CN109265458B

CN109265458B - Matrine-based antitumor drug compound

Info

Publication number: CN109265458B
Application number: CN201811337066.4A
Authority: CN
Inventors: 邹忠梅; 尚海; 李凌宇
Original assignee: Institute of Medicinal Plant Development of CAMS and PUMC
Current assignee: Institute of Medicinal Plant Development of CAMS and PUMC
Priority date: 2018-11-12
Filing date: 2018-11-12
Publication date: 2020-09-15
Anticipated expiration: 2038-11-12
Also published as: CN109265458A

Abstract

The invention discloses matrine-based antitumor drugA compound, wherein the active pharmaceutical compound consists of a compound shown in formula (I) prepared by reacting a substituted cinnamyl group and analogues thereof with a matrine compound, or a stereoisomer, a geometric isomer, a tautomer, a nitroxide compound, a hydrate, a solvate, a metabolite, a pharmaceutically acceptable salt or a prodrug of the compound shown in formula (I); the pharmaceutical compound has high anticancer curative effect, reduces the toxic and side effects of the traditional anticancer drugs by developing the pharmaceutical compound, improves the clinical treatment safety index, and has better clinical application prospect.

Description

Matrine-based antitumor drug compound

Technical Field

The invention belongs to the technical field of chemical medicines, and particularly relates to an antitumor drug compound based on matrine.

Background

Cancer is a major disease seriously threatening the health and life safety of people, and new cancer patients and the death caused by cancer in China all appear a rapid rising trend. Another problem that is common in cancer therapy is the severe toxicity associated with most antineoplastic agents. Although conventional antitumor agents, such as gemcitabine and paclitaxel, have drug resistance and severe toxicity, these drugs are still very important in cancer treatment because they can reduce tumors. Therefore, the research and development of safe and effective antitumor drugs with small toxic and side effects is a major subject related to the people's county.

Combinations of anticancer agents have now proven to be a significant advance in cancer treatment regimens, but there remains some unmet need and room for drug therapy to improve cancer for the treatment of cancers that are difficult to treat or that exhibit resistance to conventional antitumor agents as monotherapies. For example, in clinical treatment, tumor resistance ultimately leads to a poor prognosis of pancreatic cancer, with median survival of only 3-6 months and 5-year survival of less than 5%, despite gemcitabine-based combination chemotherapy regimens. Therefore, it would be a significant advance in the art to develop novel combinatorial approaches for delivering known anticancer agents with different mechanisms of action. While regimens involving combinations of anticancer agents with different mechanisms of action may work in the context of certain combinations, the same approach may not work for other combinations of anticancer agents, and such combinations may not always result in combinations with beneficial therapeutic effects.

Despite the great progress made in the combination of anticancer agents, many compositions are still based on gemcitabine, and although the use amount of gemcitabine can be reduced by using the combination, a small amount of gemcitabine still has the drug resistance and toxicity, and a certain degree of side effect can still be generated after long-term use.

For example, chinese patent application 2017114902705 discloses a pharmaceutical composition for treating tumor, which comprises an active ingredient and pharmaceutically acceptable excipients, and is characterized in that: the active ingredients are gemcitabine and diaza shown as a formula I

And carbazole compounds or pharmaceutically acceptable salts thereof,

gemcitabine and diazepine in the active ingredient

The mass ratio of the dicarbazole compound or the pharmaceutically acceptable salt thereof is (2-8): 1. the pharmaceutical composition has good anticancer effect and low adverse side effect; since diaza

The sensitivity of the carbazole compound to gemcitabine and the combined use of the carbazole compound and gemcitabine produce a synergistic effect, so that the clinical use dose of the traditional antitumor drug is reduced, the toxic and side effects caused by using the traditional antitumor drug in a large dose are reduced, the clinical treatment safety index is improved, and the carbazole compound has a good clinical application prospect. The pharmaceutical composition still contains a certain degree of gemcitabine, and still generates a certain degree of toxic and side effects.

For another example, chinese patent application 2014108361046 discloses an anti-tumor pharmaceutical composition, and the present invention relates to a pharmaceutical composition, which comprises: active pharmaceutical compounds, diluents, disintegrants, binders, lubricants; the compound is a compound of the following formula I or a pharmaceutically acceptable salt thereofAcceptable salts, solvates, polymorphs and the like

The invention also relates to a coated tablet prepared from the pharmaceutical composition. The compositions of the present invention have superior pharmaceutical properties, are primarily useful for treating diseases and disease conditions mediated by aberrant VEGFR, PDGFR, raf, p38, and/or flt-3 kinase signaling, and have certain limitations.

Therefore, the development of the anticancer drug taking traditional Chinese medicines or natural products as active ingredients of the drug is significant for treating or preventing tumors, thereby reducing the use of the traditional anticancer drugs, reducing the toxic and side effects to the maximum extent and improving the clinical treatment safety index.

The Chinese medicinal radix Sophorae Flavescentis contains various alkaloids, such as matrine, oxymatrine, sophocarpine, sophoridine, and N-oxysophocarpine. In recent years, the antitumor activity of matrine has been receiving wide attention. Research shows that the matrine plays an anti-tumor role through multiple ways of inducing apoptosis, cell cycle retardation, inducing autophagy of cells, inhibiting tumor cell metastasis and invasion, inhibiting angiogenesis, inhibiting NF-kB expression and the like, and can perform synergistic action with chemotherapeutic drugs.

The natural product of the cinnamic acid has the function of selectively inhibiting the proliferation of tumor cells. At present, the ferulic acid and the p-hydroxycinnamic acid are reported to play an anti-tumor role through a plurality of mechanisms, including reducing MAPK (mitogen-activated protein kinase) signals, inhibiting NF-kB (nuclear transcription factor) activation, inhibiting nitric oxide synthase, eliminating oxygen free radicals and the like, thereby influencing tumor cell apoptosis, invasion, metastasis and the like.

In view of this, matrine compounds (including matrine, sophoridine, sophocarpine, etc.) are combined with substituted cinnamyl and analogues thereof having antitumor activity through chemical conversion, and structural-activity relationship is analyzed, so that compounds having stronger antitumor activity and lower toxicity are found, and the method has important significance for treating tumor diseases.

Disclosure of Invention

In order to solve the defects in the prior art, the invention provides an antitumor drug compound based on matrine.

Wherein the compound is a compound shown in formula (I), or a stereoisomer, a geometrical isomer, a tautomer, a nitrogen oxide compound, a hydrate, a solvate, a metabolite, a pharmaceutically acceptable salt or a prodrug of the compound shown in formula (I); .

Wherein:

y is O, S or NH;

n is 1 to 4;

R₁and R₂May be selected from the following substituents: H. hydroxy, halogen, nitro, cyano, methoxy or C1-6 alkyl;

R₃may be selected from the following substituents: H. one or more of hydroxy, cyano, amino, halogen, nitro, alkyl, chloroalkyl, mercapto, alkoxy, alkylamino, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, cycloalkyl, and cycloalkylalkyl;

represents a single bond or a double bond.

The pharmaceutical compound may be

Wherein:

R₃₁、R₃₂、R₃₃、R₃₄each independently selected from the following substituents: H. hydroxy, cyano, amino, halogen, nitro, alkyl, chloroalkyl, mercapto, alkoxy, alkylamino, aryl, arylalkyl, heteroOne or more of aryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, cycloalkyl, and cycloalkylalkyl.

Wherein the salt comprises an acid or base inorganic salt;

the acid inorganic salt comprises hydrochloride, sulfate, phosphate, nitrate, carbonate, borate, sulfamate or hydrobromide;

the basic salt comprises sodium salt, potassium salt, lithium salt, magnesium salt, calcium salt or ammonium salt; or an organic salt comprising an acetate, propionate, butyrate, tartrate, maleate, hydroxymaleate, fumarate, citrate, lactate, mucate, gluconate, benzoate, succinate, oxalate, phenylacetate, methanesulfonate, p-toluenesulfonate, benzenesulfonate, p-aminosalicylate, aspartate, glutamate, edetate, stearate, palmitate, oleate, laurate, pantothenate, tannate, ascorbate, valerate or alkylammonium salt.

Wherein, the compound is prepared by the reaction of substituted cinnamyl and analogues thereof with matrine compounds;

the substituted cinnamyl and the analogue thereof are one of cinnamic acid, 2-chlorocinnamic acid, 3-chlorocinnamic acid, 4-chlorocinnamic acid, 3, 4-dichlorocinnamic acid, 3-bromocinnamic acid, 4-trifluoromethyl cinnamic acid, 4-nitrocinnamic acid, 4-methyl cinnamic acid, 4-methoxy cinnamic acid, 3, 4-methylene dioxy cinnamic acid or 3,4, 5-trimethoxy cinnamic acid;

the matrine compound is one of matrine, sophoridine and sophocarpine.

One synthetic route for the compounds is: taking matrine as an initial raw material, carrying out ring-opening esterification under the condition of hydrochloric acid-methanol to obtain methyl matrinate 1, and then carrying out Boc protection on 12-site amino group under the alkaline condition to obtain an intermediate 2; the intermediate 2 is reduced under the action of lithium aluminum hydride to obtain an alcoholic hydroxyl product 3, and then reacts with substituted or unsubstituted cinnamic acid under the action of 4-dimethylaminopyridine/carbodiimide (DMAP/EDCI) to obtain an intermediate 4; finally, the Boc protecting group is removed from the intermediate 4 under acidic condition, and the hydrochloride product 5 is obtained under the condition of hydrogen chloride-ether solution.

Another synthetic route for the compounds is: taking sophoridine as an initial raw material, and carrying out ring-opening esterification under the condition of hydrochloric acid-methanol to obtain sophoridine acid methyl ester 6; then carrying out Boc protection on 12-site amino group under alkaline condition to obtain an intermediate 7; the intermediate 7 is reduced under the action of lithium aluminum hydride to obtain an alcoholic hydroxyl product 8, and then reacts with substituted or unsubstituted cinnamic acid under the action of 4-dimethylaminopyridine/carbodiimide (DMAP/EDCI) to obtain an intermediate 9; finally, the intermediate 9 is stripped of Boc protecting group under acidic condition, and hydrochloride product 10 is obtained under hydrogen chloride-ether solution condition.

The third synthetic route for the compounds is: taking sophocarpine as an initial raw material, carrying out oxidation ring opening under the action of potassium permanganate and 10% sulfuric acid, and then taking methanol as a solvent, and carrying out esterification under the action of 2mol/L hydrochloric acid to obtain methyl sophorate 11; protecting 12-site amino group by Boc to obtain an intermediate 12, and then reducing by lithium aluminum hydride to obtain an alcoholic hydroxyl intermediate 13; condensing the intermediate 13 with substituted or unsubstituted cinnamic acid under the action of 4-dimethylaminopyridine/carbodiimide (DMAP/EDCI) to obtain an intermediate 14; finally, the Boc protecting group was removed under acidic conditions and the hydrochloride product 15 was obtained under HCl-ether conditions.

An antitumor pharmaceutical composition based on matrine comprises the antitumor pharmaceutical compound, and pharmaceutically acceptable carrier, diluent, binder or their combination.

