CN105859823A

CN105859823A - Application of ilicis routundae cortex acid ester derivatives in preparation of anti-tumor drugs

Info

Publication number: CN105859823A
Application number: CN201610416603.9A
Authority: CN
Inventors: 南敏伦; 李伟; 赫玉芳; 赵昱玮; 吕娜; 李雪; 赵全成
Original assignee: Jilin Academy of Traditional Chinese Medicine
Current assignee: Jilin Academy of Traditional Chinese Medicine
Priority date: 2016-06-14
Filing date: 2016-06-14
Publication date: 2016-08-17

Abstract

The invention discloses a series of derivatives prepared by extracting, separating and purifying a traditional Chinese medicine (Ilicis routundae cortex) to obtain ilicis routundae cortex acid serving as a lead compound and performing corresponding chemical reactions. Pharmacological experiments prove that the ilicis routundae cortex acid derivatives have anti-tumor activity and have the inhibition effect on prostate cancer cells, lung cancer cells, liver cancer cells, cervical cancer cells and gastric cancer cells, and the inhibition effect of the derivatives is better than that of the mother nucleus compound ilicis routundae cortex acid.

Description

The application in preparing antitumor drug of the rotundicacid esters derivative

Technical field

The present invention relates to the derivative preparation method of a kind of rotundicacid obtained from natural Chinese medicine, be specifically one Plant with Chinese medicine extraction chemical composition as lead compound, prepare a series of derivant, carry out structure with anti-tumor activity for index Modify, belong to field of medicaments.

Background technology

Rotundicacid is extracted from Chinese medicine Cortex Ilicis Rotundae (Ilicis routundae cortex), purification, separation, alkali A kind of pentacyclic triterpenoid degraded and obtain.Our research has been proven that rotundicacid has prevention and the treatment heart The pharmacologically active (patent publication No.: CN101856357A) of cerebrovascular disease, have the pharmacological actions such as blood fat reducing (patent is public simultaneously The number of opening: CN101849950A)；Meanwhile, the present inventor's research demonstrates rotundicacid amino acid derivativges and resists in preparation swollen Application (patent publication No.: CN102391352B) in the medicine of tumor；Rotundicacid acylated derivatives and preparing antitumor Medicine in application (patent publication No.: CN102127142B)；The present inventor also studies and demonstrates rotundicacid and succinic acid The derivant that anhydride reactant generates has the effect (patent publication No.: CN102140126B) of prevention and cure of cardiovascular disease.According to document report Some pentacyclic triterpenoids of road, also have stronger anti-tumor activity (China National is among the people such as ursolic acid, oleanolic acid etc. Medicine, 2009,12 phases, 14-15), therefore, it is an object of the invention to rotundicacid compound is carried out structure of modification, system Standby a series of derivants, have the new pentacyclic triterpene of higher inhibitory action to derive tumor cell line growth to finding Thing.According to China's kinds of tumor morbidity disease, have selected the tumor cells such as carcinoma of prostate, pulmonary carcinoma, hepatocarcinoma, cervical cancer, gastric cancer, enter Row anti-tumor activity is tested, thus carries out pharmacodynamic evaluation.

Before the present invention completes, in addition to the present inventor studies, the derivant of document report rotundicacid is not also had to have The effect of suppression growth of tumour cell, does not finds that the derivant of rotundicacid is at the report preparing antitumor drug application yet.This Inventor is in order to find more effectively treatment antineoplastic noval chemical compound, thus rotundicacid is carried out a series of derivatization and grinds Study carefully.

Summary of the invention

The invention reside in that offer is a kind of a series of has rescuing of antitumor action with rotundicacid for lead compound synthesis Acid derivative must be answered.

It is an object of the invention to be achieved by the following technical programs:

A kind of rotundicacid derivant with anti-tumor activity, its structural formula is as follows:

Wherein: R₁For H or CH₃Or CH₂CH₃, R₂For

A kind of rotundicacid derivant with anti-tumor activity is obtained by following methods:

1, ethanol extraction Chinese medicine Cortex Ilicis Rotundae (Ilicis routundae cortex), purified, separate, alkaline degradation and obtain The compound 1 (3 β, 19 α, 24-trihydroxy-Ursane-12-alkene-28-carboxylic acid) obtained, i.e. rotundicacid.

