CN102432663B

CN102432663B - Celastrol derivative and preparation method thereof and application of celastrol derivative to preparation of antitumor medicine

Info

Publication number: CN102432663B
Application number: CN201110331302.3A
Authority: CN
Inventors: 单伟光; 占扎君; 张丽雯; 温彦涛; 陈艳; 王建伟; 唐岚; 于海宁; 侯晓蓉
Original assignee: Zhejiang University of Technology ZJUT
Current assignee: Zhejiang University of Technology ZJUT
Priority date: 2011-10-27
Filing date: 2011-10-27
Publication date: 2014-12-03
Anticipated expiration: 2031-10-27
Also published as: CN102432663A

Abstract

本发明涉及雷公藤红素及衍生物和雷公藤红素衍生物的制备方法，还涉及以雷公藤红素衍生物为原料制备的生物盐，以及雷公藤红素衍生物在治备抗肿瘤药物中的应用，所述雷公藤红素衍生物具有如式I所示的结构：其中，R₁为H、C1-C6直链或支链烷基、苄基及苯环上有取代苄基；R₂或R₃各自独立为C5-C6的环状烃基、C1-C6的链状烷基、苯基及取代苯基，或者R₂、R₃与N成环，所述的环为含N或O的六元环状杂环，所述雷公藤红素衍生物具有良好的抗肿瘤活性、稳定性及水溶性。且可与医学上可接受的无机酸(如盐酸、硫酸和磷酸)或有机酸(柠檬酸、肉桂酸和琥珀酸等)的一种成盐，该盐具有很好的水溶性。The present invention relates to the preparation method of tripterygne and its derivatives and tripteryne derivatives, also relates to the biological salt prepared from tripteryne derivatives as raw materials, and tripteryne derivatives in the preparation of anti-tumor drugs For the application in , the tripterine derivative has a structure as shown in formula I: wherein, R ₁ is H, C1-C6 straight chain or branched chain alkyl, benzyl and substituted benzyl on the benzene ring; R ₂ or R ₃ are each independently a C5-C6 cyclic hydrocarbon group, a C1-C6 chain alkyl group, a phenyl group and a substituted phenyl group, or R ₂ , R ₃ and N form a ring, and the ring contains N Or a six-membered cyclic heterocycle of O, the tripterine derivatives have good antitumor activity, stability and water solubility. And it can form a salt with a medically acceptable inorganic acid (such as hydrochloric acid, sulfuric acid and phosphoric acid) or organic acid (citric acid, cinnamic acid and succinic acid, etc.), and the salt has good water solubility.

Description

Tripterine derivate and preparation thereof and the application in preparing antitumor drug

(1) technical field

The present invention relates to the preparation method of Tripterine and derivative and tripterine derivate, also relate to and take the biogenic salt that tripterine derivate prepared as raw material and the application of tripterine derivate in controlling standby antitumor drug.

(2) background technology

Malignant tumour serious harm people's health according to WHO statistics, is died from every year on average malignant tumour person and is reached 6,900,000 people in the whole world more than 50 hundred million populations, and new cases are 8,700,000 examples, and numeral is also increasing year by year.Antitumor drug treatment is one of main method of cancer therapy, natural product is the treasure-house of finding new type antineoplastic medicine, the lead compound of many antitumor drugs is all acquisition from natural, as vinealeucoblastine(VLB), vincristine(VCR), camptothecine, taxol etc., but these natural compoundss are often due to toxic side effect, solvability, stability etc., need to carry out structural modification or transformation, just can become the antitumor drug of clinical use.

Tripterine (Celastrol), has another name called celastrin, for deriving from the pentacyclic triterpene compound of Chinese herb triperygium wilfordii root skin.Tripterine pharmacologically active is abundant, acts on extensively, especially anti-tumor aspect in traditional Chinese medicine.Recent research shows that its antitumor action target spot may be new vessel endotheliocyte, heat shock protein(HSP) Hsp90 and proteasome etc.New vessel endotheliocyte: growth and the transfer of many primary tumo(u)rs depend on blood vessel.Tumour entity both can obtain nutrition and oxygen from host by new vessel, can to host, carry transitional cell continuously by new vessel again, and formed at other position continued growths and the induction of vascular of body, caused metastases.The new life of blood vessel depends on the balance of the different kinds of molecules of host and tumour cell release, by raising the stimulating factor of blood vessel generation and lowering the coefficient result of angiopoietic supressor.This take tumor endothelial cell for angiogenesis inhibitor provides theoretical foundation as action target spot.Compare other antitumor drugs, angiogenesis inhibitor has following advantage: (1) produces resistance hardly: tumour cell has compared with high mutagenicity makes it be easy to produce resistance, angiogenesis inhibitor be take endotheliocyte as target spot, and this cell is almost without sudden change, just can not produce drug-resistant effect yet; (2) dosage is little; (3) toxic side effect is little: in normal human, except wound healing and the reproductive cycle, nearly all new vessel is all pathologic, as rheumatic arthritis, tumour and diabetes etc.Angiogenesis inhibitor directly acts on new vessel, naturally can not produce toxic side effecties such as stomach reaction, renal toxicity; (4) drug combination is effective: the origin cause of formation of tumour is extremely complicated, and the pharmacological agent difficulty of single target spot reaches expection, and desirable treatment plan is the medication of many target drugs of associating.Neovascularization inhibitor specific action is in the blood vessel away from tumour cell, and the medicine medication as combination tumor cell target spot, will obtain better curative effect.Proteasome: be a kind of multivalence catalytic protein enzyme complex of high conservative, be extensively present in eukaryotic core and in tenuigenin.This mixture energy catalytic hydrolysis is connected with the albumen of poly ubiquitin chain, is the main ingredient of Ubiquitin-proteasome path.Ubiquitin-proteasome path mainly exerts an influence to malignant tumour by following number of ways: the propagation of (1) cell needs cyclin to regulate and control, different steps in the cell cycle relies on special cyclin, and unwanted cyclin is degraded in time, they are the substrate of proteasome just.(2) NF-kB transcription factor and tumour generation, growth, transfer and chemicotherapy opposing are closely related.Under standard state, NF-kB and its arrestin IkB combination in tenuigenin, exist with the composite form of non-activity, after proteasome hydrolysis IkB, just can discharge the activated NF-kB of tool.(3) p53 tumor-inhibiting factor low expression level in normal cell, can be degraded by proteasome very soon, suppresses the expression level that the latter can improve p53.Heat shock protein 90 (HSP90): be extensively present in cell, account for 1%～2% of normal cell total protein, its participates in the activation of client's albumen, the process such as folding, brings into play the effect of its molecular chaperones.Client's albumen of Hsp90 comprises antitumor target spot and some other albumen relevant to intracellular signal transduction pathway of the clinical confirmations such as Tyrosylprotein kinase (Her2, EGFR), metastable signal protein (Akt, Raf-1 and IKK), jump signal albumen (p53, v-Src), chimeric signal protein (Bcr-Abl), cyclin (Cdk4, Cdk6), steroid receptor (androgen receptor, estrogen receptor and progesterone).Suppress Hsp90 and can cause client's protein conformation abnormal, and paramorph albumen can be got rid of by Ubiquitin-proteasome path fast by cell.The major advantage of Hsp90 inhibitor is that they can block a plurality of paths that cancer cells forms simultaneously, and while having avoided a certain path of single inhibition, tumour cell easily produces the drawback of resistance.Hsp90 mainly exists with potential non-complexes state in normal cell, and in tumour cell, Hsp90 and multiple auxiliary molecular chaperones form mixture, with active state, exist, and make Hsp90 inhibitor have specificity and selectivity.Compare with conventional cell possible mechanism of its cytotoxicity, the antitumor drug Main Function of this class target spot inhibitor is in key molecule and the signal transduction pathway thereof of the huge regulating cell proliferate of difference in normal cell and tumour cell, and the toxicity that improved selectivity to tumour, has reduced normal tissue is because of very good to application prospect.

