CN109369649B - Matrine amide derivatives and preparation method and use thereof - Google Patents

Abstract

The invention discloses a matrine amide derivative, a preparation method and application thereof, and belongs to the field of medicinal chemistry. A series of new compounds synthesized by the invention have anti-cancer activity. The method uses matrine as raw material, hydrolyzes ring-opening under alkaline conditions, undergoes esterification reaction, hydrolysis reaction, and amide coupling reaction, and finally obtains a series of new matrine base derivatives. Since matrine itself has certain pharmacological effects, the invention uses MTT colorimetry to test the anticancer activity of the synthesized new derivative, and the anticancer activity of the new derivative is obviously better than that of matrine.

Description

Matrine amide derivatives and preparation method and use thereof

technical field

The invention relates to the synthesis of a series of new derivatives of matrine amide, and the biological activity test of the synthesized compounds belongs to the field of medicinal chemistry.

Background technique

Tumor is a common clinical disease at present, the refractory of tumor lies in tumor recurrence and metastasis, and tumor angiogenesis is one of the important conditions for tumor recurrence and metastasis. Therefore, controlling the development of tumor diseases by inhibiting tumor angiogenesis has become a hot spot of anti-tumor therapy today. Matrine (Matrine, formula 1) is an active ingredient isolated from the traditional medicinal plant Sophora flavescens, and has attracted much attention due to its wide range of biological activities, such as antitumor, anti-inflammatory, and antiviral. In China, matrine injection is clinically used to treat hepatitis and liver cancer, and matrine suppository can be used to treat vaginitis and chronic cervicitis. However, so far, matrine derivatives or analogs have not become anticancer drugs because of their moderate antitumor activity. Therefore, this method modifies the structure of matrine to synthesize a series of derivatives, in order to discover compounds that may become drug candidates. This method is based on the modification and modification of the parent structure of matrine.

Matrine is ring-opened under alkaline conditions to form matrine, and then the 16-position N is protected by benzyl group, and then hydrolyzed to obtain N-benzyl matrine, and finally the carboxyl group is supplied with NO through a linking group. 13 NO-donating matrine derivatives (Formula 2) were obtained by coupling with nitrofuran nitroxides, and their anticancer activity was superior to matrine (L.Q.He, et al., Chinese Chemical Letters, 2010, 381– 384).

In 1957, Tsuda et al. (K.Tsuda, et al., J.Org.Chem., 1958, 23(8), 1179-1183) hydrolyzed matrine in potassium hydroxide to obtain potassium matrine; Matrine is reduced by lithium aluminum hydride or catalytic hydrogenation to obtain matrine, and then the two are respectively methylated to prepare two kinds of quaternary ammonium salts of matrine derivatives (formula 3), but the activity is not seen report.

SUMMARY OF THE INVENTION

In the present invention, a series of new matrine amide derivatives are synthesized through matrine hydrolysis ring-opening, esterification and amide coupling reactions, and the anticancer activity of these compounds is tested. The synthesized matrine amide derivatives It has anticancer activity and can be used to inhibit the proliferation of cancer cells.

The structural formula of matrine amide derivatives is:

Wherein, the structural formula R ₁ is 3-fluorophenyl, 3-chlorophenyl, 3-bromophenyl, 3-methylphenyl, 3-methoxyphenyl, 3-nitrophenyl, 2-naphthyl or 2-pyridine.

The synthetic route of the new derivative of matrine amide is as follows:

The synthetic method of matrine amide derivative, the concrete synthetic steps are:

(1) Under the action of a base, add matrine to a solvent, heat under reflux for reaction, and post-process to obtain product compound 2; the base is sodium hydroxide or potassium hydroxide; the solvent is tetrahydrofuran or water.

(2) adding thionyl chloride to the solvent, stirring the reaction at low temperature, then adding compound 2, stirring at low temperature and heating to reflux, and post-processing to obtain product compound 3; the solvent is methanol, dichloromethane, chloroform or tetrahydrofuran; The low temperature stirring temperature is -20-0°C; the reflux temperature is 50-70°C.

(3) Compound 3 and 4-dimethylaminopyridine were added to the dichloromethane solution, and the reaction was stirred at room temperature, then di-tert-butyl dicarbonate was dissolved in the dichloromethane solution and added thereto, and the reaction was stirred at room temperature to obtain 16 -Boc methyl matrine, compound 4; firstly protect the 16-position N with a tert-butoxycarbonyl group, and then carry out the amide coupling reaction, which can effectively improve the reaction selectivity, so that the 16-Boc matrine is preferentially combined with 16-Boc matrine in the coupling reaction. The carboxyl group of the acid reacts; then the de-Boc reaction is carried out, the reaction operation is simple and the side reaction of the system is less.

(4) Under alkaline conditions, add 16-Boc methyl matrine to a solvent, and stir to react at room temperature to obtain compound 5; the solvent in step (4) is water, tetrahydrofuran or methanol.

(5) Dissolve compound 5 and amide coupling reagent in N,N-dimethylformamide, add N,N-diisopropylethylamine under ice bath after stirring at room temperature, then continue stirring at room temperature, add aromatic The amine compound was stirred at room temperature for 18 h to obtain the amide compound 6.

The amide coupling reagent is O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroborate (TBTU), hexafluorophosphate benzotriazole-1-yl- Oxytripyrrolidinophosphorus (PyBOP), 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) or O-benzotriazole One of triazole-tetramethylurea hexafluorophosphate (HBTU). 2-(7-Benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) is preferred as the amide coupling reagent, because the reaction is in HATU as the coupling reagent When the reagents are combined, the side reactions of the system are less and the yield is higher.

(6) Dissolving the amide compound 6 in dichloromethane, adding an acid, and stirring the reaction to obtain the matrine amide compound 7; the acid is hydrogen chloride gas or trifluoroacetic acid.