2, compound 1 reacts to obtain 3 β, 19 α, 24-trihydroxy-Ursane-12-alkene-28-respectively with iodomethane, iodoethane Carboxylic acid ester compound 2-3.

Wherein R₁For methyl or ethyl

3, compound 1-3 at room temperature reacts with aromatic acid compound, obtains a series of compound.Concrete structure formula is shown in Table 1.

The concrete each compound structure of table 1

Above-described a kind of rotundicacid derivant with anti-tumor activity, can be used for preparation treatment tumor disease Medicine.

The invention have the characteristics that the rotundicacid to extracting from Cortex Ilicis Rotundae is that lead compound is chemically modified, To a series of derivants similar to rotundicacid structure, prove there is suppression obvious to tumor cell and make through pharmacological evaluation With, and activity is better than parent compound rotundicacid.

Rotundicacid and the derivant inhibitory action to tumor cell, these pharmacological actions, by following pharmacodynamics test Example is confirmed.

Cell human cervical carcinoma cell (Hela), human small cell lung carcinoma (NCI-H446), human breast cancer cell (MCF-7), people 6 kinds of cell lines such as stomach cancer cell (SGC-7901), human liver cancer cell (BEL-7402), prostate gland cancer cell (DU 145) are purchased from Tumour hospital of Jilin Province.

Medicine and reagent rotundicacid and each derivant are the self-control of this laboratory；Tetramethyl azo azoles salt (MTT), penicillium sp Element G potassium salt, streptomycin sulfate, HEPES, DMSO are Sigma Products；Pancreatin is Difco company；New-born calf serum (FBS) it is Heng Shengma biotechnology research institute of Beijing unit product；RPMI-1640 culture medium is Hyclone Products.

Instrument CK2TRC-3 fluorescence inverted microscope, Japan's OLYMPUS product；D-63450-CO₂Incubator, Germany Hera Products；FCANF129004 microplate reader, Australia's TECAN Products.

The tumor cell line subculture according to a conventional method of 6 kinds of people of experimental technique mtt assay, trophophase cell of taking the logarithm is used In experiment.By cell with 1 × 10⁵Individual/mL concentration is inoculated in 96 orifice plates (100 μ L/ hole), and after cultivating 12h, experimental group addition is rescued must Answering sour and each derivant (or 5-fluorouracil is as positive controls) 100 μ L/ hole, negative control group adds culture fluid 100 μ L/ Hole, often organizes and sets 4 parallel holes, and sets blank well (the most only adding medicinal liquid and culture fluid, be not added with cell) respectively with zeroing.To rescue Each derivant group must cultivate 44h, add 5mg/mLMTT 20 μ L, continue to cultivate 4h, abandon supernatant, every hole adds DMSO 150 μ L, mixing, microplate reader (λ=492nm) measures absorbance (A₄₉₂), utilize SPSS statistical software, calculate cell mortality, ask for IC₅₀。

Experimental result

Result shows, through SPSS software statistics, rotundicacid and derivant to Hela, NCI-H446, MCF-7, SGC- 7901,6 kinds of tumor cells such as BEL-7402, DU 145 all have inhibitory action in various degree, rotundicacid and derivant to 6 kinds The tumour inhibiting rate of tumor cell, IC₅₀With the 5-FU IC to each cell₅₀Value refers to table 2 and 3.It can be seen that change from table 2 and table 3 The growth inhibited effect of 5,6,9,11,15,18,20,24,29 pairs of tumor cells of compound is relatively strong, and compound 1,3,8,17,26 is right The sensitivity of tumor cell is poor.Each derivant is both greater than rotundicacid to the inhibitory action of tumor cell.In addition to rotundicacid, The IC of remaining compound₅₀It is respectively less than positive control drug 5-FU, illustrates that each derivant is respectively provided with the strongest suppression tumor cell Effect.