The researchs such as Nakanishi show in Tripterine molecular structure, and C-3 hydroxyl, in conjugated system, has slightly acidic as phenolic hydroxyl group, oxidizable, and contact soda acid all can produce complicated rearrangement reaction, causes this compound stability poor.Because Alpha-hydroxy quinone methyl structural is very sensitive to acid, during the administration of prompting oral administration, acid hydrochloric acid in gastric juice can destroy part Tripterine, causes drug effect to reduce simultaneously.On the other hand, Tripterine belongs to triterpene compound, is fat-soluble component, almost insoluble in water, and bioavailability is low in body, and is difficult to directly with the administration of intravenous injection mode this very big possibility that must limit its exploitation patent medicine.

Along with going deep into of research, the viewpoint wide-scale distribution of the height toxicity of Tripterine, becomes the large difficult point that antitumor drug is developed.The researchs such as Bao Yixiao show that Tripterine can induce HMC-1 apoptosis, and apoptosis mainly occurs in the S phase.Tripterine poor selectivity is found in the research such as Sifeng Wang, and directly administration easily produces larger side effect, can affect the growth of the zebrafish embryo under normal development state under micro-molar concentration.

Publication number is that the Chinese patent of CN101041684A discloses a kind of Tripterine meglumine salt compound and preparation method thereof; The Chinese patent that publication number is CN101279995A provides a kind of tripterine salt, is specially Tripterine and medically acceptable basic aminoacids or a kind of the become salt of organic amine or mineral alkali; Publication number is composition, the preparation method and its usage that the Chinese patent of CN101496804A discloses a kind of Tripterine and arginic acid salt, above three pieces of Chinese patents only improve water-soluble in the mode of salify, the anti-tumor activity of Tripterine and toxicity are not done to modify and improve.Publication number is that the Chinese patent of CN101311187A discloses a kind of celastrol long-chain alcohol ester and preparation method thereof and purposes, this long-chain alcohol ester has significant prostate gland targeting, and antitumor and anti-inflammatory activity is better than Tripterine, but this patent has been ignored the water-soluble problem of Tripterine, in patent medicine problem, do not do relevant consideration.Publication number is that the Chinese patent of CN101352444A discloses the application of Tripterine in the medicine of the cancer of preparation treatment expression CIP2A albumen, described trypterygine have following characteristics: do not have the specific remarkable downward CIP2A albumen of clone, in the reduction lung cancer of energy time and dose-dependently, liver cancer, mammary cancer, cancer of the stomach, the expression of cancer protein CIP2A, is with a wide range of applications.The research of current domestic patent mainly concentrate on Tripterine by the mode of simple salify, improve water-soluble, to become the mode of ester to improve activity, and pharmacology activity research aspect.

(3) summary of the invention

The preparation method who the object of this invention is to provide a kind of Tripterine and derivative and tripterine derivate, also relates to and take the biogenic salt that tripterine derivate prepared as raw material and the application of tripterine derivate.

A kind of Tripterine and derivative as shown in formula I.

Wherein, R ₁for having substituent benzyl on H, C1-C6 straight or branched alkyl, benzyl and phenyl ring, the substituting group on phenyl ring is fluorine, chlorine, methoxyl group or nitro; R ₂or R ₃independent is separately the cyclic hydrocarbon group of C5-C6, the chain-like alkyl of C1-C6, phenyl and substituted-phenyl, and the substituting group of described substituted-phenyl is fluorine, chlorine, methoxyl group or nitro, or R ₂, R ₃become ring with N, described ring is the six-membered cyclic heterocycle that contains N or contain N and O.

Described R ₁for methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, n-pentyl or n-hexyl.

