CN115650981A

CN115650981A - Logenoid indole alkaloid based on loganin, synthetic method and application thereof, and medicine for preventing and treating diabetes or diabetic nephropathy

Info

Publication number: CN115650981A
Application number: CN202211283730.8A
Authority: CN
Inventors: 王福生; 秦子康; 杨宗斌; 江世智; 张兴宇
Original assignee: Dali University
Current assignee: Dali University
Priority date: 2022-10-20
Filing date: 2022-10-20
Publication date: 2023-01-31
Anticipated expiration: 2042-10-20
Also published as: CN115650981B

Abstract

The invention provides logenoid indole alkaloid based on loganin, a synthesis method and application thereof, and a medicament for preventing and treating diabetes or diabetic nephropathy, and belongs to the technical field of medicinal chemistry. The monoterpenoid indole alkaloid provided by the invention is derived from a natural product loganin, and compared with loganin with very high polarity, the monoterpenoid indole alkaloid has low polarity and good fat solubility, and is beneficial to absorption in vivo. The monoterpenoid indole alkaloid provided by the invention has high-efficiency anti-diabetic and diabetic nephropathy activity, and can be used for preparing a medicament for preventing and treating diabetes or diabetic nephropathy. The invention provides a synthetic method of the monoterpene indole alkaloid, and the method provided by the invention is simple to operate and suitable for large-scale production.

Description

Logenoid indole alkaloid based on loganin, synthetic method and application thereof, and medicine for preventing and treating diabetes or diabetic nephropathy

Technical Field

The invention relates to the technical field of medicinal chemistry, in particular to logenoid monoterpene indole alkaloid based on loganin, a synthesis method and application thereof, and a medicament for preventing and treating diabetes or diabetic nephropathy.

Background

Diabetes Mellitus (DM) is a syndrome of disturbances of sugar, fat and protein metabolism due to insufficient secretion or utilization of insulin. One treatment to prevent postprandial hyperglycemia is by inhibiting enzymes (e.g., alpha-amylase and alpha-glucosidase) to delay the digestion and absorption of carbohydrates in the gastrointestinal tract. The presence of chronic postprandial hyperglycemia impairs endogenous antioxidant defense functions, pancreatic β -cell destruction caused by oxidative stress, produces uncontrolled free radicals such as ROS, and leads to various macrovascular and microvascular complications. The alpha-amylase hydrolyzes the compound polysaccharide to generate oligosaccharide and disaccharide, then the oligosaccharide and the disaccharide are hydrolyzed into monosaccharide by the alpha-glycosidase and are absorbed by small intestine to enter hepatic portal vein, so that the postprandial blood sugar is increased. Inhibitors of alpha-amylase and alpha-glucosidase delay digestion and subsequent carbohydrate absorption, thereby lowering postprandial glucose levels. Since 1990, three α -glucosidase inhibitors have been known clinically, namely acarbose, voglibose and miglitol.

Diabetic Nephropathy (DN) is one of the common complications in DM patients, and is the leading cause of end-stage renal disease and death in DM patients, with approximately 30-40% of DM patients developing DN and increasing incidence as the course of disease increases. The clinical manifestations of early DN include increased glomerular filtration rate, increased microalbumin in urine, increased plasma urea nitrogen and creatinine, and finally progression to chronic renal insufficiency. The main pathological features of the early stage of DN are glomerular hypertrophy, thickening of glomerular and tubular basement membranes and progressive accumulation of extracellular matrix in the mesangial region; the later stage is fibrosis of the glomerulus, the tubulointerstities, and ultimately, proteinuria and renal failure.

At present, DM is mainly treated by reducing blood sugar, DN is mainly treated by combined medication, and medicaments for comprehensively reducing blood sugar, blood pressure, blood fat and the like are mainly used for treating, so that the development of DN is mainly delayed, and the effect of radical treatment cannot be achieved. Currently, hormone and cytotoxic drugs are clinically used for treating DN, and although a certain curative effect is achieved, adverse reactions such as easy relapse and easy generation of hormone dependence exist. Angiotensin converting enzyme inhibitors (e.g., captopril, enalapril, etc.) have therapeutic effects on DN. However, angiotensin converting enzyme inhibitors have a large side effect and cause symptoms such as hypotension, hyperkalemia, renal insufficiency, and cough. Therefore, aiming at DM and DN, the problem to be solved is urgently needed to find a high-efficiency and low-toxicity prevention and treatment medicine.

Disclosure of Invention

The monoterpenoid indole alkaloid provided by the invention is derived from a natural product loganin, has good lipid solubility and low toxicity compared with loganin with very high polarity, has high-efficiency activity on resisting diabetes and diabetic nephropathy, and can be used for preparing medicines for preventing and treating diabetes or diabetic nephropathy.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides a monoterpene indole alkaloid which has a structure shown in a formula I or a formula II:

the R is ¹ Is hydrogen atom, halogen atom, methyl, ethyl, hydroxyl, methoxyl, formyloxy, acetoxyl, substituted benzyloxy or unsubstituted benzyloxy; at least one substituent group is arranged on a benzene ring in the substituted benzyloxy, and the substituent group is a halogen atom, a methyl group or a nitro group;

the R is ² Is hydrogen atom, carboxyl, hydroxymethyl, methoxy formyl, ethoxy formyl, methylamine formyl, ethylamine formyl, propylamine formyl, benzylamine formyl and dimethylamine formylDiethylamidoyl or dipropamidoformyl;

the R is ³ Is hydrogen atom, methyl, ethyl, formyl, allyl, methoxy formyl or ethoxy formyl.

Preferably, the halogen atom is-F, -Cl or-Br.

Preferably, the monoterpenoid indole alkaloid is any one of the following compounds:

the invention provides a synthesis method of monoterpene indole alkaloid in the technical scheme, which comprises the following steps:

carrying out a first Pictet-Spengerer (P-S) reaction on loganin aglycone and a reaction reagent Ma under the catalysis of a first Lewis acid to obtain monoterpenoid indole alkaloid with a structure shown in a formula I;

carrying out a second P-S reaction on loganin aglycone and a reaction reagent Mb under the catalysis of a second Lewis acid to obtain monoterpenoid indole alkaloid with a structure shown in a formula II;

the loganin aglycone has a structure shown in a formula 1, the reaction reagent Ma has a structure shown in a formula 2, and the reaction reagent Mb has a structure shown in a formula 3:

the R is ¹ 、R ² And R ³ As defined in formula I and formula II.

Preferably, the first lewis acid and the second lewis acid are independently one or more of zinc trifluoromethanesulfonate, trifluoroacetic acid, aluminum trichloride, ferric trichloride, boron trifluoride, potassium trifluoromethanesulfonate and lithium trifluoromethanesulfonate.

Preferably, the temperature of the first P-S reaction and the second P-S reaction is independently 40-45 ℃ and the time is independently 1-30 h.

Preferably, the first P-S reaction and the second P-S reaction are carried out in the presence of an organic solvent, and the organic solvent required by the first P-S reaction and the second P-S reaction is one or more of dichloromethane, N-dimethylformamide, dimethyl sulfoxide and hexafluoroisopropanol independently.

Preferably, the loganin aglycone is obtained by hydrolysis reaction of loganin in the presence of beta-glucosidase; the temperature of the hydrolysis reaction is 25-50 ℃, and the time is 24-48 h; the hydrolysis reaction is carried out under the condition that the pH value is 5.0-7.0.

The invention provides application of the monoterpenoid indole alkaloid in preparation of a medicament for preventing and treating diabetes or diabetic nephropathy.

The invention provides a medicine for preventing and treating diabetes or diabetic nephropathy, which takes monoterpenoid indole alkaloid as an active ingredient.

The monoterpenoid indole alkaloid provided by the invention has high-efficiency activity on resisting diabetes and diabetic nephropathy, and can be used for preparing a medicament for preventing and treating diabetes or diabetic nephropathy.

Specifically, the evaluation mode of the anti-diabetic and diabetic nephropathy activity of the monoterpenoid indole alkaloids in the test example of the invention comprises the following steps: detecting the inhibition effect of the monoterpene indole alkaloid on alpha-glucosidase by adopting an alpha-glucosidase activity inhibition model; detecting whether the monoterpene indole alkaloid has toxicity on rat HBZY-1 cells by adopting a CKK-8 method; detecting the inhibition effect of the monoterpenoid indole alkaloid on the abnormal proliferation of the high-sugar cultured HBZY-1 cells by adopting a high-sugar cultured rat HBZY-1 cell model; and simultaneously determining the influence of the monoterpene indole alkaloids on the contents of superoxide dismutase (SOD) and Malondialdehyde (MDA) in high-sugar-induced HBZY-1 cells. In vitro pharmacological activity experiments show that the compound has no obvious toxicity to HBZY-1 cells within the range of 0.01-100 mu mol/L; when the glucose is concentratedThe cell proliferation effect is obvious when the culture time is 48h and the degree is 30mmoL/L, and the results of drug treatment experiments carried out under the condition show that the compounds a1, a2, b3, b4, b5, b6, c1, c2 and d3 have obvious inhibition effect on abnormal cell proliferation; the compounds a2, b6, c1 and d3 can obviously improve the SOD activity in the HBZY-1 cell and reduce the content of MDA, and have the effect of relieving the oxidative stress of the HBZY-1 cell in a certain concentration range; the inhibition rates of the compounds b4, b6, c and d3 on alpha-glucosidase are all higher than 50%, which shows that the compounds have the function of inhibiting the alpha-glucosidase; wherein IC of compound b4 ₅₀ The value is smaller than that of the positive control acarbose, which indicates that the acarbose has a remarkable hypoglycemic effect.

The invention provides a synthetic method of the monoterpene indole alkaloid, and the method provided by the invention is simple to operate and suitable for large-scale production.

Specifically, the invention takes commercial and easily available tryptamine, L-tryptophan or derivatives obtained by derivatization of substitutes thereof as reaction reagents (namely reaction reagent Ma and reaction reagent Mb), takes loganin aglycone obtained by hydrolyzing natural active ingredients loganin as a synthesis module, and carries out biomimetic synthesis through specific reactions (Mannich reaction and Pictet-Spengler reaction) to obtain series monoterpene indole alkaloids. Compared with the total synthesis of natural products, the method provided by the invention has the characteristic of diversity-oriented synthesis, and the reaction reagent and loganin aglycone are reacted in one step to synthesize the monoterpene indole alkaloid, so that the method has the characteristics of mild reaction conditions, simplicity in operation, rapidness and high efficiency in reaction, reaction raw material saving, and no harsh reaction conditions of high temperature, high pressure and strong acid and strong base. The reaction raw materials adopted by the invention are natural compounds which are easy to obtain and have abundant quantity, and the method is suitable for large-scale industrial production.

Drawings

FIG. 1 is a graph showing the effect of different glucose concentrations and different culture times on HBZY-1 cells;

FIG. 2 is a bar graph of the effect of compounds on HBZY-1 cell viability;

FIG. 3 is a bar graph of the effect of compounds on the viability of high-sugar-induced HBZY-1 cells.

Detailed Description

the R is ² Is hydrogen atom, carboxyl, hydroxymethyl, methoxy formyl, ethoxy formyl, methylamine formyl, ethylamine formyl, propylamine formyl, benzylamine formyl, dimethylamine formyl, diethylamine formyl or dipropamine formyl;

said R is ³ Is hydrogen atom, methyl, ethyl, formyl, allyl, methoxy formyl or ethoxy formyl.

In the present invention, the dotted double bond in the formula II represents a carbon-carbon single bond or a carbon-carbon double bond.

In the present invention, the halogen atom is preferably-F, -Cl or-Br; the halogen atom mentioned here may be specifically R ¹ Or an optional class of substituents in the substituted benzyloxy group.

In the present invention, the monoterpenoid indole alkaloid is preferably any one of the following compounds:

carrying out a first P-S reaction on loganin aglycone and a reaction reagent Ma under the catalysis of a first Lewis acid to obtain monoterpenoid indole alkaloid with a structure shown in a formula I;

in the present invention, unless otherwise specified, the starting materials used are commercially available or prepared by methods well known to those skilled in the art.

The invention takes commercial tryptamine, L-tryptophan or derivatives obtained by derivatization of substitutes thereof as reaction reagents (namely reaction reagent Ma and reaction reagent Mb), takes loganin aglycone obtained by hydrolyzing natural active ingredients loganin as a synthesis module, and obtains series monoterpene indole alkaloids by specific reaction (Mannich reaction and P-S reaction biomimetic synthesis.