Table 2 concentration is the 10.00 each compounds of the μ g/mL tumour inhibiting rates to 6 kinds of tumor cells

The each derivant of table 3 rotundicacid and the 5-FU IC to 6 kinds of tumor cells₅₀(μg/mL)

Detailed description of the invention

Below in conjunction with embodiment, the present invention is described in further detail.But following embodiment is only the letter of the present invention Easily example, does not represent or limits the scope of the present invention, and protection scope of the present invention is as the criterion with claims.

The preparation of example 1 compound 1:

Medical material with 70% alcohol reflux 3 times, each 2 hours, united extraction liquid, add extracting liquid volume 2% medicinal Activated carbon, filters, and filtrate is concentrated into every 1ml medical material Han 1g；Adding sodium hydroxide solution 2 times amount of 1mol/l, 100 DEG C of hydrolysis 4 are little Time, letting cool, adjust pH value to be 3 with hydrochloric acid, filter, discard hydrolyzed solution, filter cake is washed with deionized water to neutrality, then the ethanol with 85% Being heated to boiling, the medicinal carbon of the 1% of addition medicine liquid volume, filter while hot, filtrate reduced in volume is recycled to without alcohol taste, adds water To every 1ml medical material Han 1g, place, filter, washing, it is dried, then recrystallization 2 times, obtain compound 1.Yield is 4.2%.

The preparation of example 2 compound 2:

Compound 1 takes 5g, is dissolved in DMF 150ml, adds Anhydrous potassium carbonate 5g, stirs 30 minutes, adds iodomethane 2.5ml, stirs 6h under room temperature, add water 650ml, stirring, filter, and filter cake washes with water, and by ethyl alcohol recrystallization 3 times, obtaining Compound 2.Productivity is 96.5%.

The preparation of example 3 compound 3:

Compound 1 takes 5g, is dissolved in DMF 150ml, adds Anhydrous potassium carbonate 5g, stirs 30 minutes, adds iodoethane 2.5ml, stirs 6h under room temperature, add water 650ml, stirring, filter, and filter cake washes with water, and by ethyl alcohol recrystallization 3 times, obtaining Compound 3.Productivity is 97.8%.

The preparation of example 4 compound 4-12:

Compound 1 takes 0.7g, is dissolved in dichloromethane 50ml, acetone 50ml, adds p-methylbenzoic acid 0.7g, DMAP 0.35g, is stirred to dissolve, and adds DCC 1.0g, room temperature reaction 24 hours, filters, and filtrate is evaporated, and residue silicagel column separates, Obtaining compound 4, prepare compound 5-12 with method, the productivity of each compound is not less than 65%.

The preparation of example 5 compound 13-21:

Compound 2 takes 0.7g, is dissolved in dichloromethane 100ml, adds p-methylbenzoic acid 0.7g, DMAP 0.35g, stirs Mixing and make dissolving, add DCC 1.0g, room temperature reaction 24 hours, filter, filtrate is evaporated, and residue silicagel column separates, and obtains chemical combination Thing 13, prepares compound 14-21 with method, and the productivity of each compound is not less than 79%.

The preparation of example 6 compound 22-30:

Compound 3 takes 0.7g, is dissolved in dichloromethane 100ml, adds p-methylbenzoic acid 0.7g, DMAP 0.35g, stirs Mixing and make dissolving, add DCC 1.0g, room temperature reaction 24 hours, filter, filtrate is evaporated, and residue silicagel column separates, and obtains chemical combination Thing 22, prepares compound 23-30 with method, and the productivity of each compound is not less than 82%.

Claims

1. a rotundicacid derivant, its architectural feature is:

Wherein: R₁For H or CH₃Or CH₂CH₃, R₂For

Rotundicacid derivant the most according to claim 1, it is characterised in that have suppression growth of tumour cell effect, Can be in the application in preparing antitumor drug.