Described R ₁for there being the benzyl of replacement on H, benzyl and phenyl ring, the substituting group that has the benzyl of replacement on described phenyl ring is fluorine, chlorine, methoxyl group or nitro.

Described R ₂, R ₃become ring with N, described ring is morpholinyl, piperazinyl, N-substituted piperazinyl, piperidyl.

Described R ₂or R ₃independent is separately methyl, ethyl, n-propyl, phenyl or substituted-phenyl, and the substituting group of described substituted-phenyl is fluorine, chlorine, methoxyl group or nitro.

The preparation method of the tripterine derivate of a kind of formula as shown in (I), described tripterine derivate is prepared as follows: triphosgene is dissolved in to methylene dichloride and is made into the triphosgene solution that concentration is 0.1～0.5mol/L, after dripping concentration under ice bath be the nitrogenous compound dichloromethane solution and triethylamine of 0.1～0.5mol/L in triphosgene solution, after stirring at room 1～2h, drip concentration and be 0.1～0.5mol/L as shown in formula II containing Tripterine compound dichloromethane solution, continue to stir 20～48h, point plate detects, stopped reaction after the Tripterine compound as formula II has reacted, the deionized water that adds 3～6 times of reaction solution A volumes in reaction solution A, be extracted with ethyl acetate again, get organic layer, with saturated NaCl solution washing, use anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude product through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, point plate detects, collection contains under 254nm aobvious strong ultraviolet and polarity and is greater than accordingly as the elutriant of the compound of formula II, merges elutriant, and the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate after being dried, described Tripterine compound: triphosgene: nitrogenous compound: the amount of substance of triethylamine is than being 1:1.2～1.5:1.4～2.0:3～5, described nitrogenous compound is the cyclic amine of C5-C6, on the chain amine of C1-C6, phenyl ring, have the aniline of replacement, and the substituting group on described phenyl ring is fluorine, chlorine, methoxyl group or nitro, or containing N or containing the hexa-member heterocycle of N and O, described hexa-member heterocycle is morpholine, piperazine, N-substituted-piperazinyl, piperidines,

Wherein, R ₁during for H, described Tripterine compound is Tripterine;

R ₁while being not H, described Tripterine compound is Tripterine ester.

Method of the present invention, described Tripterine ester is prepared as follows: described Tripterine ester is prepared as follows: the Tripterine shown in formula III is dissolved in N, dinethylformamide is made the trypterygine cellulose solution that concentration is 0.1～0.3mol/L, take sodium bicarbonate as catalyzer, add halohydrocarbon R ₁x, described X is Br or I, described R ₁for having substituent benzyl on H, C1-C6 straight or branched alkyl, benzyl and phenyl ring, substituting group on described phenyl ring is fluorine, chlorine, methoxyl group or nitro, stirring at room 12～24 hours, the deionized water that adds 3～6 times of reaction solution B volumes in reaction solution B, ethyl acetate extraction, gets organic layer, with saturated sodium thiosulfate, washs, then use saturated NaCl solution washing, then use anhydrous Na ₂sO ₄dry, suction filtration, organic phase concentrates to obtain red oil, through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, the elutriant that collection contains target Tripterine ester, merges elutriant, and the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester; Described Tripterine: halohydrocarbon: the amount of substance of sodium bicarbonate is than being 1:1.2～2.0:3.0～5.0.

Preferably, the preparation method of the tripterine derivate of a kind of formula as shown in (I), described tripterine derivate is prepared as follows: triphosgene is dissolved in to methylene dichloride and is made into the triphosgene solution that concentration is 0.12mol/L, after dripping concentration under ice bath be the nitrogenous compound dichloromethane solution and triethylamine of 0.35mol/L in triphosgene solution, after stirring at room 1h, drip concentration and be 0.2mol/L as shown in formula II containing Tripterine compound dichloromethane solution, continue to stir 24h, point plate detects, stopped reaction after the Tripterine compound as formula II has reacted, the deionized water that adds 3.75 times of reaction solution A volumes in reaction solution A, be extracted with ethyl acetate again, get organic layer, with saturated NaCl solution washing, use anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude product through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, point plate detects, collection contains under 254nm aobvious strong ultraviolet and polarity and is greater than accordingly as the elutriant of the compound of formula II, merges elutriant, and the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate after being dried, described Tripterine compound: triphosgene: nitrogenous compound: the amount of substance of triethylamine is than being 1:1.2:1.75:3.75, described nitrogenous compound is the cyclic amine of C5-C6, on the chain amine of C1-C6, phenyl ring, have the aniline of replacement, and the substituting group on described phenyl ring is fluorine, chlorine, methoxyl group or nitro, or containing N or containing the hexa-member heterocycle of N and O, described hexa-member heterocycle is morpholine, piperazine, N-substituted-piperazinyl, piperidines.

Preferably, described Tripterine ester is prepared as follows: the Tripterine shown in formula III is dissolved in DMF and makes the trypterygine cellulose solution that concentration is 0.15mol/L, take sodium bicarbonate as catalyzer, adds halohydrocarbon R ₁x, described X is Br or I, described R ₁for having substituent benzyl on H, C1-C6 straight or branched alkyl, benzyl and phenyl ring, substituting group on described phenyl ring is fluorine, chlorine, methoxyl group or nitro, stirring at room 12h, the deionized water that adds 3～6 times of reaction solution B volumes in reaction solution B, ethyl acetate extraction, gets organic layer, with saturated sodium thiosulfate, washs, then use saturated NaCl solution washing, then use anhydrous Na ₂sO ₄dry, suction filtration, organic phase concentrates to obtain red oil, through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, the elutriant that collection contains Tripterine ester, merges elutriant, and the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester; Described Tripterine: halohydrocarbon: the amount of substance of sodium bicarbonate is than being 1:1.6:5.0.

