CN112939991B - Polysubstituted spiro tetrahydrocarbazole and derivative thereof, and synthesis method and application thereof - Google Patents

Abstract

The invention discloses a synthesis method of polysubstituted spiro tetrahydrocarbazole and derivatives thereof, which takes diazo compounds and substituted indolone as raw materials, rh ₂ (OAc) ₄ Is used as a catalyst and is dissolved in an organic solvent,

Description

A kind of multi-substituted spirotetrahydrocarbazole and its derivatives and its synthesis method and application

technical field

The invention belongs to the field of synthetic medicine and chemical industry, and specifically relates to a synthesis method and application of multi-substituted spirotetrahydrocarbazole and derivatives thereof.

Background technique

The tetrahydrocarbazole skeleton occupies an important position in natural products and active drugs. The existing synthetic methods are not perfect, the substrate synthesis is complicated, the reaction conditions are harsh, the energy consumption is high, the yield is low, and it is impossible to synthesize multi-substituted Tetrahydrocarbazole and its derivatives. It is an urgent scientific problem to develop a new method to synthesize tetrahydrocarbazole derivatives. ^[1-3]

Contents of the invention

The present invention overcomes the defects of low yield, complex substrate, harsh conditions, and large reaction energy consumption in the preparation method of multi-substituted spirotetrahydrocarbazole derivatives in the prior art. The invention provides a multi-substituted spirotetrahydrocarbazole derivative. A synthesis method of spirotetrahydrocarbazole and its derivatives, the method has high atom economy and step economy, good regioselectivity and diastereoselectivity, wide substrate applicability, mild reaction conditions and high yield , simple and safe operation and other beneficial effects. The multi-substituted spirotetrahydrocarbazole and derivatives thereof prepared by the invention can be used as important medicine and chemical intermediates, and have broad application prospects in the pharmaceutical field.

The present invention proposes a multi-substituted spirotetrahydrocarbazole and derivatives thereof, the structural formula of which is shown in formula (I):

In formula (I),

^R is alkyl, aryl, halogen substituted phenyl, alkyl substituted phenyl, alkoxy substituted phenyl, trifluoromethyl substituted phenyl, nitro substituted phenyl, ester, substituted the ester group;

^R is alkyl, benzyl, phenyl, Boc, Cbz, halogen-substituted benzyl, alkoxy-substituted benzyl, alkyl-substituted benzyl, trifluoromethyl-substituted benzyl, nitro-substituted Benzyl, halogen substituted phenyl, alkyl substituted phenyl, alkoxy substituted phenyl, trifluoromethyl substituted phenyl, nitro substituted phenyl, heterocyclic substituted phenyl, aromatic heterocycle Substituted phenyl;

R ³ is H, halogen, alkyl, alkoxy, nitro, trifluoromethyl;

^R4 is alkyl, benzyl, halogen-substituted benzyl, alkoxy-substituted benzyl, alkyl-substituted benzyl, trifluoromethyl-substituted benzyl, nitro-substituted benzyl, phenyl, halogen Substituted phenyl, alkyl substituted phenyl, alkoxy substituted phenyl, trifluoromethyl substituted phenyl, nitro substituted phenyl, heterocyclic substituted phenyl, aromatic heterocyclic substituted phenyl .

Preferably,

R ¹ is C1-C10 alkyl, phenyl, halogen substituted phenyl, C1-C10 alkyl substituted phenyl, C1-C10 alkoxy substituted phenyl, trifluoromethyl substituted phenyl, nitro Substituted phenyl groups, ester groups, substituted ester groups;

R ² is C1-C10 alkyl, benzyl, phenyl, Boc, Cbz, halogen substituted benzyl, C1-C10 alkoxy substituted benzyl, C1-C10 alkyl substituted benzyl, trifluoromethyl Substituted benzyl, nitro-substituted benzyl, halogen-substituted phenyl, C1-C10 alkyl-substituted phenyl, C1-C10 alkoxy-substituted phenyl, trifluoromethyl-substituted phenyl, nitro Substituted phenyl, heterocyclic substituted phenyl, aromatic heterocyclic substituted phenyl;

R ³ is H, F, Cl, Br, C1-C10 alkyl, C1-C10 alkoxy, nitro, trifluoromethyl;

R ⁴ is C1-C10 alkyl, benzyl, bromo-substituted benzyl, C1-C10 alkoxy-substituted benzyl, C1-C10 alkyl-substituted benzyl, trifluoromethyl-substituted benzyl, nitro Substituted benzyl, phenyl, halogen substituted phenyl, C1-C10 alkyl substituted phenyl, C1-C10 alkoxy substituted phenyl, trifluoromethyl substituted phenyl, nitro substituted phenyl , Heterocyclic substituted phenyl, aromatic heterocyclic substituted phenyl.

Further preferably, R ^is methyl, n-butyl, phenyl, phenyl substituted with p-methoxy, phenyl substituted with trifluoromethyl;

R ² is methyl, benzyl;

R ³ is H, F, Cl, Br, methyl, methoxy;

^R4 is methyl, benzyl, 4-bromobenzyl, 4-methoxybenzyl and the like.

In the present invention, the "ester group, substituted ester group" includes methyl ester group, ethyl ester group, isopropyl ester group, phenyl ester group, halogen substituted phenyl ester group.

In the present invention, the "heterocyclic substituted phenyl" includes three-membered heterocyclic substituted phenyl, four-membered heterocyclic substituted phenyl, five-membered heterocyclic substituted phenyl, six-membered heterocyclic substituted phenyl .

In the present invention, the "phenyl heterocyclic ring substituted" includes phenyl substituted by furan, phenyl substituted by pyridine and phenyl substituted by pyrrole.