In the invention, the loganin aglycone is preferably obtained by hydrolyzing loganin in the presence of beta-glucosidase, and the reaction formula is as follows:

in the present invention, the molar ratio of loganin to β -glucosidase is preferably 1: (0.09 to 0.12), more preferably 1: (0.10-0.11). In the present invention, the hydrolysis reaction is preferably carried out at a pH of 5.0 to 7.0, more preferably 6.0; the pH is preferably provided by a citric acid-sodium citrate buffer solution. In the present invention, the hydrolysis reaction is preferably carried out inThe method is carried out in a protective atmosphere, the type of the protective gas for providing the protective atmosphere is not particularly limited, and the protective atmosphere can be N ₂ . In the present invention, the temperature of the hydrolysis reaction is preferably 25 to 50 ℃, more preferably 35 to 45 ℃; the time is preferably 24 to 48 hours, more preferably 36 to 48 hours. After the hydrolysis reaction, the product system is preferably extracted by ethyl acetate, and the organic phase is extracted by anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating the filtrate to dryness by rotary evaporation to obtain loganin aglycone.

In the present invention, the reactive agent Ma has a structure represented by formula 2:

in the present invention, the synthesis method and conditions of some kinds of the reactive reagent Ma are as follows:

the following describes in detail the synthesis methods of the compounds Mb1, mb2, mb3, mc1, mc2 and Mc 3.

In the present invention, the method for synthesizing the compound Mb1 preferably includes the steps of: l-tryptophan, thionyl chloride and methanol were mixed and subjected to an esterification reaction to obtain a hydrochloride of the compound Mb 1. In the present invention, the molar ratio of L-tryptophan to thionyl chloride is preferably 1; the reaction A is preferably carried out under reflux conditions, and the reflux temperature is preferably 45 ℃; the time of the esterification reaction is preferably 1 hour. After the esterification reaction, the invention preferably carries out rotary evaporation on the obtained product system to remove methanol and excessive thionyl chloride, and the obtained white solid is the L-tryptophan methyl ester hydrochloride.

In the present invention, the method for synthesizing the compound Mb2 preferably includes the steps of: mixing compound Mb1 and compound NH ₂ R ⁴ And ethanol, and performing an amine transesterification reaction to obtain a compound Mb2. In the present inventionSaid compound NH ₂ R ⁴ In R ⁴ Preferably methyl, ethyl or propyl; said compound Mb1 and compound NH ₂ R ⁴ On the basis of ensuring that the compound Mb1 is fully reacted; the temperature of the amine transesterification reaction is preferably room temperature, and the time is preferably 2 hours. After the amine transesterification, the product system obtained is preferably subjected to rotary evaporation to remove the solvent, and then passed through a silica gel column (the reagent used is preferably ethyl acetate) to obtain the compound Mb2.

In the present invention, the method for synthesizing the compound Mb3 preferably includes the steps of: mixing L-tryptophan and LiAlH ₄ And Tetrahydrofuran (THF) was mixed and subjected to a reduction reaction to obtain a compound Mb3. In the present invention, the L-tryptophan is reacted with LiAlH ₄ Is preferably 100; the reduction reaction is preferably carried out under reflux conditions, and the reflux temperature is preferably 70 ℃; the time for the reduction reaction is preferably 12 hours. After the reduction reaction, the invention preferably drops saturated ammonium chloride solution into the obtained product system under stirring till no bubbles are generated, the obtained feed liquid is filtered by a short silica gel column of 5cm and then transferred into a separating funnel containing water, ethyl acetate is used for extraction, and the obtained organic phase is extracted by anhydrous Na ₂ SO ₄ Drying, filtering, and evaporating the solvent in the filtrate under reduced pressure to obtain the compound Mb3 (i.e. tryptophanol).

In the present invention, it is preferred to prepare compounds Mc1, mc2 and Mc3 according to the synthetic methods for compounds Mb1, mb2 and Mb3, respectively, except that the starting material L-tryptophan is replaced with 5-hydroxy L-tryptophan.

In the present invention, the reactive agent Mb has a structure represented by formula 3:

in the present invention, the synthesis method and conditions of the partial species reaction reagent Mb are as follows:

the following describes in detail the synthesis method of the compounds Mb4, mc4, md1 and Md 2.

In the present invention, the method for synthesizing the compound Mb4 preferably comprises the following steps:

mixing a compound Mb1, dichloromethane, triethylamine and di-tert-butyl dicarbonate, and carrying out amino Boc protection reaction to obtain a first intermediate;

mixing the N-Boc-L-tryptophan methyl ester, dimethyl sulfoxide (DMSO), KOH and methyl iodide, and performing alkylation reaction to obtain a second intermediate;

mixing the second intermediate, dichloromethane and trifluoroacetic acid, and carrying out amino de-Boc protection reaction to obtain a compound Mb4;

the structural formulas of the first intermediate and the second intermediate are sequentially as follows:

according to the invention, a compound Mb1, dichloromethane, triethylamine and di-tert-butyl dicarbonate are mixed and subjected to a D reaction to obtain a first intermediate. In the present invention, the molar ratio of the compound Mb1, triethylamine and di-tert-butyl dicarbonate is preferably 1; the temperature of the amino Boc protection reaction is preferably room temperature, and the time is preferably 1.5h. After the amino Boc protection reaction, the obtained product system is preferably mixed with water, dichloromethane is used for extracting a water phase after liquid separation, an organic phase obtained by extraction is dried by anhydrous sodium sulfate and then filtered, filtrate is subjected to vacuum concentration, and the obtained brown yellow viscous liquid is a first intermediate and can be directly used for the next reaction without further purification.

After the first intermediate is obtained, the first intermediate, dimethyl sulfoxide (DMSO), KOH and methyl iodide are mixed to carry out alkylation reaction, and a second intermediate is obtained. In the present invention, the molar ratio of the first intermediate, KOH and methyl iodide is preferably 1; the temperature of the alkylation reaction is preferably room temperature, and the time is preferably 2h. After the alkylation reaction, the obtained product system is preferably mixed with a saturated ammonium chloride solution, ethyl acetate is then added for extraction, an organic phase obtained by extraction is dried by anhydrous sodium sulfate and then filtered, the filtrate is concentrated in vacuum, and the residue is separated and purified by silica gel column chromatography (in terms of volume ratio, ethyl acetate: petroleum ether = 1).

After the second intermediate is obtained, the second intermediate, dichloromethane and trifluoroacetic acid are mixed to carry out amino de-Boc protection reaction, and the compound Mb4 is obtained. In the present invention, the molar ratio of the second intermediate to trifluoroacetic acid is preferably 1: (8-9); the temperature of the amino de-Boc protection reaction is preferably room temperature, and the time is preferably 1.5h. After the time of the amino de-Boc protection reaction is reached, preferably adding saturated sodium bicarbonate solution to quench the reaction, extracting the obtained material by using dichloromethane, drying an organic phase obtained by extraction by using anhydrous sodium sulfate, filtering, and carrying out rotary evaporation and reduced pressure concentration on the filtrate to obtain a white solid, namely the compound Mb4.

The present invention preferably prepares compound Mc4 according to the synthetic method of compound Mb4, except that the starting material L-tryptophan is replaced with 5-hydroxy L-tryptophan.

The present invention preferably prepares compound Md1 according to the synthetic method of compound Mb4, except that compound Mb1 is replaced with tryptamine.

The present invention preferably prepares the compound Md2 according to the synthetic method of the compound Mb4, except that the compound Mb1 is replaced with 5-methoxytryptamine.

The synthesis of monoterpenoid indole alkaloids of the present invention is described below.

In the invention, the synthesis method of the monoterpenoid indole alkaloid with the structure shown in the formula I comprises the following steps:

carrying out a first P-S reaction on loganin aglycone and a reaction reagent Ma under the catalysis of a first Lewis acid to obtain the monoterpenoid indole alkaloid with the structure shown in formula I.

In the present invention, the molar ratio of loganin aglycone to the reactant Ma is preferably 1: (1 to 1.3), more preferably 1: (1.2-1.3). In the invention, the first lewis acid is preferably one or more of zinc trifluoromethanesulfonate, trifluoroacetic acid, aluminum trichloride, ferric trichloride, boron trifluoride, potassium trifluoromethanesulfonate and lithium trifluoromethanesulfonate, and is more preferably zinc trifluoromethanesulfonate or trifluoroacetic acid; the molar ratio of loganin aglycone to the first Lewis acid is preferably 1: (0.09 to 0.11), more preferably 1:0.1. in the invention, the first P-S reaction is preferably carried out in the presence of an organic solvent, the organic solvent required by the first P-S reaction is preferably one or more of dichloromethane, N-dimethylformamide, dimethyl sulfoxide and hexafluoroisopropanol, and more preferably dichloromethane or hexafluoroisopropanol; the invention does not specially limit the dosage of the organic solvent, and the first P-S reaction is ensured to be carried out smoothly.

The loganin aglycone, the reaction reagent Ma, the first Lewis acid and the organic solvent are preferably mixed to carry out the first P-S reaction. In the present invention, the temperature of the first P-S reaction is preferably 35 to 45 ℃; the time is preferably 1-30 h, and specifically can be 1h, 2h, 3h, 4h, 10h, 20h or 30h; the progress of the reaction is preferably monitored by TLC according to the invention. In the present invention, the first P — S reaction is preferably carried out under stirring conditions. After the first P-S reaction, the invention preferably performs silica gel column chromatography purification on the obtained product system to obtain the monoterpenoid indole alkaloid with the structure shown in the formula I. In the present invention, the reagent used for the silica gel column chromatography purification is preferably selected according to the specific kind of the target compound; in the embodiment of the invention, a chloroform-methanol mixed reagent or a petroleum ether-ethyl acetate mixed reagent is specifically adopted, and the volume ratio of chloroform to methanol in the chloroform-methanol mixed reagent is preferably (10-15): 1, more preferably (12 to 13): 1; the volume ratio of the petroleum ether to the ethyl acetate in the petroleum ether-ethyl acetate mixed reagent is preferably (0.5-6): 1, more preferably (1 to 5): 1.

in the invention, the synthesis method of the monoterpenoid indole alkaloid with the structure shown in formula II comprises the following steps:

carrying out a second P-S reaction on loganin aglycone and a reaction reagent Mb under the catalysis of a first Lewis acid to obtain the monoterpenoid indole alkaloid with the structure shown in the formula II.

In the present invention, the molar ratio of loganin aglycone to the reactive agent Mb is preferably 1: (1 to 1.3), more preferably 1: (1.2-1.3). In the present invention, the selectable species and the amount of the second lewis acid are preferably the same as those of the first lewis acid, and are not described herein again. In the present invention, the second P-S reaction is preferably performed in the presence of an organic solvent, and the kind and the amount of the organic solvent required for the second P-S reaction are preferably the same as those of the organic solvent required for the first P-S reaction, and thus are not described herein again.

The loganin aglycone, the reaction reagent Mb and the second Lewis acid are preferably mixed with the organic solvent to carry out the second P-S reaction. In the present invention, the conditions of the second P-S reaction are preferably the same as those of the first P-S reaction, and thus, the description thereof is omitted. After the second P-S reaction, the invention preferably performs silica gel column chromatography purification on the obtained product system to obtain the monoterpenoid indole alkaloid with the structure shown in the formula II. In the present invention, the reagent used for the silica gel column chromatography purification is preferably selected according to the specific kind of the target compound; in the embodiment of the invention, a petroleum ether-ethyl acetate mixed reagent is specifically adopted, and the volume ratio of petroleum ether to ethyl acetate in the petroleum ether-ethyl acetate mixed reagent is preferably (0.5-3): 1, more preferably (1 to 2): 1.

the invention provides application of monoterpene indole alkaloids in preparation of a medicine for preventing and treating diabetes or diabetic nephropathy.

The invention provides a medicine for preventing and treating diabetes or diabetic nephropathy, which takes monoterpenoid indole alkaloid as an active ingredient. In the invention, the medicine preferably also comprises pharmaceutically acceptable auxiliary materials, and in the invention, the medicinal auxiliary materials preferably comprise one or more of diluent, excipient, filler, humectant, disintegrant, sodium absorption enhancer, surfactant, adsorption carrier, lubricant, flavoring agent and sweetening agent; the diluent preferably comprises water; the excipient preferably comprises one or more of water, mannitol, magnesium stearate, starch and cyclodextrin; the filler preferably comprises starch and/or sucrose; the mixing agent preferably comprises one or more of cellulose derivatives, alginate, gelatin and polyvinylpyrrolidone; the humectant preferably comprises glycerin; the disintegrating agent preferably comprises one or more of agar, calcium carbonate and sodium bicarbonate; the sodium absorption enhancer preferably comprises a quaternary ammonium compound; the surfactant preferably comprises cetyl alcohol and/or sodium carboxymethyl cellulose; the adsorption carrier preferably comprises kaolin and/or bentonite; the lubricant preferably comprises one or more of talcum powder, calcium stearate, magnesium stearate and polyethylene glycol; the flavoring agent and the sweetener are not particularly limited in the present invention, and those known to those skilled in the art can be used.