A biogenic salt shown in the formula IV being prepared by tripterine derivate as claimed in claim 1, biogenic salt shown in formula IV is made soluble derivative by the Tripterine as described in formula I and derivative and medicinal acceptable mineral acid or organic acid salify; Described mineral acid is hydrochloric acid, sulfuric acid and phosphoric acid; Described organic acid is citric acid, oxysuccinic acid and Fu Ma acid;

The invention still further relates to the application of described tripterine derivate in preparing antitumor drug.

Application of the present invention, is characterized in that the application of described tripterine derivate in preparing Hepatoma therapy or lung-cancer medicament.

The invention has the beneficial effects as follows:

1. by the modification to C-3 position hydroxyl, destroyed this hydroxyl and the carbonyl intramolecular hydrogen bond effect of C-2 position, this derivative is greatly improved water-soluble.On the hydroxyl of C-3 position, introduce nitrogenous hydrophilic radical simultaneously, not only greatly must improve the water-soluble of this derivative, and make it can further improve with the salify of medically acceptable mineral acid (example hydrochloric acid, sulfuric acid and phosphoric acid) or organic acid (citric acid, oxysuccinic acid and Fu Ma acid etc.) that it is water-soluble, compare with compound Tripterine, its bioavailability improves significantly, and increases the selectivity of preparation simultaneously.

2. by the modification to C-3 position hydroxyl, the enol of poor stability in Tripterine structure is protected, improved the stability of compound, be more conducive to the control of quality.

3. the tripterine derivate inside and outside anti-tumor activity evaluation the present invention relates to shows: external anti-tumor activity shows that wherein most of tripterine derivate shows stronger anti tumor activity in vitro to people's liver cancer, lung cancer cell line, but all lower than Tripterine, and activity is better than Tripterine in vivo.This explanation the present invention prompting, after structural modification, has reduced the cytotoxicity of Tripterine on the one hand, improves drug safety; On the other hand, on its C-3 hydroxyl, introduce some hydrophilic radicals, significantly improved its medicine for character, improve bioavailability, metabolism goes out former medicine in vivo simultaneously, has the character of prodrug, and this had both improved the bioavailability of medicine, the systemic side effects that can reduce medicine simultaneously, has improved security.The protecting group of 3 hydroxyls of tripterine derivate that the present invention relates to is in addition isocyanic ester.This compounds and 3 ester derivatives are more easily metabolized to former medicine, thereby have better antitumor action.

(4) embodiment

Below in conjunction with specific embodiment, the present invention is described further, but protection scope of the present invention is not limited in this:

Embodiment 1: synthetic (synthetic route chart as shown in Figure 1) of tripterine derivate CLS-1

72.8mg triphosgene (BTC) (0.24mmol) is dissolved in 2mL methylene dichloride (DCM), under ice bath, drip the morpholine that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines (0.99mmol), stirring at room 1h, drip again the Tripterine that 1mL concentration is 0.2mol/L (0.2mmol) dichloromethane solution, continue to stir 24h, stopped reaction, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, get filtrate concentrated the crude product of garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=2:1; Rf value 0.38), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-1 after being dried is 98.0mg, productive rate 87%.CLS-1: ¹hNMR (CDCl ₃, 500MHz): 7.01 (1H, dd, J=1.5,7.5Hz, H-6), 6.51 (1H, d, J=1.5Hz, H-1), 6.35 (1H, d, J=7.5Hz, H-7), 3.37-3.71 (8H, m, H-morpholine ring), 2.21 (3H, s, 4-CH ₃), 1.47 (3H, s, 9-CH ₃), 1.31 (3H, s, 13-CH ₃), 1.28 (3H, s, 14-CH ₃), 1.03 (3H, s, 17-CH ₃), 0.77 (3H, s, 20-CH ₃) .MS (ESI) [M+H] ⁺m/z564.3.

The different groups of table 1 tripterine derivate

The synthetic route of tripterine derivate CLS-1～CLS-10:

Synthetic (synthetic route chart as shown in Figure 1) of embodiment 2 tripterine derivate CLS-2

72.8mg BTC (0.24mmol) is dissolved in 2mL methylene dichloride (DCM), under ice bath, drip the N methyl piperazine that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines (0.99mmol), stirring at room 1h, adding 1mL concentration is Tripterine (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=2:1; Rf value 0.30), collection contains under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-2 after being dried is 101.6mg, productive rate 88%.CLS-2: ¹H?NMR(CDCl ₃,500MHz):3.51-3.63(2H,m,H-2′),3.36-3.46(2H,m,H-3′),2.45(2H,br?s,H-4′),2.42(2H,br?s,H-5′),2.31(3H,s,N-CH ₃).MS(ESI)[M+H] ⁺m/z577.4.

Synthetic (synthetic route chart as shown in Figure 1) of embodiment 3 tripterine derivate CLS-3

72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride (DCM), under ice bath, drip the NEP that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, drip again the Tripterine that 1mL concentration is 0.2mol/L (0.2mmol) dichloromethane solution, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=2:1; Rf value 0.33), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-3 after being dried is 99.2mg, productive rate 84%.CLS-3: ¹H?NMR(CDCl ₃,500MHz):3.53-3.60(2H,m,H-2′),3.39-3.45(2H,m,H-3′),2.39-2.50(4H,m,H-4′,5′,6′),1.07(3H,t,J=6.0,6′-CH ₃).MS(ESI)[M+H] ⁺m/z591.4.

Embodiment 4 tripterine derivate CLS-4's is synthetic

72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride (DCM), under ice bath, drip the acetylpiperazine that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution, triethylamine (100 μ L), stirring at room 1h, drip again the Tripterine that 1mL concentration is 0.2mol/L (0.2mmol) dichloromethane solution, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=2:1; Rf value 0.38), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-4 after being dried is 99.0mg, productive rate 82%.CLS-4: ¹h NMR (CDCl ₃, 500MHz): 3.36-3.61 (8H, m, H-piperazine ring), 2.11 (3H, s, 6 '-CH ₃) .MS (ESI) [M+H] ⁺m/z605.4.

