yield 71%. 1 H NMR (500 MHz, CDCl 3)δ8.35(t,1H),8.28(m,2H),7.96(m,2H),7.44(m,J=14.9Hz,4H),6.75(t,1H),2.91(dd,1H),2.73–2.54(m,4H),2.48–2.33(m ,5H),2.23(m,J=3.7Hz,2H),2.03(t,1H),1.85–1.61(m,3H),1.58–1.42(m,6H),1.41–1.21(m,5H),1.13(s,1H),1.11(m,1H). 13 C NMR (126 MHz, CDCl 3 )δ176.15(s),174.30(s),167.71(s),154.82(s),132.27(s),130.91(s),128.81(s),65.56(s),55.21(s),43.30(s),42.92(s),35.34(s) ),30.54(s),29.63(d,J=9.5Hz),25.55(s),25.02(s),22.21(s),19.16(s),14.72(s),13.71(s).δ172.80(s),158.51(s),132.73(s),13 1.32(s),129.20(s),128.39(s),126.86(s),125.90(s),125.69(s),120.96(s),118.96(s),65.58(s),57.82(s),53.09(s),48.20(s),4 1.73(s),35.91(s),33.59(s),32.72(s),32.21(s),28.68(s),28.48 (s),26.47(s),23.42(s),23.23(s),20.44(s).m / z:521.3733(M+1).

[0081] G15-Chemical Formula: C 30 H 37 N 3 O 2

[0082] Yellow-brown oil, yield 71%. 1 H NMR (500 MHz, CDCl 3)δ7.92(m,1H),7.90(m,J=5.0Hz,2H),7.75–7.65(m,3H),7.45(ddd,1H),6.34(t,1H),2.98(dd,1H),2.55(q,J=38.3Hz,3H),2.45 (m,4H),2.43–2.36(m,4H),2.04(s,1H),1.84–1.65(m,3H),1.53(m,J=0.9Hz,4H),1.48–1.34(m,6H),1.20-109(m,J=1.0Hz,3H). 13 CNMR (126MHz, CDCl 3 )δ176.15(s),174.30(s),167.71(s),154.82(s),132.27(s),130.91(s) ),128.81(s),65.56(s),55.21(s),43.30(s),42.92(s),35.34(s),30. 54(s),29.63(d,J=9.5Hz),25.55(s),25.02(s),22.21(s),19.16(s),1 4.72(s),13.71(s).δ172.80(s),158.51(s),135.42(s),135.18(s),13 3.47(s),129.20(s),128.84(s),127.83(s),126.78(s),126.35(s),12 6.09(s),125.05(s),123.32(s),65.58(s),57.82(s),53.09(s),48.20 (s),41.73(s),35.91(s),33.59(s),32.72(s),32.21(s),28.68(s),28 .48(s),26.47(s),23.42(s),23.23(s),20.44(s).m / z:471.1596(M+1).

[0083] Table 1 shows the structural formulas of oxazolone-type matrine derivatives.

[0084] Table 1 Structural formula of oxazolone matrine derivatives

[0085]

[0086]

[0087]

[0088] Example 2

[0089] The in vitro antitumor activity of the prepared oxazolone matrine derivatives was studied. The tumor cells were liver cancer cells HepG2, lung cancer cells A549, cervical cancer cells Hela, and breast cancer cells MCF-7, all of which were purchased.

[0090] Experimental method: Take cancer cell lines, revive cells and perform subculture. After 24 hours, grow to the logarithmic growth phase, digest cells with trypsin and prepare cell suspension. Inoculate 100 μL per well into a 96-well plate, set up a blank group, a control group and a sample group, and set up 3 parallel wells in each group. Place the 96-well cell culture plate at 37°C and 5% CO 2 Incubate for 24 hours, aspirate the culture medium, add different gradient compounds (dissolve the oxazolone matrine derivatives in dimethyl sulfoxide (DMSO) and dilute them with culture medium to the required solution concentration) for 48 hours. Add 10uL, 5mg / mL MTT to each well, incubate in the incubator for 4 hours, discard the supernatant, add 100uLDMSO, shake and mix, test the absorbance of each well at 490nm with a microplate reader, and calculate the cell inhibition rate.

[0091]

[0092] The above experiment was repeated 3 times, and the IC was calculated by Blies method. 50 value.

[0093] Experimental results:

[0094] Table 2 Inhibition of proliferation of HepG2 cells, A549 cells, Hela cells and MCF-7 cells by target compounds

[0095]

[0096]

[0097] Experimental data show that most of the synthesized series of matrine derivatives have good anti-tumor activity. When matrine is used as a control drug and the synthetic target compounds are administered, their inhibition rates are compared and it is found that G4, G5, G6, and G11 in the compounds have a strong inhibitory effect on tumor cells and have high anti-tumor inhibitory activity.

[0098] The above are only preferred specific implementations of the present application, but the protection scope of the present application is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed in the present application should be included in the protection scope of the present application. Therefore, the protection scope of the present application should be based on the protection scope of the claims.

Claims

1. An oxazolone-type matrine derivative, characterized in that: Having a structure as shown in general formula Ⅰ, general formula Ⅱ or general formula Ⅲ: In the general formula I, R1 = H, 9-fluorenylmethoxycarbonyl, p-tert-butylbenzenesulfonyl, tert-butyloxycarbonyl, R2 = α-naphthyl ring, β-naphthyl ring, anthracene ring, 3-indolyl, N,N-dimethylphenyl, 3,5-dichlorophenyl; In the general formula II, R1=H, tert-butyloxycarbonyl, R2=N,N-dimethylphenyl, 3-indolyl; In the general formula III, R1 = H, R2 = α-naphthalene ring, anthracene ring.