The dosage form of the medicine is not particularly limited, and the medicine can be prepared by the dosage forms known by the technicians in the field, such as tablets, granules, capsules, oral liquid, injection, freeze-dried injection or powder injection; the preparation of the tablet, the granule, the capsule, the oral liquid, the injection, the freeze-dried injection and the powder injection is not particularly limited, and the preparation method which is well known by the technical personnel in the field can be adopted.

The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Preparation example 1 preparation of Compound Mb1

A round-bottomed flask equipped with a reflux condenser and an alkaline absorber was charged with 200mg (0.98 mmol) of L-tryptophan and SOCl at a concentration of 2mol/L ₂ MeOH solution (containing 0.98mmol of SOCl) ₂ ) Heating to 45 ℃ and refluxing for 1h; after the reaction is finished, the methanol and the excessive thionyl chloride are removed by rotary evaporation to obtain whiteThe tryptophan solid was L-tryptophan methyl ester hydrochloride (i.e., hydrochloride salt of compound Mb 1), and the yield was 199.5mg, 80%. ¹ H-NMR(CDCl ₃ ,400MHz)δ3.12(d,J＝9.7Hz,1H),3.32(d,J＝5.2Hz,1H),3.65(S,3H),4.23(dd,J＝9.8,5.3Hz,1H),6.98(t,J＝8.0Hz,1H),7.08(ddd,J＝8.2Hz,1H),7.23(dt,J＝8.2Hz,1H),7.49(dt,J＝7.9Hz,1H)ppm。

Preparation example 2 preparation of Compound Mb2 (specifically, compounds Mb2a, mb2b, mb2 c)

Adding 50mg (0.2291 mmol) of the compound Mb1 and 3mL of a 0.092mol/L methylamine ethanol solution into a round-bottom flask, stirring the mixture at room temperature (25 ℃) for reaction for 2h, monitoring the reaction by TLC, and supplementing the methylamine ethanol solution if the compound Mb1 does not disappear until the compound Mb1 is completely reacted; after completion of the reaction, the solvent was removed by rotary evaporation, and the reaction mixture was passed through a silica gel column (ethyl acetate was used as a reagent) to obtain Compound Mb2a in a yield of 48.3mg and 93.4%. ¹ H-NMR(CDCl ₃ ,400MHz)δ1.62(bs,1H),2.53(m,3H),2.85(m,1H),3.14(m,1H),3.45(m,1H),6.91(s,1H),6.97-7.24(m,3H),7.33(d,1H),7.5(d,1H),7.8(bs,1H),10.8.8(bs,1H)ppm。

Compound Mb2b was prepared according to the above procedure except that the ethanol solution of methylamine was replaced with ethanol solution of ethylamine, resulting in a yield of compound Mb2b of 90.8%. ¹ H-NMR(MeOD,400MHz)δ0.96(t,3H),3.12(m,4H),3.71(t,1H),7.01(td,1H),7.09(td,1H),7.11(s,1H),7.33(dt,1H),7.33(d,1H),7.55(dt,1H),ppm。

Compound Mb2c was prepared according to the above procedure except that the ethanol solution of methylamine was replaced with an ethanol solution of propylamine, resulting in a yield of compound Mb2c of 92.7%. ¹ H-NMR(MeOD,400MHz)δ0.87(t,3H),1.48(s,2H),3.71(t,1H),2.89(dd,1H),3.21(m,2H),3.37(dd,1H),3.69(dd,1H),7.02(d,1H),7.09(m,1H),7.18(m,1H),7.32(t,1H),7.36(d,1H),7.64(d,1H),8.72(s,1H)ppm。

Preparation example 3 preparation of Compound Mb3

Weighing 100 mgL-tryptophan, dissolving in tetrahydrofuran, placing into round bottom flask, and gradually adding 22.3mg LiAlH under ice bath ₄ Stirring for 30min, heating to 70 ℃, refluxing, stirring and reacting for 12h; after the reaction is finished, inAdding saturated ammonium chloride solution dropwise into the obtained product system under stirring until no bubbles are generated, filtering the obtained material liquid with 5cm short silica gel column, transferring into separating funnel containing water, extracting with ethyl acetate for 3 times, mixing organic phases, and adding anhydrous Na ₂ SO ₄ Drying, filtering, and evaporating solvent from the filtrate under reduced pressure to obtain tryptophanol (compound Mb 3) with yield of 73.8mg and yield of 80%. ^l H-NMR(DMSO-d6,400MHz):δ2.05-2.37(mp,3H,H-2,H-3).2.77(dd,lH,J＝6.6,10.2Hz,H-1),2.90(dd,lH,J＝4.8,10.2Hz,H-1),6.48-6.63(mp,2H,H-5,H-6)6.88(d.lH,J＝8.0Hz,H-7).7.09(d,1H,J＝7.7Hz,H-4),10.37(s,1H,H-NH)。

Preparation example 4 preparation of Compound Mb4

Synthesis of N-Boc-L-tryptophan methyl ester: placing 160mg (0.6282 mmol) of compound Mb1 in a round-bottom flask, adding 0.8mL of dichloromethane to dissolve, dropwise adding 262 mu L (1.8846 mmol) of triethylamine, stirring for 10min at room temperature, cooling to 0 ℃ by using an ice-water bath, dropwise adding 289 mu L (1.2563 mmol) of di-tert-butyl dicarbonate, stirring for reacting for 1.5h at room temperature after dropwise adding is finished, and monitoring the reaction by TLC; after the reaction, 10mL of water was added, liquid separation was performed, the aqueous phase (20 mL. Times.2) was extracted with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated in vacuo to obtain a brown yellow viscous liquid, i.e., N-Boc-L-tryptophan methyl ester, with a yield of 221mg, which was used directly in the next reaction without further purification.

Synthesis of N-Boc-1-methyl-L-tryptophan methyl ester: dissolving 120mg (0.3769 mmol) of N-Boc-L-tryptophan methyl ester in 1.2mL of DMSO, adding KOH (21.1 mg) in an ice-water bath, stirring for 5min, dropwise adding 470 mu L of methyl iodide, heating to room temperature after dropwise adding, stirring for reacting for 2h, and monitoring the reaction by TLC; after the reaction, 10mL of saturated ammonium chloride solution was added, ethyl acetate was then added for extraction (20 mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated in vacuo, and the residue was separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether =1.

Synthesis of compound Mb 4: adding N-Boc-1-methyl-L-tryptophanDissolving 116mg (0.3490 mmol) of methyl ester in 1mL of dichloromethane, then dropwise adding 219 mu L (2.9503 mmol) of trifluoroacetic acid, stirring at room temperature after dropwise adding, reacting for 1.5h, then adding saturated sodium bicarbonate solution, quenching the reaction, extracting the obtained material (10 mL multiplied by 5) by using dichloromethane, combining organic phases, drying by using anhydrous sodium sulfate, filtering, and carrying out rotary evaporation and reduced pressure concentration on the filtrate to obtain a white solid, namely the compound Mb4, wherein the yield is 74mg and 91.2%. ¹ H-NMR(CDCl ₃ ,400MHz)δ3.13(dd,J＝7.5,3.3Hz,1H),3.33(dd,J＝15.0,5.2Hz,1H),3.66(s,3H),3.69(s,3H),4.20(dd,J＝10.8,5.3Hz,1H),6.91(s,1H),6.97(ddd,J＝8.0,7.0,1.0Hz,1H),7.09(ddd,J＝8.2,7.0,1.2Hz,1H),7.25(d,J＝8.3Hz,1H),7.48(dt,m,1H)ppm。

Preparation example 5 preparation of Compounds Mc1 to Mc4

Compound Mc1 was prepared according to the synthesis of compound Mb1, except that the starting material L-tryptophan was replaced with 5-hydroxy L-tryptophan, resulting in a yield of compound Mc1 of 93%.

Compound Mc2 (specifically, compounds Mc2a, mc2b, mc2 c) is prepared according to the synthesis method of compound Mb2, except that the starting material L-tryptophan is replaced with 5-hydroxy L-tryptophan, and the final yields of the compounds Mc2a, mc2b, mc2c are 93.4%, 90.8%, and 91.5%, respectively.

Compound Mc3 was prepared according to the synthesis of compound Mb3, except that the starting material L-tryptophan was replaced with 5-hydroxy L-tryptophan, resulting in a yield of compound Mc3 of 76%.

Compound Mc4 was prepared according to the synthesis of compound Mb4, except that the starting material L-tryptophan was replaced with 5-hydroxy L-tryptophan, resulting in a yield of compound Mc4 of 89.6%.

Preparation example 6 preparation of compounds Md1, md 2:

compound Md1 was prepared according to the procedure of preparation 4, except that compound Mb1 was replaced with tryptamine, and the final yield of compound Md1 was 32.5% based on the total yield of three steps.

Compound Md2 was prepared according to the procedure of preparation 4, except that compound Mb1 was replaced with 5-methoxytryptamine, and the final yield of compound Md2 was 57.1% based on the total yield of three steps.

Preparation example 7

Weighing loganin 3g, placing into a hard test tube, adding beta-glucosidase 200mg, dissolving with 50mL citric acid-sodium citrate buffer solution (pH = 6.0), and replacing the ambient atmosphere with N ₂ Then stirring and reacting for 2 days at the temperature of 45 ℃; then pouring the obtained product system into a separating funnel, adding ethyl acetate for extraction for three times, combining organic phases, and adding anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating the filtrate to dryness by rotary evaporation to obtain 1.6g of loganin aglycone with the yield of 92%.

The reaction formula for preparing loganin aglycone is shown as follows:

example 1

Weighing loganin aglycone 14.8mg (0.0648 mmol), tryptamine 20.8mg (0.1296 mmol) and zinc triflate 2.4mg (0.0065 mmol), placing into sealed tube, and vacuum replacing to N ₂ Then 2mL of dichloromethane is added, the mixture is stirred for reaction for 4h under the condition of oil bath at 45 ℃, and the reaction is monitored by TLC; after the reaction is finished, transferring the obtained product system into a separating funnel filled with water, and dropwise adding saturated NaHCO ₃ Extracting the solution with ethyl acetate for 3 times until no bubbles are formed, mixing the organic phases, and purifying with anhydrous Na ₂ SO ₄ After drying, filtration was performed, the solvent in the filtrate was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform: methanol =12 by volume) to obtain compound a1 (yield 18.3mg, yield 79.6%).

The compound a1 is yellow oil; molecular formula of C ₂₁ H ₂₄ N ₂ O ₃ ；

Is-52.5 (c =0.1,meoh); ¹ H-NMR(400MHz,CDCl ₃ )δ：8.30(br.s,1H),7.45(d,J＝7.6Hz,1H),7.36(s,1H),7.32(d,J＝7.6Hz,1H),7.15(t,J＝7.6Hz,1H),7.10(t,J＝7.6Hz,1H),4.39(br.s,1H),4.24(t,J＝5.6Hz,1H),3.73(dd,J＝13.8,5.4Hz,1H),3.62(s,3H),3.46(td,J＝13.8,5.8Hz,1H),2.95-3.00(m,1H),2.95-2.90(m,1H),2.77(dd,J＝15.1,5.4Hz,1H),2.39-2.43(m,1H),2.24(dd,J＝14.7,8.0Hz,1H),2.08(m,1H),1.80(dt,J＝14.7,5.6Hz,1H),1.20(d,J＝6.9Hz,3H)； ¹³ C NMR(100MHz,CDCl ₃ )δ:168.6,144.7,135.8,133.4,127.1,121.9,119.6,118.0,111.0,108.6,104.3,74.5,52.8,51.5,50.6,44.9,42.5,41.5,30.9,22.2,12.8。

example 2

Weighing loganin aglycone 21.9mg (0.0960 mmol), 5-methoxytryptamine 21.9mg (0.1151 mmol) and zinc triflate 3.5mg (0.0096 mmol), placing into a sealed tube, and vacuum replacing to N ₂ Then 2mL of dichloromethane is added, the mixture is stirred for reaction for 4h under the condition of oil bath at 45 ℃, and the reaction is monitored by TLC; after the reaction is finished, transferring the obtained product system into a separating funnel filled with water, and dropwise adding saturated NaHCO ₃ Until no bubble is generated, extracting with ethyl acetate for 3 times, mixing organic phases, and passing through anhydrous Na ₂ SO ₄ After drying, filtration was performed, the solvent in the filtrate was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform: methanol =13 by volume ratio = 1) to obtain compound a2 (yield 29mg, yield 79.0%).