Embodiment 5 tripterine derivate CLS-5's is synthetic

72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride (DCM), under ice bath, dripping 1mL concentration is propionyl piperazine (0.35mmol) dichloromethane solution and the 100 μ L triethylamines of 0.35mol/L, stirring at room 1h, drip again the Tripterine that 1mL concentration is 0.2mol/L (0.2mmol) dichloromethane solution, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=2:1; Rf value 0.40), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-5 after being dried is 84.0mg, productive rate 68%.CLS-5: ¹h NMR (CDCl ₃, 500MHz): 3.37-3.64 (8H, m, H-piperazine ring), 1.17 (3H, t, J=6.0Hz, 7 '-CH ₃) .MS (ESI) [M+H] ⁺m/z619.4.

Synthetic (synthetic route chart as shown in Figure 1) of embodiment 6 tripterine derivate CLS-6

72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride (DCM), under ice bath, drip the benzyl diethylenediamine that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, drip again the Tripterine that 1mL concentration is 0.2mol/L (0.2mmol) dichloromethane solution, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=2:1; Rf value 0.44), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-6 after being dried is 70.4mg, productive rate 54%.CLS-6: ¹h NMR (CDCl ₃, 500MHz): 7.28-7.33 (5H, m, H-phenyl ring), 3.78 (2H, brs, H-6 '), 3.52 (2H, brs, H-2 '), 3.39 (2H, brs, H-3 '), 2.41-2.45 (4H, br s, H-4 ', 5 ') .MS (ESI) [M+H] ⁺m/z653.4.

Synthetic (synthetic route chart as shown in Figure 1) of embodiment 7 tripterine derivate CLS-7

72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride (DCM), under ice bath, drip dichloromethane solution and the 100 μ L triethylamines of the piperidines that 1mL concentration is 0.35mol/L (0.35mmol), stirring at room 1h, drip again the Tripterine that 1mL concentration is 0.2mol/L (0.2mmol) dichloromethane solution, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=2:1; Rf value 0.42), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-7 after being dried is 94.4mg, productive rate 84%.CLS-7: ¹H?NMR(CDCl ₃,500MHz):3.49(2H,m,H-2′),3.33(2H,m,H-3′).MS(ESI)[M+H] ⁺m/z562.3.

Synthetic (synthetic route chart as shown in Figure 1) of embodiment 8 tripterine derivate CLS-8

72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride (DCM), under ice bath, drip the pipecoline that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, drip again the Tripterine that 1mL concentration is 0.2mol/L (0.2mmol) dichloromethane solution, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=2:1; Rf value 0.47), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-8 after being dried is 74.8mg, productive rate 65%.CLS-8: ¹H?NMR(CDCl ₃,500MHz):3.39(1H,m,H-2′),3.29(1H,m,H-2′),1.26(3H,d,J=7.5Hz,2′-CH ₃).MS(ESI)[M+H] ⁺m/z576.3.

Synthetic (synthetic route chart as shown in Figure 1) of embodiment 9 tripterine derivate CLS-9

72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride (DCM), under ice bath, drip the diethylamine that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, drip again the Tripterine that 1mL concentration is 0.2mol/L (0.2mmol) dichloromethane solution, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=2:1; Rf value 0.38), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-9 after being dried is 59.6mg, productive rate 49%.CLS-9: ¹H?NMR(CDCl ₃,500MHz):3.34(2H,m,H-2′),3.19(2H,m,H-3′),1.27(3H,m,2′-CH ₃),1.16(3H,m,3′-CH ₃).MS(ESI)[M+H] ⁺m/z550.3.

Synthetic (synthetic route chart as shown in Figure 1) of embodiment 10 tripterine derivate CLS-10

72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride (DCM), under ice bath, drip dichloromethane solution and the 100 μ L triethylamines of the methylphenylamine that 1mL concentration is 0.35mol/L (0.35mmol), stirring at room 1h, drip again the Tripterine that 1mL concentration is 0.2mol/L (0.2mmol) dichloromethane solution, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=2:1; Rf value 0.47), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-10 after being dried is 60.0mg, productive rate 51%.CLS-10: ¹h NMR (CDCl ₃, 500MHz): 7.18-7.39 (5H, m, H-phenyl ring), 3.30 (3H, s, N-CH ₃) .MS (ESI) [M+H] ⁺m/z584.3.

Synthetic (synthetic route chart as shown in Figure 2) of embodiment 11 tripterine derivate CLS-11

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.48mmol benzyl bromine, under room temperature, stir stopped reaction after 12h, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, with saturated sodium thiosulfate, wash 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester.72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip the morpholine that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, dripping 1mL concentration is Tripterine benzyl ester (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.45), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-11 after being dried is 98.0mg, productive rate 87%.CLS-11: ¹h NMR (CDCl ₃, 500MHz): 7.28-7.35 (5H, m, H-phenyl ring), 4.99 (2H, dd, J=12.5,25.5Hz, H-1 '), 3.58-3.78 (8H, m, H-morpholine ring), 2.19 (3H, s, 4-CH ₃), 0.52 (3H, s, 20-CH ₃) .MS (ESI) [M+H] ⁺m/z654.4.

The synthetic route of tripterine derivate CLS-11～CLS-30:

Synthetic (synthetic route chart as shown in Figure 2) of embodiment 12 tripterine derivate CLS-12

135mg Tripterine (0.3mmol) is dissolved in to 2mLN, in dinethylformamide (DMF), adds 126mg sodium bicarbonate (1.5mmol) and 0.48mmol benzyl bromine.Under room temperature, stir 12h, stopped reaction, adds 10mL deionized water, and ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, and then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester; 72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip dichloromethane solution and the 100 μ L triethylamines of the N methyl piperazine that 1mL concentration is 0.35mol/L (0.35mmol), stirring at room 1h, dripping 1mL concentration is Tripterine benzyl ester (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.40), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-12 after being dried is 101.2mg, productive rate 88%.CLS-12:MS(ESI)[M+H] ⁺m/z667.4.