分子筛为吸水的添加剂，反应得到式(I)所示的多取代螺四氢咔唑及其衍生物；所述合成反应如反应式(II)所示：The present invention also proposes a synthetic method of multi-substituted spirotetrahydrocarbazoles and derivatives thereof, using formula (1) diazo compounds, formula (2) substituted indolones as raw materials, and Rh ₂ (OAc) ₄ as catalyst , in an organic solvent, 4

Molecular sieve is the additive of water absorption, and reaction obtains many replacement spirotetrahydrocarbazoles and derivatives thereof shown in formula (I); Described synthetic reaction is as shown in reaction formula (II):

In formula (II),

^R is alkyl, aryl, halogen substituted phenyl, alkyl substituted phenyl, alkoxy substituted phenyl, trifluoromethyl substituted phenyl, nitro substituted phenyl, ester, substituted the ester group;

^R is alkyl, benzyl, phenyl, Boc, Cbz, halogen-substituted benzyl, alkoxy-substituted benzyl, alkyl-substituted benzyl, trifluoromethyl-substituted benzyl, nitro-substituted Benzyl, halogen substituted phenyl, alkyl substituted phenyl, alkoxy substituted phenyl, trifluoromethyl substituted phenyl, nitro substituted phenyl, heterocyclic substituted phenyl, aromatic heterocycle Substituted phenyl;

R ³ is H, halogen, alkyl, alkoxy, nitro, trifluoromethyl;

^R4 is alkyl, benzyl, halogen-substituted benzyl, alkoxy-substituted benzyl, alkyl-substituted benzyl, trifluoromethyl-substituted benzyl, nitro-substituted benzyl, phenyl, halogen Substituted phenyl, alkyl substituted phenyl, alkoxy substituted phenyl, trifluoromethyl substituted phenyl, nitro substituted phenyl, heterocyclic substituted phenyl, aromatic heterocyclic substituted phenyl .

Preferably,

R ¹ is C1-C10 alkyl, phenyl, halogen substituted phenyl, C1-C10 alkyl substituted phenyl, C1-C10 alkoxy substituted phenyl, trifluoromethyl substituted phenyl, nitro Substituted phenyl groups, ester groups, substituted ester groups;

R ² is C1-C10 alkyl, benzyl, phenyl, Boc, Cbz, halogen substituted benzyl, C1-C10 alkoxy substituted benzyl, C1-C10 alkyl substituted benzyl, trifluoromethyl Substituted benzyl, nitro-substituted benzyl, halogen-substituted phenyl, C1-C10 alkyl-substituted phenyl, C1-C10 alkoxy-substituted phenyl, trifluoromethyl-substituted phenyl, nitro Substituted phenyl, heterocyclic substituted phenyl, aromatic heterocyclic substituted phenyl;

R ³ is H, F, Cl, Br, C1-C10 alkyl, C1-C10 alkoxy, nitro, trifluoromethyl;

R ⁴ is C1-C10 alkyl, benzyl, bromo-substituted benzyl, C1-C10 alkoxy-substituted benzyl, C1-C10 alkyl-substituted benzyl, trifluoromethyl-substituted benzyl, nitro Substituted benzyl, phenyl, halogen substituted phenyl, C1-C10 alkyl substituted phenyl, C1-C10 alkoxy substituted phenyl, trifluoromethyl substituted phenyl, nitro substituted phenyl , Heterocyclic substituted phenyl, aromatic heterocyclic substituted phenyl.

Further preferably, R ^is methyl, n-butyl, phenyl, phenyl substituted with p-methoxy, phenyl substituted with trifluoromethyl;

R ² is methyl, benzyl;

R ³ is H, F, Cl, Br, methyl, methoxy;

R ⁴ is methyl, benzyl, 4-bromobenzyl, 4-methoxybenzyl.

In the present invention, the "ester group, substituted ester group" includes methyl ester group, ethyl ester group, isopropyl ester group, phenyl ester group, halogen substituted phenyl ester group.

In the present invention, the "heterocyclic substituted phenyl" includes three-membered heterocyclic substituted phenyl, four-membered heterocyclic substituted phenyl, five-membered heterocyclic substituted phenyl, six-membered heterocyclic substituted phenyl .

In the present invention, the "phenyl heterocyclic ring substituted" includes phenyl substituted by furan, phenyl substituted by pyridine and phenyl substituted by pyrrole.

In the synthetic reaction of the present invention,

Wherein, the molar ratio of the diazo compound of the formula (1) and the substituted indolinone of the formula (2) is 1.2:1-2.0:1; preferably, it is 2.0:1.0.

Wherein, the amount of the catalyst Rh ₂ (OAc) ₄ is based on the amount of the substituted indolinone, which is 0.5mol%-2.0mol% of the substituted indolinone; preferably, it is 1.0mol%.

分子筛的质量以取代吲哚酮的用量为基准，为取代吲哚酮质量的50％-200％；优选地，为100％。Among them, the additive 4

The quality of the molecular sieve is based on the amount of the substituted indolinone, which is 50%-200% of the mass of the substituted indolinone; preferably, it is 100%.

Wherein, the organic solvent is selected from one or more of DCM, DCE, EA, and toluene; preferably, it is DCM.

Wherein, the amount of the organic solvent is based on the substrate (2) substituted indolinone, so that the concentration range of the substituted indolinone in the organic solvent is 0.05M-0.15M; preferably, the substituted indolinone in the organic solvent Medium concentration range is 0.10M.

Wherein, the reaction temperature is 0°C-40°C; preferably, 25°C.

Wherein, the reaction time is 1h-5h; preferably, 2h.

分子筛溶于有机溶剂中，在25℃下，加入所述重氮化合物的有机溶剂溶液，反应得到式(I)所示的化合物。Wherein, the method specifically includes the following steps, firstly substituted indolinone, Rh ₂ (OAc) ₄ , 4

Molecular sieves are dissolved in an organic solvent, and the organic solvent solution of the diazo compound is added at 25° C. to react to obtain a compound represented by formula (I).

Wherein, the organic solvent in the organic solvent solution of the diazo compound is one or more of DCM, DCE, EA, toluene, etc.; preferably, it is DCM.

Wherein, the step of separating and purifying the compound obtained from the reaction is further included; the separating and purifying is to perform column chromatography with a solution whose volume ratio is ethyl acetate:petroleum ether=1:20～1:3.

The chemical mechanism involved in the synthetic method of the present invention is shown in the following formula (III):

Wherein, the definitions of R ¹ , R ² , R ³ and R ⁴ are the same as those in formula (II).

The invention also proposes the application of the multi-substituted spirotetrahydrocarbazole and its derivatives in the preparation of anti-tumor cell active drugs.

The tumor cells are one or more of MG63 type osteosarcoma cells, MNNG/HOS type osteosarcoma cells, HOS type osteosarcoma cells, and 143B type osteosarcoma cells.