The carrier is one of a microcapsule, a microsphere, a nanoparticle or a liposome;

the diluent comprises one or more of calcium hydrogen phosphate, kaolin, dextrin, lactose, mannitol, sucrose, microcrystalline cellulose, precipitated calcium carbonate, sorbitol, corn starch, potato starch, compressible starch, modified starch, pregelatinized starch, erythritol, xylitol and fructose;

the binder comprises one or more of hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinylpyrrolidone, polyvinyl alcohol, acacia, alginic acid, sodium alginate and gelatin.

The application also provides the matrine-based antitumor drug compound and application of the antitumor drug compound in preparation of an antitumor drug.

Preferably, the cancer includes, but is not limited to, cervical cancer, liver cancer, pancreatic cancer, colorectal cancer, and non-small cell lung cancer.

Has the advantages that: compared with the prior art, the pharmaceutical composition provided by the invention has the following advantages and remarkable progress:

(1) the compound shown in the formula (I) disclosed by the invention is prepared by reacting two compounds, namely substituted cinnamyl and analogues thereof and a matrine compound, has higher biological activity, can better inhibit the growth of tumors, and has the possibility of being developed into first-line clinical medicines.

(2) The pharmaceutical composition used in the present application does not contain any conventional anticancer drugs having toxicity, such as gemcitabine and paclitaxel, but develops a novel compound (I) for treating and preventing cancer, which has less toxic and side effects and can be safely used for a long time.

Detailed Description

Reaction reagents and conditions: (a)2mol/L HCl, MeOH, reflux; (b) boc₂O，K₂CO₃，CH₂Cl₂；(c)LiAlH₄，THF，(d)unsubstituted or substituted trans-cinnamic acid，DMAP,EDCI,CH₂Cl₂；(e)i)CF₃COOH，CH₂Cl₂，ii)HCl-Et₂O

The synthetic route is as follows: taking matrine as an initial raw material, carrying out ring-opening esterification under the condition of hydrochloric acid-methanol to obtain methyl matrinate 1, and then carrying out Boc protection on 12-site amino group under the alkaline condition to obtain an intermediate 2; the intermediate 2 is reduced under the action of lithium aluminum hydride to obtain an alcoholic hydroxyl product 3, and then reacts with substituted or unsubstituted cinnamic acid under the action of 4-dimethylaminopyridine/carbodiimide (DMAP/EDCI) to obtain an intermediate 4; finally, the Boc protecting group is removed from the intermediate 4 under acidic condition, and the hydrochloride product 5 is obtained under the condition of hydrogen chloride-ether solution.

Example 1

Preparation of methyl sophora flavescens acid (1) hydrochloride

Dissolving matrine (5.0g, 20.1mmol) in 50mL methanol, adding 2mol/L hydrochloric acid 30mL, heating to 115 deg.C, and reacting for 24 h. And (3) detecting the reaction by TLC (thin layer chromatography), stopping the reaction, cooling to room temperature, evaporating the solvent under reduced pressure to obtain a crude product, adding 20mL of acetone into a reaction bottle, stirring at room temperature for 1h, and filtering to obtain a white solid 5.4g with the yield of 76%.

Example 2

Preparation of 12N- (tert-butyloxycarbonyl) methyl sophora flavescente (2)

Compound 1(5.0g, 14.2mmol) was dissolved in 50mL of anhydrous methanol, di-tert-butyl dicarbonate (4.6g, 21.3mmol) and anhydrous potassium carbonate (5.9g, 42.6mmol) were added in this order, and after completion of addition, reaction was carried out at room temperature for 4 hours. TLC detection of the reaction was complete, reaction stopped, 100mL water was added, dichloromethane (100mL × 3) was extracted, organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, purified by silica gel column chromatography, eluent: dichloromethane-methanol ═ 40:1(V/V), giving 4.8g of a colourless oil, 89% yield.

Example 3

Preparation of 12N- (tert-butyloxycarbonyl) kurarinol (3)

Compound 2(4.8g, 12.6mmol) was dissolved in 30mL tetrahydrofuran and cooled to 0. Mixing LiAlH₄(574mg, 15.1mmol) was dissolved in 5mL of tetrahydrofuran, and the solution was added dropwise to the reaction mixture at 0 deg.CAfter dropping, the mixture is stirred and reacted for 6h at room temperature, TLC detects that the reaction is complete, the reaction is stopped, 5mL of acetone is added, 5mL of saturated ammonium chloride is added, the mixture is stirred for 30min, filtered, washed by ethyl acetate, concentrated, 100mL of water is added, ethyl acetate (100mL × 3) is extracted, organic phases are combined, dried by anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography, and an eluent, namely dichloromethane-methanol, is 40:1(V/V) is obtained, so that 3.2g of colorless oily matter is obtained, and the yield is 72%.

Example 4

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O-cinnamoyl kurarinol (4a)

Compound 3(200mg, 0.6mmol) was dissolved in 5mL of dichloromethane, and DMAP (98mg, 0.8mmol), EDCI (154mg, 0.8mmol) and cinnamic acid (118mg, 0.8mmol) were added in this order, and after the addition was completed, the reaction was carried out at room temperature. TLC detection of the reaction was complete, reaction stopped, 100mL water was added, dichloromethane (100mL × 3) was extracted, organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, purified by silica gel column chromatography, eluent: dichloromethane-methanol-40: 1(V/V) gave 198mg of a colorless oil in 69% yield.

Example 5

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (2-chlorocinnamoyl) kurarinol (4b)

The compound 3 and 2-chlorocinnamic acid are used as raw materials, and the synthesis method of the compound 4a is operated to obtain colorless oily matter with the yield of 60 percent.

Example 6

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (3-chlorocinnamoyl) kurarinol (4c)

The compound 3 and the 3-chlorocinnamic acid are used as raw materials, and the synthesis method of the compound 4a is operated to obtain colorless oily substance with the yield of 55 percent.

Example 7

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (4-chlorocinnamoyl) kurarinol (4d)

The compound 3 and 4-chlorocinnamic acid are used as raw materials, and the synthesis method of the compound 4a is operated to obtain colorless oily matter with the yield of 50 percent.

Example 8

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (3, 4-dichlorocinnamoyl) kurarinol (4e)

The compound 3 and the 3, 4-dichlorocinnamic acid are used as raw materials, and the synthesis method of the compound 4a is operated to obtain colorless oily substance with the yield of 42 percent.

Example 9

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (3-bromocinnamoyl) kurarinol (4f)

The compound 3 and 3-bromocinnamic acid are used as raw materials, and the synthesis method of the compound 4a is operated to obtain colorless oily substance with the yield of 43 percent.

Example 10

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (4-trifluoromethylcinnamoyl) kurarinol (4g)

The compound 3 and the 4-trifluoromethyl cinnamic acid are used as raw materials, and the synthesis method of the compound 4a is operated to obtain colorless oily substance with the yield of 56 percent.

Example 11

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (4-nitrocinnamoyl) kurarinol (4h)

The compound 3 and the 4-nitrocinnamic acid are used as raw materials, and the synthesis method of the compound 4a is operated to obtain colorless oily matter with the yield of 48 percent.

Example 12

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (4-methylcinnamoyl) kurarinol (4i)

The compound 3 and the 4-methyl cinnamic acid are used as raw materials, and the synthesis method of the compound 4a is operated to obtain colorless oily substance with the yield of 48 percent.

Example 13

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (4-methoxycinnamoyl) kurarinol (4j)

The compound 3 and the 4-methoxy cinnamic acid are used as raw materials, and the synthesis method of the compound 4a is operated to obtain colorless oily matter with the yield of 65 percent.

Example 14

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (3, 4-methylenedioxycinnamoyl) kurarinol (4k)

The compound 3 and the 3, 4-methylene dioxy cinnamic acid are used as raw materials, and the synthesis method of the compound 4a is operated to obtain colorless oily matter with the yield of 71 percent.

Example 15

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (3,4, 5-trimethoxycinnamoyl) kurarinol (4l)

The compound 3 and the 3,4, 5-trimethoxy cinnamic acid are used as raw materials, and the synthesis method of the compound 4a is operated to obtain colorless oily matter with the yield of 68 percent.

Example 16

Preparation of 4' -O-cinnamoyl sophora flavescens alkoxide (5a)

Compound 4(198mg) was dissolved in 3mL of dichloromethane, 600uL of trifluoroacetic acid was added, and after the addition was completed, the reaction was carried out at room temperature. TLC checked the reaction was complete, stopped, added saturated sodium bicarbonate, extracted with dichloromethane (50mL × 3), combined organic phases, dried over anhydrous sodium sulfate, filtered, and concentrated to give an oily product. Adding a small amount of dichloromethane for dissolution, adding 5mL of 2mol/L hydrogen chloride-ether solution, stirring at room temperature for 10min, and evaporating under reduced pressure to remove the solvent to obtain a white solid 162mg, wherein the yield is as follows: 87 percent.

Melting point (mp) of 156.0-157.2 deg.C;

¹H-NMR(600MHz,CD₃OD):7.70(d,J＝16.0Hz,1H),7.62-7.59(m,2H),7.42-7.38(m,3H),6.54(d,J＝16.0Hz,1H),4.28-4.23(m,2H),4.11-4.04(m,1H),3.84(t,J＝13.6Hz,1H),3.70-3.64(m,1H),3.45-3.37(m,2H),3.27(dd,J＝13.4,4.6Hz,1H),3.09-3.00(m,2H),2.52-2.44(m,1H),2.26-2.20(m,1H),2.12-1.91(m,5H),1.90-1.74(m,6H),1.73-1.53(m,3H)；

¹³C-NMR(150MHz,CD₃OD):168.6,146.3,135.7,131.6,130.1,129.3,118.8,65.0,62.7,56.6,56.6,53.6,43.8,37.9,32.9,30.9,29.5,25.5,24.5,21.9,19.6,19.5；

HRMS m/z calculated for C₂₄H₃₅N₂O₂[M+H]⁺:383.2699；

found:383.2691。

example 17

Preparation of 4' -O- (2-chlorocinnamyl) sophora flavescens alkoxide (5b)

The synthesis method of the compound 5a was performed using the compound 4b as a starting material to obtain a white solid with a yield of 95%.

Melting point (mp) 157.8-158.6 deg.C;

¹H-NMR(600MHz,CD₃OD):8.07(d,J＝16.0Hz,1H),7.79(dd,J＝7.7,1.7Hz,1H),7.47(dd,J＝7.9,1.3Hz,1H),7.39(td,J＝7.4,1.7Hz,1H),7.35(td,J＝7.4,1.0Hz,1H),6.58(d,J＝16.0Hz,1H),4.28(t,J＝6.4Hz,2H),4.07(ddd,J＝12.2,7.3,3.4Hz,1H),3.84(t,J＝13.6Hz,1H),3.69-3.64(m,1H),3.45-3.38(m,2H),3.27(dd,J＝13.3,4.6Hz,1H),3.09-3.00(m,2H),2.51-2.44(m,1H),2.25-2.20(m,1H),2.11-1.91(m,5H),1.90-1.75(m,6H),1.74-1.53(m,3H)；

¹³C-NMR(150MHz,CD₃OD):168.0,141.4,135.8,133.6,132.7,131.2,129.0,128.7,121.8,65.2,62.7,56.6,56.6,53.6,43.8,37.9,32.9,30.9,29.5,25.5,24.5,21.9,19.6,19.5；

HRMS m/z calculated for C₂₄H₃₄ClN₂O₂[M+H]⁺:417.2309；

found:417.2302。

example 18

Preparation of 4' -O- (3-chlorocinnamyl) sophora flavescens alkoxide (5c)

The synthesis method of the compound 5a was performed using the compound 4c as a starting material to obtain a white solid with a yield of 92%.