Synthetic (synthetic route chart as shown in Figure 2) of embodiment 13 tripterine derivate CLS-13

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.48mmol benzyl bromine, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester; 72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip dichloromethane solution and the 100 μ L triethylamines of the NEP that 1mL concentration is 0.35mol/L (0.35mmol), stirring at room 1h, dripping 1mL concentration is Tripterine benzyl ester (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.42), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-13 after being dried is 100.0mg, productive rate 84%.CLS-13:MS(ESI)[M+H] ⁺m/z681.4.

Synthetic (synthetic route chart as shown in Figure 2) of embodiment 14 tripterine derivate CLS-14

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.48mmol benzyl bromine, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester; 72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip the acetylpiperazine that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, dripping 1mL concentration is Tripterine benzyl ester (0.2mmol) dichloromethane solution of 0.35mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.49), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-14 after being dried is 99.2mg, productive rate 82%.CLS-14:MS(ESI)[M+H] ⁺m/z695.4.

Synthetic (synthetic route chart as shown in Figure 2) of embodiment 15 tripterine derivate CLS-15

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.48mmol benzyl bromine, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester; 72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, dripping 1mL concentration is propionyl piperazine (0.35mmol) dichloromethane solution and the 100 μ L triethylamines of 0.35mol/L, stirring at room 1h, dripping 1mL concentration is Tripterine benzyl ester (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.50), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-15 after being dried is 84.0mg, productive rate 68%.CLS-15:MS(ESI)[M+H] ⁺m/z709.4.

Embodiment 16 prepares synthetic (synthetic route chart as shown in Figure 2) of tripterine derivate CLS-16

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.48mmol benzyl bromine, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester; 72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip the benzyl diethylenediamine that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, dripping 1mL concentration is Tripterine benzyl ester (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.55), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-16 after being dried is 70.6mg, productive rate 54%.CLS-16:MS(ESI)[M+H] ⁺m/z743.4.

Embodiment 17 prepares synthetic (synthetic route chart as shown in Figure 2) of tripterine derivate CLS-17

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.48mmol benzyl bromine, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester; 72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, dripping 1mL concentration is the piperidines (25.6mg of 0.35mol/L, dichloromethane solution 0.35mmol) and 100 μ L triethylamines, stirring at room 1h, dripping 1mL concentration is Tripterine benzyl ester (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.47), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-17 after being dried is 94.6mg, productive rate 84%.CLS-17:MS(ESI)[M+H] ⁺m/z652.4.

Embodiment 18 prepares synthetic (synthetic route chart as shown in Figure 2) of tripterine derivate CLS-18

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.48mmol benzyl bromine, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester.72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip the pipecoline that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, dripping 1mL concentration is Tripterine benzyl ester (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.43), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-18 after being dried is 75.0mg, productive rate 65%.CLS-18:MS(ESI)[M+H] ⁺m/z666.4。

Synthetic (synthetic route chart as shown in Figure 2) of embodiment 19 tripterine derivate CLS-19

135mg Tripterine (0.3mmol) is dissolved in to 2mLN, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.48mmol benzyl bromine, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 3 times, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester.72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip dichloromethane solution and the 100 μ L triethylamines of the diethylamine that 1mL concentration is 0.35mol/L (0.35mmol), stirring at room 1h, dripping 1mL concentration is Tripterine benzyl ester (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.38), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-19 after being dried is 45.0mg, productive rate 49%.CLS-19:MS(ESI)[M+H] ⁺m/z640.4。

Synthetic (synthetic route chart as shown in Figure 2) of embodiment 20 tripterine derivate CLS-20

135.0mg Tripterine (0.3mmol) is dissolved in to 2mLN, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.48mmol benzyl bromine, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester.72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip dichloromethane solution and the 100 μ L triethylamines of the methylphenylamine that 1mL concentration is 0.35mol/L (0.35mmol), stirring at room 1h, dripping 1mL concentration is Tripterine benzyl ester (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.38), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-19 after being dried is 60.0mg, productive rate 51%.CLS-20:MS(ESI)[M+H] ⁺m/z674.4。

Synthetic (synthetic route chart as shown in Figure 2) of embodiment 21 tripterine derivate CLS-21

135.0mg Tripterine (0.3mmol) is dissolved in to 2mLN, in dinethylformamide (DMF), adds 126mg sodium bicarbonate (1.5mmol), add methyl iodide (0.6mmol).Under room temperature, stir stopped reaction after 12h, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merging organic layer, washes 4mL*3 time with saturated sodium thiosulfate, and then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester.72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip dichloromethane solution and the 100 μ L triethylamines of the N methyl piperazine that 1mL concentration is 0.35mol/L (0.35mmol), stirring at room 1h, drip again the pristimerin that 1mL concentration is 0.2mol/L (0.2mmol) dichloromethane solution, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.36), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-21 after being dried is 101.2mg, productive rate 78%.CLS-21: ¹H?NMR(CDCl ₃,500MHz):3.68(3H,s,1’-CH ₃).MS(ESI)[M+H] ⁺m/z591.4.

Embodiment 22 prepares synthetic (synthetic route chart as shown in Figure 2) of tripterine derivate CLS-22

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.6mmol monobromethane, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester.72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip the N methyl piperazine that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, dripping 1mL concentration is Tripterine ethyl ester (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.38), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-22 after being dried is 101.2mg, productive rate 76%.CLS-22:MS(ESI)[M+H] ⁺m/z605.4.