(7) Compound 7 was dissolved in dichloromethane, a base was added, then benzoyl chloride was added dropwise, the reaction was stirred at room temperature, concentrated, and purified by column chromatography to obtain the matrine amide derivative 8a; the base was triethylamine , ethylenediamine or potassium hydroxide.

Or dissolving compound 7 and triethylamine in a solvent, adding benzaldehyde dropwise, stirring and refluxing for the reaction, then slowly adding a reducing agent to the reaction solution in batches, continuing to reflux, cooling to room temperature, concentrating extraction, and purifying by column chromatography , the matrine amide derivative 8b was obtained. The solvent is 1,2-dichloroethane or dichloromethane; the reducing agent is sodium triacetoxyborohydride or sodium borohydride.

The beneficial effects of the present invention are:

The advantage of this method is that a series of new derivatives of matrine amides are prepared, and the biological activity test results show that the anticancer activity of the new derivatives is obviously better than matrine.

Detailed ways

The present invention will now be further described with reference to examples.

Step 1: Preparation of 4-(decahydro-1H,4H-pyrido[1,6]naphthyridin-1-yl)butanoic acid (compound 2)

Example 1: Compound 1 (20 g, 80.5 mmol) was dissolved in water (456 mL), sodium hydroxide (9.1 g, 228 mmol) was continuously added to dissolve, and the mixture was heated to reflux at 90° C. for 12 h. After the reaction was completed, the pH was adjusted to 5-7 with 2N hydrochloric acid solution, the solvent was concentrated, 460 mL of ethanol was added, stirred at room temperature for 30 min, and 21 g of white solid was obtained by suction filtration with a yield of 99%. ¹ H NMR (300 MHz, D ₂ O): δ 1.61-1.80 (m, 9H), 1.83-2.01 (m, 4H), 2.16-2.20 (t, 1H), 2.26-2.34 (m, 4H), 2.90 _-2.97 (q, J1 ₌ 12.6Hz, J2=4.5Hz, 1H), 2.71(s, 1H), 3.23-3.31(t, J=12.9Hz, 1H), 3.37-3.41(q, 1H). ¹³ C NMR (75MHz, D ₂ O): δ 22.00, 22.26, 23.16, 27.27, 28.33, 32.52, 35.32, 39.68, 40.25, 46.18, 58.64, 58.70, 64.32. HRMS (ESI): m/z [MH] ^- calcd for C ₁₅ H ₂₅ N ₂ O ₂ : 265.1922; found: 265.1924.

Example 2: Other conditions were the same as in Example 1, except that the base used was changed from sodium hydroxide to potassium hydroxide, and the yield of the obtained product was 70%.

Example 3: Other conditions were the same as in Example 1, except that the reaction solvent was changed from water to tetrahydrofuran, and the yield of the obtained product was 70%.

Step 2: Preparation of methyl 4-(decahydro-1H,4H-pyrido[1,6]naphthyridin-1-yl)butanoate (compound 3)

Example 4: SOCl ₂ (29.5 mL, 40.7 mmol) was dissolved in MeOH (336 mL) at 0 °C and stirred for 1 h. Afterwards, a mixture of compound 2 (13 g, 48.8 mmol) dissolved in MeOH (168 mL) was added thereto, reacted in an ice bath for 2 h, and then refluxed for 3 h. After the reaction was completed, it was cooled to room temperature, 356 mL of CHCl ₃ and 50 g of NaHCO ₃ were added, stirred at room temperature for 0.5 h, suction filtered, and the mother liquor was concentrated. 9 g of white solid were obtained in 66% yield. ¹ H NMR (300 MHz, CD ₃ OD): δ 1.44-1.64 (m, 5H), 1.71-1.64 (m, 5H), 1.74-1.80 (m, 3H), 1.91-1.94 (d, 1H), 2.00 -2.08(m, 3H), 2.26(s, 1H), 2.26(s, 1H), 2.41-2.37(m, 2H), 2.37-2.41(m, 2H), 2.94-2.98(dd, J ₁ =6.2 Hz, J ₂ =2.1Hz, 1H), 3.40-3.46(t, J=12.8Hz, 1H), 3.54-3.60(m, 1H), 3.65-3.63(d, 3H). ¹³ CNMR(75MHz, CD ₃ OD):δ20.44,20.63,25.63,26.66,30.47,33.68,33.90,38.51,44.51,48.15,49.85,49.85,53.92,57.27,62.93,175.06.HRMS(ESI):m/z[M+Na] ⁺ calcd for C ₁₆ H ₂₈ O ₂ Na: 303.2043; found: 303.2039.

Example 5: Other conditions were the same as in Example 4, except that the reaction solvent was changed from methanol to dichloromethane, and the yield of the obtained product was 50%.

Example 6: Other conditions were the same as in Example 4, except that the reaction solvent was changed from methanol to tetrahydrofuran, and the yield of the obtained product was 55%.

Example 7: Other conditions were the same as in Example 4, except that the reaction temperature was changed from 0°C to -10°C, and the yield of the obtained product was 45%.

Example 8: Other conditions were the same as in Example 4, except that the reaction temperature was changed from 0°C to -20°C, and the yield of the obtained product was 54%.

Example 9: Other conditions were the same as in Example 4, except that the reflux temperature was changed from 70°C to 50°C, and the yield of the obtained product was 48%.