The beneficial effect of the present invention is that: the present invention provides a kind of synthesis method of multi-substituted spirotetrahydrocarbazole and its derivatives, the method has high atom economy and step economy, regioselectivity and diastereoselectivity Good (dr>85:15), wide substrate applicability, mild reaction conditions, high yield (83%-99%), simple and safe operation and other beneficial effects provide a new way for efficient construction of such compounds. The multi-substituted spirotetrahydrocarbazole and derivatives thereof prepared by the invention can be used as important medicine and chemical intermediates, and have broad application prospects in the pharmaceutical field.

Description of drawings

FIG. 1 is a schematic diagram of ¹ H NMR of the product obtained in Example 1.

FIG. 2 is a schematic diagram of ¹³ C NMR of the product obtained in Example 1.

FIG. 3 is a schematic diagram of ¹ H NMR of the product obtained in Example 2.

FIG. 4 is a schematic diagram of ¹³ C NMR of the product obtained in Example 2.

FIG. 5 is a schematic diagram of ¹ H NMR of the product obtained in Example 3.

FIG. 6 is a schematic diagram of ¹³ C NMR of the product obtained in Example 3.

FIG. 7 is a schematic diagram of ¹ H NMR of the product obtained in Example 4.

FIG. 8 is a schematic diagram of ¹³ C NMR of the product obtained in Example 4.

FIG. 9 is a schematic diagram of ¹ H NMR of the product obtained in Example 5.

FIG. 10 is a schematic diagram of ¹³ C NMR of the product obtained in Example 5.

FIG. 11 is a schematic diagram of ¹ H NMR of the product obtained in Example 6.

FIG. 12 is a schematic diagram of ¹³ C NMR of the product obtained in Example 6.

FIG. 13 is a schematic diagram of ¹ H NMR of the product obtained in Example 7.

FIG. 14 is a schematic diagram of ¹³ C NMR of the product obtained in Example 7.

Fig. 15 is a schematic diagram of ¹ H NMR of the product obtained in Example 8.

FIG. 16 is a schematic diagram of ¹³ C NMR of the product obtained in Example 8.

Fig. 17 is a schematic diagram of ¹⁹ F NMR of the product obtained in Example 8.

Figure 18 is a schematic diagram of ¹ H NMR of the product obtained in Example 9

FIG. 19 is a schematic diagram of ¹³ C NMR of the product obtained in Example 9.

20 is a schematic diagram of ¹⁹ F NMR of the product obtained in Example 9.

Fig. 21 is a schematic diagram of ¹ H NMR of the product obtained in Example 10.

22 is a schematic diagram of ¹³ C NMR of the product obtained in Example 10.

Fig. 23 is a schematic diagram of ¹ H NMR of the product obtained in Example 11.

Fig. 24 is a schematic diagram of ¹³ C NMR of the product obtained in Example 11.

Detailed ways

The present invention will be described in further detail in conjunction with the following specific examples and accompanying drawings, and the protection content of the present invention is not limited to the following examples. Without departing from the spirit and scope of the inventive concept, changes and advantages conceivable by those skilled in the art are all included in the present invention, and the appended claims are the protection scope.

Example 1

3-Hydroxy-1',3,9-trimethyl-1,2,3,9-tetrahydrospiro[carbazole-4,3'-indoline]-2'-one

分子筛(100％)和Rh₂(OAc)₄(1.0mol％)溶解在装有1.0mL二氯甲烷的试管中，于特定温度(25℃)下搅拌30分钟，再将溶有重氮(0.4mmol)的二氯甲烷(1.0mL)溶液通过蠕动泵加入到前述混合溶液中，一小时加完。将混合物进行柱层析纯化(乙酸乙酯：石油醚＝1:20～1:3)得到一对非对映异构体，结构如上。合计产率：99％，dr>20:1。The substituted indolinone (0.2mmol), 4

Molecular sieves (100%) and Rh ₂ (OAc) ₄ (1.0mol%) were dissolved in a test tube containing 1.0 mL of dichloromethane, stirred at a specific temperature (25° C.) for 30 minutes, and dissolved diazo (0.4 mmol) in dichloromethane (1.0 mL) was added to the aforementioned mixed solution through a peristaltic pump, and the addition was completed in one hour. The mixture was purified by column chromatography (ethyl acetate:petroleum ether=1:20~1:3) to obtain a pair of diastereoisomers with the above structures. Total yield: 99%, dr>20:1.

¹ H NMR (400MHz, Chloroform-d) δ7.32–7.26(m,1H),7.21(d,J=8.2Hz,1H),7.08–6.92(m,4H),6.83(t,J=7.6Hz ,1H),6.58(d,J=8.0Hz,1H),4.59(s,1H),3.68(s,3H),3.47(s,3H),3.22(dt,J=16.7,8.7Hz,1H) ,2.89(d,J＝16.6Hz,1H),2.40–2.27(m,2H),1.02(s,3H).

¹³ C NMR(100MHz,Chloroform-d)δ179.72,143.25,137.28,136.66,132.71,128.12,125.16,124.97,123.11,120.82,119.18,117.10,108.89,108.32,72.95,32.41,29.29,26.53,25.09,19.16.

HRMS (ESI) Calcd. for C ₂₂ H ₂₂ N ₂ O ₂ Na (M+Na) ⁺ 369.1588, found: 369.1579.

Example 2

1'-Benzyl-3-hydroxy-3,9-dimethyl-1,2,3,9-spirotetrahydro[carbazole-4,3'-indoline]-2'-one

分子筛(100％)和Rh₂(OAc)₄(1.0mol％)溶解在装有1.0mL二氯甲烷的试管中，于特定温度(25℃)下搅拌30分钟，再将溶有重氮(0.4mmol)的二氯甲烷(1.0mL)溶液通过蠕动泵加入到前述混合溶液中，一小时加完。将混合物进行柱层析纯化(乙酸乙酯：石油醚＝1:20～1:3)得到一对非对映异构体，结构如上。合计产率：92％，dr>20:1。The substituted indolinone (0.2mmol), 4

Molecular sieves (100%) and Rh ₂ (OAc) ₄ (1.0mol%) were dissolved in a test tube containing 1.0 mL of dichloromethane, stirred at a specific temperature (25° C.) for 30 minutes, and dissolved diazo (0.4 mmol) in dichloromethane (1.0 mL) was added to the aforementioned mixed solution through a peristaltic pump, and the addition was completed in one hour. The mixture was purified by column chromatography (ethyl acetate:petroleum ether=1:20~1:3) to obtain a pair of diastereoisomers with the above structures. Total yield: 92%, dr>20:1.