Embodiment 23 prepares synthetic (synthetic route chart as shown in Figure 2) of tripterine derivate CLS-23

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.6mmol N-PROPYLE BROMIDE, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester.72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip dichloromethane solution and the 100 μ L triethylamines of the N methyl piperazine that 1mL concentration is 0.35mol/L (0.35mmol), stirring at room 1h, dripping 1mL concentration is the positive butyl ester of Tripterine (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.44), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-23 after being dried is 101.2mg, productive rate 69%.CLS-23:MS(ESI)[M+H] ⁺m/z619.4

Synthetic (synthetic route chart as shown in Figure 2) of embodiment 24 tripterine derivate CLS-24

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.6mmol bromo propane, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester.72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip the pipecoline that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, dripping 1mL concentration is Tripterine isopropyl ester (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.42), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-24 after being dried is 74.0mg, productive rate 65%.CLS-24:MS(ESI)[M+H] ⁺m/z619.4

Synthetic (synthetic route chart as shown in Figure 2) of embodiment 25 tripterine derivate CLS-25

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.5mmol n-butyl bromide, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester.72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip the pipecoline that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, dripping 1mL concentration is Tripterine n-pentyl ester (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.44), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-25 after being dried is 74.0mg, productive rate 65%.CLS-25:MS(ESI)[M+H] ⁺m/z633.4.

Embodiment 26 prepares synthetic (synthetic route chart as shown in Figure 2) of tripterine derivate CLS-26

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.6mmol isobutane bromide, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester.72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, dripping 1mL concentration is the morpholine (30.5mg of 0.35mol/L, 0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, dripping 1mL concentration is Tripterine isobutyl ester (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.42), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-26 after being dried is 98.0mg, productive rate 77%.CLS-26:MS(ESI)[M+H] ⁺m/z633.4.

Embodiment 27 prepares synthetic (synthetic route chart as shown in Figure 2) of tripterine derivate CLS-27

135mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.6mmol bromo pentane silane, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester.72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip the morpholine that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, dripping 1mL concentration is the just own ester of Tripterine (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.48), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-27 after being dried is 100.0mg, productive rate 76%.CLS-27:MS(ESI)[M+H] ⁺m/z647.4.

Embodiment 28 prepares synthetic (synthetic route chart as shown in Figure 2) of tripterine derivate CLS-28

135.0mg Tripterine (0.3mmol) is dissolved in to 2mLN, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.4mmol4-nitrobenzyl bromine, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester.72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip dichloromethane solution and the 100 μ L triethylamines of the NEP that 1mL concentration is 0.35mol/L (0.35mmol), stirring at room 1h, dripping 1mL concentration is Tripterine 4 p-Nitrobenzyls (0.2mmol) dichloromethane solution of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.33), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-28 after being dried is 101.4mg, productive rate 84%.CLS-28:MS(ESI)[M+H] ⁺m/z712.4.

Synthetic (synthetic route chart as shown in Figure 2) of embodiment 29 tripterine derivate CLS-29

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.4mmol4-methyl benzyl bromine, under room temperature, stir stopped reaction after 12h, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, with saturated sodium thiosulfate, wash 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester.72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip the NEP that 1mL concentration is 0.35mol/L (0.35mmol) dichloromethane solution and 100 μ L triethylamines, stirring at room 1h, dripping 1mL concentration is Tripterine 4 methyl benzyl ester (0.2mmol) dichloromethane solutions of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.40), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-29 after being dried is 100.2mg, productive rate 80%.CLS-29:MS(ESI)[M+H] ⁺m/z681.4.

Synthetic (synthetic route chart as shown in Figure 2) of embodiment 30 tripterine derivate CLS-30

135.0mg Tripterine (0.3mmol) is dissolved in to 2mL N, in dinethylformamide (DMF), add 126mg sodium bicarbonate (1.5mmol) and 0.5mmol3-bromine chloride, under room temperature, stir 12h, stopped reaction, add 10mL deionized water, ethyl acetate extraction 4mL*3 time, merges organic layer, with saturated sodium thiosulfate, washes 4mL*3 time, then saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, concentrates to obtain red oil.Through normal hexane: ethyl acetate volume ratio is that 3:1 is the rapid column chromatography method wash-out of eluent, point plate detects, being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the red solid that pressure reducing and steaming eluting solvent obtains is Tripterine ester; 72.8mg triphosgene BTC (0.24mmol) is dissolved in 2mL methylene dichloride, under ice bath, drip dichloromethane solution and the 100 μ L triethylamines of the piperidines that 1mL concentration is 0.35mol/L (0.35mmol), stirring at room 1h, dripping 1mL concentration is Tripterine 3 benzyl chloride ester (0.2mmol) dichloromethane solutions of 0.2mol/L again, stopped reaction after continuation stirring 24h, add 15mL deionized water, ethyl acetate extraction 4mL*3 time, merge organic layer, saturated NaCl solution is washed 4mL*3 time, anhydrous Na ₂sO ₄dry, suction filtration, the crude product of concentrated garnet oily, then by crude extract through normal hexane: the rapid column chromatography method wash-out that the mixed solvent that acetone volume ratio is 3:1 is eluent, some plate detects (developping agent sherwood oil: acetone=3:1; Rf value 0.38), being collected under 254nm aobvious strong ultraviolet and polarity is greater than corresponding as the elutriant of the compound of formula II, merge elutriant, the product that pressure reducing and steaming eluting solvent obtains obtains tripterine derivate CLS-30 after being dried is 97.3mg, productive rate 75%.CLS-30:MS(ESI)[M+H] ⁺m/z701.4.

The anti tumor activity in vitro experiment of the tripterine derivate that embodiment 31 the present invention are synthetic

By Bel cell (3 * 10 ⁴cell/ml) be inoculated in respectively (100ul/ hole) in 96 well culture plates, be placed in 37 ℃, 5%CO ₂cultivate 4 hours in incubator, add containing the tripterine derivate of serial ratio of two term weaker concn and positive control drug Etoposide, carboplatin.Be placed in 37 ℃, 5%CO ₂with in the incubator of saturated humidity, cultivate 48 hours.