Melting point (mp) is 159.6-160.5 ℃;

¹H-NMR(600MHz,CD₃OD):7.68-7.63(m,2H),7.55(ddd,J＝6.3,2.1,1.8Hz,1H),7.42-7.37(m,2H),6.58(d,J＝16.1Hz,1H),4.30-4.22(m,2H),4.08(ddd,J＝12.2,7.1,3.1Hz,1H),3.85(t,J＝13.6Hz,1H),3.69-3.65(m,1H),3.45-3.38(m,2H),3.28(dd,J＝13.4,4.7Hz,1H),3.09-3.00(m,2H),2.52-2.46(m,1H),2.26-2.20(m,1H),2.12-1.91(m,5H),1.88-1.75(m,6H),1.74-1.53(m,3H)；

¹³C-NMR(150MHz,CD₃OD):168.1,144.5 137.8,136.0,131.6,131.3,128.9,127.6,120.6,65.1,62.7,56.6,56.6,53.6,43.8,37.9,32.9,30.9,29.5,25.5,24.5,21.9,19.6,19.5；

HRMS m/z calculated for C₂₄H₃₄ClN₂O₂[M+H]⁺:417.2309；

found:417.2303。

example 19

Preparation of 4' -O- (4-chlorocinnamyl) sophora flavescens alkoxide (5d)

The synthesis method of compound 5a was performed using compound 4d as a starting material to obtain a white solid with a yield of 91%.

Melting point (mp) 151.3-152.2 deg.C;

¹H-NMR(600MHz,CD₃OD):7.67(d,J＝16.0Hz,1H),7.61(d,J＝8.5Hz,2H),7.41(d,J＝8.5Hz,2H),6.55(d,J＝16.0Hz,1H),4.30-4.20(m,2H),4.10-4.03(m,1H),3.85(t,J＝13.6Hz,1H),3.68-3.64(m,1H),3.45-3.37(m,2H),3.27(dd,J＝13.3,4.4Hz,1H),3.08-3.00(m,2H),2.51-2.45(m,1H),2.25-2.20(m,1H),2.11-1.90(m,5H),1.89-1.75(m,6H),1.74-1.53(m,3H)；

¹³C-NMR(150MHz,CD₃OD):168.3,144.8,137.3,134.5,130.7,130.2,119.7,65.1,62.7,56.6,56.6,53.6,43.8,38.0,32.9,30.9,29.5,25.5,24.5,21.9,19.6,19.5；

HRMS m/z calculated for C₂₄H₃₄ClN₂O₂[M+H]⁺:417.2309；

found:417.2302。

example 20

Preparation of 4' -O- (3, 4-dichlorocinnamoyl) sophora flavescens alkoxide (5e)

The synthesis method of compound 5a was performed using compound 4e as the starting material to obtain a white solid with a yield of 87%.

Melting point (mp) is 152.1-153.4 deg.C;

¹H-NMR(600MHz,CD₃OD):7.81(d,J＝1.5Hz,1H),7.64(d,J＝16.0Hz,1H),7.58-7.54(m,2H),6.60(d,J＝16.0Hz,1H),4.30-4.21(m,2H),4.07(ddd,J＝12.2,7.1,3.1Hz,1H),3.85(t,J＝13.6Hz,1H),3.69-3.64(m,1H),3.45-3.38(m,2H),3.28(dd,J＝13.4,4.6Hz,1H),3.09-3.00(m,2H),2.51-2.45(m,1H),2.26-2.20(m,1H),2.12-1.89(m,5H),1.88-1.75(m,6H),1.74-1.52(m,3H)；

¹³C-NMR(150MHz,CD₃OD):168.0,143.4,136.3,135.0,134.0,132.1,131.0,128.7,121.2,65.2,62.7,56.6,56.6,53.6,43.8,37.9,32.9,30.9,29.5,25.5,24.5,21.9,19.6,19.5；

HRMS m/z calculated for C₂₄H₃₃Cl₂N₂O₂[M+H]⁺:451.1919；

found:451.1918。

example 21

Preparation of 4' -O- (3-bromocinnamoyl) sophora flavescens alkoxide (5f)

The synthesis method of the compound 5a was performed using the compound 4f as a starting material to obtain a white solid with a yield of 98%.

Melting point (mp) of 158.1-159.3 deg.C;

¹H-NMR(600MHz,CD₃OD):7.81-7.79(m,1H),7.65(d,J＝16.1Hz,1H),7.60(d,J＝7.7Hz,1H),7.57-7.54(m,1H),7.33(t,J＝7.9Hz,1H),6.57(d,J＝16.1Hz,1H),4.30-4.22(m,2H),4.07(ddd,J＝12.2,7.1,3.2Hz,1H),3.84(t,J＝13.6Hz,1H),3.68-3.64(m,1H),3.45-3.38(m,2H),3.27(dd,J＝13.4,4.7Hz,1H),3.08-3.00(m,2H),2.52-2.44(m,1H),2.25-2.20(m,1H),2.11-1.90(m,5H),1.88-1.74(m,6H),1.73-1.53(m,3H)；

¹³C-NMR(150MHz,CD₃OD):168.1,144.5,138.1,134.3,131.9,131.8,128.0,124.0,120.6,65.1,62.7,56.6,56.6,53.6,43.8,37.9,32.9,30.9,29.5,25.5,24.5,21.9,19.6,19.5；

HRMS m/z calculated for C₂₄H₃₄BrN₂O₂[M+H]⁺:461.1804；

found:461.1804。

example 22

Preparation of 4' -O- (4-trifluoromethylcinnamoyl) Sophorae Flavescentis alkoxide (5g)

The same synthesis as for compound 5a was carried out using 4g of the compound as the starting material to give a white solid with a yield of 95%.

Melting point (mp) 155.7-156.8 deg.C;

¹H-NMR(600MHz,CD₃OD):7.81(d,J＝8.2Hz,2H),7.75(d,J＝16.1Hz,1H),7.70(d,J＝8.2Hz,2H),6.68(d,J＝16.1Hz,1H),4.30-4.25(m,2H),4.08(ddd,J＝12.2,7.1,3.1Hz,1H),3.85(t,J＝13.6Hz,1H),3.69-3.64(m,1H),3.46-3.37(m,2H),3.28(dd,J＝13.4,4.7Hz,1H),3.09-2.98(m,2H),2.52-2.45(m,1H),2.26-2.20(m,1H),2.12-1.91(m,5H),1.90-1.75(m,6H),1.74-1.53(m,3H)；

¹³C-NMR(150MHz,CD₃OD):168.0,144.2,139.5,132.7(q,J＝32.4Hz),129.7,126.9(q,J＝3.7Hz),125.4(q,J＝269.4Hz),121.8,65.2,62.7,56.6,56.6,53.7,43.8,38.0,32.9,30.9,29.5,25.5,24.5,21.9,19.6,19.5；

HRMS m/z calculated for C₂₅H₃₄F₃N₂O₂[M+H]⁺:451.2572；

found:451.2568。

example 23

Preparation of 4' -O- (4-nitro cinnamoyl) sophora flavescens alkoxide (5h)

The synthesis method of the compound 5a was operated using the compound 4h as a starting material to obtain a white solid with a yield of 95%.

Melting point (mp) 162.1-163.4 deg.C;

¹H-NMR(600MHz,CD₃OD):8.26(d,J＝8.8Hz,2H),7.87(d,J＝8.8Hz,2H),7.77(d,J＝16.1Hz,1H),6.70(d,J＝16.1Hz,1H),4.32-4.24(m,2H),4.08(ddd,J＝12.2,7.1,3.1Hz,1H),3.85(t,J＝13.

6Hz,1H),3.69-3.63(m,1H),3.47-3.38(m,2H),3.28(dd,J＝13.3,4.6Hz,1H),3.09-2.99(m,2H),2.51-2.45(m,1H),2.25-2.19(m,1H),2.12-1.91(m,5H),1.88-1.75(m,6H),1.74-1.55(m,3H)；

¹³C-NMR(150MHz,CD₃OD):167.7,150.0,143.4,142.0,130.2,125.1,123.3,65.3,62.7,56.7,56.6,53.7,43.8,38.0,32.9,30.9,29.5,25.5,24.5,21.9,21.5,19.6,19.5；

HRMS m/z calculated for C₂₅H₃₆N₃O₄[M+H]⁺:428.2549；

found:428.2541。

example 24

Preparation of 4' -O- (4-methyl cinnamoyl) sophora flavescens alkoxide (5i)

The synthesis method of the compound 5a was performed using the compound 4i as a starting material to obtain a white solid with a yield of 94%.

Melting point (mp) is 150.8-151.5 deg.C;

¹H-NMR(600MHz,CD₃OD):7.66(d,J＝16.0Hz,1H),7.49(d,J＝8.0Hz,2H),7.22(d,J＝7.9Hz,2H),6.47(d,J＝16.0Hz,1H),4.28-4.21(m,2H),4.06(ddd,J＝12.0,7.0,2.7Hz,1H),3.84(t,J＝13.6Hz,1H),3.67-3.62(m,1H),3.45-3.37(m,2H),3.27(dd,J＝13.3,4.4Hz,1H),3.08-2.99(m,2H),2.50-2.42(m,1H),2.35(s,3H),2.44-2.18(m,1H),2.12-1.90(m,5H),1.87-1.75(m,6H),1.73-1.52(m,3H)；

¹³C-NMR(150MHz,CD₃OD):168.8,146.4,142.3,132.9,130.7,129.3,117.7,64.9,62.7,56.7,56.6,53.6,43.8,38.0,32.9,30.9,29.5,25.5,24.5,21.9,21.5,19.6,19.5；

HRMS m/z calculated for C₂₅H₃₇N₂O₂[M+H]⁺:397.2855；

found:397.2848。

example 25

Preparation of 4' -O- (4-methoxy cinnamoyl) sophora flavescens alkoxide (5j)

The synthesis method of the compound 5a is operated by taking the compound 4j as a raw material to obtain a white solid with the yield of 90 percent.

Melting point (mp) 157.6-158.1 deg.C;

¹H-NMR(600MHz,CD₃OD):7.65(d,J＝16.0Hz,1H),7.55(d,J＝8.7Hz,2H),6.95(d,J＝8.7Hz,2H),6.38(d,J＝16.0Hz,1H),4.27-4.20(m,2H),4.10-4.03(m,1H),3.84(t,J＝13.6Hz,1H),3.82(s,3H),3.68-3.62(m,1H),3.46-3.38(m,2H),3.27(dd,J＝13.4,4.4Hz,1H),3.08-2.98(m,2H),2.49-2.42(m,1H),2.24-2.18(m,1H),2.11-1.90(m,5H),1.86-1.75(m,6H),1.73-1.52(m,3H)；

¹³C-NMR(150MHz,CD₃OD):169.0,163.2,146.2,131.0,128.2,116.0,115.4,64.8,62.7,56.7,56.6,55.9,53.7,43.8,38.0,32.9,30.9,29.6,25.5,24.5,21.9,19.6,19.5；

HRMS m/z calculated for C₂₅H₃₇N₂O₃[M+H]⁺:413.2804；

found:413.2802。

example 26

Preparation of 4' -O- (3, 4-methylenedioxycinnamoyl) sophora flavescens alkoxide (5k)

The synthesis method of compound 5a was performed using compound 4k as the starting material to obtain a white solid with a yield of 84%.