¹ H NMR (400MHz, CDCl3) 7.50(d, J=7.4Hz, 2H), 7.41–7.29(m, 3H), 7.20(d, J=8.2Hz, 2H), 7.08–7.00(m, 2H), 6.94(dd, J=21.8,7.8Hz,2H),6.72(t,J=7.7Hz,1H),6.42(d,J=8.0Hz,1H),5.24(d,J=15.3Hz,1H), 5.02(d,J=15.2Hz,1H),4.31(s,1H),3.68(s,2H),3.29–3.16(m,1H),2.89(d,J=17.0Hz,1H),2.50(d ,J＝14.1Hz,1H),2.33–2.21(m,2H),1.10(s,3H).

¹³ C NMR(100MHz,CDCl3)δ179.69,142.72,137.29,136.68,135.76,132.40,128.85,128.11,128.07,127.90,125.40,125.17,122.92,120.78,119.02,117.56,109.24,108.82,106.78,99.98,77.34, 77.03, 76.71, 73.32, 56.17, 44.38, 32.51, 29.30, 25.27, 19.35.

HRMS (ESI) Calcd. for C ₂₈ H ₂₇ N ₂ O ₂ (M+H) ⁺ 423.2079, found: 423.2073.

Example 3

1'-(4-bromobenzyl)-3-hydroxy-3,9-dimethyl-1,2,3,9-spirotetrahydro[carbazole-4,3'-indoline]-2' -ketone

分子筛(100％)和Rh₂(OAc)₄(1.0mol％)溶解在装有1.0mL二氯甲烷的试管中，于特定温度(25℃)下搅拌30分钟，再将溶有重氮(0.4mmol)的二氯甲烷(1.0mL)溶液通过蠕动泵加入到前述混合溶液中，一小时加完。将混合物进行柱层析纯化(乙酸乙酯：石油醚＝1:20～1:3)得到一对非对映异构体，结构如上。合计产率：90％，dr＝86:14。The substituted indolinone (0.2mmol), 4

Molecular sieves (100%) and Rh ₂ (OAc) ₄ (1.0mol%) were dissolved in a test tube containing 1.0 mL of dichloromethane, stirred at a specific temperature (25° C.) for 30 minutes, and dissolved diazo (0.4 mmol) in dichloromethane (1.0 mL) was added to the aforementioned mixed solution through a peristaltic pump, and the addition was completed in one hour. The mixture was purified by column chromatography (ethyl acetate:petroleum ether=1:20~1:3) to obtain a pair of diastereoisomers with the above structures. Total yield: 90%, dr=86:14.

¹ H NMR (400MHz, Chloroform-d) δ7.53–7.46(m,2H),7.37(d,J=7.9Hz,2H),7.24–7.18(m,2H),7.08–7.02(m,2H) ,6.97–6.89(m,2H),6.72(t,J=7.7Hz,1H),6.32(d,J=8.0Hz,1H),5.15(d,J=15.4Hz,1H),4.97(d, J＝15.3Hz, 1H), 4.06(s, 1H), 3.68(s, 3H), 3.21(dt, J＝16.0, 7.3Hz, 1H), 2.94–2.85(m, 1H), 2.59–2.50(m ,1H),2.24(dt,J=14.1,7.1Hz,1H),1.11(s,3H).

¹³ C NMR(100MHz,Chloroform-d)δ179.66,142.67,136.68,129.59,128.02,127.86,125.30,122.89,120.77,118.98,117.57,114.18,109.22,108.79,73.24,56.05,55.31,43.79,32.51,29.29, 25.24, 19.29.

HRMS (ESI) Calcd. for C ₂₈ H ₂₅ N ₂ O ₂ NaBr (M+Na) ⁺ 523.1001, found: 523.0997.

Example 4

3-Hydroxy-1'-(4-methoxybenzyl)-3,9-dimethyl-1,2,3,9-spirotetrahydro[carbazole-4,3'-indoline]-2 '-ketone

分子筛(100％)和Rh₂(OAc)₄(1.0mol％)溶解在装有1.0mL二氯甲烷的试管中，于特定温度(25℃)下搅拌30分钟，再将溶有重氮(0.4mmol)的二氯甲烷(1.0mL)溶液通过蠕动泵加入到前述混合溶液中，一小时加完。将混合物进行柱层析纯化(乙酸乙酯：石油醚＝1:20～1:3)得到一对非对映异构体，结构如上。合计产率：99％，dr＝85:15。The substituted indolinone (0.2mmol), 4

Molecular sieves (100%) and Rh ₂ (OAc) ₄ (1.0mol%) were dissolved in a test tube containing 1.0 mL of dichloromethane, stirred at a specific temperature (25° C.) for 30 minutes, and dissolved diazo (0.4 mmol) in dichloromethane (1.0 mL) was added to the aforementioned mixed solution through a peristaltic pump, and the addition was completed in one hour. The mixture was purified by column chromatography (ethyl acetate:petroleum ether=1:20~1:3) to obtain a pair of diastereoisomers with the above structures. Total yield: 99%, dr=85:15.

¹ H NMR (400MHz, Chloroform-d) δ7.44 (d, J=8.1Hz, 2H), 7.23–7.17 (m, 2H), 7.07–6.98 (m, 3H), 6.94–6.87 (m, 3H) ,6.71(t,J=7.7Hz,1H),6.40(d,J=8.0Hz,1H),5.15(d,J=15.1Hz,1H),4.98(d,J=15.1Hz,1H),4.41 (s,1H),3.81(s,3H),3.68(s,3H),3.22(dt,J=15.5,7.5Hz,1H),2.93–2.84(m,1H),2.46(d,J=13.9 Hz,1H),2.33–2.21(m,1H),1.07(s,3H).

¹³ C NMR(100MHz,CDCl3)δ179.66,159.27,142.67,136.68,129.59,128.02,127.86,125.30,125.18,122.89,120.77,118.98,117.57,114.18,109.22,108.79,77.34,77.03,76.71,73.24,56.05, 55.31, 43.79, 32.51, 29.29, 25.24, 19.29.

