CN107540528B - A kind of fat-soluble emodin derivates and its preparation method and application - Google Patents

Abstract

The invention discloses a kind of fat-soluble emodin derivates and its preparation method and application, the compound concrete structure formula are as follows:

Description

A kind of fat-soluble emodin derivates and its preparation method and application

Technical field

The invention belongs to field of medicinal chemistry, and in particular to a kind of emodin derivates and its preparation method and application.

Background technique

Leukaemia is a kind of Clonal malignant disease for being derived from candidate stem cell, is typically characterised by proliferation out of control, causes A large amount of leukaemia cell's accumulation, infiltrate marrow and its adjacent tissue and lead to marrow normal hematopoiesis dysfunction.Therefore, how to control The unlimited multiplication capacity of tumour cell processed, inducing its apoptosis is one of the critical issue in oncotherapy.Bone-marrow transplantation is traditional The treatment methods such as chemicotherapy cannot fully meet the needs for the treatment of, chemicotherapy not only offer limited effectiveness, but also erious adverse reaction, marrow Transplanting further relates to graft-versus-host reaction (GVHD) and relapse after transplantation.Currently, being closed based on the design of natural products skeleton structure The hot spot of leukemia treating area research is had become at more target therapeutic agents.

Rheum emodin is a kind of derivative of hydroxy-anthraquione, is one of effective component of rheum officinale, be present in rheum officinale, the fleece-flower root, In a variety of medicinal materials such as cassia seed, the vine of multiflower knotweed, polygonum cuspidate, pharmacological action is extensive, has preferable clinical value.In recent years The study found that rheum emodin has inhibiting effect to kinds of tumors such as lung cancer, gastric cancer, cancer of pancreas, breast cancer, mechanism and inhibition are swollen Tumor cell proliferation promotes the number of ways such as its apoptosis, anti-angiogenesis related.Know that rheum emodin has inactivation HL60(people at present Promyelocytic leukemia cell), but due to its activity be not it is highly desirable, selected using rheum emodin as primer and to it It is chemically modified to improve its anti-tumor activity.

The present invention obtains 4,5- dihydroxy -7- methyl -9,10- diketone -9,10- anthraquinone -2- quinhydrones -2- by two-step reaction Valproic acid ester, preparation method is simple, and reaction condition is mild, and low energy consumption, high income, and product purity is high, practical, and is closed At 4,5- dihydroxy -7- methyl -9,10- diketone -9,10- anthraquinone -2- quinhydrones -2- valproic acid ester be a kind of targeting HL60(people Promyelocytic leukemia cell) small molecule compound, relative molecular mass is 500 hereinafter, external anticancer with higher is living Property, it is expected to be used for more target therapeutic agents of preparation leukaemia.

Summary of the invention

The purpose of the present invention is to provide a kind of fat-soluble emodin derivates and its preparation method and application, the compounds Can selective depression HL60 cell, there is certain pharmacological activity, and preparation method is simple, and experiment condition is mild, should not The harsh conditions such as high temperature and pressure, strong acid and strong base are sought, reaction yield is high.

To achieve the above object, the present invention adopts the following technical scheme:

A kind of emodin derivates, concrete structure formula are as follows:

。

The preparation approach of the compound is as follows:

Its specific preparation step includes:

1) using valproic acid and oxalyl chloride as starting material, using methylene chloride as solvent, reaction is obtained under condition of ice bath 2- propyl valeric chloride；

2) under condition of ice bath, 1,3,8- trihydroxy -6- methyl -9,10- anthraquinone is dissolved in methylene chloride, is sequentially added 2- propyl valeric chloride, triethylamine, are stirred to room temperature, continue to be stirred to react, purify, and 4,5- dihydroxy -7- methyl-is made 9,10- diketone -9,10- anthraquinone -2- quinhydrones -2- valproic acid ester.

More specifically, preparation step are as follows:

1) 10 mmol valproic acids are dissolved in 20mL methylene chloride, 12 mmol oxalyl chlorides is then added.By this Mixed solution is placed under condition of ice bath and stirs 12h, through in TLC detection architecture without raw material after stop reaction (expansion system :), obtain Obtain crude product 2- propyl valeric chloride (compound 1)；

2) under condition of ice bath, 0.4 mmol rheum emodin (1,3,8- trihydroxy -6- methyl -9,10- anthraquinone) is dissolved in 2mL In methylene chloride, it is then gradually adding the obtained 2- propyl valeric chloride of 0.48mmol step 1), 1.0mmol triethylamine, stirring It mixes to room temperature, continues to stir 4h.Through in TLC detection architecture without raw material after stop reaction (expansion system: PE/EtOAc= 8:1), 4,5- of purified acquisition dihydroxy -7- methyl -9,10- diketone -9,10- anthraquinone -2- quinhydrones -2- valproic acid ester (compound 2)；

The purifying are as follows: reaction solution is diluted with ethyl acetate, then successively uses NaHCO₃Solution extracting and washing 2 times, go from Sub- water washing 1 time, saturated common salt water washing 1 time, each 5mL, liquid separation obtains organic layer, and organic layer adds appropriate anhydrous sodium sulfate, It removes water within stirring 2-3 minutes, then reaction solution is filtered, wash sodium sulphate with ethyl acetate, collect cleaning solution and filtrate, warp Crude product is obtained after Rotary Evaporators revolving；Then with petroleum ether and the methylene chloride eluant, eluent that 2:1 is mixed to prepare by volume It being rinsed, collection obtains target product, and it is rotated with Rotary Evaporators and removes solvent, then pump a small amount of residual solvent with oil pump, Finally obtain derivative.