Melting point (mp) is 152.3-153.8 ℃;

¹H-NMR(600MHz,CD₃OD):7.60(d,J＝15.9Hz,1H),7.16(d,J＝1.5,1H),7.09(dd,J＝8.0,1.6Hz,1H),6.84(d,J＝8.0Hz,1H),6.36(d,J＝15.9Hz,1H),6.00(s,2H),4.26-4.20(m,2H),4.07(ddd,J＝12.2,7.2,3.2Hz,1H),3.85(t,J＝13.6Hz,1H),3.69-3.64(m,1H),3.45-3.37(m,2H),3.27(dd,J＝13.3,4.6Hz,1H),3.09-3.00(m,2H),2.52-2.44(m,1H),2.26-2.20(m,1H),2.12-1.89(m,5H),1.88-1.75(m,6H),1.74-1.52(m,3H)；

¹³C-NMR(150MHz,CD₃OD):168.9,151.3,149.9,146.2,130.1,125.9,116.6,109.5,107.4,103.1,64.9,62.7,56.6,56.6,53.6,43.8,37.9,32.9,30.9,29.5,25.5,24.5,21.9,19.6,19.5；

HRMS m/z calculated for C₂₅H₃₅N₂O₄[M+H]⁺:427.2597；

found:427.2590。

example 27

Preparation of 4' -O- (3,4, 5-trimethoxycinnamoyl) sophora flavescens alkoxide (5l)

The synthesis method of compound 5a was performed using 4l of compound as a starting material to obtain a white solid with a yield of 85%.

Melting point (mp) is 171.2-172.6 deg.C;

¹H-NMR(600MHz,CD₃OD):7.64(d,J＝15.9Hz,1H),6.94(s,2H),6.50(d,J＝15.9Hz,1H),4.30-4.20(m,2H),4.12-4.04(m,1H),3.87(s,6H),3.84(t,J＝13.4Hz,1H),3.78(s,3H),3.68-3.62(m,1H),3.45-3.37(m,2H),3.27(dd,J＝13.4,4.1Hz,1H),3.10-2.98(m,2H),2.52-2.44(m,1H),2.27-2.18(m,1H),2.12-1.90(m,5H),1.89-1.74(m,6H),1.73-1.54(m,3H)；

¹³C-NMR(150MHz,CD₃OD):168.7,154.8,146.4,141.3,131.6,118.2,106.7,64.9,62.7,61.2,56.8,56.7,56.6,53.7,43.8,38.0,32.9,30.9,29.5,25.5,24.5,22.0,19.6,19.5；

HRMS m/z calculated for C₂₇H₄₁N₂O₅[M+H]⁺:473.3015；

found:473.3011。

reaction reagents and conditions (a)2mol/L HCl, MeOH, reflux; (b) boc₂O，K₂CO₃，CH₂Cl₂；(c)LiAlH₄，THF；(d)unsubstituted or substituted trans-cinnamic acid，DMAP，EDCI，CH₂Cl₂；(e)i)CF₃COOH，CH₂Cl₂，ii)HCl-Et₂O

The synthetic route is as follows: taking sophoridine as an initial raw material, carrying out ring-opening esterification under the condition of hydrochloric acid-methanol to obtain sophoridine methyl ester 6, and carrying out Boc protection on 12-site amino group under the alkaline condition to obtain an intermediate 7; the intermediate 7 is reduced under the action of lithium aluminum hydride to obtain an alcoholic hydroxyl product 8, and then reacts with substituted or unsubstituted cinnamic acid under the action of 4-dimethylaminopyridine/carbodiimide (DMAP/EDCI) to obtain an intermediate 9; finally, the intermediate 9 is stripped of Boc protecting group under acidic condition, and hydrochloride product 10 is obtained under hydrogen chloride-ether solution condition.

Example 28

Preparation of sophoridine acid methyl ester (6) hydrochloride

Sophoridine (5.0g, 20.1mmol) was dissolved in 50mL of methanol, 30mL of hydrochloric acid (2mol/L) was added, and the temperature was raised to 115 ℃ for reaction for 24 h. TLC detects that the reaction is complete, stops the reaction, cools to room temperature, evaporates under reduced pressure to remove the solvent to obtain a crude product, adds 20mL of acetone into a reaction bottle, stirs for 1h at room temperature, and filters to obtain 5.2g of white solid with 73% yield.

Example 29

Preparation of 12N- (tert-butoxycarbonyl) sophoridinate methyl ester (7)

Compound 6(1.0g, 2.8mmol) was dissolved in 10mL of anhydrous methanol, di-tert-butyl dicarbonate (916mg, 4.2mmol) and anhydrous potassium carbonate (1.2g, 8.4mmol) were added in this order, and after completion of addition, reaction was carried out at room temperature for 4 hours. The reaction was stopped by TLC detection, 100mL of water was added, dichloromethane (100 mL. times.3) was extracted, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give 1.0g of a colorless oil in 91% yield.

Example 30

Preparation of 12N- (tert-butyloxycarbonyl) sophoridine alcohol (8)

Compound 7(1.0g, 2.6mmol) was dissolved in 15mL tetrahydrofuran and cooled to 0 ℃. Mixing LiAlH₄(122mg, 3.2mmol) is dissolved in 3mL tetrahydrofuran, and the solution is added dropwise at 0 ℃ to the reaction solution after dropping, the reaction is stirred at room temperature for 6h, the TLC detection reaction is complete, the reaction is stopped, 3mL acetone is added, 3mL saturated ammonium chloride is added, the solution is stirred for 30min, suction filtration is carried out, ethyl acetate is used for washing, filtrate is concentrated, 100mL water is added, ethyl acetate (100mL × 3) is used for extraction, organic phases are combined, anhydrous sodium sulfate is used for drying, filtration and concentration are carried out, silica gel column chromatography purification is carried out, and an eluent, namely dichloromethane-methanol, is 20:1(V/V), 0.9g of colorless oily matter is obtained, and the yield is 98%.

Example 31

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O-cinnamoyl sophoridine (9a)

Compound 8(493mg, 1.4mmol) was dissolved in 5mL of dichloromethane, and DMAP (220mg, 1.8mmol), EDCI (345mg, 1.8mmol) and cinnamic acid (266mg, 1.8mmol) were added successively, and after the addition was completed, the reaction was carried out at room temperature. TLC detection of the reaction was complete, reaction stopped, 100mL water was added, dichloromethane (100mL × 3) was extracted, organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, purified by silica gel column chromatography, eluent: dichloromethane-methanol 40:1(V/V) gave 572mg of a colorless oil in 85% yield.

Example 32

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (2-chlorocinnamoyl) sophoridine alcohol (9b)

The synthesis method of the compound 9a is operated by taking the compound 8 and the 2-chlorocinnamic acid as raw materials to obtain g of colorless oily matter with the yield of 60 percent.

Example 33

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (3-chlorocinnamoyl) sophoridine alcohol (9c)

The compound 8 and the 3-chlorocinnamic acid are used as raw materials, and the synthesis method of the compound 9a is operated to obtain colorless oily substance with the yield of 78 percent.

Example 34

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (4-chlorocinnamoyl) sophoridine alcohol (9d)

The compound 8 and the 4-chlorocinnamic acid are used as raw materials, and the synthesis method of the compound 9a is operated to obtain colorless oily substance with the yield of 58 percent.

Example 35

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (3, 4-dichlorocinnamoyl) sophoridin alcohol (9e)

The compound 8 and 3, 4-dichlorocinnamic acid are used as raw materials, and the synthesis method of the compound 9a is operated to obtain colorless oily substance with the yield of 57 percent.

Example 36

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (3-bromocinnamoyl) sophoridin alcohol (9f)

The compound 8 and the 3-bromocinnamic acid are used as raw materials, and the synthesis method of the compound 9a is operated to obtain colorless oily substance with the yield of 63 percent.

Example 37

Preparation of 12N- (tert-Butoxycarbonyl) -4' -O- (4-trifluoromethylcinnamoyl) sophoridin alcohol (9g)

The compound 8 and the 4-trifluoromethyl cinnamic acid are used as raw materials, and the synthesis method of the compound 9a is operated to obtain colorless oily substance with the yield of 45 percent.

Example 38

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (4-nitrocinnamoyl) sophoridin alcohol (9h)

The compound 8 and the 4-nitrocinnamic acid are used as raw materials, and the synthesis method of the compound 9a is operated to obtain colorless oily matter with the yield of 55 percent.

Example 39

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (4-methylcinnamoyl) sophoridine alcohol (9i)

The compound 8 and the 4-methyl cinnamic acid are used as raw materials, and the synthesis method of the compound 9a is operated to obtain colorless oily substance with the yield of 54 percent.

Example 40

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (4-methoxycinnamoyl) sophoridin alcohol (9j)

The compound 8 and the 4-methoxy cinnamic acid are used as raw materials, and the synthesis method of the compound 9a is operated to obtain colorless oily substance with the yield of 51 percent.

EXAMPLE 41

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (3, 4-methylenedioxycinnamoyl) sophoridin alcohol (9k)

The compound 8 and the 3, 4-methylene dioxy cinnamic acid are used as raw materials, and the synthesis method of the compound 9a is operated to obtain colorless oily matter with the yield of 40 percent.

Example 42

Preparation of 4' -O-cinnamoyl sophoridine hydrochloride (10a)

Compound 9a (572mg) was dissolved in 3mL of dichloromethane, and 600uL of trifluoroacetic acid was added to react at room temperature. TLC checked the reaction was complete, stopped, added saturated sodium bicarbonate, extracted with dichloromethane (50mL × 3), combined organic phases, dried over anhydrous sodium sulfate, filtered, and concentrated to give an oily product. Adding a small amount of dichloromethane for dissolution, adding 5mL of 2mol/L hydrogen chloride-ether solution, stirring at room temperature for 10min, and evaporating under reduced pressure to remove the solvent to obtain 470mg of white solid, wherein the yield is as follows: 87 percent.

Melting point (mp) 171.5-173.0 deg.C;

¹H-NMR(600MHz,CD₃OD):7.70(d,J＝16.0Hz,1H),7.65-7.57(m,2H),7.44-7.35(m,3H),6.56(d,J＝16.0Hz,1H),4.26(t,J＝6.4Hz,2H),3.83(dd,J＝12.1,4.2Hz,1H),3.66-3.61(m,1H),3.53(dd,J＝8.9,5.6Hz,1H),3.41-3.32(m,2H),3.24(dd,J＝12.7,4.7Hz,1H),3.19-3.13(m,1H),3.02(t,J＝12.6Hz,1H),2.70-2.60(m,1H),2.45-2.38(m,1H),2.10-1.90(m,5H),1.89-1.76(m,5H),1.73-1.66(m,1H),1.65-1.59(m,1H),1.58-1.50(m,1H),1.49-1.42(m,1H)；

¹³CNMR(150MHz,CD₃OD):168.6,146.3,135.7,131.6,130.1,129.3,118.9,65.1,59.3,58.0,54.1,46.4,43.6,35.8,29.5,29.1,26.9,26.5,23.9,23.8,22.8,18.7；

HRMS m/z calculated for C₂₄H₃₅N₂O₂[M+H]⁺:383.2699；

found:383.2699。

example 43

Preparation of 4' -O- (2-chlorocinnamyl) sophoridine alkoxide (10b)

The synthesis method of the compound 10a was performed using the compound 9b as a starting material to obtain a white solid with a yield of 92%.