HRMS(ESI)Calcd.for:C ₂₉ H ₂₉ N ₂ O ₃ (M+H) ⁺ 453.2175, found: 453.2178.

Example 5

1'-Benzyl-3-hydroxy-5'-methoxy-3,9-dimethyl-1,2,3,9-spirotetrahydro[carbazole-4,3'-indoline]- 2'-keto

分子筛(100％)和Rh₂(OAc)₄(1.0mol％)溶解在装有1.0mL二氯甲烷的试管中，于特定温度(25℃)下搅拌30分钟，再将溶有重氮(0.4mmol)的二氯甲烷(1.0mL)溶液通过蠕动泵加入到前述混合溶液中，一小时加完。将混合物进行柱层析纯化(乙酸乙酯：石油醚＝1:20～1:3)得到一对非对映异构体，结构如上。合计产率：89％，dr>20:1。The substituted indolinone (0.2mmol), 4

Molecular sieves (100%) and Rh ₂ (OAc) ₄ (1.0mol%) were dissolved in a test tube containing 1.0 mL of dichloromethane, stirred at a specific temperature (25° C.) for 30 minutes, and dissolved diazo (0.4 mmol) in dichloromethane (1.0 mL) was added to the aforementioned mixed solution through a peristaltic pump, and the addition was completed in one hour. The mixture was purified by column chromatography (ethyl acetate:petroleum ether=1:20~1:3) to obtain a pair of diastereoisomers with the above structures. Total yield: 89%, dr>20:1.

¹ H NMR (400MHz, Chloroform-d) δ7.50 (d, J = 7.4Hz, 2H), 7.42–7.29 (m, 3H), 7.21 (d, J = 8.1Hz, 1H), 7.04 (t, J ＝7.8Hz, 1H), 6.86(d, J＝8.6Hz, 1H), 6.77–6.67(m, 2H), 6.64(s, 1H), 6.49(d, J＝8.0Hz, 1H), 5.23(d ,J=15.2Hz,1H),5.00(d,J=15.3Hz,1H),4.59(s,1H),3.68(d,J=2.2Hz,3H),3.64(d,J=2.2Hz,3H ),3.22(dt,J＝16.6,8.0Hz,1H),2.93–2.83(m,1H),2.46–2.36(m,1H),2.35–2.22(m,1H),1.09(s,3H).

¹³ C NMR(100MHz,CDCl3)δ179.42,156.12,137.28,136.66,136.11,135.80,134.09,128.85,128.16,127.91,125.21,120.78,119.04,117.53,113.21,111.81,109.47,108.81,106.79,77.35,77.03, 76.71, 73.14, 56.31, 55.62, 44.50, 32.43, 29.29, 25.31, 19.20.

HRMS(ESI)Calcd.for:C ₂₉ H ₂₉ N ₂ O ₃ (M+H) ⁺ 453.2175, found: 453.2178.

Example 6

9-Phenyl-3-hydroxy-1'-methyl-3-phenyl-1,2,3,9-spirotetrahydro[carbazole-4,3'-indoline]-2'-one

分子筛(100％)和Rh₂(OAc)₄(1.0mol％)溶解在装有1.0mL二氯甲烷的试管中，于特定温度(25℃)下搅拌30分钟，再将溶有重氮(0.4mmol)的二氯甲烷(1.0mL)溶液通过蠕动泵加入到前述混合溶液中，一小时加完。将混合物进行柱层析纯化(乙酸乙酯：石油醚＝1:20～1:3)得到一对非对映异构体，结构如上。合计产率：87％，dr>20:1。The substituted indolinone (0.2mmol), 4

Molecular sieves (100%) and Rh ₂ (OAc) ₄ (1.0mol%) were dissolved in a test tube containing 1.0 mL of dichloromethane, stirred at a specific temperature (25° C.) for 30 minutes, and dissolved diazo (0.4 mmol) in dichloromethane (1.0 mL) was added to the aforementioned mixed solution through a peristaltic pump, and the addition was completed in one hour. The mixture was purified by column chromatography (ethyl acetate:petroleum ether=1:20~1:3) to obtain a pair of diastereoisomers with the above structures. Total yield: 87%, dr>20:1.

¹ H NMR (400MHz, Chloroform-d) δ7.45 (d, J = 7.5Hz, 1H), 7.32 (q, J = 8.5, 7.9Hz, 4H), 7.20 (d, J = 8.3Hz, 1H), 7.15–7.03(m,8H),7.00(t,J=7.7Hz,1H),6.72(t,J=7.6Hz,1H),6.60(d,J=7.8Hz,1H),6.10(d,J ＝8.0Hz, 1H), 5.39(dd, J＝3.8Hz, 2H), 3.87(q, J＝9.4Hz, 1H), 3.17–3.08(m, 2H), 2.76(s, 3H), 2.62(s ,1H),2.13(d,J=13.4Hz,1H).

¹³ C NMR(100MHz,CDCl3)δ176.43,144.05,141.15,137.54,137.31,136.89,128.92,128.52,127.44,127.40,127.28,126.92,126.18,125.89,122.06,121.22,119.25,118.58,109.31,107.29,106.73, 77.33,77.22,77.02,76.70,75.87,59.18,46.66,29.48,25.86,18.84,0.01.

HRMS (ESI) Calcd. for C ₃₃ H ₂₉ N ₂ O ₂ (M+H) ⁺ 485.2250, found: 485.2229.

Example 7

3-n-Butyl-3-hydroxy-1',9-methyl-1,2,3,9-spirotetrahydro[carbazole-4,3'-indoline]-2'-one

分子筛(100％)和Rh₂(OAc)₄(1.0mol％)溶解在装有1.0mL二氯甲烷的试管中，于特定温度(25℃)下搅拌30分钟，再将溶有重氮(0.4mmol)的二氯甲烷(1.0mL)溶液通过蠕动泵加入到前述混合溶液中，一小时加完。将混合物进行柱层析纯化(乙酸乙酯：石油醚＝1:20～1:3)得到一对非对映异构体，结构如上。合计产率：91％，dr>20:1。The substituted indolinone (0.2mmol), 4

Molecular sieves (100%) and Rh ₂ (OAc) ₄ (1.0mol%) were dissolved in a test tube containing 1.0 mL of dichloromethane, stirred at a specific temperature (25° C.) for 30 minutes, and dissolved diazo (0.4 mmol) in dichloromethane (1.0 mL) was added to the aforementioned mixed solution through a peristaltic pump, and the addition was completed in one hour. The mixture was purified by column chromatography (ethyl acetate:petroleum ether=1:20~1:3) to obtain a pair of diastereoisomers with the above structures. Total yield: 91%, dr>20:1.

