Matrine amine thiazole derivative as well as preparation method and application thereof

Abstract

The invention discloses a matrine amine thiazole derivative as well as a preparation method and application thereof, the matrine amine thiazole derivative has a structural formula as shown in Figure 1, and in the structural formula, R is any one of 4-chlorphenyl, 4-nitrophenyl, 2, 4-difluorophenyl, 4-bromophenyl, 4-methoxyphenyl, 4-phenylphenyl, naphthyl and 4-iodophenyl. The method comprises the following steps: firstly, synthesizing thiourea matrine by taking sophocarpine and thiourea as initial substrates and N, N-dimethylformamide as a solvent, and then synthesizing a series of matrine derivatives by taking thiourea matrine and alpha-bromo-R-ethyl ketone as reactants and taking ethanol as a solvent. The anti-tumor activity of the matrine derivative is enhanced, the preparation method is simple and convenient, and reaction conditions are easy to control.

Description

Technical Field

[0001] The invention belongs to the technical field of pesticide synthesis, and specifically relates to a matrine amine thiazole derivative and a preparation method and application thereof. Background Art

[0002] Matrine is an alkaloid extracted from the dried roots of Sophora flavescens, Sophora flavescens, etc. Matrine-type alkaloids are quinolizidine analogues. Matrine-type alkaloids include matrine, allomatine, sophoracarpine, sophoridine, oxymatrine, sophoramine, sophorol, etc. Matrine has effective effects in antibacterial, anti-inflammatory, anti-allergic, antiarrhythmic, immune and biological regulation.

[0003] Matrine has poor chemical stability and is toxic to the central nervous system, so its bioavailability is low. Its structure needs to be modified to obtain matrine amine thiazole derivatives with high biological activity and safety. Summary of the invention

[0004] Purpose of the invention: To address the problem of low bioavailability of matrine, the present invention provides an anti-tumor matrine amine thiazole derivative.

[0005] Technical solution: To achieve the above purpose, the technical solution adopted by the present invention is:

[0006] A matrine amine thiazole derivative, the structural formula of the matrine amine thiazole derivative is as follows:

[0007]

[0008] Wherein, R is any one of phenyl, 4-nitrophenyl, naphthyl, 4-iodophenyl and 4-nitrilephenyl.

[0009] Another object of the present invention is to provide a method for preparing a matrine amine thiazole derivative, comprising the following steps:

[0010] Step 1, preparing thiourea matrine intermediate 3, thiourea matrine intermediate 3 is prepared by the following reaction equation:

[0011]

[0012] Step 2, prepare the matrine amine thiazole derivative 5 by the following reaction formula:

[0013]

[0014] Wherein, R is any one of phenyl, 4-nitrophenyl, naphthyl, 4-iodophenyl and 4-nitrilephenyl.

[0015] Preferably: firstly, NaH, DMF solution and thiourea are added to the reaction container in sequence and stirred to mix; secondly, sophoracarpine is added to the reaction container to react; after the reaction is completed, the reaction is quenched; then, extraction, concentration and drying, column chromatography separation and purification are performed in sequence to obtain a thiourea matrine intermediate.

[0016] Preferably: firstly, the thiourea matrine intermediate and ethanol are poured into a reactor, and then α-bromo R-ethyl ketone is added to react, and a reaction solution is obtained after the reaction is completed. Finally, the reaction solution is concentrated, dried, and separated by column chromatography to obtain the matrine amine thiazole derivative 5.

[0017] Preferably, during the reaction, the reaction process is monitored by TLC plate.

[0018] Preferably: in step 2, the thiourea matrine intermediate 3 is reacted with α-bromoacetophenone, α-bromo-4-nitroacetophenone, α-bromonaphthylacetophenone, α-bromo-4-iodoacetophenone, and α-bromo-4-nitrileacetophenone to obtain matrine-C13 aminothiazole derivative 5-1, matrine-C13 aminothiazole derivative 5-4, matrine-C13 aminothiazole derivative 5-15, matrine-C13 aminothiazole derivative 5-16, and matrine-C13 aminothiazole derivative 5-17.