Step 3: tert-butyl-1-(4-methoxy-4-oxobutyl)octahydro-1H,4H-pyrido[1,6]naphthalene-2(3H)-carboxylic acid ( Preparation of compound 4)

Example 10: Compound 3 (6.5 g, 23.2 mmol) and 4-dimethylaminopyridine (650 mg) were dissolved in dichloromethane (116 mL), followed by di-tert-butyl dicarbonate (7.6 g, 34.8 mmol) in Dichloromethane (23 mL) solution was dissolved in it, and the reaction was stirred at room temperature for 16 h. The reaction was monitored by TLC and concentrated. Dichloromethane:methanol=50:1 column chromatography to obtain 7.3 g of yellow viscous liquid with a yield of 83%. ¹ H NMR (300 MHz, CDCl ₃ ): δ 1.42-1.34 (m, 3H), 1.45 (s, 11H), 1.61-1.58 (m, 2H), 1.83-1.63 (m, 9H), 1.96-1.95 ( t,J=4.1Hz,1H),2.31-2.38(m,2H),2.67-2.71(t,J=6.8Hz,2H),3.26-3.33(dd,J1 ₌ 9.8Hz,J2= _18.2Hz , 1H), 3.50-3.55 (dd, 1H, J ₁ =10.9Hz, J ₂ =18.0Hz), 3.65 (s, 3H), 3.78-3.86 (m, 1H). ¹³ C NMR (75MHz, CDCl ₃ ) :δ20.81,20.97,21.67,27.95,28.22,28.90,31.15,33.50,34.70,40.14,43.89,51.04,53.81,56.48,56.67,63.01,78.53,155.45,173.71.HRMS(ESI):m/z[z M+H] ⁺ calcd for C ₂₁ H ₃₇ N ₂ O ₄ : 381.2748; found: 381.2749.

Step 4: 4-((3aS, 3a1S, 10aR)-2-(tert-butoxycarbonyl)decahydro-1H,4H-pyrido[1,6]naphthyridin-1-yl)butanoic acid (Compound 5) preparation

Example 11: Compound 4 (5 g, 13.1 mmol) was dissolved in methanol (59 mL), 4N sodium hydroxide solution (13.2 mL) was added, and the reaction was stirred at room temperature for 4 h. TLC monitoring, concentrated. Dichloromethane:methanol=10:1 was passed through the column to obtain 4 g of white solid in a yield of 85%. ¹ H NMR (400 MHz, CDCl ₃ ): δ 1.26-1.22 (m, 1H), 1.30 (s, 10H), 1.36 (m, 2H), 1.40-1.52 (m, 5H), 1.66-1.68 (m, 2H), 1.75-1.78(d, 3H), 1.91(s, 1H), 2.02-2.07(t, J=5.9Hz, 2H), 2.18-2.20(m, 1H), 2.30(m, 1H), 2.48 (s, 1H), 3.12-3.20 (q, J=11.2Hz, 2H), 3.27-3.34 (m, 1H), 3.51-3.55 (dd, J ₁ =5.0Hz, J ₂ =6.8Hz, 1H), 3.66-3.73(m, 1H). ¹³ C NMR (100MHz, CDCl ₃ )δ19.90, 20.19, 21.93, 26.47, 27.37, 28.56, 29.45, 34.51, 35.92, 39.60, 47.72, 54.89, 56.09, 56.20, 65.28, 79.75, 156.12, 177.82. HRMS(ESI): m/z[M+H] ⁺ calcd for C ₂₀ H ₃₅ N ₂ O ₄ : 367.2591; found: 367.2593.

Example 12: Other conditions were the same as in Example 11, except that the reaction solvent was replaced with water, and the yield was 54%.

Example 13: Other conditions were the same as those in Example 11, except that the reaction solvent was changed to tetrahydrofuran, and the yield was 32%.

Step 5: Preparation of 16-Boc Matrine Derivative (Compound 6)

Example 14: tert-Butyl(1R,3aS,3a1S,10aR)-1-(4-((3-fluorophenyl)amino)-4-oxobutyl)octahydro-1H,4H-pyrido[ Preparation of 1,6]naphthyridine-2(3H)-carboxylate tert-butyl ester (6a)

Compound 5 (100 mg, 0.27 mmol) and HATU (124 mg) were stirred at room temperature for 30 min. Then, under ice bath condition, DIEA (67 μL) was added, stirred at room temperature for 1 h, and 3-fluoroaniline (32 μL) was added, and stirred at room temperature for 18 h. The reaction was monitored by TLC and concentrated by column chromatography. 110 mg of white solid were obtained in 88% yield. ¹ H NMR (300 MHz, CD ₃ OD): δ 1.19-1.24 (t, 1H, J=7.0 Hz), 1.32 (s, 1H), 1.52 (s, 9H), 1.66-1.78 (m, 4H), 1.85-1.88(t, 5H, J=3.7Hz), 1.91(s, 1H), 2.05(S, 1H), 2.33-2.34(d, 1H), 2.44-2.49(m, 2H), 3.38(s, 1H), 3.44-3.50(m, 3H), 3.81-3.88(dd, 1H, J ₁ =13.0Hz, J ₂ =6.8Hz), 6.80-6.87(m, 1H), 7.24-7.36(m, 2H) , 7.55-7.60 (dd, 1H, J ₁ =11.4Hz, J ₂ =2.1Hz). ¹³ C NMR (75MHz, CD ₃ OD) δ 19.81, 20.28, 23.46, 26.77, 27.92, 28.60, 32.50, 34.33, 37.38,42.27,54.86,56.05,56.36,64.14,82.30,107.80,108.15,111.15,111.43,116.32,116.36,129.66,130.57,131.07,131.20,141.56,141.70,151.70,157.28,162.60,165.82,174.12.HRMS( ESI): m/z[M+H] ⁺ calcd for C ₂₆ H ₃₉ FN ₃ O ₃ : 460.2970; found: 460.2973.