Melting point (mp) 143.0-143.5 deg.C;

¹H-NMR(600MHz,CD₃OD):8.07(d,J＝16.0Hz,1H),7.81(dd,J＝7.7,1.7Hz,1H),7.47(dd,J＝8.0,1.1Hz,1H),7.39(td,J＝7.4,1.8Hz,1H),7.35(td,J＝7.4,1.1Hz,1H),6.60(d,J＝16.0Hz,1H),4.28(t,J＝6.4Hz,2H),3.84(dd,J＝12.2,4.5Hz,1H),3.63(td,J＝12.9,3.0Hz,1H),3.53(dd,J＝9.3,5.5Hz,1H),3.43-3.32(m,2H),3.24(dd,J＝13.1,4.4Hz,1H),3.19-3.13(m,1H),3.03(t,J＝12.5,1H),2.70-2.60(m,1H),2.43(dt,J＝13.2,3.8Hz,1H),2.10-1.92(m,5H),1.89-1.78(m,5H),1.73-1.67(m,1H),1.66-1.59(m,1H),1.58-1.51(m,1H),1.50-1.42(m,1H)；

¹³C-NMR(150MHz,CD₃OD):168.1,141.4,135.8,133.6,132.7,131.2,129.0,128.7,121.8,65.3,59.3,58.0,54.1,46.4,43.6,35.8,29.4,29.0,26.9,26.5,23.8,23.7,22.8,18.7；

HRMS m/z calculated for C₂₄H₃₄ClN₂O₂[M+H]⁺:417.2309；

found:417.2313。

example 44

Preparation of 4' -O- (3-chlorocinnamyl) sophoridine hydrochloride (10c)

The synthesis method of compound 10a was performed using compound 9c as a starting material to obtain a white solid with a yield of 89%.

Melting point (mp) is 135.4-136.8 deg.C;

¹H-NMR(600MHz,CD₃OD):7.67-7.63(m,2H),7.56(ddd,J＝6.4,2.4,1.6Hz,1H),7.41-7.37(m,2H),6.60(d,J＝16.1Hz,1H),4.26(t,J＝6.4Hz,2H),3.83(dd,J＝12.2,4.4Hz,1H),3.63(td,J＝12.9,3.0Hz,1H),3.53(dd,J＝9.4,5.5Hz,1H),3.42-3.32(m,2H),3.24(dd,J＝13.1,4.4Hz,1H),3.19-3.14(m,1H),3.03(t,J＝12.7Hz,1H),2.70-2.61(m,1H),2.44(dt,J＝13.1,3.8Hz,1H),2.10-1.91(m,5H),1.89-1.77(m,5H),1.72-1.66(m,1H),1.66-1.60(m,1H),1.59-1.51(m,1H),1.50-1.41(m,1H)；

¹³C-NMR(150MHz,CD₃OD):168.2,144.5,137.8,136.0,131.6,131.2,128.9,127.6,120.7,65.3,59.3,58.0,54.1,46.4,43.6,35.7,29.4,29.1,26.9,26.5,23.8,23.8,22.8,18.7；

HRMS m/z calculated for C₂₄H₃₄ClN₂O₂[M+H]⁺:417.2309；

found:417.2310。

example 45

Preparation of 4' -O- (4-chlorocinnamyl) sophoridine hydrochloride (10d)

The synthesis method of compound 10a was performed using compound 9d as a starting material to obtain a white solid with a yield of 88%.

Melting point (mp) is 147.5-148.8 deg.C;

¹H-NMR(600MHz,CD₃OD):7.67(d,J＝16.0Hz,1H),7.62(d,J＝8.5Hz,2H),7.41(d,J＝8.4Hz,2H),6.57(d,J＝16.0Hz,1H),4.26(t,J＝6.4Hz,2H),3.82(dd,J＝12.1,4.1Hz,1H),3.62(td,J＝12.7,2.5Hz,1H),3.55-3.50(m,1H),3.42-3.32(m,2H),3.24(dd,J＝13.1,4.1Hz,1H),3.20-3.13(m,1H),3.02(t,J＝12.7Hz,1H),2.69-2.58(m,1H),2.45-2.36(m,1H),2.15-1.90(m,5H),1.88-1.75(m,5H),1.73-1.66(m,1H),1.65-1.58(m,1H),1.58-1.50(m,1H),1.50-1.41(m,1H)；

¹³C-NMR(150MHz,CD₃OD):168.4,144.8,137.3,134.5,130.8,130.3,119.7,65.2,59.3,58.0,54.1,46.4,43.6,35.8,29.5,29.1,26.9,26.5,23.9,23.7,22.8,18.7；

HRMS m/z calculated for C₂₄H₃₄ClN₂O₂[M+H]⁺:417.2309；

found:417.2310。

example 46

Preparation of 4' -O- (3, 4-dichlorocinnamoyl) sophoridine hydrochloride (10e)

The synthesis method of compound 10a was performed using compound 9e as the starting material to obtain a white solid with a yield of 72%.

Melting point (mp) is 138.8-139.4 deg.C;

¹H-NMR(600MHz,CD₃OD):7.82(d,J＝1.8Hz,1H),7.63(d,J＝16.0Hz,1H),7.59-7.53(m,2H),6.62(d,J＝16.0Hz,1H),4.26(t,J＝6.4Hz,2H),3.83(dd,J＝12.2,4.4Hz,1H),3.63(td,J＝12.9,3.0Hz,1H),3.53(dd,J＝9.5,5.5Hz,1H),3.41-3.32(m,2H),3.24(dd,J＝13.2,4.4Hz,1H),3.19-3.14(m,1H),3.03(t,J＝12.5Hz,1H),2.69-2.60(m,1H),2.44(dt,J＝12.8,3.9Hz,1H),2.10-1.92(m,5H),1.90-1.76(m,5H),1.72-1.67(m,1H),1.66-1.59(m,1H),1.58-1.51(m,1H),1.50-1.41(m,1H)；

¹³C-NMR(150MHz,CD₃OD):168.0,143.4,136.3,135.0,134.0,132.1,131.1,128.7,121.3,65.3,59.3,58.0,54.1,46.4,43.6,35.7,29.4,29.1,26.9,26.5,23.9,23.7,22.8,18.7；

HRMS m/z calculated for C₂₄H₃₃Cl₂N₂O₂[M+H]⁺:451.1919；

found:451.1927。

example 47

Preparation of 4' -O- (3-bromocinnamyl) sophoridine hydrochloride (10f)

The same synthesis as that of compound 10a was carried out using compound 9f as the starting material to give a white solid with a yield of 79%.

Melting point (mp) is 146.0-148.0 deg.C;

¹H-NMR(600MHz,CD₃OD):7.80(s,1H),7.64(d,J＝16.0Hz,1H),7.61(d,J＝7.8Hz,1H),7.55(d,J＝7.9Hz,1H),7.33(t,J＝7.9Hz,1H),6.59(d,J＝16.0Hz,1H),4.26(t,J＝6.4Hz,2H),3.83(dd,J＝12.1,2.9Hz,1H),3.63(td,J＝12.8,2.5Hz,1H),3.53(dd,J＝9.3,5.5Hz,1H),3.41-3.32(m,2H),3.24(dd,J＝13.2,4.4Hz,1H),3.20-3.14(m,1H),3.03(t,J＝12.6Hz,1H),2.70-2.60(m,1H),2.47-2.40(m,1H),2.11-1.92(m,5H),1.90-1.77(m,5H),1.72-1.67(m,1H),1.66-1.60(m,1H),1.59-1.51(m,1H),1.50-1.42(m,1H)；

¹³C-NMR(150MHz,CD₃OD):168.2,144.4,138.1,134.2,132.0,131.8,128.0,123.9,120.7,65.3,59.3,58.0,54.1,46.4,43.6,35.7,29.4,29.1,26.9,26.5,23.8,23.8,22.8,18.7；

HRMS m/z calculated for C₂₄H₃₄BrN₂O₂[M+H]⁺:461.1804；

found:461.1813。

example 48

Preparation of 4' -O- (4-trifluoromethylcinnamoyl) sophoridine hydrochloride (10g)

The same synthesis as that of compound 10a was carried out using 9g of the compound as a starting material to obtain a white solid with a yield of 92%.

Melting point (mp) is 131.3-132.0 deg.C;

¹H-NMR(600MHz,CD₃OD):7.82(d,J＝8.2Hz,2H),7.74(d,J＝16.1Hz,1H),7.70(d,J＝8.3Hz,2H),6.71(d,J＝16.0Hz,1H),4.27(t,J＝6.4Hz,2H),3.83(dd,J＝12.2,4.4Hz,1H),3.63(td,J＝12.9,3.0Hz,1H),3.54(dd,J＝9.4,5.4Hz,1H),3.42-3.32(m,2H),3.24(dd,J＝13.1,4.4Hz,1H),3.19-3.13(m,1H),3.03(t,J＝12.7Hz,1H),2.70-2.60(m,1H),2.45(dt,J＝13.0,3.8Hz,1H),2.12-1.93(m,5H),1.89-1.76(m,5H),1.73-1.67(m,1H),1.66-1.60(m,1H),1.59-1.51(m,1H),1.50-1.42(m,1H)；

¹³C-NMR(150MHz,CD₃OD):168.0,144.2,139.5,132.7(q,J＝32.2Hz),129.8,126.9(q,J＝3.5Hz),125.4(q,J＝271.5Hz),121.9,65.3,59.3,58.0,54.1,46.4,43.6,35.7,29.4,29.1,26.9,26.5,23.8,23.7,22.8,18.7；

HRMS m/z calculated for C₂₅H₃₄F₃N₂O₂[M+H]⁺:451.2572；

found:451.2576。

example 49

Preparation of 4' -O- (4-nitrocinnamoyl) sophoridine hydrochloride (10h)

The synthesis method of the compound 10a was performed using the compound 9h as a starting material to obtain a white solid with a yield of 92%.