¹ H NMR (400MHz, Chloroform-d) δ7.30–7.26(m,1H),7.20(d,J=8.2Hz,1H),7.06–6.92(m,4H),6.82(t,J=7.7Hz ,1H),6.58(d,J=8.0Hz,1H),4.54(s,1H),3.68(s,3H),3.46(s,3H),3.18(dt,J=16.7,8.0Hz,1H) ,2.87(dd,J＝17.1,5.6Hz,1H),2.49(dd,J＝14.7,6.0Hz,1H),2.23–2.13(m,1H),1.47–1.01(m,7H),0.79(t ,J＝7.4Hz,3H).

¹³ C NMR(100MHz,CDCl3)δ180.12,143.13,137.22,136.77,132.71,128.02,125.15,123.16,120.75,119.17,117.01,108.90,108.36,106.83,77.35,77.24,77.04,76.72,74.97,56.21,36.78, 29.27, 29.00, 26.52, 25.19, 23.31, 18.98, 14.14.

HRMS (ESI) Calcd. for C ₃₃ H ₂₉ N ₂ O ₂ (M+H) ⁺ 485.2250, found: 485.2229.

Example 8

3-Hydroxy-1',9-dimethyl-3-(4-(trifluoromethoxy)phenyl)-1,2,3,9-spirotetrahydro[carbazole-4,3'-ind Indoline]-2'-one

分子筛(100％)和Rh₂(OAc)₄(1.0mol％)溶解在装有1.0mL二氯甲烷的试管中，于特定温度(25℃)下搅拌30分钟，再将溶有重氮(0.4mmol)的二氯甲烷(1.0mL)溶液通过蠕动泵加入到前述混合溶液中，一小时加完。将混合物进行柱层析纯化(乙酸乙酯：石油醚＝1:20～1:3)得到一对非对映异构体，结构如上。合计产率：99％，dr>20:1The substituted indolinone (0.2mmol), 4

Molecular sieves (100%) and Rh ₂ (OAc) ₄ (1.0mol%) were dissolved in a test tube containing 1.0 mL of dichloromethane, stirred at a specific temperature (25° C.) for 30 minutes, and dissolved diazo (0.4 mmol) in dichloromethane (1.0 mL) was added to the aforementioned mixed solution through a peristaltic pump, and the addition was completed in one hour. The mixture was purified by column chromatography (ethyl acetate:petroleum ether=1:20~1:3) to obtain a pair of diastereoisomers with the above structures. Total yield: 99%, dr>20:1

¹ H NMR (400MHz, Chloroform-d) δ7.28(s,1H),7.20–7.06(m,5H),6.95(t,J=7.7Hz,1H),6.86(t,J=7.6Hz,1H ),6.62(d,J=8.0Hz,1H),6.56(d,J=7.9Hz,1H),6.01(s,1H),3.76(s,3H),3.39(p,J=9.1Hz,1H ), 3.12–2.99(m,5H), 2.37(dd,J＝14.3,5.9Hz,1H).

¹³ C NMR(100MHz,CDCl3)δ178.90,146.63,142.51,137.23,136.78,131.52,δ129.64–128.46(m),128.49,126.28,125.21,125.04,123.57,123.53,123.00,121.10,119.39,117.04,109.04 ,108.19,105.76,77.34,77.03,76.71,76.21,56.85,30.80,29.41,26.11,19.31.

¹⁹ F NMR (400MHz, CDCl3) δ-62.49.

HRMS (ESI) Calcd. for C ₂₈ H ₂₄ N ₂ O ₂ F ₃ (M+H) ⁺ 477.1788, found: 477.1790.

Example 9

5'-fluoro-3-hydroxy-1',3,9-trimethyl-1,2,3,9-spirotetrahydro[carbazole-4,3'-indoline]-2'-one

分子筛(100％)和Rh₂(OAc)₄(1.0mol％)溶解在装有1.0mL二氯甲烷的试管中，于特定温度(25℃)下搅拌30分钟，再将溶有重氮(0.4mmol)的二氯甲烷(1.0mL)溶液通过蠕动泵加入到前述混合溶液中，一小时加完。将混合物进行柱层析纯化(乙酸乙酯：石油醚＝1:20～1:3)得到一对非对映异构体，结构如上。合计产率：83％，dr>20:1The substituted indolinone (0.2mmol), 4

Molecular sieves (100%) and Rh ₂ (OAc) ₄ (1.0mol%) were dissolved in a test tube containing 1.0 mL of dichloromethane, stirred at a specific temperature (25° C.) for 30 minutes, and dissolved diazo (0.4 mmol) in dichloromethane (1.0 mL) was added to the aforementioned mixed solution through a peristaltic pump, and the addition was completed in one hour. The mixture was purified by column chromatography (ethyl acetate:petroleum ether=1:20~1:3) to obtain a pair of diastereoisomers with the above structures. Total yield: 83%, dr>20:1

¹ H NMR (400MHz, Chloroform-d) δ7.23(d, J=8.1Hz, 1H), 7.07(t, J=8.0Hz, 1H), 6.97(t, 1H), 6.95–6.89(m, 1H ),6.86(t,J=7.7Hz,1H),6.80(d,J=8.3Hz,1H),6.57(d,J=8.0Hz,1H),4.58(s,1H),3.69(s,3H ),3.46(s,3H),3.22(dt,J＝16.7,7.8Hz,1H),2.89(dd,1H),2.36(dd,J＝14.4,5.9Hz,1H),2.31–2.20(m, 1H), 1.04(s, 3H).

¹³ C NMR (100MHz, Chloroform-d) δ179.32, 141.82, 137.29, 136.77, 134.48, 128.35 (d, J=43.0Hz), 125.19 (d, J=42.1Hz), 121.00, 119.32, 116.88, 109.24, 109.03, 72.90, 56.15, 32.42, 29.33, 26.66, 25.06, 19.06.