Example 15: tert-Butyl(1R,3aS,3a1S,10aR)-1-(4-((3-chlorophenyl)amino)-4-oxobutyl)octahydro-1H,4H-pyrido[ Preparation of 1,6]naphthyridine-2(3H)-carboxylate tert-butyl ester (6b)

Other conditions were the same as in Example 14, except that 3-fluoroaniline was replaced with 3-chloroaniline, and the yield was 78%. ¹ H NMR (300MHz, CD ₃ OD): δ 1.29-1.32(d, 1H), 1.49(s, 9H), 1.59-1.76(m, 4H), 1.81-1.94(m, 8H), 2.03(s) ,1H),2.28(s,1H),2.44-2.48(t,2H,J＝6.9Hz),2.87-3.03(m,3H),3.37(s,1H),3.44-3.47(t,1H,J = 3.8Hz), 3.49-3.50(d, 1 1H), 3.52-3.53(d, 1H), 3.83-3.90(m, 1H), 7.07-7.11(m, 1H), 7.26-7.31(t, 1H, J=8.1Hz), 7.42-7.46 (m, 1H), 7.81-7.82 (t, 1H, J=2.0Hz). ¹³ CNMR (75MHz, CD ₃ OD) δ 19.86, 20.18, 23.35, 26.71, 27.52, 28.61, 32.17, 33.49, 37.20, 39.01, 43.41, 54.99, 56.12, 56.44, 64.47, 82.18, 119.00, 120.79, 124.76, 131.07, 135.28, 141.31, 157.29, 174.19.HRMS(EMH:m/z[SIMH+HRMS]) ] ⁺ calcd for C ₂₆ H ₃₉ ClN ₃ O ₃ : 476.2674; found: 476.2678.

Example 16: tert-Butyl(1R,3aS,3a1S,10aR)-1-(4-((3-bromophenyl)amino)-4-oxobutyl)octahydro-1H,4H-pyrido[ Preparation of 1,6]naphthyridine-2(3H)-carboxylate tert-butyl ester (6c)

Other conditions were the same as in Example 14, except that 3-fluoroaniline was replaced with 3-bromoaniline and the yield was 85%. ¹ H NMR (300MHz, CD ₃ OD): δ 1.49(s, 9H), 1.56-1.87(m, 12H), 1.95(s, 2H), 2.23(s, 1H), 2.42-2.47(m, 2H) ), 2.78-2.86(m, 2H), 3.23(s, 2H), 3.26(s, 1H), 3.45-3.48(d, 2H), 3.81-3.88(m, 1H), 7.22-7.24(m, 2H ),7.45-7.49(m,1H),7.95-7.99(m,1H) ^.13C NMR(75MHz, _CD3OD )δ20.12,20.51,23.44,17.19,27.98,28.64,33.14,33.83,37.32, 39.28,42.95,54.92,56.29,56.55,64.22,81.99,119.43,123.21,123.70,127.75,141.41,157.28,174.17.HRMS(ESI):m/z[M+H] ⁺ calcd for C ₂₆ H ₃₉ BrN ₃ O ₃ :520.2169;found:520.2164.

Example 17: tert-Butyl(1R,3aS,3a1S,10aR)-1-(4-((3-methoxyphenyl)amino)-4-oxobutyl)octahydro-1H,4H-pyridine Preparation of [1,6]naphthyridine-2(3H)-carboxylate tert-butyl ester (6d)

Other conditions were the same as in Example 14, except that 3-fluoroaniline was replaced with 3-methoxyaniline, and the yield was 88%. ¹ H NMR (400MHz, CD ₃ OD): δ 1.47(s, 9H), 1.52-1.59(m, 1H), 1.67-1.89(m, 9H), 1.94-2.01(m, 3H), 2.16(s) ,1H),2.41-2.54(m,2H),2.92-3.00(dd,2H,J ₁ =20.8Hz,J ₂ =9.4Hz),2.32-3.36(m,2H),3.47(s,1H), 3.51-3.64(m, 2H), 3.77(s, 3H), 3.90-3.96(dd, 1H, J ₁ =15.0Hz, J ₂ =9.0Hz), 6.64-6.67(m, 1H), 7.16-7.22( m, 2H), 7.38(s, 1H). ¹³ C NMR (100MHz, CD ₃ OD) δ19.92, 19.95, 23.23, 26.53, 26.68, 31.85, 34.41, 36.99, 39.19, 45.55, 55.16, 55.70, 56.23, _found ^_ _{_ _ _} :472.3171.

Example 18: tert-Butyl(1R,3aS,3a1S,10aR)-1-(4-((3-methylphenyl)amino)-4-oxobutyl)octahydro-1H,4H-pyrido Preparation of [1,6]naphthyridine-2(3H)-carboxylate tert-butyl ester (6e)

Other conditions were the same as in Example 14, except that 3-fluoroaniline was replaced with 3-methylaniline, and the yield was 95%. ¹ H NMR (300MHz, CDCl ₃ ): δ1.20-1.24(d,1H), 1.29-1.34(m,1H), 1.42(s,9H), 1.62-1.65(d,7H), 1.77-1.80( d, 2H), 1.87(s, 2H), 1.96-2.04(m, 2H), 2.28(s, 3H), 2.34-2.42(dd, 1H, J ₁ =14.9Hz, J ₂ =7.4Hz), 2.45 -2.54(m, 1H), 2.62-2.65(d, 2H), 3.07-3.17(m, 3H), 3.32-3.40(m, 1H), 3.50-3.56(m, 1H), 3.59-3.65(m, 1H), 6.88-6.90(d, 1H), 7.12-7.17(t, 1H, J=7.7Hz), 7.30-7.33(d, 1H), 7.37(s, 1H), 8.26(s, 1H). ¹³ C NMR (75MHz, CD ₃ OD): δ19.40, 21.48, 26.02, 26.21, 28.38, 31.10, 33.44, 36.50, 38.47, 44.34, 53.57, 54.07, 55.59, 64.31, 80.88, 118.06, 121.62, 128.740 137.91, 138.78, 155.73, 172.42. HRMS(ESI): m/z[M+H] ⁺ calcd for C ₂₇ H ₄₂ N ₃ O ₃ : 456.3221; found: 456.3219.