Melting point (mp) of 148.0-150.0 deg.C;

¹H-NMR(600MHz,CD₃OD):8.25(d,J＝8.3Hz,2H),7.87(d,J＝8.7Hz,2H),7.75(d,J＝15.9Hz,1H),6.75(d,J＝16.1Hz,1H),4.27(t,J＝6.2Hz,2H),3.83(dd,J＝12.1,4.1Hz,1H),3.63(td,J＝13.0,3.0Hz,1H),3.54(dd,J＝9.2,5.3,1H),3.43-3.32(m,2H),3.25(dd,J＝13.1,4.3Hz,1H),3.20-3.14(m,1H),3.03(t,J＝12.6Hz,1H),2.70-2.60(m,1H),2.49-2.42(m,1H),2.10-1.93(m,5H),1.89-1.76(m,5H),1.73-1.67(m,1H),1.66-1.61(m,1H),1.60-1.51(m,1H),1.50-1.42(m,1H)；

¹³C-NMR(150MHz,CD₃OD):167.8,149.9,143.3,142.0,130.2,125.1,123.4,65.4,59.3,58.0,54.1,46.4,43.6,35.7,29.4,29.1,26.9,26.5,23.8,23.7,22.8,18.7；

HRMS m/z calculated for C₂₄H₃₄N₃O₄[M+H]⁺:428.2549；

found:428.2547。

example 50

Preparation of 4' -O- (4-methylcinnamoyl) sophoridine hydrochloride (10i)

The synthesis method of compound 10a was performed using compound 9i as a starting material to obtain a white solid with a yield of 90%.

Melting point (mp) is 129.7-130.5 deg.C;

¹H-NMR(600MHz,CD₃OD):7.65(d,J＝16.0Hz,1H),7.50(d,J＝8.2Hz,2H),7.22(d,J＝7.9Hz,2H),6.49(d,J＝16.0Hz,1H),4.24(t,J＝6.4Hz,2H),3.83(dd,J＝12.2,4.5Hz,1H),3.63(td,J＝12.9,3.0Hz,1H),3.53(dd,J＝9.3,5.5,1H),3.42-3.31(m,2H),3.24(dd,J＝13.2,4.4Hz,1H),3.19-3.13(m,1H),3.02(t,J＝12.7Hz,1H),2.70-2.60(m,1H),2.47-2.40(m,1H),2.35(s,3H),2.10-1.92(m,5H),1.88-1.76(m,5H),1.72-1.66(m,1H),1.66-1.59(m,1H),1.59-1.51(m,1H),1.50-1.42(m,1H)；

¹³C-NMR(150MHz,CD₃OD):168.8,146.3,142.2,132.9,130.7,129.3,117.8,65.1,59.3,58.0,54.1,46.4,43.6,35.8,29.5,29.1,26.9,26.5,23.8,23.8,22.8,21.5,18.7；

HRMS m/z calculated for C₂₅H₃₇N₂O₂[M+H]⁺:397.2855；

found:397.2856。

example 51

Preparation of 4' -O- (4-methoxycinnamoyl) sophoridine hydrochloride (10j)

The synthesis method of compound 10a was performed using compound 9j as a starting material to obtain a white solid with a yield of 81%.

Melting point (mp) is 169.0-170.0 deg.C;

¹H-NMR(600MHz,CD₃OD):7.64(d,J＝15.9Hz,1H),7.56(d,J＝8.8Hz,2H),6.95(d,J＝8.8Hz,2H),6.40(d,J＝16.0Hz,1H),4.24(t,J＝6.4Hz,2H),3.85-3.80(m,4H),3.63(td,J＝12.9,2.6Hz,1H),3.53(dd,J＝9.2,5.5Hz,1H),3.42-3.32(m,2H),3.24(dd,J＝13.1,4.4Hz,1H),3.19-3.13(m,1H),3.02(t,J＝12.7Hz,1H),2.70-2.60(m,1H),2.46-2.39(m,1H),2.10-1.92(m,5H),1.87-1.77(m,5H),1.72-1.66(m,1H),1.65-1.59(m,1H),1.58-1.51(m,1H),1.50-1.42(m,1H)；

¹³C-NMR(150MHz,CD₃OD):169.1,163.2,146.1,131.0,128.2,116.1,115.4,65.0,59.3,58.0,55.9,54.1,46.4,43.6,35.8,29.5,29.1,26.9,26.5,23.8,23.8,22.8,18.7；

HRMS m/z calculated for C₂₅H₃₇N₂O₃[M+H]⁺:413.2804；

found:413.2806。

example 52

Preparation of 4' -O- (3, 4-methylenedioxycinnamoyl) sophoridine hydrochloride (10k)

The synthesis method of compound 10a was performed using compound 9k as the starting material to obtain a white solid with a yield of 95%.

Melting point (mp) is 142.7-143.5 deg.C;

¹H-NMR(600MHz,CD₃OD):7.60(d,J＝15.9Hz,1H),7.17(s,1H),7.09(dd,J＝8.0,1.3Hz,1H),6.84(d,J＝8.0Hz,1H),6.38(d,J＝15.9Hz,1H),6.00(s,2H),4.23(t,J＝6.4Hz,2H),3.83(dd,J＝12.0,3.8Hz,1H),3.63(td,J＝12.8,2.3Hz,1H),3.53(dd,J＝9.1,5.4Hz,1H),3.42-3.32(m,2H),3.24(dd,J＝13.1,4.2Hz,1H),3.19-3.13(m,1H),3.02(t,J＝12.6Hz,1H),2.70-2.60(m,1H),2.44-2.40(m,1H),2.12-1.91(m,5H),1.88-1.75(m,5H),1.71-1.67(m,1H),1.66-1.59(m,1H),1.58-1.51(m,1H),1.50-1.42(m,1H)；

¹³C-NMR(150MHz,CD₃OD):168.9,151.3,149.9,146.1,130.1,125.9,116.7,109.5,107.4,103.1,65.0,59.3,58.0,54.1,46.4,43.6,35.8,29.5,29.1,26.9,26.5,23.8,23.8,22.8,18.7；

HRMS m/z calculated for C₂₅H₃₅N₂O₄[M+H]⁺:427.2597；

found:427.2599。

reaction reagents and conditions (b) Boc₂O，K₂CO₃，CH₂Cl₂；(c)LiAlH₄，THF；(d)unsubstituted orsubstituted trans-cinnamic acid，DMAP，EDCI，CH₂Cl₂；(e)i)CF₃COOH，CH₂Cl₂，ii)HCl-Et₂O；(f)i)KMnO₄，10％H₂SO₄ii)2mol/L HCl，MeOH。

The synthetic route is as follows: taking sophocarpine as an initial raw material, carrying out oxidation ring opening under the action of potassium permanganate and 10% sulfuric acid, and then taking methanol as a solvent, and carrying out esterification under the action of 2mol/L hydrochloric acid to obtain methyl sophorate 11; protecting 12-site amino group by Boc to obtain an intermediate 12, and then reducing by lithium aluminum hydride to obtain an alcoholic hydroxyl intermediate 13; condensing the intermediate 13 with substituted or unsubstituted cinnamic acid under the action of 4-dimethylaminopyridine/carbodiimide (DMAP/EDCI) to obtain an intermediate 14; finally, the Boc protecting group was removed under trifluoroacetic acid and the hydrochloride product 15 was obtained under HCl-ether conditions.

Example 53

Preparation of Sophora fruit acid (11)

Placing sophocarpine (2.0g, 8.1mmol) in a 100mL reaction bottle, adding 10% sulfuric acid 20mL, slowly adding potassium permanganate (1.9g,12.2mmol) under ice bath, heating to 115 ℃ after adding, and reacting for 24 h. And (4) detecting the reaction by TLC (thin layer chromatography), stopping the reaction, cooling to room temperature, adding 200ml of methanol, stirring for 10min, performing suction filtration, and concentrating the filtrate to obtain a white solid. The white solid was placed in a 100mL reaction flask, 20mL of methanol and 20mL of hydrochloric acid (2mol/L) were added, and the temperature was raised to 115 ℃ for reaction for 24 hours. And (3) detecting the reaction is complete by TLC, stopping the reaction, cooling to room temperature, adjusting the pH to 6-7, evaporating the solvent under reduced pressure, adding 100mL of methanol, filtering, concentrating the filtrate, and purifying by silica gel column chromatography, wherein an eluent: dichloromethane-methanol 30:1(V/V) gave 1.8g of white solid in 68% yield.

Example 54

Preparation of 12N- (tert-butyloxycarbonyl) methyl sophorate (12)

Compound 11(1.8g, 5.5mmol) was dissolved in 20mL of anhydrous methanol, di-tert-butyl dicarbonate (1.8g, 8.3mmol) and anhydrous potassium carbonate (2.3g, 16.6mmol) were added in this order, and after completion of addition, reaction was carried out at room temperature for 4 hours. TLC detection of the reaction was complete, reaction stopped, 100mL water was added, dichloromethane (100mL × 3) was extracted, organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, purified by silica gel column chromatography, eluent: dichloromethane-methanol 50:1(V/V) gave 885mg of a colorless oil in 46% yield.

Example 55

Preparation of 12N- (tert-butyloxycarbonyl) sophocarpol (13)

Compound 12(885mg, 2.5mmol) was dissolved in 20mL tetrahydrofuran and cooled to 0 ℃. Mixing LiAlH₄(114.0mg, 3.0mmol) is dissolved in 3mL tetrahydrofuran, and the solution is dripped into the reaction solution at 0 ℃, after dripping, the reaction is stirred at room temperature for 6h, TLC detects that the reaction is complete, the reaction is stopped, 3mL acetone is added, 3mL saturated ammonium chloride is added, the solution is stirred for 30min, suction filtration is carried out, ethyl acetate is used for washing, filtrate is concentrated, 100mL water is added, ethyl acetate (100mL × 3) is used for extraction, organic phases are combined, anhydrous sodium sulfate is used for drying, filtration, concentration and silica gel column chromatography purification are carried out, and an eluent, namely dichloromethane-methanol, is 40:1(V/V), 651mg of colorless oily matter is obtained, and the yield is 80%.

Example 56

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O-cinnamoyl sophocarpol (14a)

Compound 13(200mg, 0.6mmol) was dissolved in 5mL of dichloromethane, and DMAP (98mg, 0.8mmol), EDCI (154mg, 0.8mg) and cinnamic acid (118mg, 0.8mmol) were added in this order, and after the addition was completed, the reaction was carried out at room temperature. TLC detection of the reaction was complete, reaction stopped, 100mL water was added, dichloromethane (100mL × 3) was extracted, organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, purified by silica gel column chromatography, eluent: dichloromethane-methanol 50:1(V/V) gave 261mg of a colorless oil in 96% yield.

Example 57

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (2-chlorocinnamoyl) sophocarpol (14b)

The compound 13 and 2-chlorocinnamic acid are used as raw materials, and the synthesis method of the compound 14a is operated to obtain colorless oily substance with the yield of 92 percent.

Example 58

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (3-chlorocinnamoyl) sophocarpol (14c)

The compound 13 and 3-chlorocinnamic acid are used as raw materials, and the synthesis method of the compound 14a is operated to obtain colorless oily substance with the yield of 75 percent.

Example 59

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (4-chlorocinnamoyl) sophocarpol (14d)

The compound 13 and 4-chlorocinnamic acid are used as raw materials, and the synthesis method of the compound 14a is operated to obtain colorless oily substance with the yield of 68 percent.

Example 60

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (3, 4-dichlorocinnamoyl) sophocarpol (14e)

The compound 13 and the 3, 4-dichlorocinnamic acid are used as raw materials, and the synthesis method of the compound 14a is operated to obtain colorless oily substance with the yield of 65 percent.

Example 61

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (3-bromocinnamoyl) sophocarpol (14f)

The compound 13 and 3-bromocinnamic acid are used as raw materials, and the synthesis method of the compound 14a is operated to obtain colorless oily substance with the yield of 45 percent.