¹⁹ F NMR (400MHz, CDCl3) δ-119.73.

HRMS (ESI) Calcd. for C ₂₂ H ₂₁ N ₂ O ₂ NaF (M+Na) ⁺ 387.1504, found: 387.1485.

Example 10

1'-Phenyl-3-hydroxy-3,5,'9-trimethyl-1,2,3,9-spirotetrahydro[carbazole-4,3'-indoline]-2'-one

分子筛(100％)和Rh₂(OAc)₄(1.0mol％)溶解在装有1.0mL二氯甲烷的试管中，于特定温度(25℃)下搅拌30分钟，再将溶有重氮(0.4mmol)的二氯甲烷(1.0mL)溶液通过蠕动泵加入到前述混合溶液中，一小时加完。将混合物进行柱层析纯化(乙酸乙酯：石油醚＝1:20～1:3)得到一对非对映异构体，结构如上。合计产率：80％，dr>20:1The substituted indolinone (0.2mmol), 4

Molecular sieves (100%) and Rh ₂ (OAc) ₄ (1.0mol%) were dissolved in a test tube containing 1.0 mL of dichloromethane, stirred at a specific temperature (25° C.) for 30 minutes, and dissolved diazo (0.4 mmol) in dichloromethane (1.0 mL) was added to the aforementioned mixed solution through a peristaltic pump, and the addition was completed in one hour. The mixture was purified by column chromatography (ethyl acetate:petroleum ether=1:20~1:3) to obtain a pair of diastereoisomers with the above structures. Total yield: 80%, dr>20:1

¹ H NMR (400MHz, Chloroform-d) δ7.50(d, J=7.3Hz, 2H), 7.40–7.29(m, 3H), 7.21(d, J=8.3Hz, 1H), 7.04(t, J =7.4Hz, 2H), 6.98(d, J=8.1Hz, 1H), 6.85(d, J=9.0Hz, 2H), 6.73(t, J=7.3Hz, 2H), 6.47(d, J=7.9 Hz,1H),5.23(d,J=15.2Hz,1H),5.00(d,J=15.3Hz,1H),4.54(s,1H),3.69(s,3H),3.23(dt,J=16.5 ,8.0Hz,1H),2.94–2.85(m,1H),2.46–2.37(m,1H),2.35–2.26(m,1H),2.17(s,3H),1.08(s,3H).

¹³ C NMR(101MHz,CDCl ₃ )δ179.68,140.24,137.25,136.67,135.85,132.59,132.43,128.82,128.38,128.17,127.87,126.03,125.23,120.74,119.01,117.60,108.99,108.79,106.93,77.35,77.03 ,76.71,73.18,56.04,44.40,32.51,29.30,25.33,21.18,19.24.

HRMS (ESI) Calcd. for C ₂₉ H ₂₈ N ₂ O ₂ (M+H) ⁺ 437.2201, found: 437.2229.

Example 11

1'-Phenyl-5'-chloro-3-hydroxy-3,9-dimethyl-1,2,3,9-spirotetrahydro[carbazole-4,3'-indoline]-2' -ketone

分子筛(100％)和Rh₂(OAc)₄(1.0mol％)溶解在装有1.0mL二氯甲烷的试管中，于特定温度(25℃)下搅拌30分钟，再将溶有重氮(0.4mmol)的二氯甲烷(1.0mL)溶液通过蠕动泵加入到前述混合溶液中，一小时加完。将混合物进行柱层析纯化(乙酸乙酯：石油醚＝1:20～1:3)得到一对非对映异构体，结构如上。合计产率：95％，dr>20:1The substituted indolinone (0.2mmol), 4

Molecular sieves (100%) and Rh ₂ (OAc) ₄ (1.0mol%) were dissolved in a test tube containing 1.0 mL of dichloromethane, stirred at a specific temperature (25° C.) for 30 minutes, and dissolved diazo (0.4 mmol) in dichloromethane (1.0 mL) was added to the aforementioned mixed solution through a peristaltic pump, and the addition was completed in one hour. The mixture was purified by column chromatography (ethyl acetate:petroleum ether=1:20~1:3) to obtain a pair of diastereoisomers with the above structures. Total yield: 95%, dr>20:1

¹ H NMR (400MHz, Chloroform-d) δ7.47 (d, J = 7.4Hz, 2H), 7.41–7.32 (m, 3H), 7.23 (d, J = 8.1Hz, 1H), 7.16 (d, J =8.5Hz, 1H), 7.06(t, J=7.8Hz, 1H), 7.01(s, 1H), 6.88(d, J=8.3Hz, 1H), 6.75(t, J=7.6Hz, 1H), 6.42(d, J=7.9Hz, 1H), 5.22(d, J=15.3Hz, 1H), 5.00(d, J=15.3Hz, 1H), 4.26(s, 1H), 3.69(s, 3H), 3.22(dt,J=16.3,7.8Hz,1H),2.90(d,J=16.8Hz,1H),2.49(d,J=13.7Hz,1H),2.23(dt,J=14.0,7.6Hz,1H ),1.11(s,3H).

¹³ C NMR(100MHz,CDCl ₃ )δ179.30,141.25,137.30,136.79,135.30,134.24,128.95,128.41,128.08,125.72,125.00,125.00,120.96,119.19,117.35,110.17,108.96,106.13,77.34,77.22,77.02 ,76.79,76.70,73.27,56.38,44.51,32.56,29.34,25.24,19.25.

HRMS (ESI) Calcd. for C ₂₈ H ₂₅ ClN ₂ O ₂ (M+H) ⁺ 457.1655, found: 457.1683.

Example 12 Anti-tumor cell activity test experiment:

1. Cell inoculation:

1) Prepare a single cell suspension with DMEM culture medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, inoculate 2500 MG63 osteosarcoma cells per well into a 96-well cell culture plate, with a volume of 100 μL per well .

2) Prepare a single cell suspension with DMEM culture medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, and inoculate 2500 MNNG/HOS type osteosarcoma cells into a 96-well cell culture plate in each well. Volume 100 μL.

3) Prepare a single cell suspension with 1640 culture medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, inoculate 2500 HOS type osteosarcoma cells into a 96-well cell culture plate with a volume of 100 μl per well .