Example 19: tert-Butyl(1R,3aS,3a1S,10aR)-1-(4-((3-nitrophenyl)amino)-4-oxobutyl)octahydro-1H,4H-pyrido Preparation of [1,6]naphthyridine-2(3H)-carboxylate tert-butyl ester (6f)

Other conditions were the same as in Example 14, except that 3-fluoroaniline was replaced with 3-nitroaniline, and the yield was 77%. ¹ H NMR (400MHz, CD ₃ OD): δ 1.49(s, 10H), 1.63-1.77(m, 5H), 1.80-1.91(m, 8H), 2.04-2.05(d, 1H), 2.31(s) ,1H),2.47-2.51(m,2H),2.94-3.02(m,2H),3.43-3.45(t,1H,J=3.6Hz),3.47-3.49(dd,2H,J ₁ =8.7Hz, J ₂ =3.9Hz),3.82-3.88(dd,1H,J ₁ =14.0Hz,J ₂ =6.7Hz),7.52-7.56(t,J=8.2Hz),7.83-7.85(m,1H),8.67 -8.68 (t, 1H, J=1.9Hz), 7.91-7.94 (m, 1H). ¹³ C NMR (100 MHz, CD ₃ OD): δ 19.84, 20.26, 23.28, 26.76, 27.78, 28.59, 32.74, 34.01 ,37.28,38.93,42.69,54.91,56.07,56.38,64.23,82.22,115.21,119.24,126.36,130.84,141.23,149.76,157.29,174.36.HRMS(ESI):m/z[M+H] ⁺ calcd for C _26H39N4O5 : _{487.2915 ;} found: _487.2919 .

Example 20: tert-Butyl(1R,3aS,3a1S,10aR)-1-(4-oxo-4-(pyridin-2-ylamino)butyl)octahydro-1H,4H-pyrido[1, 6] Preparation of naphthyridine-2(3H)-carboxylate tert-butyl ester (6g)

Other conditions were the same as in Example 14, except that 3-fluoroaniline was replaced with 2-aminopyridine, and the yield was 97%. ¹ H NMR (400MHz, CD ₃ OD): δ 1.49 (s, 10H), 1.68-1.75 (m, 5H), 1.79-1.86 (m, 8H), 2.01 (s, 1H), 2.31 (s, 1H) ), 2.47-2.52(dd, 2H, J ₁ =7.5Hz, J ₂ =6.8Hz), 2.91-2.98(m, 2H), 3.39(s, 1H), 3.44-3.46(d, 2H), 3.79- 3.84 (m, 1H), 7.09-7.12 (m, 1H), 7.74-7.78 (t, 1H, J=7.8Hz), 8.07-8.09 (d, 1H). ¹³ C NMR (100 MHz, CD ₃ OD): delta :m/z[M+H] ⁺ calcd for C ₂₅ H ₃₉ N ₄ O ₃ : 443.3017; found: 443.3017.

Example 21: tert-Butyl(1R,3aS,3a1S,10aR)-1-(4-(naphthalen-2-ylamino)4-oxobutyl)octahydro-1H,4H-pyrido[1,6 Preparation of ]naphthyridine-2(3H)-carboxylate tert-butyl ester (6h)

Other conditions were the same as in Example 14, except that 3-fluoroaniline was replaced with 2-naphthylamine, and the yield was 81%. ¹ H NMR (400MHz, CD ₃ OD): δ 1.27-1.46 (m, 1H), 1.50 (s, 9H), 1.62-1.65 (d, 2H), 1.71-1.85 (m, 11H), 2.16 (s) ,1H),2.46-2.52(m,2H),2.81-2.89(m,2H),3.29(s,1H),3.35-3.41(m,2H),3.77-3.81(t,1H,J=6.7Hz ), 7.40-7.52 (m, 2H), 7.61-7.65 (dd, 1H, J ₁ =8.8 Hz, J ₂ =2.0 Hz), 7.79-7.88 (dd, 3H, J ₁ =15.1 Hz, J ₂ =8.7 Hz). ¹³ C NMR (75MHz, CD ₃ OD): δ19.65, 20.13, 23.55, 26.61, 27.77, 28.55, 32.25, 34.30, 37.44, 38.71, 42.02, 54.65, 55.88, 56.19, 63.88, 82.18, 117.76, _found ^_ _{_ _ _} :492.3224.

Example 22: Other conditions were the same as those in Example 14, except that the coupling reagent was replaced by HBTU, and the yield was 20%.

Example 23: Other conditions were the same as in Example 14, except that the coupling reagent was replaced with TBTU, and the yield was 54%.

Example 24: Other conditions were the same as those in Example 14, except that the coupling reagent was changed to PyBOP, and the yield was 60%.

Step 6: Preparation of 16-H matrine amide

Example 25: 4-((1R,3aS,3a1S,10aR)-decahydro-1H,4H-pyrido[1,6]naphthyridin-1-yl)-N-(3-fluorophenyl)butanamide Preparation of (7a)

At 0°C, compound 6a (1 g, 2.73 mmol) was dissolved in dichloromethane (10 mL), and hydrogen chloride gas was passed into it until the reaction was completed, and 10% sodium carbonate solution was added to adjust to pH=7-8, and the reaction solution was filtered. Concentration gave 95% yield. ¹ H NMR (300 MHz, CD ₃ OD): δ 1.44-1.58 (m, 4H), 1.66-1.67 (m, 5H), 1.73-1.87 (m, 3H), 1.90-2.10 (m, 5H), 2.25 (s, 1H), 2.47-2.52 (m, 2H), 2.82-2.88 (dd, J ₁ =12.1Hz, J ₂ =4.7Hz), 3.35-3.41 (m, 3H), 6.82-6.89 (m, 1H _The ^_ ,45.99,53.24,58.15,58.24,64.54,107.79,108.14,111.11,111.40,116.32,116.36,121.04,128.41,131.05,131.17,141.61,144.76,148.73,162.59,165.80,174.HRMS(ESI):m/ z[M+H] ⁺ calcd for C ₂₁ H ₃₁ FN ₃ O: 360.2446; found: 360.2451.