[0019] Another object of the present invention is to provide a use of the matrine amine thiazole derivative in the preparation of tumor inhibitors.

[0020] Preferably, the tumor inhibitor is a lung cancer cell inhibitor, a cervical cancer cell inhibitor, or a colon cancer cell inhibitor.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] The matrine aminothiazole derivatives of the present invention form matrine derivatives with an aminothiazole structure while completely retaining the matrine structure, thereby enhancing the anti-tumor activity of the matrine derivatives. The molecular structures thereof are novel and all of them are new compounds. They have distinct chemical structure characteristics, and the preparation method is not only simple, but also the reaction conditions are easily controlled, and the products can be obtained by column chromatography. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 Schematic diagram of the structure of matrine amine thiazole derivatives in the embodiments of the present invention.

[0024] Figure 2 This is the hydrogen nuclear magnetic resonance spectrum of Example Compound 5-1 of the present invention.

[0025] Figure 3 This is the carbon nuclear magnetic resonance spectrum of Example Compound 5-1 of the present invention.

[0026] Figure 4 This is the hydrogen nuclear magnetic resonance spectrum of Example Compound 5-4 of the present invention.

[0027] Figure 5 This is the carbon nuclear magnetic resonance spectrum of Example Compound 5-4 of the present invention.

[0028] Figure 6 This is the hydrogen nuclear magnetic resonance spectrum of Example Compound 5-15 of the present invention.

[0029] Figure 7 The carbon nuclear magnetic resonance spectrum of Example Compound 5-15 of the present invention.

[0030] Figure 8 The hydrogen nuclear magnetic resonance spectrum of Example Compound 5-16 of the present invention.

[0031] Fig. 9 The carbon nuclear magnetic resonance spectrum of Example Compound 5-16 of the present invention.

[0032] Fig.10 This is the hydrogen nuclear magnetic resonance spectrum of Example Compound 5-17 of the present invention.

[0033] Fig.11 The carbon nuclear magnetic resonance spectrum of Example Compound 5-7 of the present invention. DETAILED DESCRIPTION

[0034] The present invention is further explained below in conjunction with the accompanying drawings and specific embodiments. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. After reading the present invention, various equivalent forms of modifications to the present invention by those skilled in the art all fall within the scope defined by the claims attached to this application.

[0035] Example 1

[0036] A matrine amine thiazole derivative, the structural formula of the matrine amine thiazole derivative is as follows: Figure 1 As shown, Figure 1 In the above, R is as follows, which is any one of phenyl 5-1, 4-nitrophenyl 5-4, naphthyl 5-15, 4-iodophenyl 5-16 and 4-nitrilephenyl 5-17.

[0037]

[0038] Example 2

[0039] This embodiment provides a method for preparing a matrine amine thiazole derivative, comprising the following steps:

[0040] Step 1, preparing thiourea matrine intermediate 3 by reacting sophocarpine 1 with thiourea 2, using the lone pair of electrons on the nitrogen atom of thiourea to attack the C13 position in matrine, and synthesizing thiourea matrine intermediate 3 by nucleophilic addition.

[0041] First, 60% pure NaH, DMF solution, and thiourea were added to the reaction vessel in order, and stirred at room temperature for 10 minutes. Then, sophoracarpine was added to the reaction vessel for reaction. During the reaction, the reaction process was monitored by TLC plate. After the reaction was completed, an appropriate amount of acetic acid was added to quench the reaction, and then extraction, concentration and drying, column chromatography separation (dichloromethane: methanol = 10: 1-8: 1) were performed in order to purify to obtain thiourea matrine intermediate 3.

[0042] The reaction formula for synthesizing thiourea matrine intermediate 3 is:

[0043]

[0044] Step 2, prepare matrine amine thiazole derivative 5 via thiourea matrine intermediate 3 and α-bromo R-ethyl ketone.