4) Prepare a single cell suspension with 1640 culture medium containing 10% fetal bovine serum, 1% penicillin and streptomycin, inoculate 2500 143B type osteosarcoma cells into a 96-well cell culture plate with a volume of 100 μl per well .

2. Administration: The compounds of formula (I) synthesized in the foregoing examples of the present invention were formulated with DMSO into a mother solution with a final concentration of 25 mM, diluted to 1 μM with complete medium, and acted on cells, and three replicate wells were set.

3. Cultivation: 5% CO ₂ , 37°C saturated humidity incubator for 72 hours.

4. Color development: After culturing for 72 hours, aspirate the medium, add 100 μL of complete medium and 10 μL CCK8 to each well, and incubate at 37°C for 4 hours.

5. Colorimetry: select wavelengths of 620 and 450 nm, measure the optical density (OD) value of each well on a microplate reader, and record the results.

6. The 450nm absorbance value-620nm absorbance value (background absorbance value) of the same well is used as the final absorbance, which is substituted into the following formula.

7. Cell proliferation activity (%)=[A (drug added)-A (blank)]/[A (no drug added)-A (blank)]×100%.

A (dosing): with osteosarcoma cells, CCK8 solution (Cell Counting Kit-8, is a WST-8 (chemical name: 2-(2-methoxy-4-nitrophenyl)-3-( 4-nitrophenyl)-5-(2,4-disulfobenzene)-2H-tetrazolium monosodium salt) rapid and high-sensitivity detection reagent for cell proliferation and cytotoxicity) and drug solution (the drug solution refers to the The absorbance of the wells of the compound sample DMSO solution)

A (Blank): Absorbance of wells with culture medium and CCK8 solution without cells

A (no drug added): absorbance of wells with cells, CCK8 solution and no drug solution

8. The experimental results show that: under the action of the compound of formula (I) of the present invention at a concentration of 1 μM, the survival rates of 4 different subtypes of osteosarcoma cells are shown in Table 1 below. This shows that, among these 11 compounds, compound 3h, 3b show better inhibitory activity to osteosarcoma cell MG63; Compound 3h, 3b, 3j show better inhibitory activity to osteosarcoma cell MNNG/HOS; Compound 3l, 3k, 3h, 3b, 3i, 3j showed better inhibitory activity on osteosarcoma cell HOS; compounds 3l, 3k, 3m, 3h, 3i showed better inhibitory activity on osteosarcoma cell 143B.

Table 1

references:

1.Carl Djerassi et al.Helv.Chim.Acta 1957,40,1189.Nat.Prod.Rep.2013,30,694.

2.A) Ross.Widenhoefer et al.J.Am.Chem.Soc.2006,128,9066.B)Hyun-Ji Songet al.J.Am.Chem.Soc.2005,127,8942.

3.A) Shuli You et al.Angew.Chem.Int.Ed.2012,51,1680.B)Beeraiah Baire et al.Org.Lett.2018,20,1118.

Claims (8)

1. a polysubstituted spirotetrahydrocarbazole derivative is characterized in that the structure is shown as follows,

2. a method for synthesizing a polysubstituted spirotetrahydrocarbazole derivative as claimed in claim 1, characterized in that the diazo compound represented by formula (1) and the substituted indolone represented by formula (2) are used as raw materials, rh ₂ (OAc) ₄ To catalyzeThe agent is prepared by reacting, in an organic solvent,

the molecular sieve is an additive and reacts to obtain a polysubstituted spiro tetrahydrocarbazole derivative shown in the formula (I); the reaction process is shown as a reaction formula (II):

wherein,

R ¹ is methyl, R ² Is methyl, R ³ Is H, R ⁴ Is benzyl;

or, R ¹ Is methyl, R ² Is methyl, R ³ Is H, R ⁴ Is a bromo-substituted benzyl group;

or, R ¹ Is methyl, R ² Is methyl, R ³ Is H, R ⁴ Is a methoxy substituted benzyl group;

or, R ¹ Is methyl, R ² Is methyl, R ³ Is methoxy, R ⁴ Is benzyl;

or, R ¹ Is butyl, R ² Is methyl, R ³ Is H, R ⁴ Is methyl;

or, R ¹ Is methyl, R ² Is methyl, R ³ Is methyl, R ⁴ Is benzyl;

or, R ¹ Is methyl, R ² Is methyl, R ³ Is chlorine, R ⁴ Is benzyl.

3. The synthesis method of claim 2, wherein the reaction temperature is 0 ℃ to 40 ℃; and/or the reaction time is 1h-5h.

4. The synthesis method according to claim 2, wherein the diazo compound represented by formula (1) and the substituted indolone represented by formula (2) are present in a molar ratio of 1.2:1-2.0:1; and/or, the catalyst Rh ₂ (OAc) ₄ The dosage of the compound is 0.5mol percent to 2.0mol percent of the substituted indolone based on the substituted indolone; and/or, the additive

The dosage of the molecular sieve is 50-200% of the mass of the substituted indolone based on the substituted indolone.

5. The synthesis method according to claim 2, wherein the organic solvent is selected from one or more of DCM, DCE, EA, and toluene; and/or the amount of the organic solvent is based on the substrate (2) substituted indolone, so that the concentration of the substituted indolone in the organic solvent is in a range of 0.05M-0.15M.

6. The synthesis method according to claim 2, characterized in that it comprises in particular the following steps: firstly, the substituted indolone shown in the formula (2) and Rh are ₂ (OAc) ₄ 、

Dissolving a molecular sieve in the organic solvent, adding the organic solvent solution of the diazo compound shown in the formula (1) at 25 ℃, and reacting to obtain the polysubstituted spirotetrahydrocarbazole derivative shown in the formula (I).

7. The synthesis method according to claim 6, characterized in that the polysubstituted spirotetrahydrocarbazole derivative obtained by the reaction is further separated and purified; the separation and purification are carried out by using a solvent comprising ethyl acetate, petroleum ether =1:20 to 1: and 3, carrying out column chromatography on the solution.

8. The use of a polysubstituted spirotetrahydrocarbazole derivative according to claim 1 for the preparation of a medicament having an active anti-tumor cell activity, wherein said tumor cell is one or more of an MG63 type osteosarcoma cell, an MNNG/HOS type osteosarcoma cell, an HOS type osteosarcoma cell, and a 143B type osteosarcoma cell.

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