Example 26: 4-((1R,3aS,3a1S,10aR)-decahydro-1H,4H-pyrido[1,6]naphthyridin-1-yl)-N-(3-chlorophenyl)butanamide Preparation of (7b)

Other conditions were the same as in Example 25, except that compound 6a was replaced by compound 6b, and the yield was 97%. ¹ H NMR (400 MHz, CD ₃ OD): δ 1.44-1.72 (m, 9H), 1.75-1.88 (m, 4H), 1.92-1.97 (m, 1H), 2.04-2.06 (d, 3H), 2.27 (s,1H),2.46-2.49(t,2H,J=6.5Hz),2.80-2.86(m,2H),2.99-3.03(dd,1H,J1 ₌ 12.3Hz,J2= _4.2Hz ), 3.42-3.49(t,1H,J=12.8Hz),3.59-3.63(m,1H),7.05-7.07(dd,1H,J1 ₌ 7.9Hz,J2=1.2Hz ₎ ,7.24-7.28(t, 1H, J=8.1Hz), 7.39-7.41 (m, 1H), 7.78-7.79 (t, 1H, J=1.9Hz). ¹³ C NMR (100MHz, CD ₃ OD): δ 20.84, 21.17, 26.66, 27.64,30.62,34.39,36.91,39.26,44.97,54.12,57.74,57.81,63.12,116.52,119.05,119.43,120.86,124.81,131.04,135.28,141.22,173.75.HRMS(ESI): ] ⁺ calcd for C ₂₁ H ₃₁ ClN ₃ O: 376.2150; found: 376.2155.

Example 27: 4-((1R,3aS,3a1S,10aR)-decahydro-1H,4H-pyrido[1,6]naphthyridin-1-yl)-N-(3-bromophenyl)butanamide Preparation of (7c)

Other conditions were the same as in Example 25, except that compound 6a was replaced by compound 6c, and the yield was 95%. ¹ H NMR (400MHz, d ₆ -DMSO): δ1.35(s, 3H), 1.43-1.52(m, 5H), 1.71-1.83(m, 7H), 1.98-2.01(d, 2H), 2.31- 2.39(m, 2H), 2.68-2.71(d, 2H), 2.87-2.90(d, 1H), 3.22-3.28(t, 1H, J=3.1Hz), 3.44-3.48(m, 3H), 7.17- 7.25(m, 2H), 7.49-7.51(d. 1H), 7.97(s, 1H), 10.31(s, 1H). ¹³ C NMR (100MHz, CD ₃ OD): δ 20.17, 21.08, 26.51, 27.56 ,29.81,30.91,35.39,36.81,40.25,45.25,52.22,57.20,57.29,63.16,118.73,122.50,123.13,127.08,130.29,139.74,172.55.HRMS(ESIcalc):m/z[M ⁺ H] for C ₂₁ H ₃₁ BrN ₃ O: 420.1645; found: 420.1640.

Example 28: 4-((1R,3aS,3a1S,10aR)-decahydro-1H,4H-pyrido[1,6]naphthyridin-1-yl)-N-(3-methoxyphenyl) Preparation of butanamide (7d)

Other conditions were the same as in Example 25, except that compound 6a was replaced by compound 6d, and the yield was 98%. ¹ H NMR (400MHz, CD ₃ OD): δ 1.19-1.31 (m, 1H), 1.41-1.49 (m, 3H), 1.51-1.55 (m, 1H), 1.59-1.62 (m, 3H), 1.66 -1.70(m, 2H), 1.73-1.87(m, 3H), 1.92-2.04(m, 4H), 2.20(s, 1H), 2.29(s, 3H), 2.42-2.45(m, 2H), 2.76 -2.88(m, 4H), 3.34-3.35(d, 1H), 3.40-3.45(m, 1H), 6.87-6.90(d, 1H), 7.14-7.18(t, 1H, J=2.0Hz), 7.30 -7.38(m, 2H). ¹³ C NMR (100MHz, d ₆ -DMSO): δ19.73, 19.98, 20.29, 25.51, 26.44, 29.08, 32.43, 36.02, 52.39, 55.01, 56.37, 56.46, 61.56, 105.11, 108.49, 111.56, 115.72, 118.69, 129.53, 140.60, 171.05. HRMS(ESI): m/z[M+H] ⁺ calcd for C ₂₂ H ₃₄ N ₃ O ₂ : 372.2646; found: 372.2651.

Example 29: 4-((1R,3aS,3a1S,10aR)-decahydro-1H,4H-pyrido[1,6]naphthyridin-1-yl)-N-(3-methylphenyl)butane Preparation of Amide (7e)

Other conditions were the same as in Example 25, except that compound 6a was replaced by compound 6e, and the yield was 95%. ¹ H NMR (400MHz, d ₆ -DMSO): δ 1.35-1.39 (d, 3H), 1.45-1.56 (m, 5H), 1.66-1.76 (m, 4H), 1.80-1.87 (m, 3H), 1.99-2.01(d, 2H), 2.28-2.40(m, 2H), 2.72(s, 2H), 2.87-2.91(m, 1H), 3.23-3.28(t, 2H, J=2.5Hz), 3.70( s, 3H), 6.58-6.60(m, 1H), 7.13-71.8(m, 2H), 7.30-7.31(d, 1H). ¹³ C NMR (100MHz, CD ₃ OD): δ 21.50, 21.58, 21.79 ,27.01,28.13,31.52,35.51,37.31,40.33,45.64,53.57,58.01,58.10,64.02,118.39,121.81,129.59,139.65,173.88.HRMS(ESI):m/z[M+H] ⁺ calcd for C _22H34N3O : _356.2696 ; found: _356.2696 .