The compound a2 is yellow oil; molecular formula of C ₂₂ H ₂₆ N ₂ O ₄ ；

Is-55 (c =0.1,meoh); ¹ H NMR(400MHz,CDCl ₃ )δ:1.06(d,J＝6.9Hz,3H),2.00(ddd,J＝9.6,7.2,5.5Hz,1H),4.06(td,J＝5.6,2.1Hz,1H),1.72(dt,J＝14.3,5.8Hz,1H),2.07(dd,J＝14.2,7.7,2.1Hz,1H),2.81(q,J＝7.5Hz,1H),7.37(s,J＝1.0Hz,1H),3.69-3.63(m,1H),3.35(ddd,J＝12.9,11.6,4.5Hz,1H),2.63-2.60(m,1H),2.72(ddd,J＝11.6,5.9,2.4Hz,1H),6.79(d,J＝2.4Hz,1H),6.64(dd,J＝8.7,2.5Hz,1H),7.14(d,J＝8.1Hz,1H),4.32(dt,J＝3.9,1.8Hz,1H),2.30(ddd，J＝9.5,7.8,4.1Hz,1H),3.53(s,3H),3.71(s,3H)； ¹³ C NMR(100MHz,CDCl ₃ )δ：170.88,155.10,147.04,135.72,132.97,128.67,112.73,112.07,108.55,104.42,100.70,74.70,56.16,54.57,51.16,46.68,43.06,42.38,32.31,30.78,23.44,13.44。

example 3

Weighing 22.8mg (0.0999 mmol) of loganin aglycone, 21.1mg (0.1199 mmol) of 5-hydroxytryptamine and 3.7mg (0.0099 mmol) of zinc trifluoromethanesulfonate, filling into a sealed tube, adding 1.5mL of hexafluoroisopropanol, stirring for reaction at 45 ℃ in a water bath for 2h, and monitoring the reaction by TLC; after the reaction was completed, the solvent in the system was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =1 by volume) to obtain compound a3 (yield 14.6mg, yield 39.6%).

The compound a3 is yellow solid with a molecular formula of C ₂₁ H ₂₄ N ₂ O ₄ ； ¹ H NMR(400MHz,Methanol-d ₄ )δ7.41(d,J＝1.1Hz,1H,H-5),7.22(d,J＝8.6Hz,1H,H-9),7.12(d,J＝2.3Hz,1H,H-12),6.95(dd,J＝8.6,2.4Hz,1H,H-11),4.32(p,J＝1.7Hz,1H,H-13b),4.18–4.06(m,1H,H-2),3.39(dd,J＝13.1,5.8Hz,1H,H-7),3.34(s,3H,-OCH ₃ ),3.15–3.09(m,1H,H-7)，3.09–3.03(m,1H,H-8),2.76(td,J＝9.4,8.2,3.3Hz,1H,H-3a),2.64–2.50(m,1H,H-8),2.40–2.33(m,1H,H-1),2.33–2.27(m,1H,H-3),1.96(ddd,J＝10.2,7.0,4.9Hz,1H,H-13c),1.80(dt,J＝14.0,5.5Hz,1H,H-3),1.35(d,J＝6.9Hz,3H,-CH ₃ ). ¹³ C NMR(100MHz,Pyridine-d ₅ )δ168.97(-COO-),153.03(C-10),145.74(C-5),136.58(C-14),132.52(C-13a),129.75(C-8b),112.86(C-11),112.80(C-12),107.88(C-8a),105.81(C-9),103.95(C-4),73.90(C-2),54.32(C-7),52.23(C-13b),50.91(-OCH ₃ ),46.04(C-13c),44.02(C-3),42.44(C-1),32.07(C-3a),23.30(C-8),14.07(-CH ₃ )。

Example 4

Weighing loganin aglycone 45mg (0.1972 mmol), 5-fluorotryptamine 42mg (0.2366 mmol) and zinc trifluoromethanesulfonate 7.2mg (0.0197 mmol), putting into a sealed tube, adding 1.5mL hexafluoroisopropanol, stirring in a water bath at 35 ℃ for reaction for 1h, and monitoring the reaction by TLC; after the reaction was completed, the solvent in the obtained product system was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =2 by volume ratio).

The compound a4 is a yellow solid; molecular formula C ₂₁ H ₂₃ N ₂ O ₃ F； ¹ H NMR(400MHz,CDCl ₃ )δ7.34(d,J＝1.0Hz,1H,H-5),7.20(dd,J＝8.7,4.3Hz,1H,H-9),7.06(dd,J＝9.5,2.5Hz,1H,H-12),6.85(td,J＝9.1,2.5Hz,1H,H-11),4.38(dt,J＝3.7,1.9Hz,1H,H-13b),4.23(t,J＝5.0Hz,1H,H-2),3.77–3.67(m,1H,H-7),3.60(s,3H,-OCH ₃ ),3.44(ddd,J＝13.2,11.8,4.7Hz,1H,H-7),2.93(d,J＝7.6Hz,1H,H-8),2.91–2.83(m,1H,H-3a),2.75–2.66(m,1H,H-8),2.48(ddd,J＝11.0,7.9,3.5Hz,1H,H-1),2.25(ddd,J＝14.8,8.1,1.2Hz,1H,H-3),2.07–1.95(m,1H,H-13c),1.79(dt,J＝14.8,5.6Hz,1H,H-3),1.15(d,J＝6.9Hz,3H). ¹³ C NMR(100MHz,CDCl ₃ )δ168.82(-COO-),144.73(C-5),135.62(C-14),132.43(C-13a),127.58(C-8b),111.79(C-10),111.69(C-11),110.06(C-9),109.80(C-12),108.56(C-8a),104.83(C-4),74.72(C-2),52.87(C-13b),51.54(-OCH ₃ ),44.47(C-13c),42.39(C-3),41.45(C-1),30.72(C-3a),22.26(C-8),12.83(-CH ₃ )。

The compound c5 is a yellow solid; molecular formula C ₂₁ H ₂₅ N ₂ O ₃ F； ¹ H NMR(400MHz,CDCl ₃ )δ7.32(d,J＝0.9Hz,1H,H-5),7.31–7.27(m,1H,H-9),7.19(dd,J＝9.5,2.5Hz,1H,H-12),7.01(d,J＝2.4Hz,1H,H-14),6.94(td,J＝9.1,2.5Hz,1H,H-11),4.16–4.06(m,1H,H-2),3.64(s,3H,-OCH ₃ ),3.41(t,J＝7.3Hz,2H,H-7),3.11(dd,J＝12.2,4.8Hz,1H,H-13b),3.07–3.01(m,1H,H-13b),2.96–2.93(m,1H,H-8),2.93–2.91(m,1H,H-3a),2.74(dd,J＝12.2,6.9Hz,1H,H-8),2.22(ddd,J＝14.1,7.3,1.6Hz,1H,H-1),1.92–1.85(m,1H,H-3),1.67(td,J＝7.3,5.0Hz,1H,H-13c),1.54(ddd,J＝13.8,8.4,5.1Hz,1H,H-3),1.01(d,J＝7.1Hz,3H,-CH ₃ )； ¹³ C NMR(100MHz,CDCl ₃ )δ169.54(-COO-),146.13(C-5),132.89(C-13a),127.61(C-8b),124.13(C-14),112.61(C-10),112.17(C-11),112.08(C-9),110.76(C-12),110.49(C-8a),103.35(C-4),74.51(C-2),56.06(C-7),50.67(-OCH ₃ ),47.46(C-13b),43.17(C-13c),41.97(C-3),41.84(C-1),32.50(C-3a),25.14(C-8),13.23(-CH ₃ )。

Example 5

Weighing 25.2mg (0.1104 mmol) of loganin aglycone, 25.8mg (0.1325 mmol) of 5-chlorotryptamine and 4.1mg (0.0110 mmol) of zinc trifluoromethanesulfonate, filling into a sealed tube, adding 1.5mL of hexafluoroisopropanol, stirring for reacting for 2h under the condition of water bath at 45 ℃, and monitoring the reaction by TLC; after the reaction was completed, the solvent in the obtained product system was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =2 by volume ratio = 1) to obtain compound a5 (yield 14mg, yield 32.7%).

The compound a5 is a yellow solid; molecular formula of C ₂₁ H ₂₃ N ₂ O ₃ Cl； ¹ H NMR(400MHz,CDCl ₃ )δ7.40(d,J＝2.0Hz,1H,H-5),7.33(d,J＝1.2Hz,1H,H-9),7.22(d,J＝8.6Hz,1H,H-12),7.08(dd,J＝8.6,2.0Hz,1H,H-11),4.39(s,1H,H-13b),4.24(t,J＝5.0Hz,1H,H-2),3.81–3.68(m,1H,H-7),3.62(s,3H,H-OCH ₃ ),3.44(ddd,J＝13.4,11.7,4.7Hz,1H,H-7),3.01–2.93(m,1H,H-8),2.93–2.83(m,1H,H-3a),2.71(dd,J＝15.4,4.7Hz,1H,H-8),2.48–2.39(m,1H,H-1),2.23(dd,J＝14.8,8.0Hz,1H,H-3),2.10(s,1H,H-13c)1.80(dt,J＝14.8,5.6Hz,1H,H-3),1.18(d,J＝6.9Hz,3H,H-CH ₃ )； ¹³ C NMR(100MHz,CDCl ₃ )δ168.68(-COO-),144.57(C-5),135.13(C-14),134.25(C-13a),128.36(C-8b),125.46(C-10),122.16(C-11),117.72(C-9),112.11(C-12),108.42(C-8a),105.02(C-4),74.79(C-2),60.59(C-7),52.80(C-13b),51.47(-OCH ₃ ),44.61(C-13c),42.42(C-3),41.45(C-1),30.80(C-3a),22.16(C-8),12.79(-CH ₃ )。

Example 6

Weighing loganin aglycone (34.6 mg, 0.1516mmoL), L-tryptophan (37mg, 0.1819mmoL) and zinc trifluoromethanesulfonate (111mg, 0.0303mmoL) into a 15mL sealed tube, adding 2mL dimethyl sulfoxide for dissolving, reacting at 35 ℃ in an oil bath for 24h, and monitoring the reaction by TLC; after the disappearance of the raw materials, water quenching was added to the reaction, the resulting system was extracted 5 times with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =1 by volume) to obtain compound b1 (yield 11.6mg, yield 20%).

The compound b1 is light yellow powder; molecular formula C ₂₂ H ₂₄ N ₂ O ₅ ；HRESIMS m/z 395.1665[M-H] ^- (calcd 395.1665)。UV(CH ₃ OH)λ _max ＝286.50nm、226.20nm；IR(KBr)ν _max 3459,3317,2960,2924,1710,1658,1591,1415,1344,1366,1239,1217,1195,1135,749cm ^-1 ； ¹ H NMR(400MHz,DMSO-d ₆ )δ：0.96(d,J＝7.2Hz,1H,H-CH ₃ ),2.39(m,1H,H-1),4.04(m,1H,H-2),1.45-1.27(m,1H,H-3a),2.21(dd,J＝12.5,6.1Hz,1H,H-3b),3.07-3.01(m,1H,H-3a),7.59(s,1H,H-5),4.80(d,J＝5.8Hz,1H,H-7β),2.97(dd,J＝14.2,5.2Hz,1H,H-8a),3.25(d,J＝15.1Hz,1H,H-8b),7.43(d,J＝8.0Hz,1H,H-9),6.98(t,J＝7.4Hz,1H,H-10),7.06(t,J＝7.5Hz,1H,H-11),7.35(d,J＝8.1Hz,1H,H-12),4.20(d,J＝8.4Hz,1H,H13b),1.77-1.61(m,1H,H-13c),3.57(s,3H,H-OCH ₃ )； ¹³ C NMR(100MHz,DMSO)δ：14.90(C-CH ₃ ),41.37(C-1),71.42(C-2),43.33(C-3),33.63(C-3a),100.45(C-4),146.90(C-5),61.45(C-7),23.87(C-8),106.08(C-8a),126.30(C-8b),117.59(C-9),118.71(C-10),121.07(C-11),111.38(C-12),132.99(C-13a),51.58(C-13b),47.42(C-13c),136.50(C-14),50.21(-OCH ₃ ),167.88(C＝O),172.88(-COOH)。

Example 7

23.5mg (0.1030 mmol) of loganin aglycone, 1mg (0.1236 mmol) of compound Mb and 2.6mg (0.0071 mmol) of zinc trifluoromethanesulfonate were weighed and placed in a sealed tube, and replaced with N under vacuum ₂ Then 2mL of dichloromethane is added, the mixture is stirred for reaction for 4h under the condition of oil bath at 45 ℃, and the reaction is monitored by TLC; after the reaction is finished, transferring the obtained product system into a separating funnel filled with water, and dropwise adding saturated NaHCO ₃ Until no bubble is generated, extracting the obtained system with ethyl acetate for 3 times, combining organic phases, and purifying with anhydrous Na ₂ SO ₄ After drying, filtration was performed, the solvent in the filtrate was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =2 by volume ratio = 1) to obtain compound b2 (yield 18mg, yield 42.8%).