[0045] A series of matrine derivatives were synthesized using thiourea matrine and α-bromo R-ethyl ketone as reactants and ethanol as solvent. Specifically, thiourea matrine intermediate 3 and solvent ethanol were first poured into a reactor, and thiourea matrine intermediate 3 was dissolved by ethanol. α-bromo R-ethyl ketone was then added, and the molar ratio of thiourea matrine intermediate 3 to α-bromo R-ethyl ketone was mmol:mmol=0.3:0.36. The reaction was carried out at room temperature, and the reaction process was monitored by TLC plate during the reaction, and the reaction time was 1h. The reaction solution was obtained after the reaction was completed, and finally the reaction solution was concentrated and dried, and separated by column chromatography (dichloromethane:methanol=30:1-20:1) to obtain matrine amine thiazole derivative 5.

[0046] The reaction formula for synthesizing matrine amine thiazole derivative 5 is:

[0047]

[0048] Here, R is any one of phenyl 5-1, 4-nitrophenyl 5-4, naphthyl 5-15, 4-iodophenyl 5-16, and 4-nitrilephenyl 5-17.

[0049] In this embodiment, thiourea matrine intermediate is first synthesized using sophoracarpine and thiourea as starting substrates and N,N-dimethylformamide as solvent, and then a series of matrine amine thiazole derivatives are synthesized using thiourea matrine intermediate and α-bromo R-ethyl ketone as reactants and ethanol as solvent. While the structure of matrine is completely retained, the matrine amine thiazole derivatives with amine thiazole structure are formed, thereby enhancing the antibacterial activity of the matrine amine thiazole derivatives.

[0050] Example 3

[0051] The difference between this embodiment and embodiment 2 is that the matrine-C13 aminothiazole derivative 5-1 is synthesized by reacting the thiourea matrine intermediate 3 with α-bromoacetophenone.

[0052] Add thiourea matrine intermediate 3 into a round-bottom flask, add solvent ethanol to dissolve, then add α-bromoacetophenone, the molar ratio of thiourea matrine intermediate 3 to α-bromoacetophenone is mmol:mmol:0.3:0.36; react at room temperature, monitor the reaction with a TLC plate until the reaction is completed, reaction time: 1h. The reaction solution is concentrated and dried, and separated by column chromatography (dichloromethane:methanol=30:1~20:1) to obtain the product matrine-C13 aminothiazole derivative 5-1:

[0053]

[0054] like Figure 2 , 3 As shown, the spectrum data analysis of matrine-C13 aminothiazole derivative 5-1:

[0055] White solid, R f :0.65 (DCM:MeOH=10:1), 126.6mg (99.5% yield).

[0056] 1 H NMR (400 MHz, CDCl 3 )δ7.81(d,J=7.1Hz,2H),7.37(t,J=7.6Hz,2H),7.27(t,J=7.3Hz,1H),6.68(s,1H),6.05(d,J =6.8Hz,1H),4.41(dd,J=12.7,4.4Hz,1H),4.26(d,J=6.2Hz,1H),4.16–4.05(m,1H),3.18(t,J =12.7Hz,1H),2.84(t,J=11.9Hz,2H),2.76(dd,J=17.1,4.9Hz,1H),2.57(dd,J=17.2,6.0Hz, 1H),2.39–2.85(m,1H),2.20(s,1H),2.07–1.90(m,4H),1.82–1.52(m,6H),1.51–1.37(m,3H).

[0057] 13 C NMR (101 MHz, CDCl 3)δ167.4,167.1,151.1,135.0,128.5(2C),127.5,126.0(2C),101.0,63.8,57.2,57. 1,50.4,46.6,42.5,41.8,38.3,35.5,30.6,27.6,26.4,21.0,20.5.HRMS(ESI)Calcd for C 24 H 30 N 4 OS[M+H] + m / z 423.2213, found 423.2215.

[0058] Example 4

[0059] The difference between this embodiment and embodiment 2 is that the matrine-C13 aminothiazole derivative 5-4 is synthesized by reacting the thiourea matrine intermediate 3 with α-bromo-4-nitroacetophenone, as shown below:

[0060]

[0061] like Figure 4 , 5 As shown, the spectrum data analysis of matrine-C13 aminothiazole derivative 5-4:

[0062] Yellow solid, R f :0.65 (DCM:MeOH=10:1), 138.9mg (99.0% yield).