2. A method for preparing the oxazolone matrine derivative according to claim 1, characterized in that: The oxazolone matrine derivative has a structure of general formula I and R1 is not H, comprising the following steps: 1) Mix matrine and potassium hydroxide solution, stir and reflux, cool to room temperature after the reaction, adjust to pH=7-8, filter and dry to obtain a white solid; dissolve the white solid in methanol, filter, concentrate the filtrate under reduced pressure and precipitate with acetone to obtain intermediate 1 potassium salt of matrine, which is directly used in the next reaction without purification; the structural formula of the intermediate 1 is as follows: 2) The intermediate 1 obtained in step 1) and SOCl2 are placed in methanol, refluxed under nitrogen protection, and then the solvent is removed under reduced pressure to obtain the intermediate 2 methyl berberine; the structural formula of the intermediate 2 is as follows: 3) The intermediate 2 and K2CO3 are dissolved in anhydrous acetonitrile, and a substituted acyl or sulfonyl group is added and stirred for reaction; after the reaction is completed, the mixture is filtered, the filtrate is concentrated, extracted, and the organic phase is dried to obtain a residue; the residue is eluted and purified to obtain an intermediate 3; the structural formula of the intermediate 3 is as follows: Wherein, R1=9-fluorenylmethoxycarbonyl, p-tert-butylbenzenesulfonyl or tert-butyloxycarbonyl; 4) placing the intermediate 3 in an HCl solution, reflux, and cooling to room temperature after the reaction is completed, adjusting the pH to 5-6, and then extracting, drying the organic phase to obtain a residue; the residue is eluted and purified to obtain an intermediate 4; the structural formula of the intermediate 4 is as follows: 5) The intermediate 4 is dissolved in dichloromethane, and dichlorothionyl and dimethylformamide are added. After the reaction is completed, the solvent is removed in vacuo to obtain the intermediate 5; the structural formula of the intermediate 5 is as follows: 6) The intermediate 5 is added to a sodium hydroxide solution of glycine in portions, stirred, cooled, and acidified to obtain a white solid, which is washed, dried, and recrystallized to obtain an intermediate 6; the structural formula of the intermediate 6 is as follows: Wherein, n=1; 7) The intermediate 6 is mixed with aldehyde, anhydrous sodium acetate and acetic anhydride, refluxed, filtered, washed and recrystallized to obtain the target product 7, i.e., the oxazolone matrine derivative having the structure of general formula I and R1 not being H.

3. The preparation method according to claim 2, characterized in that: In step 1), the mass fraction of the potassium hydroxide solution is 10%, the material-liquid ratio of matrine to potassium hydroxide solution is 10 mmol:40 mL; the stirring reflux temperature is 120° C., and the time is 8 h; and the pH is adjusted in an ice bath.

4. The preparation method according to claim 2, characterized in that: In step 2), the material-liquid ratio of the intermediate 1, SOCl2 and methanol is 10 mmol: 2.4 mL: 30 mL; and the reflux time is 3 h.

5. The preparation method according to claim 2, characterized in that: In step 3), the solid-liquid ratio of the intermediate 2, K2CO3, anhydrous acetonitrile and substituted acyl or sulfonyl is 10mmol:35mmol:30mL:10mmol; the extraction solvent is a mixture of ethyl acetate and water in a volume ratio of 2:1; the drying desiccant is Na2SO4; the elution eluent is a mixture of CH2Cl2 and CH3OH in a volume ratio of 40:1; and the purification is silica gel column chromatography purification.

6. The preparation method according to claim 2, characterized in that: In step 4), the concentration of the HCl solution is 3 mol / L, the solid-liquid ratio of the intermediate 3 and the HCl solution is 10 mmol: 20 mL; the reflux temperature is 70°C and the time is 3 h; the extraction agent is a mixture of ethyl acetate and water in a volume ratio of 2: 1; the drying desiccant is Na2SO4; the elution eluent is a mixture of CH2Cl2 and CH3OH in a volume ratio of 40: 1; and the purification is silica gel column chromatography purification.

7. The preparation method according to claim 2, characterized in that: In step 5), the material-liquid ratio of the intermediate 4, dichloromethane, dichlorothionyl and dimethylformamide is 50 mmol: 50 mL: 75 mmol: 0.05 mmol; and the reaction time is 1 h.

8. The preparation method according to claim 2, characterized in that: In step 6), the preparation method of the sodium hydroxide solution of glycine is: dissolving 10 mmol of glycine in 30 mL of a 10% NaOH solution; the mass ratio of glycine to intermediate 5 is 1:2; the stirring time is 1 hour; and the recrystallization is recrystallization from ethanol.

9. The preparation method according to claim 2, characterized in that: In step 7), the solid-liquid ratio of the intermediate 6, aldehyde, anhydrous sodium acetate and acetic anhydride is 10mmol:20mmol:10mmol:30mmol; the reflux reaction time is 2h, and the temperature is 95-100°C; the recrystallization is recrystallization from ethanol.

10. Use of the oxazolone matrine derivative according to claim 1 in the preparation of drugs for liver cancer, lung cancer, cervical cancer and breast cancer.

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