The compound b2 is a yellow oil; molecular formula of C ₂₁ H ₂₄ N ₂ O ₃ ；HRESIMS m/z353.2196[M+H] ⁺ (calcd353.2193)。UV(CH ₃ OH)λ _max ＝281.7nm、223.50nm； ¹ H NMR(400MHz,DMSO-d ₆ )δ：0.95(d,J＝7.0Hz,3H,H-CH ₃ ),2.39(td,J＝12.7,6.5Hz,1H,H-1),4.03(m,1H,H-2),1.36(m,1H,H-3a),2.18(dd,J＝12.8,6.5Hz,1H,H-3b),3.04-2.98(m,1H,H-3a),7.54(s,1H,H-5),4.17-4.09(m,2H,H-7),2.92(dd,J＝15.0,6.1Hz,1H,H-8a),3.24(d,J＝15.1Hz,1H,H-8b),7.39(d,J＝7.8Hz,1H,H-9),6.95(t,J＝7.9Hz,1H,H-10),7.03(t,J＝8.1Hz,1H,H-11),7.34(d,J＝7.9Hz,1H,H-12),10.82(s,1H,H-13),4.26(d,J＝8.5Hz,1H,H-13b),1.71(m,1H,H-13c),3.55(s,3H,H-OCH ₃ )； ¹³ C NMR(100MHz,DMSO)δ：14.88(C-CH ₃ ),41.31(C-1),71.38(C-2),43.28(C-3),99.96(C-4),33.50(C-3a),147.13(C-5),67.42(C-7),23.26(C-8),106.20(C-8a),126.38(C-8b),117.48(C-9),118.54(C-10),120.86(C-11),111.36(C-12),133.18(C-13a),51.48(C-13b),47.35(C-13c),136.50(C-14),50.12(-OCH ₃ ),167.89(C＝O)。

Example 8

30mg (0.1314 mmol) of loganin aglycone, 29mg (0.1512 mmol) of tryptanthrin and 7mg (0.0202 mmol) of zinc trifluoromethanesulfonate were weighed into a sealed tube and replaced by N under vacuum ₂ Then 1.5mL Tetrahydrofuran (THF) is added, the reaction is stirred for 4h under the condition of oil bath temperature of 45 ℃, and the reaction is monitored by TLC; after the reaction is finished, transferring the obtained product system into a separating funnel filled with water, and dropwise adding saturated NaHCO ₃ Until no bubble is generated, extracting the obtained system with ethyl acetate for 3 times, combining organic phases, and purifying with anhydrous Na ₂ SO ₄ After drying, filtration was performed, the solvent in the filtrate was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =1.5 by volume) to obtain compound b3 (yield 26mg, purity 68.4%).

The compound b3 is a yellow oil; molecular formula C ₂₂ H ₂₆ N ₂ O ₄ ；

Is-186 (c =0.1,meoh); UV lambda _max (MeOH)221,292nm；IR(KBr)cm ^-1 :3754,3413,1656,1605,1226 and 746; ¹ H NMR(400MHz,CD ₃ OD)δ：7.50(d,J＝7.8Hz,1H),7.30(d,J＝8.1Hz,2H),7.06(ddd,J＝8.2,6.9,1.2Hz,1H),6.99(d,J＝8.0Hz,1H),4.11–4.00(m,2H),4.00–3.90(m,2H),3.84(dd,J＝9.0,6.3Hz,1H),3.55(s,3H),3.27(p,J＝1.7Hz,1H),3.21(dt,J＝13.3,7.5Hz,1H),2.92(dd,J＝14.1,6.0Hz,1H),2.83(dd,J＝14.2,7.9Hz,1H),2.31–2.19(m,1H),2.06(dtd,J＝11.8,7.2,4.4Hz,1H),1.95(d,J＝11.0Hz,2H),1.40(td,J＝8.8,4.0Hz,1H),1.19(t,J＝7.1Hz,1H),1.10(d,J＝7.3Hz,3H)； ¹³ C NMR(100MHz,CD ₃ OD)δ：171.29,141.20,138.12,128.57,124.66,122.49,119.87,119.27,111.35,100.80,91.997,74.43,71.81,61.54,59.49,51.08,48.23,44.60,44.31,35.83,32.33,15.00。

example 9

Weighing loganin aglycone 30mg (0.1314 mmol), compound Mb2a 34mg (0.1517 mmol) and zinc triflate 9.5mg (0.0263 mmol), placing into sealed tube, and vacuum replacing to N ₂ Then 2mL of dichloromethane is added, the mixture is stirred and reacted for 4h at the temperature of 45 ℃ in an oil bath, and the reaction is monitored by TLC; after the reaction is finished, transferring the obtained product system into a separating funnel filled with water, and dropwise adding saturated NaHCO ₃ Until no bubble is generated, extracting the obtained system with ethyl acetate for 3 times, combining organic phases, and purifying with anhydrous Na ₂ SO ₄ After drying, filtration was performed, the solvent in the filtrate was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =4 by volume ratio = 1) to obtain compound b4 (yield 19.4mg, yield 36.0%).

The compound b4 is yellow oil; molecular formula C ₂₃ H ₂₇ N ₃ O ₄ ； ¹ H-NMR(400MHz,CD ₃ OD)δ:7.53(d,J＝1.1Hz,1H),7.37(dd,J＝7.8,1.1Hz,1H),7.29–7.25(m,1H),7.02(ddd,J＝8.2,7.1,1.3Hz,1H),6.94(ddd,J＝8.0,7.1,1.1Hz,1H),4.45(d,J＝5.8Hz,1H),4.32(dt,J＝7.0,1.5Hz,1H),4.14(td,J＝5.1,1.4Hz,1H),3.60(s,3H),3.40(dt,J＝15.4,1.4Hz,1H),3.07(d,J＝8.0Hz,1H),2.97(ddd,J＝15.5,6.2,2.0Hz,1H),2.60(s,3H),2.33–2.21(m,2H),1.93(d,J＝7.4Hz,1H),1.63(ddd,J＝13.9,9.2,4.9Hz,1H),1.00(d,J＝7.1Hz,3H)； ¹³ C-NMR(100MHz,CD ₃ OD)δ:173.53,171.07,148.08,138.11,133.75,128.11,122.37,119.98,118.58,112.05,107.16,104.18,74.44,64.61,52.80,51.27,48.24,43.48,42.97,34.51,26.62,23.96,14.28。

Example 10

Weighing loganin aglycone 23mg (0.1008 mmol), compound Mb2b 37mg (0.1598 mmol) and zinc triflate 9.5mg (0.0266 mmol), placing into sealed tube, and vacuum replacing to N ₂ Then 2mL of methylene chloride was added, the reaction was stirred at 45 ℃ in an oil bath for 4h and monitored by TLC. After the reaction is finished, transferring the obtained product system into a separating funnel filled with water, and dropwise adding saturated NaHCO ₃ Until no bubble is generated, extracting the obtained system with ethyl acetate for 3 times, combining organic phases, and purifying with anhydrous Na ₂ SO ₄ After drying, filtration was performed, the solvent in the filtrate was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =2 by volume).

The compound b5 is a yellow oil; molecular formula C ₂₄ H ₂₉ N ₃ O ₄ ； ¹ H-NMR(400MHz,CD ₃ OD)δ:7.50(d,J＝1.2Hz,1H),7.36(dt,J＝7.8,1.0Hz,1H),7.25(dd,J＝8.0,1.0Hz,1H),7.03–6.96(m,1H),6.93(ddd,J＝8.1,7.1,1.1Hz,1H),4.42(d,J＝6.0Hz,1H),4.33(dd,J＝6.6,1.5Hz,1H),4.12(td,J＝5.2,1.5Hz,1H),3.58(s,3H),3.37(dt,J＝15.5,1.4Hz,1H),3.10(dd,J＝13.3,7.3Hz,1H),3.07–2.99(m,1H),2.94(ddd,J＝15.5,6.3,1.9Hz,1H),2.33–2.17(m,2H),1.99(dt,J＝13.4,6.7Hz,1H),1.64(ddd,J＝13.7,8.7,5.0Hz,1H),1.00(d,J＝7.1Hz,3H),0.92(t,J＝7.2Hz,3H)； ¹³ C-NMR(100MHz,CD ₃ OD)δ:172.57,171.02,148.05,138.07,133.85,128.19,122.33,119.06,118.55,112.04,107.16,104.42,74.46,64.61,52.68,51.27,47.97,43.41,42.80,35.47,34.23,23.97,14.93,14.22。

Example 11

30mg (0.1314 mmol) of loganin aglycone, 38.7mg (0.1577 mmol) of compound Mb2c and 9.5mg (0.0263 mmol) of zinc trifluoromethanesulfonate were weighed and chargedIn sealed tube, vacuum replacement to N ₂ Then 2mL of dichloromethane is added, the mixture is stirred and reacted for 4h at the temperature of 45 ℃ in an oil bath, and the reaction is monitored by TLC; after the reaction is finished, transferring the obtained product system into a separating funnel filled with water, and dropwise adding saturated NaHCO ₃ Until no bubbles are generated, extracting the obtained system with ethyl acetate for 3 times, combining organic phases, and treating with anhydrous Na ₂ SO ₄ After drying, filtration was performed, the solvent in the filtrate was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =2 by volume).

The compound b6 is a yellow oil; molecular formula C ₂₅ H ₃₁ N ₃ O ₄ ； ¹ H-NMR(400MHz,CD ₃ OD)δ:7.53(d,J＝1.2Hz,1H),7.37(dd,J＝7.7,1.2Hz,1H),7.26(d,J＝8.1Hz,1H),7.01(ddd,J＝8.2,7.1,1.3Hz,1H),6.93(td,J＝7.5,1.1Hz,1H),4.44(dd,J＝6.1,1.4Hz,1H),4.33(d,J＝7.0Hz,1H),4.13(td,J＝5.1,1.5Hz,1H),3.60(s,3H),3.38(d,J＝15.4Hz,1H),3.11–3.02(m,1H),3.01–2.97(m,1H),2.95(dd,J＝6.2,1.8Hz,1H),2.34–2.29(m,1H),2.25(ddd,J＝13.6,6.7,1.6Hz,1H),1.93(s,3H),1.62(ddd,J＝13.9,9.3,4.9Hz,1H),1.29(q,J＝7.2Hz,2H),0.99(d,J＝7.1Hz,3H),0.64(t,J＝7.4Hz,3H)； ¹³ C-NMR(100MHz,CD ₃ OD)δ:172.83,171.10,148.22,138.13,133.83,128.15,122.35,119.96,118.55,112.03,107.21,103.97,74.40,64.74,53.05,51.26,48.38,43.50,42.96,42.31,34.55,24.27,23.68,14.40,11.45。

Example 12

34.4mg (0.1507 mmol) of loganin aglycone, 40mg (0.2261 mmol) of 1-methyltryptamine and 10.9mg (0.0301 mmol) of zinc trifluoromethanesulfonate were weighed and placed in a sealed tube, and vacuum-replaced with N ₂ Then 2mL of dichloromethane is added, the mixture is stirred for reaction for 4h under the condition of oil bath at 45 ℃, and the reaction is monitored by TLC; after the reaction is finished, transferring the obtained product system into a separating funnel filled with water, and dropwise adding saturated NaHCO ₃ Until no bubble is generated, extracting the obtained system with ethyl acetate for 3 times, combining organic phases, and purifying with anhydrous Na ₂ SO ₄ Drying, filtering, evaporating solvent from the filtrate under reduced pressure, and subjecting the residue to silica gel column chromatography (by volume, petroleum ether): ethyl acetate =1.5: 1) Purification gave compound c1 (yield 26.2mg, 47.2%).

The compound c1 is a yellow oil; molecular formula C ₂₂ H ₂₈ N ₂ O ₃ ；

Is 55 (c =0.1,meoh); UV lambda _max (MeOH)224,294nm；IR(KBr)cm ^-1 :3735,3438,2926,2357,1649,1510 and 739; ¹ H-NMR(400MHz,CD ₃ OD)δ:7.50(dt,J＝7.9,0.9Hz,1H),7.27(d,J＝8.2Hz,1H),7.22(s,1H),7.11(ddd,J＝8.2,6.9,1.1Hz,1H),7.00(ddd,J＝8.0,7.0,1.0Hz,1H),6.90(s,1H),3.95(td,J＝5.3,1.7Hz,1H),3.68(s,3H),3.42(td,J＝6.9,2.7Hz,2H),3.06(dd,J＝12.4,4.8Hz,1H),2.93(dt,J＝7.0,3.3Hz,2H),2.87(d,J＝8.0Hz,1H),2.71(dd,J＝12.4,6.8Hz,1H),2.08(ddd,J＝13.9,7.4,1.8Hz,1H),1.74(qd,J＝7.5,4.8Hz,1H),1.59–1.48(m,1H),1.39(ddd,J＝13.7,8.2,5.3Hz,1H),0.89(d,J＝7.0Hz,3H)； ¹³ C-NMR(100MHz,CD ₃ OD)δ:171.54,148.27,138.69,129.06,128.51,122.50,119.75,119.47,111.96,110.28,99.71,74.97,57.31,50.91,48.28,43.75,43.46,42.94,33.45,32.64,25.99,13.59。

example 13

Weighing loganin aglycone 30.4mg (0.1316 mmol), 1-methyl-5-methoxytryptamine 37mg (0.1974 mmol) and zinc triflate 10mg (0.0263 mmol), placing into sealed tube, and vacuum replacing to N ₂ Then 2mL of dichloromethane is added, the mixture is stirred for reaction for 4h under the condition of oil bath at 45 ℃, and the reaction is monitored by TLC; after the reaction is finished, transferring the obtained product system into a separating funnel filled with water, and dropwise adding saturated NaHCO ₃ Until no bubble is generated, extracting the obtained system with ethyl acetate for 3 times, combining organic phases, and purifying with anhydrous Na ₂ SO ₄ After drying, filtration was performed, the solvent in the filtrate was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =1 by volume) to obtain compound c2 (yield 28.6mg, yield 54.6%).