Example 30: 4-((1R,3aS,3a1S,10aR)-decahydro-1H,4H-pyrido[1,6]naphthyridin-1-yl)-N-(3-nitrophenyl)butane Preparation of amide (7f)

Other conditions were the same as in Example 25, except that compound 6a was replaced by compound 6f, and the yield was 94%. ¹ H NMR (400MHz, CD ₃ OD): δ 1.52-1.67 (m, 9H), 1.82-1.94 (m, 5H), 2.19 (s, 3H), 2.51 (s, 3H), 2.93 (s, 2H) ),3.04-3.07(m,1H),3.44-3.52(m,1H),3.63-3.68(m,1H),7.46-7.50(t,1H,J＝2.0Hz),7.85-7.90(t,2H) , J＝1.4Hz), 8.61(s, 1H). ¹³ C NMR (100MHz, CD ₃ OD): δ20.87, 30.46, 33.87, 36.97, 38.79, 44.69, 49.84, 53.89, 57.43, 63.05, 113.55, 115.43 , 116.46, 119.35, 122.26, 126.77, 130.68, 149.48, 174.30. HRMS(ESI): m/z[M+H] ⁺ calcd for C ₂₁ H ₃₁ N ₄ O ₃ : 387.2391; found: 387.2401.

Example 31: 4-((1R,3aS,3a1S,10aR)-decahydro-1H,4H-pyrido[1,6]naphthyridin-1-yl)-N-(pyridin-2-yl)butanamide Preparation of (7g)

Other conditions were the same as in Example 25, except that compound 6a was replaced by compound 6g, and the yield was 90%. ¹ H NMR (400 MHz, d ₆ -DMSO): δ 1.34-1.37 (dd, 4H, J ₁ =6.0 Hz, J ₂ =5.5 Hz), 1.50-1.56 (m, 6H), 1.63-1.89 (m, 8H), 2.02(s, 2H), 2.38-2.46(m, 2H), 2.73-2.79(d, 2H), 2.89-2.93(t, 1H, J＝10.0Hz), 7.04-7.07(m, 1H) , 7.72-7.76 (m, 1H), 8.05-8.07 (d, 1H), 8.27-8.28 (dd, 1H, J ₁ =3.8Hz, J ₂ =0.9Hz). ¹³ C NMR (100MHz, CD ₃ OD) :δ26.46,28.01,29.90,35.95,38.54,41.85,45.27,46.60,52.89,61.82,63.76,65.83,71.06,75.52,125.57,128.84,130.32,147.68,157.63),161. z[M+H] ⁺ calcd for C ₂₀ H ₃₁ N ₄ O: 343.2492; found: 343.2494.

Example 32: 4-((1R,3aS,3a1S,10aR)-decahydro-1H,4H-pyrido[1,6]naphthyridin-1-yl)-N-(naphthalen-2-yl)butanamide Preparation of (7h)

Other conditions were the same as in Example 25, except that compound 6a was replaced by compound 6h, and the yield was 94%. ¹ H NMR (400MHz, CD ₃ OD): δ 1.13 (s, 1H), 1.32-1.49 (m, 8H), 1.66-1.89 (m, 8H), 2.09 (s, 1H), 2.63-2.69 (m ,2H),2.85-2.87(d,1H),3.17-3.28(m,1H),3.48(s,1H),7.24-7.30(dd,2H,J ₁ =4.1Hz,J ₂ =1.8Hz), 7.41-7.43(d, 1H), 7.61-7.65(m, 3H), 8.08(s, 1H). ¹³ C NMR (100MHz, CD ₃ OD): δ 20.93, 21.21, 21.47, 26.71, 27.67, 30.71, 34.49,36.95,39.36,44.99,54.17,57.76,57.85,63.12,117.84,121.26,126.00,127.47,128.49,128.57,129.53.HRMS(ESI):m/z[M+H] ⁺ calcd for C ₂₅ H ₃₄ N ₃ O: 392.2696; found: 392.2694.

Example 33: Compound 6a (500 mg, 1.4 mmol) was dissolved in dichloromethane (10 mL) at room temperature, trifluoroacetic acid (2 mL) was added dropwise, and the mixture was stirred at room temperature for 2 h. After the reaction was completed, 10% sodium carbonate solution was added to adjust pH=7-8, filtered and concentrated, and the yield was 78%.

Example 34: Other conditions were the same as in Example 33, except that compound 6a was replaced by compound 6b, and the yield was 60%.

Example 35: Other conditions were the same as in Example 33, except that compound 6a was replaced by compound 6c, and the yield was 75%.

Example 36: Other conditions were the same as those of Example 33, except that compound 6a was replaced by compound 6d, and the yield was 45%.

Example 37: Other conditions were the same as those in Example 33, except that compound 6a was replaced by compound 6e, and the yield was 50%.

Example 38: Other conditions were the same as those in Example 33, except that compound 6a was replaced by compound 6f, and the yield was 47%.

Example 39: Other conditions were the same as those of Example 33, except that compound 6a was replaced by compound 6g, and the yield was 65%.

Example 40: Other conditions were the same as those in Example 33, except that compound 6a was replaced by compound 6h, and the yield was 60%.

Step 7: 4-((1R, 3aS, 3a1S, 10aR)-2-benzoyldecahydro-1H,4H-pyrido[1,6]naphthyridin-1-yl)-N-(m-tolyl) Preparation of butanamide (8a)

Example 41: Compound 7e (400 mg, 1.13 mmol) was dissolved in 10 mL of dichloromethane, triethylamine (117 μL) was added, then benzoyl chloride (156 μL) was added dropwise, and the mixture was stirred at room temperature for 1 h. Concentration and purification by column chromatography gave 450 mg of a white solid in 87% yield. ¹ H NMR (400 MHz, CDCl ₃ ): δ 1.16-1.19 (t, 1H, J=1.8 Hz), 1.36 (s, 1H), 1.44-1.53 (m, 4H), 1.58-1.70 (m, 3H) ,1.77-1.91(m,4H),2.11(s,1H),2.25(s,3H),2.29-2.34(t,2H,J＝2.5Hz),2.46-2.49(m,2H),2.98-3.03 (q, 1H, J=1.8Hz), 3.07-3.12 (t, 2H, J=2.5Hz), 3.41-3.46 (dd, 1H, J1 ₌ 3.5Hz, J2=1.7Hz), _3.61-3.67 ( m, 1H), 4.18(s, 1H), 6.84-6.86(s, 1H), 7.10-7.13(t, 1H, J=2.0Hz), 7.35-7.50(m, 7H). ¹³ C NMR(100MHz, CDCl ₃ ):δ1.14,8.67,19.96,21.59,22.31,26.82,27.09,35.58,36.39,38.46,46.66,56.31,63.98,117.04,120.58,124.72,127.23,128.31,128.809,128.31,128.809, 132.07, 136.66, 138.57, 138.69, 172.14, 173.53. HRMS(ESI): m/z[M+H] ⁺ calcd for C ₂₉ H ₃₈ N ₃ O ₂ : 460.2959; found: 460.2960.