[0063] 1 H NMR (400 MHz, CDCl 3 )δ8.19(d,J=8.4Hz,2H),7.92(d,J=8.4Hz,2H),6.89(s,1H),6.33(d,J=6.6Hz,1H),4.40(dd,J= 12.8,4.3Hz,1H),4.30(t,J=6.2Hz,1H),4.17–4.04(m,1H),3.19(t,J=12.8Hz,1H),2.96–2.80(m ,3H),2.73(dd,J=17.1,4.8Hz,1H),2.59(dd,J=17.3,5.3Hz,1H),2.45–2.34(m,1H),2.23(s,1H) ,2.08–1.96(m,3H),1.93–1.84(m,1H),1.81–1.73(m,1H),1.71–1.54(m,4H),1.50–1.38(m,3H).

[0064] 13C NMR (101 MHz, CDCl 3 )δ167.5,166.7,149.1,146.8,140.7,126.4(2C),124.0(2C),105.2,63.7,57.2,50. 2,46.9,42.8,41.8,38.2,35.4,30.8,29.7,27.6,26.5,21.1,20.6.HRMS(ESI)Calcd for C 24 H 29 N 5 O 3 S[M+H] + m / z 468.2064, found 468.2061.

[0065] Example 5

[0066] The difference between this embodiment and embodiment 2 is that the matrine-C13 aminothiazole derivative 5-15 is synthesized by reacting the thiourea matrine intermediate 3 with α-bromonaphthaleneacetone, as shown below:

[0067]

[0068] like Figure 6 , 7 As shown, the spectrum data analysis of matrine-C13 position thiazole derivatives 5-15:

[0069] White solid, R f :0.65 (DCM:MeOH=10:1), 124.7 mg (88.0% yield).

[0070] 1 H NMR (400 MHz, CDCl 3 )δ8.35(s,1H),7.93–7.79(m,4H),7.52–7.44(m,2H),6.85(s,1H),5.37(d,J=6.9Hz,1H), 4.44(dd,J=12.7,4.4Hz,1H),4.33–4.23(m,1H),4.16–4.08(m,1H),3.19(t,J=12.7Hz,1H ),2.89–2.78(m,3H),2.61(dd,J=17.0,6.2Hz,1H),2.38–2.29(m,1H),2.20(s,1H),2.14– 1.96(m,4H),1.84–1.72(m,3H),1.71–1.64(m,2H),1.63–1.52(m,2H),1.51–1.42(m,2H).

[0071] 13 C NMR (151 MHz, CDCl 3 )δ167.2,166.6,151.4,133.7,133.0,132.2,128.3,128.1,127.7,126.2,125.9,125.0,124.1,101.8 ,63.8,57.2,57.2,50.5,47.0,42.6,41.9,38.5,35.6,30.8,27.7,26.6,21.1,20.6.HRMS(ESI)Calcd for C 28 H 32 N 4 OS[M+H] + m / z 473.2370, found 473.2365.

[0072] Example 6

[0073] The difference between this embodiment and embodiment 2 is that the matrine-C13 aminothiazole derivative 5-16 is synthesized by reacting the thiourea matrine intermediate 3 with α-bromo-4-iodoacetophenone:

[0074]

[0075] like Figure 8 , 9 As shown, the spectrum data analysis of matrine-C13 aminothiazole derivatives 5-16:

[0076] White solid, R f :0.65 (DCM:MeOH=10:1), 156.3 mg (95.0% yield).

[0077] 1 H NMR (400 MHz, CDCl 3)δ7.69(d,J=8.1Hz,2H),7.54(d,J=8.1Hz,2H),6.70(s,1H),5.86(s,1H),4.44(dd,J=1 3.0,4.0Hz,1H),4.23(s,1H),4.19–4.08(m,1H),3.21(t,J=12.8Hz,1H),3.00–2.91(m,2 H),2.74(dd,J=17.2,4.7Hz,1H),2.61(dd,J=17.4,5.5Hz,1H),2.37–2.28(m,2H),2.12 –1.92(m,4H),1.87–1.78(m,2H),1.74–1.56(m,4H),1.53–1.40(m,3H).HRMS(ESI)Calcd for C 24 H 29 IN 4 OS[M+H] + m / z 549.1180, found 549.1171.