The compound c2 is a yellow oil; molecular formula C ₂₃ H ₃₀ N ₂ O ₄ ；

52.4 (c =0.1,meoh); UV lambda _max (MeOH)223,295nm；IR(KBr)cm ^-1 :3438,1681,1498,1175 and 783; ¹ H-NMR(400MHz,CD ₃ OD)δ:7.24–7.22(m,1H),7.14(d,J＝8.8Hz,1H),6.95(d,J＝2.4Hz,1H),6.85(s,1H),6.75(dd,J＝8.8,2.4Hz,1H),3.94–3.88(m,1H),3.75(s,3H),3.63(s,3H),3.50(s,3H),3.38(td,J＝6.8,5.5Hz,2H),3.02(dd,J＝12.4,4.8Hz,1H),2.87(td,J＝6.8,3.2Hz,3H),2.66(dd,J＝12.4,6.9Hz,1H),2.05(ddd,J＝13.9,7.4,1.8Hz,1H),1.70(qd,J＝7.5,4.9Hz,1H),1.49(td,J＝7.2,5.3Hz,1H),1.35(ddd,J＝13.7,8.3,5.3Hz,1H),0.86(d,J＝7.0Hz,3H)； ¹³ C-NMR(100MHz,CD ₃ OD)δ:171.53,155.14,148.28,134.08,129.37,129.12,112.61,111.60,111.05,101.48,99.64,74.95,57.28,56.25,50.92,48.43,43.74,43.42,42.94,33.44,32.81,25.98,13.61。

example 14

Weighing loganin aglycone 30mg (0.1314 mmol), compound Mb 4mg (0.1577 mmol) and zinc trifluoromethanesulfonate 9.5mg (0.0263 mmol), placing into a sealed tube, and vacuum replacing to N ₂ Then 2mL of dichloromethane is added, the mixture is stirred and reacted for 4h at the temperature of 45 ℃ in an oil bath, and the reaction is monitored by TLC; after the reaction is finished, transferring the obtained product system into a separating funnel filled with water, and dropwise adding saturated NaHCO ₃ Until no bubbles are generated, extracting the obtained system with ethyl acetate for 3 times, combining organic phases, and treating with anhydrous Na ₂ SO ₄ After drying, filtration was performed, the solvent in the filtrate was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =1.5 by volume) to obtain a compound c3 (yield 24.4mg, yield 43.5%) and a compound c4 (yield 29.2mg, yield 52.4%), respectively.

The compound c3 is a yellow oil; molecular formula of C ₂₄ H ₃₀ N ₂ O ₅ ；

57 (c =0.1,meoh); UV lambda _max (MeOH)221,290nm；IR(KBr)cm ^-1 ：3438,2945,1732,1675,1611,1169 and 739; ¹ H-NMR(400MHz,CD ₃ OD)δ:7.50–7.46(m,1H),7.30(s,1H),7.25(d,J＝8.3Hz,1H),7.09(ddd,J＝8.2,7.0,1.2Hz,1H),6.97(ddd,J＝8.0,7.0,1.0Hz,1H),6.91(s,1H),4.20(dd,J＝10.8,5.3Hz,1H),3.75–3.71(m,1H),3.69(s,3H),3.66(s,3H),3.52(s,3H),3.33(dd,J＝15.0,5.3Hz,1H),3.19–3.15(m,1H),3.13(dd,J＝7.5,3.3Hz,1H),2.79(q,J＝7.8Hz,1H),2.61(dd,J＝12.2,4.7Hz,1H),2.00–1.93(m,1H),1.72(dt,J＝9.2,4.6Hz,1H),1.36–1.27(m,1H),1.11–1.01(m,1H),0.58(d,J＝6.9Hz,3H)； ¹³ C-NMR(100MHz,CD ₃ OD)δ:173.14,171.18,146.49,138.58,129.12,128.93,122.60,119.93,119.40,110.37,110.37，102.98,75.46,67.99,52.79,51.09,47.32,43.88,43.65,41.86,33.73,32.70,27.14,12.68。

the compound c4 is a yellow oil; molecular formula C ₂₄ H ₂₈ N ₂ O ₅ ；

Is-46 (c =0.1,meoh); UV lambda _max (MeOH)220,286nm；IR(KBr)cm ^-1 :3741,3432,2926,1738,1542,1125 and 739; ¹ H-NMR(400MHz,CD ₃ OD)δ:7.49(dt,J＝7.9,1.0Hz,1H),7.23(dt,J＝8.2,0.9Hz,1H),7.08(ddd,J＝8.2,5.1,1.2Hz,1H),7.00–6.96(m,1H),6.95(dt,J＝2.6,1.1Hz,1H),6.91(s,1H),5.70(dt,J＝2.6,1.4Hz,1H),4.23(dd,J＝9.8,5.3Hz,1H),4.23(dd,J＝9.8,5.3Hz,1H),4.00(td,J＝6.1,3.0Hz,1H),3.68(s,3H),3.65(s,3H),3.51(s,3H),3.47(s,1H),3.32(d,J＝5.2Hz,1H),3.12(dd,J＝14.9,9.7Hz,1H),2.52(ddt,J＝7.3,5.6,1.7Hz,1H),2.31–2.22(m,1H),1.61(ddd,J＝13.1,10.1,6.3Hz,1H),0.94(d,J＝7.3Hz,3H)； ¹³ C-NMR(100MHz,CD ₃ OD)δ:172.57,170.73,143.39,138.46,129.20,128.99,128.19,122.54,120.86,119.92,119.46,110.24,109.93,100.86,73.51,66.87,52.93,51.23,44.12,42.06,34.50,32.72,27.71,13.16。

example 15

Weighing 16mg (0.0701 mmol) of loganin aglycone, 19.7mg (0.0841 mmol) of 5-hydroxytryptophan and 2.6mg (0.0072 mmol) of zinc trifluoromethanesulfonate into a sealed tube, adding 1.5mL of hexafluoroisopropanol, stirring for reaction at 45 ℃ in a water bath for 10h, and monitoring the reaction by TLC; after the reaction was completed, the solvent in the obtained product system was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =1 by volume) to obtain compound d2 (yield 13.5mg, yield 41.8%).

The compound d2 is a yellow solid; molecular formula C ₂₃ H ₂₇ N ₂ O ₆ ； ¹ H NMR(400MHz,Methanol-d ₄ )δ7.59(d,J＝1.1Hz,1H,H-5),7.12(d,J＝8.6Hz,1H,H-9),6.76(d,J＝2.3Hz,1H,H-12),6.61(dd,J＝8.7,2.4Hz,1H,H-11),4.60(dd,J＝5.8,1.7Hz,1H,H-7),4.20(d,J＝9.1Hz,1H,H-13b),4.15(t,J＝5.0Hz,1H,H-2),3.63(s,3H,-COOCH ₃ ),3.51(s,3H,-COOCH ₃ ),3.24(s,1H,H-8),3.20–3.11(m,1H,H-3a),3.00(ddd,J＝15.0,5.6,1.7Hz,1H,H-8),2.44–2.35(m,1H,H-1),2.34(dd,J＝13.3,6.3Hz,1H,H-3),1.68(td,J＝8.5,5.2Hz,1H,H-1,H-13c),1.54–1.39(m,1H,H-3),1.02(d,J＝7.1Hz,3H,-CH ₃ ). ¹³ C NMR(100MHz,Methanol-d ₄ )δ173.45(-COO-),171.31(-COO-),151.54(C-10),148.73(C-5),134.32(C-14),133.15(C-13a),128.50(C-8b),112.55(C-11),112.38(C-9),106.86(C-8a),102.91(C-12),102.11(C-4),74.19(C-2),63.95(C-7),53.73(C-13b),52.85(-OCH ₃ ),51.22(-OCH ₃ ),48.36(C-13c),44.19(C-3),43.51(C-1),35.54(C-3a),25.17(C-8),15.04(-CH ₃ )。

Example 16

Weighing 20mg (0.0876 mmol) of loganin aglycone, 25mg (0.1072 mmol) of 5-hydroxytryptophan formamide and 3.2mg (0.0088 mmol) of zinc trifluoromethanesulfonate, filling into a sealed tube, adding 1.5mL of hexafluoroisopropanol, stirring for reaction at 45 ℃ in a water bath for 20h, and monitoring the reaction by TLC; after the reaction was completed, the solvent in the obtained product system was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =1 by volume) to obtain compound d3 (yield 15.7mg, yield 42.1%).

The compound d3 is a yellow solid; molecular formula C ₂₃ H ₂₈ N ₃ O ₅ ； ¹ H NMR(400MHz,Acetone-d ₆ )δ9.88(s,1H,-NH),7.47–7.40(m,1H,H-5),7.16(d,J＝8.6Hz,1H,H-9),6.84(d,J＝2.3Hz,1H,H-12),6.64(dd,J＝8.6,2.4Hz,1H,H-11),4.52(d,J＝6.7Hz,1H,H-7),4.47–4.39(m,1H,H-13b),4.12(t,J＝4.2Hz,1H,H-2),3.54(d,J＝1.0Hz,3H,-OCH ₃ ),3.47(dd,J＝15.4,1.3Hz,1H,H-8),3.02(d,J＝7.3Hz,1H,H-3a),2.95–2.86(m,1H,H-8),2.69(d,J＝4.7Hz,3H,-NHCH ₃ ),2.24(d,J＝3.9Hz,1H,H-1),2.21–2.16(m,1H,H-3),1.88–1.81(m,1H,H-13c),1.61(ddd,J＝13.8,7.5,5.1Hz,1H,H-3),1.11–1.04(m,3H,-CH ₃ )； ¹³ C NMR(100MHz,Acetone-d ₆ )δ171.07(-CONH-),168.63(-COO-),151.74(C-10),146.30(C-5),134.68(C-14),132.13(C-13a),128.88(C-8b),112.30(C-11),111.83(C-9),106.53(C-8a),105.59(C-12),102.95(C-4),73.71(C-1),63.98(C-7),51.25(C-13b),50.53(-OCH ₃ ),46.34(C-13c),43.58(C-3),42.20(C-1),33.09(C-3a),26.46(-NHCH ₃ ),22.70(C-8),13.80(-CH ₃ )。

Example 17

Weighing loganin aglycone 30mg (0.1314 mmol), 5-hydroxytryptophan acetamide 39mg (0.1577 mmol) and zinc trifluoromethanesulfonate 4.8mg (0.0131 mmol), putting into a sealed tube, adding 1.5mL hexafluoroisopropanol, stirring for reaction for 30h under the condition of water bath 45 ℃, and monitoring the reaction by TLC; after the reaction was completed, the solvent in the obtained product system was evaporated to dryness under reduced pressure, and the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate =1.