Gained compound is as rheum emodin, to HL60(people in loop) there are inhibitory activity, Er Qiesuo The activity for obtaining compound is higher than rheum emodin (its IC to HL60₅₀Value is the one third of rheum emodin), it is expected to be used for leukaemia More target therapeutic agents.

Remarkable advantage of the invention is:

(1) present invention only requires two-step reactions, and synthetic method is simple, and experiment condition is mild, do not require high temperature and pressure etc. Harsh conditions, yield is generally up to 70% or more；

(2) relative molecular mass of present invention gained emodin derivates belongs to small molecule compound less than 500, and raw Object cell experiment confirms the anti-HL60(people in loop that the compound has) activity is better than rheum emodin, therefore the change Closing object has the prospect for being used to prepare anticancer drug.

Specific embodiment

In order to make content of the present invention easily facilitate understanding, With reference to embodiment to of the present invention Technical solution is described further, but the present invention is not limited only to this.

Embodiment 1

Fat-soluble emodin derivates preparation

(1) preparation of 2- propyl valeric chloride (compound 1)

1. reaction: taking the 100 mL eggplant type flasks an of dried and clean, 10 mmol valproic acids are dissolved in 20 mL In methylene chloride, then 12 mmol ethanedioly chlorides are added in reaction system with spoon.This mixed solution is placed on ice bath Under the conditions of stir 12h, through in TLC detection architecture without raw material after stop reaction (expansion system: methylene chloride: methanol=20:1, body Product ratio)；

2. purifying: directly being spin-dried for obtaining crude product with Rotary Evaporators, purify up to 2- propyl valeric chloride (compound 1) ?.

¹ ₃ δ 2.84 – 2.74 (m, 1H), 1.82 – 1.66 (m, 2H), 1.57 – 1.48 (m, 2H), 1.43 – 1.32 (m, 4H), 0.93 (t, J = 7.2 Hz, 6H).¹³C NMR (101 MHz, CDCl₃) δ 177.55,56.93,34.26,20.35,13.99.(this step is reacted mainly for participating in next step in actual production process Reaction, it is not necessary to carry out excessive purifying again)；

(2) system of 4,5- dihydroxy -7- methyl -9,10- diketone -9,10- anthraquinone -2- quinhydrones -2- valproic acid (compound 2) It is standby

1. reaction: under the conditions of ice-water bath, rheum emodin (108 mg, 0.4 mmol) is added into reaction flask, is taken with dropper The methylene chloride of about 2m L is dissolved, and compound 1(78 mg, 0.48 mmol are then sequentially added) and 140 μ L three Ethamine.It is stirred to room temperature, continues to stir 4h, take in a small amount of liquid TLC detection architecture without stopping reaction after raw material.

2. purifying: a) diluting reaction solution 15mL ethyl acetate, then successively use NaHCO₃Solution extracting and washing 2 times is gone Ion water washing 1 time, saturated common salt water washing 1 time, each 5mL, liquid separation obtains organic layer, and organic layer adds appropriate anhydrous slufuric acid Sodium is stirred 2-3 minutes and is removed water, be then filtered to reaction solution, wash sodium sulphate with a small amount of ethyl acetate (3 × 2 mL), receives Collect cleaning solution and filtrate, obtains crude product after Rotary Evaporators rotate；B) use petroleum ether: methylene chloride=2:1 is eluant, eluent punching It washes, collection obtains target product, is rotated with Rotary Evaporators and removes solvent, then pumps a small amount of residual solvent with oil pump, final to obtain To 118 mg red solids, yield 74%.

Product characters: red solid (under room temperature).Fusing point: 115.2-115.9 DEG C.

: R_f=0.65(PE:EtOAc=8:1).

¹ ₃ δ 12.16 (s, 1H), 11.92 (s, 1H), 7.60 (d, J = 1.6 Hz, 1H), 7.47 (d,J = 2.3 Hz, 1H), 7.06 (s, 1H), 7.00 (d, J = 2.3 Hz, 1H), 2.68-2.60 (m, 1H), 2.44 (s, 3H), 1.81-1.71 (m, 2H), 1.62-1.53 (m, 2H), 1.48-1.40 (m, 4H), 0.98 (t, J = 7.2 Hz, 6H).