[0078] 13CNMR(101MHz,CDCl3)δ167.4,166.9,150.2,137.5(2C),134.4,127.8(2C),101.9,9 2.9,64.0,57.0(2C),50.2,46.7,42.3,41.5,38.2,35.2,30.8,27.2,26.1,20.6,20.1.

[0079] Example 6

[0080] The difference between this embodiment and embodiment 2 is that the matrine-C13 aminothiazole derivative 5-17 is synthesized by reacting the thiourea matrine intermediate 3 with α-bromo-4-nitrile acetophenone:

[0081]

[0082] like Fig.10 , 11 As shown, the spectrum data analysis of matrine-C13 aminothiazole derivatives 5-17:

[0083] White solid, R f :0.65 (DCM:MeOH=10:1), 123.5mg (92.0% yield).

[0084] 1 H NMR (400 MHz, CDCl 3)δ7.91(d,J=8.5Hz,2H),7.66(d,J=8.4Hz,2H),6.87(s,1H),5.73(d,J=6.8Hz,1H),4.43(dd,J=12 .8,4.4Hz,1H),4.33–4.23(m,1H),4.14–4.05(m,1H),3.19(t,J=12.7Hz,1H),2.86(t,J=12.5Hz,2H ),2.77(dd,J=17.1,4.8Hz,1H),2.66–2.57(m,1H),2.43–2.32(m,1H),2.23(s,1H),2.07–1.99(m,3 H),1.97–1.89(m,1H),1.85–1.75(m,2H),1.74–1.69(m,1H),1.67–1.56(m,3H),1.52–1.41(m,3H).

[0085] 13CNMR(151MHz,CDCl3)δ167.4,166.8,149.4,139.6,132.4(2C),126.4(2C),118.7,110.6,104.4, 63.7,57.2(2C),50.2,46.9,42.8,41.8,38.2,35.4,30.8,27.6,26.4,21.0,20.6.HRMS(ESI)Calcd for C 25 H 29 N 5 OS[M+H] + m / z 448.2166, found 448.2157.

[0086] The experiment explored aspects such as material ratio, catalyst, temperature, solvent, and found the best solution for synthesizing thiourea intermediates.

[0087] Example 7

[0088] This example tests the in vitro antitumor activity of matrine derivatives.

[0089] Cell lines: human lung cancer cells (A549), cervical cancer cells (Hela), human colon cancer cells (HCT-116)

[0090] Method: MTT method

[0091] The specific process is as follows:

[0092] (1) Cell culture and drug treatment. Actively growing tumor cells were digested with trypsin, centrifuged, and resuspended in fresh culture medium. The cells were diluted and counted, and then inoculated into a 96-well plate at a density of 5,000 cells / well. The plates were placed at 37°C and 5% CO. 2 The cells were cultured overnight in an incubator. On the second day, the cells were observed to be in an adherent state and the old culture medium was discarded. The samples to be tested were added to the experimental group. Five sample concentration gradients were set and the samples were diluted to a final concentration of 200, 100, 50, 25, and 12.5 μmol / L. Five replicate wells were set for each concentration, with 250 μL in each well. New culture medium was added to the negative control group and the blank group and the cells were cultured in an incubator for 48 h. The approximate concentration of 50% inhibition rate was screened out. Based on this concentration, 3-4 concentrations were tested at intervals of 10 ug / ml, as described above.