Example 62

Preparation of 12N- (tert-Butoxycarbonyl) -4' -O- (4-trifluoromethylcinnamoyl) sophocarpol (14g)

The compound 13 and the 4-trifluoromethyl cinnamic acid are used as raw materials, and the synthesis method of the compound 14a is operated to obtain colorless oily substance with the yield of 82 percent.

Example 63

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (4-nitrocinnamoyl) sophocarpol (14h)

The compound 13 and the 4-nitrocinnamic acid are used as raw materials, and the synthesis method of the compound 14a is operated to obtain colorless oily matter with the yield of 81 percent.

Example 64

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (4-methylcinnamoyl) sophocarpol (14i)

The compound 13 and the 4-methyl cinnamic acid are used as raw materials, and the synthesis method of the compound 14a is operated to obtain colorless oily substance with the yield of 85 percent.

Example 65

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (4-methoxycinnamoyl) sophocarpol (14j)

The compound 13 and 4-methoxy cinnamic acid are used as raw materials, and the synthesis method of the compound 14a is operated to obtain colorless oily substance with the yield of 81 percent.

Example 66

Preparation of 12N- (tert-butyloxycarbonyl) -4' -O- (3, 4-methylenedioxycinnamoyl) sophocarpol (14k)

The compound 13 and 3, 4-methylene dioxycinnamic acid are used as raw materials, and the synthesis method of the compound 14a is operated to obtain colorless oily matter with the yield of 81 percent.

Example 67

Preparation of 4' -O-cinnamoyl-sophocarpolate hydrochloride (15a)

Compound 14a (261mg) was dissolved in 3mL of dichloromethane, and 600uL of trifluoroacetic acid was added to react at room temperature after the addition. TLC checked the reaction was complete, stopped, added saturated sodium bicarbonate, extracted with dichloromethane (50mL × 3), combined organic phases, dried over anhydrous sodium sulfate, filtered, and concentrated to give an oily product. Dissolving with a small amount of dichloromethane, adding 5mL of 2mol/L hydrogen chloride-diethyl ether solution, stirring at room temperature for 10min, and evaporating under reduced pressure to remove the solvent to obtain 240mg of white solid with yield of 98%.

Melting point (mp) is 140.3-142.1 deg.C;

¹H-NMR(600MHz,CD₃OD):7.82(d,J＝16.1Hz,1H),7.70-7.65(m,2H),7.42-7.38(m,3H),6.66(d,J＝16.1Hz,1H),4.43(dt,J＝11.8,5.3Hz,1H),4.41-4.34(m,1H),4.34-4.27(m,1H),3.91(t,J＝13.7Hz,1H),3.74-3.68(m,1H),3.47-3.38(m,2H),3.33(dd,J＝13.3,4.6Hz,1H),3.10-3.00(m,2H),2.60-2.48(m,1H),2.45-2.37(m,1H),2.37-2.26(m,1H),2.14(d,J＝14.6Hz,1H),2.09-1.91(m,4H),1.91-1.75(m,4H)；

¹³C-NMR(150MHz,CD₃OD):168.2,146.8,135.8,131.6,130.0,129.5,118.7,62.6,61.0,56.7,56.6,51.8,43.9,38.2,32.8,30.7,25.5,24.7,19.6,19.4；

HRMS m/z calculated for C₂₂H₃₁N₂O₂[M+H]⁺:355.2386；

found:355.2389。

example 68

Preparation of 4' -O- (2-chlorocinnamyl) sophocarpolate (15b)

The synthesis of compound 15a was carried out using compound 14b as a starting material to give a white solid with a yield of 94%.

Melting point (mp) is 134.1-135.2 deg.C;

¹H-NMR(600MHz,CD₃OD):8.10(d,J＝16.0Hz,1H),7.85(dd,J＝7.7,1.7Hz,1H),7.46(d,J＝7.9Hz,1H),7.39(t,J＝7.7Hz,1H),7.35(t,J＝7.6Hz,1H),6.75(d,J＝16.0Hz,1H),4.45-4.40(m,2H),4.32-4.25(m,1H),3.90(t,J＝13.6Hz,1H),3.74-3.68(m,1H),3.47-3.39(m,2H),3.33(dd,J＝13.4,4.6Hz,1H),3.11-3.01(m,2H),2.58-2.49(m,1H),2.45-2.38(m,1H),2.36-2.28(m,1H),2.16(d,J＝14.6Hz,1H),2.09-1.91(m,4H),1.91-1.74(m,4H)；

¹³C-NMR(150MHz,CD₃OD):167.7,141.7,135.8,133.6,132.7,131.2,129.2,128.6,121.7,62.6,61.2,56.6,56.6,51.8,43.9,38.1,32.8,30.6,25.5,24.6,19.6,19.4；

HRMS m/z calculated for C₂₂H₃₀ClN₂O₂[M+H]⁺:389.1996；

found:389.1989。

example 69

Preparation of 4' -O- (3-chlorocinnamyl) sophocarpolate (15c)

The synthesis method of compound 15a was operated using compound 14c as a starting material to obtain a white solid with a yield of 91%.

Melting point (mp) 130.0-130.5 deg.C;

¹H-NMR(600MHz,CD₃OD):7.80(d,J＝16.1Hz,1H),7.73(s,1H),7.61(d,J＝6.8Hz,1H),7.42-7.37(m,2H),6.70(d,J＝16.1Hz,1H),4.47-4.41(m,1H),4.40-4.30(m,2H),3.92(t,J＝13.7Hz,1H),3.74-3.66(m,1H),3.47-3.38(m,2H),3.35-3.30(m,1H),3.10-3.01(m,2H),2.59-2.49(m,1H),2.45-2.38(m,1H),2.35-2.27(m,1H),2.14(d,J＝14.8Hz,1H),2.10-1.90(m,4H),1.90-1.75(m,4H)；

¹³C-NMR(150MHz,CD₃OD):167.8,145.0,137.9,135.9,131.5,131.3,129.1,127.8,120.4,62.6,61.0,56.6,56.6,51.7,43.9,38.2,32.8,30.7,25.5,24.7,19.6,19.3；

HRMS m/z calculated for C₂₂H₃₀ClN₂O₂[M+H]⁺:389.1996；

found:389.1995。

example 70

Preparation of 4' -O- (4-chlorocinnamyl) sophocarpolate (15d)

The synthesis method of compound 15a was operated using compound 14d as a starting material to obtain a white solid with a yield of 90%. Melting point (mp) 133.2-135.0 deg.C;

¹H-NMR(600MHz,CD₃OD):7.81(d,J＝16.1Hz,1H),7.68(d,J＝8.5Hz,2H),7.41(d,J＝8.5Hz,2H),6.67(d,J＝16.1Hz,1H),4.44(dt J＝11.8,6.2Hz,1H),4.38-4.28(m,2H),3.91(t,J＝13.7Hz,1H),3.71-3.65(m,1H),3.48-3.39(m,2H),3.32(dd,J＝13.4,4.5Hz,1H),3.10-3.00(m,2H),2.54-2.47(m,1H),2.45-2.38(m,1H),2.32-2.25(m,1H),2.15(d,J＝14.7Hz,1H),2.10-1.91(m,4H),1.90-1.75(m,4H)；

¹³C-NMR(150MHz,CD₃OD):167.9,145.3,137.4,134.6,130.9,130.2,119.4,62.6,60.9,56.7,56.6,51.6,43.9,38.3,32.9,30.7,25.5,24.7,19.6,19.4；

HRMSm/z calculated for C₂₂H₃₀ClN₂O₂[M+H]⁺:389.1996；

found:389.1992。

example 71

Preparation of 4' -O- (3, 4-dichlorocinnamoyl) sophocarpolate (15e)

The synthesis method of compound 15a was operated using compound 14e as a starting material to obtain a white solid with a yield of 93%.

Melting point (mp) of 156.2-157.1 deg.C;

¹H-NMR(600MHz,CD₃OD):7.90(d,J＝1.8Hz,1H),7.79(d,J＝16.1Hz,1H),7.63(dd,J＝8.3,2.0Hz,1H),7.55(d,J＝8.4Hz,1H),6.70(d,J＝16.1Hz,1H),4.45(dt,J＝11.9,5.3Hz,1H),4.38-4.31(m,2H),3.93(t,J＝13.7Hz,1H),3.72-3.68(m,1H),3.48-3.39(m,2H),3.33(dd,J＝13.4,4.6Hz,1H),3.10-3.01(m,2H),2.56-2.50(m,1H),2.46-2.38(m,1H),2.34-2.28(m,1H),2.14(d,J＝14.7Hz,1H),2.10-1.90(m,4H),1.90-1.76(m,4H)；

¹³C-NMR(150MHz,CD₃OD):167.6,143.9,136.4,135.0,134.0,132.1,131.2,128.9,120.9,62.6,60.9,56.7,56.6,51.6,43.9,38.3,32.8,30.7,25.5,24.7,19.6,19.3；

HRMS m/z calculated for C₂₂H₂₉Cl₂N₂O₂[M+H]⁺:423.1606；

found:423.1600。

example 72

Preparation of 4' -O- (3-bromocinnamoyl) sophocarpolate (15f)

The synthesis method of compound 15a was operated using compound 14f as a starting material to obtain a white solid with a yield of 98%.

Melting point (mp) is 164.5-165.3 deg.C;

¹H-NMR(600MHz,CD₃OD):7.87(s,1H),7.78(d,J＝16.1Hz,1H),7.66(d,J＝7.8Hz,1H),7.55(d,J＝7.9Hz,1H),7.32(t,J＝7.9Hz,1H),6.69(J＝16.1Hz,1H),4.44(dt,J＝11.9,5.3Hz,1H),4.40-4.30(m,2H),3.92(t,J＝13.7Hz,1H),3.73-3.68(m,1H),3.48-3.39(m,2H),3.33(dd,J＝13.4,4.6Hz,1H),3.10-3.01(m,2H),2.58-2.50(m,1H),2.45-2.38(m,1H),2.36-2.27(m,1H),2.14(d,J＝14.6Hz,1H),2.10-1.91(m,4H),1.90-1.75(m,4H)；

¹³C-NMR(150MHz,CD₃OD):167.7,145.0,138.2,134.3,132.1,131.7,128.2,123.9,120.4,62.6,61.0,56.6,56.6,51.7,43.9,38.2,32.8,30.7,25.5,24.7,19.6,19.4；

HRMS m/z calculated for C₂₂H₃₀BrN₂O₂[M+H]⁺:433.1491；

found:433.1483。

example 73

Preparation of 4' -O- (4-trifluoromethylcinnamoyl) Sophora fruit alkoxide (15g)

A method for synthesizing compound 15a was performed using 14g of the compound as a starting material to obtain a white solid with a yield of 98%.