The compound d4 is a yellow solid; molecular formula C ₂₄ H ₃₀ N ₃ O ₅ ； ¹ H NMR(400MHz,Acetone-d ₆ )δ9.87(s,1H,-NH),7.67(s,1H,H-5),7.15(d,J＝8.6Hz,1H,H-9),6.84(d,J＝2.4Hz,1H,H-12),6.64(dd,J＝8.6,2.4Hz,1H,H-11),4.51(d,J＝6.9Hz,1H,H-7),4.45(dd,J＝4.4,2.0Hz,1H,H-13b),4.12(d,J＝4.7Hz,1H,H-2),3.53(s,3H,-OCH ₃ ),3.47(dt,J＝15.4,1.3Hz,1H,H-8),3.30–3.16(m,2H,-NHCH ₂ -),3.16–3.13(m,1H,H-3a),2.97(d,J＝7.5Hz,1H,H-8),2.37(ddd,J＝9.4,8.0,4.5Hz,1H,H-1),2.21–2.17(m,1H,H-3),2.17–2.14(m,1H,H-13c),1.64(ddd,J＝14.0,6.7,5.2Hz,1H,H-3),1.10(d,J＝6.9Hz,3H,-CH ₃ ),1.10–1.01(m,3H,-CH ₃ )； ¹³ C NMR(100MHz,Acetone-d ₆ )δ169.97(-CONH-),168.43(-COO-),151.73(C-10),146.18(C-5),134.80(C-14),132.07(C-13a),129.00(C-8b),112.27(C-11),111.79(C-9),106.58(C-8a),106.19(C-12),102.94(C-4),73.79(C-2),63.97(C-7),50.93(C-13b),50.53(-OCH ₃ ),45.80(C-13c),43.52(C-3),41.96(C-1),34.98(C-3a),32.63(-NHCH ₂ -),22.42(C-8),15.26(-CH ₃ ),13.63(-CH ₃ )。

Example 18

Weighing loganin aglycone 30mg (0.1314 mmol), 5-hydroxytryptophan propionamide 41.2mg (0.1577 mmol) and zinc trifluoromethanesulfonate 4.8mg (0.0131 mmol), putting into a sealed tube, adding 1.5mL hexafluoroisopropanol, stirring for reaction for 30h under the condition of water bath 45 ℃, and monitoring the reaction by TLC; after the reaction was completed, the solvent in the obtained product system was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate =1 by volume) to obtain compound d5 (yield 22.1mg, yield 37.1%).

The compound d5 is a brown solid; molecular formula C ₂₅ H ₃₂ N ₃ O ₅ ； ¹ H NMR(400MHz,Acetone-d ₆ )δ9.87(s,1H,-NH),7.69(s,1H,H-5),7.15(dd,J＝8.6,0.6Hz,1H,H-9),6.84(d,J＝2.3Hz,1H,H-12),6.64(dd,J＝8.6,2.4Hz,1H,H-11),4.52(d,J＝6.7Hz,1H,H-7),4.46(dt,J＝4.8,1.8Hz,1H,H-13b),4.12(s,1H,H-2),3.54(s,3H,-OCH ₃ ),3.47(dt,J＝15.4,1.3Hz,1H,H-8),3.14(dtd,J＝14.5,7.2,5.9Hz,2H,-NHCH ₂ -),3.00(d,J＝7.6Hz,1H,H-3a),2.92–2.85(m,1H,H-8),2.40–2.27(m,1H,H-1,H-1),2.27–2.18(m,1H,H-3),2.20–2.15(m,1H,H-13c),1.63(ddd,J＝13.9,7.1,5.1Hz,1H,H-3),1.54–1.38(m,2H,-CH ₂ CH ₃ ),1.09(d,J＝6.9Hz,3H,-CH ₃ ),0.79(t,J＝7.4Hz,3H,-CH ₂ CH ₃ )； ¹³ C NMR(100MHz,Acetone-d ₆ )δ170.25(-CONH-),168.50(-COO-),151.72(C-10),146.34(C-5),134.74(C-14),132.10(C-13a),128.94(C-8b),112.26(C-11),111.80(C-9),106.59(C-8a),105.81(C-12),102.94(C-4),73.73(C-2),64.02(C-7),51.18(C-13b),50.54(-OCH ₃ ),46.13(C-13c),43.53(C-3),42.07(-NHCH ₂ -),41.93(C-1),32.83(C-3a),23.62(-CH ₂ -),22.66(C-8),13.78(-CH ₃ ),11.65(-CH ₃ )。

Test example

1. Test for inhibitory Activity of Compound on alpha-glucosidase

Alpha-glucosidase (AGI) hydrolyzes p-nitrophenyl-alpha-D-glucoside (PNPG) which is hydrolyzed into glucose and p-nitrophenol (PNP), and the PNP has maximum absorption at 405nm, by utilizing the principle, after a sample to be tested is added, the change of the absorbance is tested, the inhibition rate of the sample on the AGI is calculated, and the IC of the acarbose is positively controlled ₅₀ The values are compared and the samples are evaluated for their inhibitory effect on AGI.

1.1 Experimental reagents

TABLE 1 test reagents

Reagent	Company (SA)
		Alpha-glucosidase (AGI)	SHANGHAI YUANYE BIOTECHNOLOGY Co.,Ltd.
Acarbose	Sigma-Aldrich, USA
		p-Nitrobenzene-alpha-D-glucoside (PNPG)	Sigma-Aldrich, USA
Sodium carbonate (analytically pure)	Aladdin

1.2 Experimental groups

Sample group: the concentrations of the test compounds were 12.5nmmoL/L, 25nmmoL/L, 50nmmoL/L, 100nmmoL/L, 200nmmoL/L.

Positive control group: acarbose, at the same concentration as the test compound; sample blank, negative control group, blank group.

1.3 solvent preparation and Experimental operation

Preparing a solvent: the compounds were formulated as 12.5nmmo L/L, 25nmmo L/L, 50nmmo L/L, 100nmmo L/L, 200nmmo L/L, alpha glucosidase concentration 0.5U/mL, PNPG concentration 5mmoL/L, sodium carbonate concentration 0.2moL/L, all dissolved in PBS (pH = 6.8). The experimental procedure was as per table 2:

TABLE 2 alpha-glucosidase experiment the order of addition of reagents

Calculating the formula: inhibition I = [1- (a-b)/(d-c) ] × 100%, where a is a sample group, b is a sample control group, c is a blank group, and d is a negative control group; the experimental data of this study were analyzed using GraphPad Prism 8.0 software.

1.4 results of the experiment

As can be seen from Table 3, the inhibition ratios of the compounds b4, b6, c1 and d3 to α -glucosidase were all greater than 50%, indicating that they have the effect of inhibiting α -glucosidase. Wherein IC of compound b4 ₅₀ The value is smaller than that of the positive control acarbose, which indicates that the acarbose has a remarkable hypoglycemic effect.

Results of the alpha-glucosidase inhibitory Activity of the Compounds of Table 3

Class of compounds	Inhibition ratio (%)	IC ₅₀ (nmoL/L)
			Acarbose	94.5	65.204
Compound b4	86.7	60.124
			Compound b6	79.8	74.953
Compound c1	92.3	76.944
			Compound d3	69.3	147.3

2. Activity test experiment of compound on abnormal proliferation of HBZY-1 cells cultured with high sugar

2.1 establishment of HBZY-1 cell hyperglycosemia model

2.1.1 Experimental groups

A (control group): cell, low carbohydrate (11 mmol/L) DMEM complete medium; b (high sugar group): cells, high sugar (22.5 mmol/L, 25mmol/L, 27.5mmol/L, 30mmol/L, 32.5 mmol/L) DMEM complete medium; c (blank group): low-sugar DMEM complete medium (11 mmol/L). Each set requires five parallel replicates in a 96-well plate.

2.1.2 preparation of solutions

Complete medium with a glucose content of 22.5mmol/L (50 mL): 50mL low sugar medium +102.5mg glucose; complete medium with a glucose content of 25mmol/L (50 mL): 50mL of low-sugar medium +125mg of glucose; complete medium with a glucose content of 27.5mmol/L (50 mL): 50mL of low-sugar medium +147.5mg of glucose; complete medium with a glucose content of 30mmol/L (50 mL): 50mL of low-sugar medium +170mg of glucose; complete medium with a glucose content of 32.5mmol/L (50 mL): 50mL of low-sugar medium +192.5mg of glucose; complete medium with a glucose content of 35mmol/L (50 mL): 50mL of low-sugar medium +215mg of glucose.

2.1.3 Experimental methods

After cell digestion, 10. Mu.L of the cells were put on a cell counting plate, counted under a microscope, and counted by (four large grids/4). Times.10 according to the principle of counting cells without counting down or counting left or right ⁴ Calculating cell density by using a formula of x cell dilution factor, and diluting to make plate density reach 5 x 10 ⁴ and/mL. Performing experiments according to groups, wherein the concentration of high glucose group glucose is 25mmol/L, 27.5mmol/L, 30mmol/L, 32.5mmol/L and 35mmol/L, culturing for different culture times (24 h, 48h, 72h and 96 h), each well culture medium is 200 μ L, adding 20 μ L CCK-8 reagent for detection after corresponding culture time is reached, and detecting with CO ₂ After culturing for 15min in an incubator, measuring the absorbance at 450nm, and comparing the cell viability of the HBZY-1 cells in abnormal proliferation under different glucose concentrations and different time conditions. Calculating the formula: cell viability (%) = ((a) _B -A _C )/(A _A -A _C ) X 100% in which A is _A Represents the absorbance of group A, A _B Represents the absorbance of group B, A _C Represents the absorbance of group C.

2.1.4 data processing

The experimental data of the research are analyzed by GraphPad Prism 8.0, the experimental result is expressed by mean + -SEM, the average number comparison of multiple groups of samples is carried out by applying a single factor, and the statistical significance is shown when the p value is less than 0.05.

2.1.5 results of the experiment

FIG. 1 is a graph showing the effect of different glucose concentrations and different culture times on HBZY-1 cells (Data representation mean. + -. SEM, n =5; as compared with HG, "+" indicates that p is less than 0.05 and "+" indicates that p is less than 0.01), and the results of the in vitro pharmacological activity test shown in FIG. 1 show that when the glucose concentration is 30mmoL/L and the culture time is 48 hours, the cell proliferation is significant, and the drug intervention is performed under these conditions.

2.2 determination of the Effect of Compounds on HBZY-1 cell viability

2.2.1 Experimental groups

Blank group: contains culture medium and CCK-8 reagent, and is free of cells and drugs;

control group: contains cells, culture medium and CCK-8 reagent, and does not contain medicine;

drug group: contains cells, culture medium, CCK-8 reagent and medicine, and has administration concentration of 0.01. Mu. Mol/L, 0.1. Mu. Mol/L, 1. Mu. Mol/L, 10. Mu. Mol/L, 100. Mu. Mol/L.

2.2.2 Experimental methods

Selecting 96-well plate by CCK-8 method, grouping the plates, each group having 5 multiple wells, each well having 100 μ L cell culture solution, cell concentration of 5 × 10 ⁴ And culturing cells/mL for 24h, sucking out old culture solution, adding corresponding culture solution and drugs according to experimental groups, culturing for 48h, adding 10 mu L of CCK-8 reagent into each hole, incubating for 1-4 h in an incubator, and measuring the absorbance value at the wavelength of 490nm at intervals of 1h. And judging whether the compounds with different concentrations are toxic to HBZY-1 cells according to the cell viability.

2.2.1 results of the experiment

FIG. 2 is a bar chart of the effect of compounds on HBZY-1 cell viability, and the results show that, in the drug concentration range of 0.01-100. Mu. Mol/L, the compounds have promotion effect on HRMC cell proliferation, and a few have inhibition effect, but the statistical results show that no significant difference (p > 0.05) exists, so the compounds are judged to be non-toxic. Therefore, 0.01 to 100. Mu. Mol/L was selected as the drug concentration range for the subsequent experiments.

2.3 Effect of Compounds on the high sugar Induction of abnormal proliferation of HBZY-1 cells

2.3.1 Experimental groups

A (control group): cells, low-sugar DMEM complete medium (glucose content 11 mmol/L); b (high carbohydrate group) cells, high carbohydrate (30 mmol/L) DMEM complete medium; c (administration group): synthesis of the compound: cells, high-sugar (30 mmol/L) DMEM complete medium, different concentrations (0.01, 0.1, 1, 10, 100. Mu. Mol/L) of each drug; d (blank group): high sugar (30 mmol/L) DMEM complete medium; each set of five parallel replicates were placed in a 96-well plate.

2.3.2 Experimental methods

And (3) detecting whether the compound has an inhibition effect on the proliferation of the high-sugar induced HBZY-1 cells under different concentrations by adopting a CCK-8 method.

Taking cells cultured by normal sugar, digesting, adding culture medium, blowing uniformly, counting, regulating cell seed density to 1 × 10 ⁴ And each/mL, grouping according to experiments, taking 100 mu L of the compound, adding a complete culture medium to 200 mu L, after culturing for 24h, absorbing the old culture medium, adding 200 mu L of a serum-free culture medium into each well, administering after culturing for 24h, absorbing the old culture medium, wherein the administration volume is 200 mu L, after culturing for 48h, adding 20 mu L of CCK-8 reagent into each well, placing the mixture in an incubator for reacting for 2h, measuring the absorbance at 450nm by using a microplate reader, calculating the cell viability, and detecting whether the compound inhibits the abnormal proliferation of the high-sugar-induced HBZY-1 cells.

2.3.3 data processing

Cell viability (%) = ((a) _C -A _D )/(A _B -A _D ))×100％。

The experimental data of the research are analyzed by GraphPad Prism 8.0, and the experimental results are mean

SEM shows that when the average number of multiple groups of samples is compared by using a single factor, the p value is less than 0.05, which indicates that the statistical significance is achieved.