¹³C NMR (101 MHz, CDCl₃) δ 191.57, 181.30, 173.93, 164.06, 162.85, 157.49, 149.44, 135.07, 133.05, 124.74, 121.71, 116.92, 114.13, 113.90, 113.59, 45.57, 34.63, 22.35, 20.84, 14.16.

HRMS (ESI): calcd for C₂₃H₂₄O₆ [M - H]^- m/z 395.1500, found 395.1498.

Application Example 1

Active testing

Compound 2 and rheum emodin are to HL60(people in loop) proliferation and survival inhibiting effect

Material: HL60 cell strain (people in loop)；

Test medicine: 1 gained final compound 2 of embodiment and rheum emodin；

Cell culture processes: the HL60 cell strain frozen in liquid nitrogen is taken out, thaws in 37 DEG C of warm water, cell is hanged Liquid moves into 1.5 mL centrifuge tubes, is placed in a centrifuge, and 1500 rpm are centrifuged 5 min, discards supernatant liquid, and 1 mL is added 1640 complete culture solution of RPMI, gently piping and druming uniformly, cell suspension is added in culture dish, 3 mL RPMI 1640 are added Culture dish is placed in 5% CO by complete culture solution₂, cultivate in 37 DEG C of incubators.

Cytotoxicity experiment: by TOLEDO cell with 2 × 10⁴The density of a cells/well is inoculated into 96 well culture plates, The compound (or rheum emodin) of 10 μ Μ/L is added, places it in 37 DEG C, cultivate 48 hours in 5 % carbon dioxide incubators.It takes 10 μ L CCK-8 reagents are added in culture plate out, every hole, continue to be placed in 37 DEG C for culture plate, incubate in 5 % carbon dioxide incubators After educating 2 hours, culture is terminated, careful inhale abandons supernatant, and 150 μ L DMSO are added in every hole, and being protected from light 10 min of oscillation makes crystal Sufficiently dissolution；The trap OD at 450 nm is detected with microplate reader₄₅₀, and cell survival rate is calculated according to following formula:

Cell survival rate %=(processing group OD₄₅₀/ control group OD₄₅₀) × 100%, then using 13.0 software of SPSS to data It is handled, and calculates the half-inhibitory concentration (IC of cancer cell multiplication₅₀).HL60 can be effectively suppressed in rheum emodin and compound 2 The growing multiplication of cell strain (people in loop), but effective inhibition concentration of gained compound is the three of rheum emodin / mono-.

The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with Modification, is all covered by the present invention.

Claims (6)

1. a kind of fat-soluble emodin derivates, it is characterised in that: the derivative concrete structure formula is as follows:

。

2. a kind of preparation method of fat-soluble emodin derivates as described in claim 1, it is characterised in that: including walking as follows It is rapid:

1) using valproic acid and oxalyl chloride as starting material, using methylene chloride as solvent, reaction obtains 2- third under condition of ice bath Base valeric chloride；

2) under condition of ice bath, 1,3,8- trihydroxy -6- methyl -9,10- anthraquinone is dissolved in methylene chloride, 2- third is sequentially added Base valeric chloride, triethylamine, are stirred to room temperature, continue to be stirred to react, purify, and methyl -9 4,5- dihydroxy -7- are made, 10- diketone -9,10- anthraquinone -2- quinhydrones -2- valproic acid ester.

3. the preparation method of fat-soluble emodin derivates according to claim 2, it is characterised in that: 2- propyl in step 1) The molar ratio of valeric acid and oxalyl chloride is 1:1.2.

4. the preparation method of fat-soluble emodin derivates according to claim 2, it is characterised in that: 1,3,8- in step 2 The molar ratio of trihydroxy -6- methyl -9,10- anthraquinone, 2- propyl valeric chloride and triethylamine are as follows: 1:1.2:2.5.

5. the preparation method of fat-soluble emodin derivates according to claim 2, it is characterised in that: continue to stir in step 2 The time for mixing reaction is 4h；The purifying are as follows: reaction solution is diluted with ethyl acetate, then successively uses NaHCO₃Solution extracting and washing 2 times, deionized water washing 1 time, saturated common salt water washing 1 time, each 5mL, liquid separation obtain organic layer, and organic layer adds appropriate anhydrous Sodium sulphate, stirs 2-3 minute and removes water, be then filtered to reaction solution, wash sodium sulphate with ethyl acetate, collection cleaning solution with Filtrate obtains crude product after Rotary Evaporators rotate；Then with petroleum ether and methylene chloride, 2:1 is mixed to prepare by volume Eluant, eluent is rinsed, and collection obtains target product, is rotated with Rotary Evaporators and is removed solvent, then pumps a small amount of residual with oil pump Solvent finally obtains derivative.

6. a kind of application of fat-soluble emodin derivates as described in claim 1 on preparation leukaemia target therapeutic agent.