Example 42: Other conditions were as in Example 41, the base was replaced with ethylenediamine, and the yield was 56%.

Example 43: Other conditions were as in Example 41, the base was replaced with potassium hydroxide, and the yield was 30%.

Step 8: 4-((3aS, 3a1S, 10aR)-2-benzyldecahydro-1H,4H-pyrido[1,6]naphthyridin-1-yl)-N-(m-tolyl)butanamide ( 8b) Preparation

Example 44: Compound 7e (604 mg, 1.7 mmol) and triethylamine (236 μL) were dissolved in 1,2-dichloroethane (7 mL), benzaldehyde (260 μL) was added dropwise, the reaction solution was refluxed for 2 h, Then sodium triacetoxyborohydride (540 mg) was slowly added to the reaction solution in batches, and the reflux was continued for 6 h. Cool to room temperature. Concentrated extraction. Extract 3 times with 20 mL of ethyl acetate, wash 3 times with 15 mL of water, and 3 times with 15 mL of saturated brine, pour the organic phase into a clean conical flask, add an appropriate amount of anhydrous sodium sulfate and dry for half an hour . After purification by column chromatography, a white solid was obtained in 78% yield. ¹ H NMR (400 MHz, CDCl ₃ ): δ 0.84-0.90 (m, 1H), 1.24-1.28 (m, 2H), 1.40-1.46 (m, 3H), 1.51-1.61 (m, 4H), 1.83- 2.13(m, 6H), 2.29(s, 3H), 2.41-2.50(m, 2H), 2.55-2.59(dd, 1H, J ₁ =3.4Hz, J ₂ =0.9Hz), 2.75(s, 1H) ,2.87-2.95(d,1H),3.23-3.42(d,3H),3.67-3.70(d,1H),3.85-3.92(d,1H),6.83-6.85(d,1H),7.11-7.15( t, 1H, J=1.9Hz), 7.25-7.33(m, 3H), 7.38-7.39(d, 2H), 7.55=7.57(d, 1H), 7.67(s, 1H). ¹³ C NMR(100MHz, CDCl ₃ ):δ14.22,18.98,19.51,20.36,21.67,22.78,25.64,26.19,29.45,29.78,31.53,32.01,35.49,36.60,56.15,117.09,120.58,124.44,128.70,8 139.00, 162.65, 172.17. HRMS(ESI): m/z[M+H] ⁺ calcd for C ₂₉ H ₄₀ N ₃ O: 446.3166; found: 446.3165.

Example 45: Other conditions were as in Example 44, the solvent was changed from 1,2-dichloroethane to dichloromethane, and the yield was 60%.

Example 46: Other conditions were as in Example 44, except that sodium triacetoxyborohydride was changed to sodium borohydride, and the yield was 65%.

2. Determination of anticancer activity

HepG2 (human liver cancer cells) and A549 (human lung cancer cells) cell lines were selected, and matrine derivatives were tested for in vitro anticancer activity by MTT method, and matrine was used as the control group. Cancer cells in the log phase growth phase were taken, centrifuged, diluted with RPM1640 medium to 1×10 ⁴ /mL, and inoculated in a 96-well plate. Incubate at 37°C for 24h, add 21 samples of gradient concentration, incubate for 72h, add 50μL 10% MTT solution, incubate at 37°C for 4h, add 100μL DMSO to each well. After shaking for 30min, the 96-well plate was placed on an automatic microplate spectrophotometer, and the absorbance value was measured at 570nm, and the half-effective inhibitory concentration (IC ₅₀ ) was calculated by the Biss method. Each group of samples was tested in parallel for 3 times. The results are shown in Table 1.

Table 1 Anticancer activity of matrine amide derivatives

^a Note: ND=not determined

The proliferation-inhibitory effect of the newly synthesized 19 compounds on HepG2 and A549 cancer cells was determined by MTT method. The results are shown in Table 1. The test results show that when the substituents on the benzene ring are electron-withdrawing groups, such as Cl, Br, NO ₂ , they have obvious anti-cancer effects on both types of cancer cells; and when they are electron-donating groups, CH ₃ , they also have significant anti-cancer effects on both types of cells. It has obvious anticancer activity. Among the 16-Boc matrine derivatives, when the substituents on the benzene ring are Cl, CH ₃ , NO ₂ , it has a significant inhibitory effect on A549 cancer cells; when the substituents on the benzene ring are F, Cl, NO ₂ , the The proliferation of HepG2 cells was inhibited. In 16-H matrine derivatives, when the substituent is naphthalene ring, it has a certain inhibitory effect on the proliferation of A549 cell line; when the group on the benzene ring is Br, CH ₃ , it has inhibitory effect on A549 and HepG2 cells. Comparing the above synthesized derivatives with matrine, it shows that when the substituent on the benzene ring is an electron withdrawing group, its inhibitory effect on the proliferation of cancer cells can be improved.

Claims (1)

1. The application of the matrine amide derivative in preparing the medicine for inhibiting the lung cancer cell proliferation is characterized in that: the matrine amide derivative is one of the following structural formulas:

。