2.3.4 results of the experiment

The effect of the compounds on the activity of high-glucose-induced HBZY-1 cells was plotted on a bar graph with the administration concentration as abscissa and the cell activity as ordinate, and the results are shown in FIG. 3. As can be seen from fig. 3, compounds a1, a2, b3, b4, b5, b6, c1, c2, and d3 had significant inhibitory effects on abnormal cell proliferation.

3. Effect of compound on SOD and MDA content in high-sugar-induced HBZY-1 cells

The experimental method is carried out according to the specifications of SOD and MDA kits.

3.1 Experimental groups

Control group: cell, low carbohydrate (11 mmol/L) DMEM complete medium; high sugar group: cells, high sugar (30 mmol/L) DMEM complete medium; administration group: (1) test compound: cells, high-sugar (30 mmol/L) DMEM complete medium, different concentrations of each drug (50. Mu.g/mL, 100. Mu.g/mL, 200. Mu.g/mL, 400. Mu.g/mL, 800. Mu.g/mL), (2) compound: cells, high-sugar (30 mmol/L) DMEM complete medium, different concentrations of each drug (0.01. Mu. Mol/L, 0.1. Mu. Mol/L, 1. Mu. Mol/L, 10. Mu. Mol/L, 100. Mu. Mol/L); positive control group: cells, high-sugar complete medium (30 mmol/L), positive control group (Tempol, 100. Mu. Mol/L). Each set of three parallel multiple wells was placed in a 24-well plate.

3.2 Experimental methods

According to the operation instructions of different kits, the contents of SOD and MDA in each component cell are calculated according to the BCA method.

Cell count, adjusted cell density to 6X 10 ⁵ Taking 100 mu L of the mixture per mL, placing experimental groups, namely each drug contains 8 experimental groups (five administration groups with different concentrations, NG, HG, tempol) and each group needs to be provided with three parallel multiple wells, culturing for 24 hours, changing a complete culture medium without serum to culture for 24 hours, then changing the complete culture medium with different drug concentrations to culture for 48 hours, and then operating according to the kit instructions.

3.3BCA kit procedures

(1) Pretreatment of cells

Removing old culture medium in a 24-pore plate by suction, adding 200 muL pancreatin into each pore, placing in an incubator for digestion for 2min, adding 200 muL culture medium to stop digestion, transferring cells to a 1.5mL centrifuge tube for centrifugation (1000rmp, 5 min), removing supernatant after the centrifugation, rinsing with PBS twice, adding 1mL PBS for uniform blowing, counting cells, centrifuging again after counting, discarding supernatant to obtain cell precipitate

(2) BCA assay

Adding 70-130 mul of protein lysate into each tube, putting the tube into an ultrasonic instrument, carrying out ultrasonic treatment with 300w power in ice-water bath for 6 times and 50s each time to release the protein in the cells. Centrifuging for 5min under 10000rmp, taking 20 μ L of supernatant to a 96-well plate, adding 200 μ L of LBCA working solution, reacting for 15min at 37 ℃, measuring the absorbance value at 562nm by using an enzyme-labeling instrument, and calculating the protein concentration according to a standard curve.

Preparation of a protein standard curve: preparing a working solution: the number of standards and samples was calculated as BCA: the requirements of the Cu reagent =50 (V: V) were satisfied, and a BCA working solution was prepared and mixed well by shaking. Diluting the standard substance: mu.L of BSA standard was diluted to 100. Mu.L with PBS, i.e., to a final concentration of 0.5mg/mL. Taking 0 μ L, 2 μ L, 4 μ L, 6 μ L, 8 μ L, 12 μ L, 16 μ L and 20 μ L of standard substances respectively, adding PBS to make up to 20 μ L, sequentially adding 200 μ L of BCA working solution into each well, and standing at 37 deg.C for 15min. And measuring the absorbance of the protein at 562nm by using a microplate reader, and preparing a protein standard curve by taking the protein concentration as an abscissa and the absorbance as an ordinate.

3.4SOD kit operation

Counting cells, adding 1mL of SOD extracting solution into every 500 ten thousand cells, carrying out ultrasonic treatment to break the cells, carrying out ultrasonic treatment for 3s each time for 30 times at intervals of 10s each time, centrifuging for 10min at 8000g and 4 ℃ to obtain supernatant, namely a sample to be detected, and placing the sample on ice to be detected.

The determination step comprises:

preheating an enzyme-labeling instrument for more than 30min before measurement, and setting the wavelength to be 560nm;

before use, the reagent II needs to be sequentially centrifuged and blown; adding the reagent IV into the reagent V, dissolving by ultrasonic wave, and preparing in situ; before testing, the first, third and fifth reagents need to be bathed in water at 25 ℃ for more than 5min, and then the operations are carried out according to the following table 5:

TABLE 5 SOD reagent procedure

The reagents are added into a 96-well plate with groups according to the sequence, fully and uniformly mixed, placed in an environment with the temperature of 37 ℃ for reaction for 30min, and an enzyme-linked immunosorbent assay device is used for measuring the absorbance value of the reagents at the position of 560 nm.

3.5MDA kit manipulation

Counting cells, adding 1mL of MDA extracting solution into every 500 ten thousand cells, performing ultrasonic treatment for breaking the cells, performing ultrasonic treatment for 3s every time for 30 times at intervals of 10s every time, centrifuging for 10min at 8000g and 4 ℃ to obtain supernatant, which is a sample to be detected, and placing the sample on ice for detection.

The determination step comprises:

preheating an enzyme-labeling instrument for more than 30min before measurement;

preparing an MDA working solution: during the use, add reagent one of 15mL into reagent two, fully dissolve, can go on under water bath 70 ℃ condition during the dissolution, shake repeatedly, dissolve with the supersound if not good, place 4 ℃ refrigerator for use, later operate according to table 6:

TABLE 6 MDA reagent procedure

Reagent name (μ L)	Measuring tube	Blank tube
			MDA detection working solution	150	150
Distilled water	-	50
			Sample(s)	50	-
Reagent III	50	50

Adding all reagents into a 1.5mL EP tube, reacting in a metal bath at 100 ℃ for 60min after the addition is finished, then placing in an ice bath for cooling, centrifuging for 10min under the conditions of normal temperature and 10000g, absorbing 100 mu L of supernatant into a 96-well plate which is provided with groups, and measuring the absorbance values at 450nm, 532nm and 600nm respectively by using an enzyme labeling instrument.

3.6 calculation method

The BCA content of each group of samples was calculated from the protein standard curve.

SOD activity (U/mL) =11.11 × inhibition percentage ÷ (1-inhibition percentage) ÷ Cpr × F;

Δ a assay = a assay-a control; Δ a blank = a blank 1-a blank 2;

MDA content (nmol/mgpot) =5 × (12.9 × (Δ a532- Δ a 600) -2.58 × Δ a 450) ÷ Cpr;

ΔA450＝A450 _{measurement of} -A450 _{Blank space} ，ΔA532＝A532 _{Measurement of} -A532 _{Blank space} ，ΔA600＝A600 _{Measurement of} -A600 _{Blank space} ；

Wherein Cpr represents the sample protein concentration; f represents the dilution factor;

Δ A represents the difference between the two absorbance groups, and A represents the absorbance.

3.7 results of the experiment

In vitro pharmacological activity experiments show that the compounds a2, b6, c1 and d3 can obviously improve SOD activity in HBZY-1 cells and reduce the content of MDA, and have the effect of relieving oxidative stress of the HBZY-1 cells within a certain concentration range. Wherein the compound b6 is 0.1-100 mu mol/L, the compounds a2 and c1 are 1-10 mu mol/L, and the compound d3 is 0.01-10 mu mol/L, which has relieving effect on the oxidative stress of HBZY-1 cells induced by high sugar, and shows that the SOD activity is obviously increased, the MDA content is obviously reduced, and a concentration dependence relationship exists. The SOD activity and MDA content are shown in tables 7-9:

TABLE 7 SOD Activity and MDA content in HBZY-1 cells induced by high sugar after administration of Compound a2

TABLE 8 high sugar Induction of SOD Activity and MDA content in HBZY-1 cells after administration of Compound b6

Group (Compound b 6)	SOD activity (U/mL)	MDA content (nmol/mg pot)
			NG	4.74±0.11**	0.82±0.178**
Tempol	4.05±0.42**	1.69±0.53**
			HG	2.57±0.46	2.33±0.34
0.01μmol/L	4.33±0.58**	1.80±0.21
			0.1μmol/L	4.00±0.09**	0.88±0.25**
1μmol/L	4.64±0.25**	0.58±0.08**
			10μmol/L	6.06±0.82**	0.36±0.12**
100μmol/L	7.45±0.48**	0.37±0.08**

TABLE 9 high sugar Induction of SOD Activity and MDA content in HBZY-1 cells after administration of Compound c1

Group (Compound c 1)	SOD activity (U/mL)	MDA content (nmol/mg pot)
			NG	7.17±0.24**	0.35±0.06**
Tempol	6.44±0.27**	1.678±1.21*
			HG	3.71±1.11	3.15±0.55
0.01μmol/L	3.79±0.894	1.38±0.81*
			0.1μmol/L	4.04±0.82	1.32±0.17*
1μmol/L	5.12±0.17**	1.14±0.82**
			10μmol/L	7.86±0.40**	1.08±0.94**
100μmol/L	10.36±0.67**	2.81±1.22

TABLE 10 SOD activity and MDA content in HBZY-1 cells induced by high sugar after administration of Compound d3

Group (Compound d 3)	SOD activity (U/mL)	MDA content (nmol/mg pot)
			NG	8.07±0.47**	0.80±0.04**
Tempol	6.72±0.23**	1.24±0.22**
			HG	2.24±0.25	3.95±0.33
0.01μmol/L	4.66±0.15*	2.41±1.15**
			0.1μmol/L	4.88±0.61*	1.49±0.89**
1μmol/L	5.70±0.51*	0.93±0.20**
			10μmol/L	6.63±1.75*	0.78±0.13**
100μmol/L	33.20±4.86	1.55±0.22

Remarking: in tables 7 to 9, data representation mean ± SEM, n =5; in comparison to HG, "'means p is less than 0.05 and"' means p is less than 0.01.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims

1. A monoterpene indole-like alkaloid has a structure shown in formula I or formula II:

said R is ¹ Is hydrogen atom, halogen atom, methyl, ethyl, hydroxyl, methoxyl, formyloxy, acetoxyl, substituted benzyloxy or unsubstituted benzyloxy; at least one substituent group is arranged on a benzene ring in the substituted benzyloxy, and the substituent group is a halogen atom, a methyl group or a nitro group;

2. The monoterpenoid indole alkaloid of claim 1, wherein the halogen atom is-F, -Cl, or-Br.

3. The monoterpenoid indole alkaloid of claim 1 or 2, wherein the monoterpenoid indole alkaloid is any one of the following compounds:

4. a process for the synthesis of a monoterpenoid indole alkaloid according to any one of claims 1 to 3, comprising the steps of:

carrying out a first Pictet-Spengerer reaction on loganin aglycone and a reaction reagent Ma under the catalysis of a first Lewis acid to obtain monoterpene-like indole alkaloid with a structure shown in a formula I;

carrying out a second Pictet-Spenger reaction on loganin aglycone and a reaction reagent Mb under the catalysis of a second Lewis acid to obtain monoterpene-like indole alkaloid with a structure shown in a formula II;

said R is ¹ 、R ² And R ³ As defined in formula I and formula II.

5. The synthesis method according to claim 4, wherein the first Lewis acid and the second Lewis acid are independently one or more of zinc trifluoromethanesulfonate, trifluoroacetic acid, aluminum trichloride, ferric trichloride, boron trifluoride, potassium trifluoromethanesulfonate and lithium trifluoromethanesulfonate.

6. The synthesis process according to claim 4 or 5, wherein the first and second Pictet-spengler reactions are independently carried out at a temperature of 40 to 45 ℃ and independently carried out for a time of 1 to 30 hours.

7. The method as in claim 4, wherein the first and second Pictet-Spenger reactions are carried out in the presence of an organic solvent, and the organic solvent required for the first and second Pictet-Spenger reactions is one or more selected from the group consisting of dichloromethane, N-dimethylformamide, dimethyl sulfoxide, and hexafluoroisopropanol.

8. The synthesis method of claim 4, wherein the loganin aglycone is obtained by hydrolyzing loganin in the presence of beta-glucosidase; the temperature of the hydrolysis reaction is 25-50 ℃, and the time is 24-48 h; the hydrolysis reaction is carried out under the condition that the pH value is 5.0-7.0.

9. Use of the monoterpenoid indole alkaloids according to any one of claims 1 to 3 in the manufacture of a medicament for the prevention or treatment of diabetes or diabetic nephropathy.

10. A medicament for the prevention and treatment of diabetes or diabetic nephropathy, comprising the monoterpenoid indole alkaloid of any one of claims 1 to 3 as an active ingredient.