[0093] (2) MTT assay: 20 μL of pre-prepared MTT solution was added to each well and incubated in an incubator for 4 h. After incubation, the liquid in the well was aspirated and 200 μL of dimethyl sulfoxide was added to each well. The well was fully shaken for 10 min. The absorbance was detected at 490 nm using an ELISA reader. The data were recorded and the inhibition rate was calculated according to the formula. The IC was obtained by fitting with the Origin software. 50 value

[0094] (3) Calculation: Cell inhibition rate = [(control - background) - (drug administration - background)] / (control - background) * 100%

[0095] The measurement results are shown in the table:

[0096] Table 1 In vitro antitumor activity IC of matrine-C13 aminothiazole derivatives 50 (ug / ml)

[0097]

[0098] The in vitro antitumor activity results of the experimental group compounds and the parent matrine are shown in Table 1 above. The in vitro antitumor activity of compounds 5-1, 5-4, 5-15, 5-16, and 5-17 are significantly improved relative to the parent matrine. Among them, compound 5-15 has a good inhibitory effect on human lung cancer cells (A549) and human colon cancer cells (HCT-116). 50 The values ​​were 18.1ug / ml and 16.3ug / ml respectively; the compound 5-1 with the best effect on cervical cancer cells (Hela) had an IC 50 The value was 23.2ug / ml. The results showed that matrine derivatives have potential anti-tumor activity and can be developed as anti-tumor drugs.

[0099] The anti-tumor matrine derivatives of the present invention have obvious structural differences and distinct chemical structural characteristics, and show good effects on inhibiting tumor cells. They can be developed as anti-tumor drugs. The preparation method of the compounds is simple and the product properties are stable. The anti-tumor matrine derivatives synthesized by the present invention have good activity and medicinal value, which has certain implications for designing new anti-tumor agents from natural products.

[0100] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A matrine amine thiazole derivative, characterized in that: The structural formula of the matrine amine thiazole derivative is as follows: Wherein, R is any one of phenyl, 4-nitrophenyl, naphthyl, 4-iodophenyl and 4-nitrilephenyl.

2. A method for preparing a matrine amine thiazole derivative, characterized in that: The following steps are involved: Step 1, preparing thiourea matrine intermediate 3, thiourea matrine intermediate 3 is prepared by the following reaction equation: Step 2, prepare the matrine amine thiazole derivative 5 by the following reaction formula: Wherein, R is any one of phenyl, 4-nitrophenyl, naphthyl, 4-iodophenyl and 4-nitrilephenyl.

3. The method for preparing the matrine amine thiazole derivative according to claim 2, characterized in that: Firstly, NaH, DMF solution and thiourea are added to a reaction container in sequence and stirred and mixed. Secondly, sophoracarpine is added to the reaction container for reaction. After the reaction is completed, the reaction is quenched. Then, extraction, concentration and drying, column chromatography separation and purification are carried out in sequence to obtain a thiourea matrine intermediate.

4. The method for preparing the matrine amine thiazole derivative according to claim 3, characterized in that: First, the thiourea matrine intermediate and ethanol are poured into a reactor, and then α-bromo R-ethyl ketone is added to react. After the reaction is completed, a reaction liquid is obtained. Finally, the reaction liquid is concentrated and dried, and separated by column chromatography to obtain the matrine amine thiazole derivative 5.

5. The method for preparing the matrine amine thiazole derivative according to claim 4, characterized in that: During the reaction, the reaction progress was monitored by TLC plate.

6. The method for preparing the matrine amine thiazole derivative according to claim 5, characterized in that: In step 2, the thiourea matrine intermediate 3 is reacted with α-bromoacetophenone, α-bromo-4-nitroacetophenone, α-bromonaphthylacetophenone, α-bromo-4-iodoacetophenone, and α-bromo-4-nitrileacetophenone to obtain matrine-C13 aminothiazole derivative 5-1, matrine-C13 aminothiazole derivative 5-4, matrine-C13 aminothiazole derivative 5-15, matrine-C13 aminothiazole derivative 5-16, and matrine-C13 aminothiazole derivative 5-17.

7. Use of the matrine amine thiazole derivative as claimed in claim 1 in the preparation of tumor inhibitors.

8. The application according to claim 7, characterized in that: The tumor inhibitors are lung cancer cell inhibitors, cervical cancer cell inhibitors, and colon cancer cell inhibitors.

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