Melting point (mp) is 137.8-138.3 deg.C;

¹H-NMR(600MHz,CD₃OD):7.92-7.87(m,3H),7.70(d,J＝8.2Hz,2H),6.80(d,J＝16.1Hz,1H),4.46(dt,J＝11.9,5.3Hz,1H),4.42-4.32(m,2H),3.94(t,J＝13.7Hz,1H),3.73-3.70(m,1H),3.48-3.40(m,2H),3.34(dd,J＝13.4,4.6Hz,1H),3.11-3.01(m,2H),2.59-2.51(m,1H),2.47-2.39(m,1H),2.36-2.30(m,1H),2.16(d,J＝14.7Hz,1H),2.11-1.93(m,4H),1.91-1.76(m,4H)；

¹³C-NMR(150MHz,CD₃OD):167.6,144.7,139.6,132.7(q,J＝32.3Hz),129.9,126.8(q,J＝3.2Hz),125.4(q,J＝271.0Hz),121.6,62.6,61.0,56.7,56.6,51.6,43.9,38.3,32.8,30.7,25.5,24.7,19.6,19.4；

HRMS m/z calculated for C₂₃H₃₀F₃N₂O₂[M+H]⁺:423.2259；

found:423.2260。

example 74

Preparation of 4' -O- (4-nitrocinnamoyl) sophocarpolate (15h)

The synthesis of compound 15a was carried out using compound 14h as the starting material to give a white solid with a yield of 77%.

The melting point (mp) is 175.2-176 ℃;

¹H-NMR(600MHz,CD₃OD):8.25(d,J＝8.9Hz,2H),7.95-7.90(m,3H),6.85(d,J＝16.2Hz,1H),4.47(dt,J＝11.9,5.3Hz,1H),4.41-4.33(m,2H),3.94(t,J＝13.6Hz,1H),3.73-3.70(m,1H),3.48-3.40(m,2H),3.34(dd,J＝13.4,4.6Hz,1H),3.11-3.03(m,2H),2.58-2.53(m,1H),2.48-2.40(m,1H),2.35-2.30(m,1H),2.16(d,J＝14.7Hz,1H),2.11-1.92(m,4H),1.91-1.75(m,4H)；

¹³C-NMR(150MHz,CD₃OD):167.3,149.9,143.8,142.1,130.4,125.0,123.1,62.6,61.1,56.7,56.6,51.5,43.9,38.3,32.8,30.7,25.5,24.7,19.6,19.4；

HRMS m/z calculated for C₂₂H₃₀N₃O₄[M+H]⁺:400.2236；

found:400.2229。

example 75

Preparation of 4' -O- (4-methylcinnamoyl) sophocarpolate (15i)

The synthesis of compound 15a was carried out using compound 14i as a starting material to give a white solid with a yield of 93%.

Melting point (mp) 176.6-177.8 deg.C;

¹H-NMR(600MHz,CD₃OD):7.78(d,J＝16.1Hz,1H),7.55(d,J＝8.1Hz,2H),7.22(d,J＝8.0Hz,2H),6.60(d,J＝16.0Hz,1H),4.42(dt,J＝11.9,6.2Hz,1H),4.39-4.33(m,1H),4.30(ddd,J＝12.4,8.2,3.2Hz,1H),3.91(t,J＝13.6Hz,1H),3.71-3.68(m,1H),3.46-3.39(m,2H),3.32(dd,J＝13.6,4.8Hz,1H),3.10-3.01(m,2H),2.56-2.50(m,1H),2.44-2.37(m,1H),2.35(s,3H),2.33-2.28(m,1H),2.14(d,J＝14.7Hz,1H),2.10-1.91(m,4H),1.90-1.74(m,4H)；

¹³C-NMR(150MHz,CD₃OD):168.4,146.9,142.3,133.0,130.7,129.5,117.5,62.6,60.9,56.7,56.6,51.8,43.9,38.2,32.8,30.7,25.5,24.7,21.5,19.6,19.4；

HRMS m/z calculated for C₂₃H₃₃N₂O₂[M+H]⁺:369.2542；

found:369.2540。

example 76

Preparation of 4' -O- (4-methoxycinnamoyl) sophocarpolate (15j)

The synthesis method of compound 15a was operated using compound 14j as a starting material to obtain a white solid with a yield of 90%.

Melting point (mp) is 145.4-146.5 deg.C;

¹H-NMR(600MHz,CD₃OD):7.76(d,J＝16.0Hz,1H),7.62(d,J＝8.8Hz,2H),6.95(d,J＝8.6Hz,2H),6.50(d,J＝16.0Hz,1H),4.42(dt,J＝11.9,5.5Hz,1H),4.39-4.32(m,1H),4.29(ddd,J＝12.5,8.0,2.9Hz,1H),3.91(t,J＝13.7Hz,1H),3.82(s,3H),3.71-3.68(m,1H),3.47-3.39(m,2H),3.32(dd,J＝13.4,4.6Hz,1H),3.11-3.00(m,2H),2.57-2.50(m,1H),2.44-2.36(m,1H),2.35-2.28(m,1H),2.14(d,J＝11.5Hz,1H),2.10-1.91(m,4H),1.91-1.75(m,4H)；

¹³C-NMR(150MHz,CD₃OD):168.6,163.3,146.7,131.2,128.3,115.8,115.4,62.6,60.8,56.7,56.6,55.9,51.8,43.9,38.2,32.8,30.8,25.5,24.6,19.6,19.4；

HRMS m/z calculated for C₂₃H₃₃N₂O₃[M+H]⁺:385.2491；

found:385.2490。

example 77

Preparation of 4' -O- (3, 4-methylenedioxycinnamoyl) sophocarpolate (15k)

The synthesis of compound 15a was carried out using compound 14k as the starting material to give a white solid with a yield of 83%.

Melting point (mp) 157.8-158.5 deg.C;

¹H-NMR(600MHz,CD₃OD):7.72(d,J＝16.0Hz,1H),7.23(d,J＝1.6Hz,1H),7.16(dd,J＝8.0,1.6Hz,1H),6.84(d,J＝8.0Hz,1H),6.49(d,J＝16.0Hz,1H),6.00(s,2H),4.42(dt,J＝11.9,5.5Hz,1H),4.38-4.27(m,2H),3.91(t,J＝13.7Hz,1H),3.73-3.67(m,1H),3.47-3.38(m,2H),3.32(dd,J＝13.4,4.6Hz,1H),3.10-3.01(m,2H),2.57-2.50(m,1H),2.43-2.36(m,1H),2.34-2.28(m,1H),2.14(d,J＝12.8Hz,1H),2.10-1.91(m,4H),1.90-1.74(m,4H)；

¹³C-NMR(150MHz,CD₃OD):168.5,151.4,149.9,146.7,130.2,126.1,116.4,109.4,107.6,103.1,62.6,60.8,56.6,56.6,51.8,43.9,38.2,32.8,30.7,25.5,24.7,19.6,19.4；

HRMS m/z calculated for C₂₃H₃₁N₂O₄[M+H]⁺:399.2284；

found:399.2289。

test example 1 analysis of cell proliferation by Tetraazathiazole blue (MTT) method

Respectively inoculating cells in logarithmic growth phase into 96-well plate, culturing under conventional conditions for 24 hr, grouping, and adding diluted culture medium200 μ L of test solution for each extraction site of compounds with the same concentration (final concentration of 0.125, 0.250, 0.500, 1.000, 2.000mg/mL), and setting control group wells (adding equal volume of culture solution), each group having 6 multiple wells, placing 5% CO₂Carrying out incubator culture at 37 ℃, taking out a corresponding 96-well plate after the drugs act for 24 hours, 36 hours, 48 hours and 72 hours, respectively adding 20 mu L of MTT (methyl thiazolyl tetrazolium) of 5mg/mL into each well, continuing to culture for 4 hours, sucking and removing liquid in each well, adding 150 mu L of DMSO into each well, oscillating until blue crystals are dissolved, detecting the absorbance (A) value of each well at 570nm (reference wavelength of 630nm) by using an enzyme-labeling instrument, determining the inhibition rate of the drugs on the growth of tumor cells, calculating the cell growth inhibition rate, namely the cell growth inhibition rate (%) (1-experiment group OD value/control group OD value) × 100%, and obtaining the specific data shown in the following table 1.

TABLE 1

The spirit of the present invention is described in detail by the preferred embodiments of the present invention. It will be understood by those skilled in the art that any modification, equivalent change and modification made to the above embodiments in accordance with the technical spirit of the present invention fall within the scope of the present invention.

Claims

1. A compound characterized by: the pharmaceutical compound may be

、

Or

Wherein:

R₃₁、R₃₂、R₃₃、R₃₄each independently selected from the following substituents: H. one or more of hydroxy, cyano, amino, halogen, nitro, methyl, methoxy, or trifluoromethyl;

or, R₃₁And R₃₄Is H, R₃₂And R₃₃Together form a methylenedioxy group.

2. A process for preparing a compound of claim 1, wherein: the compound is prepared by the reaction of substituted cinnamyl and analogues thereof with a matrine compound;

the matrine compound is one of matrine, sophoridine and sophocarpine.

3. The method of claim 2, wherein: taking matrine as a starting material, and carrying out ring-opening esterification under the condition of hydrochloric acid-methanol to obtain methyl matrinate 1; then carrying out Boc protection on 12-site amino group under alkaline condition to obtain an intermediate 2; the intermediate 2 is reduced under the action of lithium aluminum hydride to obtain an alcoholic hydroxyl product 3, and then reacts with substituted or unsubstituted cinnamic acid under the action of 4-dimethylaminopyridine and carbodiimide to obtain an intermediate 4; finally, the intermediate 4 is subjected to Boc protecting group removal under acidic conditions, and a hydrochloride product 5 is obtained under the conditions of hydrogen chloride-ether solution;

。

4. the method of claim 2, wherein: taking sophoridine as an initial raw material, and carrying out ring-opening esterification under the condition of hydrochloric acid-methanol to obtain sophoridine acid methyl ester 6; then carrying out Boc protection on 12-site amino group under alkaline condition to obtain an intermediate 7; the intermediate 7 is reduced under the action of lithium aluminum hydride to obtain an alcoholic hydroxyl product 8, and then reacts with substituted or unsubstituted cinnamic acid under the action of 4-dimethylaminopyridine and carbodiimide to obtain an intermediate 9; finally, the intermediate 9 is subjected to Boc protecting group removal under acidic conditions, and a hydrochloride product 10 is obtained under the conditions of hydrogen chloride-ether solution;

。

5. the method of claim 2, wherein: taking sophocarpine as an initial raw material, carrying out oxidation ring opening under the action of potassium permanganate and sulfuric acid, and then taking methanol as a solvent, and esterifying under the action of hydrochloric acid to obtain methyl sophorate 11; protecting 12-site amino group by Boc to obtain an intermediate 12, and then reducing by lithium aluminum hydride to obtain an alcoholic hydroxyl intermediate 13; condensing the intermediate 13 with substituted or unsubstituted cinnamic acid under the action of 4-dimethylaminopyridine and carbodiimide to obtain an intermediate 14; finally, removing Boc protecting group under acidic condition, and obtaining hydrochloride product 15 under hydrogen chloride-ether condition;

。

6. an antitumor pharmaceutical composition based on matrine, comprising the compound of claim 1 or the product obtained by the preparation method of any one of claims 2-5, and a pharmaceutically acceptable carrier, diluent, binder or their combination.

7. Use of a compound according to claim 1, or a product thereof prepared by a process according to any one of claims 2 to 5, or a composition according to claim 6, in the manufacture of an anti-cancer medicament.

8. Use according to claim 7, characterized in that: the cancer is cervical cancer, liver cancer, pancreatic cancer, colorectal cancer and non-small cell lung cancer.