By A549 cell (3 * 10 ⁴cell/mL) be inoculated in respectively (100 μ L/ hole) in 96 well culture plates, be placed in 37 ℃, 5%CO ₂cultivate 4 hours in incubator, add containing the tripterine derivate of serial ratio of two term weaker concn and positive control drug Etoposide, carboplatin.Be placed in 37 ℃, 5%CO ₂with in the incubator of saturated humidity, cultivate 48 hours.The MTT (20 μ L/ hole) that adds 5mg/ml, mixes latter 37 ℃, 5%CO ₂under condition, hatch 4 hours.Add DMSO (150 μ L/ hole), dissolve hyacinthine crystallization, by microplate reader, measure (570/490nm) absorbancy (OD value).Calculate the inhibiting rate IC of compound to cell ₅₀(μ M)

The present invention get shown in the table of embodiment of the present invention gained tripterine derivate the anti-tumor activity of Bel, A549 be the results are shown in Table to 2

The anti tumor activity in vitro test of table 2 tripterine derivate

The above results shows: from external activity, find out, the tripterine derivate the present invention relates to all has good anti-tumor activity, and its activity is obviously better than positive control.Compare with Tripterine, the compound activity the present invention relates to a little less than.Consider that the compound the present invention relates to is all easily metabolized to Tripterine in vivo, thereby the compound the present invention relates to may be the prodrug of Tripterine, this point is tested and has been obtained sufficient proof in body.Thereby the tripterine derivate the present invention relates to and Tripterine relatively have following advantage: (1) this analog derivative destroys original intramolecular hydrogen bond in Tripterine structure, have simultaneously and introduce nitrogenous polar functional group, thereby water-soluble improving significantly, bioavailability can improve.Such can also make water miscible salt with sour salify simultaneously, increases the administering mode of medicine.(2) prodrug that is Tripterine due to the derivative of the Tripterine the present invention relates to, its cytotoxicity has also significantly weakened, side effect meeting to Whole Body is less, thereby by internal metabolism, becomes former medicine to bring into play its drug effect when medicine arrives target organs in vivo.

Embodiment 32 Tripterine of the present invention and the restraining effect of derivative to hepatocellular carcinoma in nude mice thereof

1. material and animal subject

1.1 materials:

Laboratory sample 1: Tripterine (6mg/kg)

Laboratory sample 2:CLS-12(high dosage 4mg/kg; Middle dosage 2mg/kg; Low dosage 1mg/kg; )

Positive control: Fluracil (20mg/kg)

1.2 animal subjects: SPF level BALB/c nude mice is purchased from the Academy of Medical Sciences, Zhejiang Experimental Animal Center, credit number: nude mice 4-6 age in week, body weight 18 ± 20g, male and female half and half.

2. experimental technique

2.1 tumour modelings: get the Bel culturing cell that growth conditions is good, with D-Hanks liquid, wash 3 times, 0.25% trysinization, centrifugal 8 minutes of 1000rpm, collecting cell, serum-free medium washed cell twice, re-suspended cell, the concentration of cell is adjusted into 5 * 10 ⁷individual/mL.Iodophor disinfection SPF level BALB/c nude mice oxter, left side and the right skin of right side inguinal region with 0.5%, with No. 5 needle inoculations, every about 0.2mL cell of nude inoculation.During injection there is skin mound in visible part, inoculates nude mice and occur that diameter is greater than the subcutaneous nodule of 5mm after a couple of days, observes transplanted tumor tumor formation rate and homogeneity.

2.2 grouping and administrations:

Model group is divided into 6 groups at random, 10 every group, is respectively solvent control group, Tripterine group (6mg/kg), trypterygine prodrug group (high dosage: 4mg/kg; Middle dosage 2mg/kg; Low dosage: 1mg/kg), positive drug control group (Fluracil, 20mg/kg).Administering mode is abdominal injection once a day, Sunday no collection.Successive administration 3 weeks.Solvent control group abdominal injection equivalent solvent.The general status of close observation nude mice during administration, comprises the mental status, activity, diet and stool and urine, records weekly nude mice body weight twice, uses vernier caliper measurement tumor volume twice weekly.

(3) draw materials and measure: administration was weighed after 20 days, nude mice, the transfer case of anatomic observation nude mice tumour are put to death in cervical vertebra dislocation.Take out tumor tissues, scales/electronic balance weighing.Vernier caliper measurement tumour major diameter (a) and minor axis (b), obtain tumor quality and gross tumor volume (TV=a * b ²/ 2), and calculate tumour inhibiting rate

3. experimental result

Table 2 Tripterine, the inhibition test of CLS-12 to hepatocellular carcinoma in nude mice

The data of associative list 1 and table 2, we can find out following aspect: (1) the present invention all has good antitumor action in vivo and in vitro to tripterine derivate.(2) tripterine derivate of the present invention in vivo its anti-tumor activity be obviously better than positive control drug Fluracil.In addition, in animals administer process, tripterine derivate administration group of the present invention, the animal mental status is good, and diet is normal, does not occur that mouse is dead, and body weight also has no and alleviates.And positive controls is in administration process, animal ill health, activity obviously reduces, and occurs that mouse is dead, and not dead mouse body weight also alleviates, and the tripterine derivate that this explanation the present invention relates to is also obviously better than positive control drug aspect security.(3) the present invention to tripterine derivate in vivo its anti-tumor activity be obviously better than Tripterine group, but its external activity is lower than Tripterine.The tripterine derivate that this explanation the present invention relates to should be to become Tripterine to bring into play drug effect by the metabolic conversion in body, because the tripterine derivate the present invention relates to and Tripterine relatively have water-solublely preferably, improved its bioavailability and produced better biological activity.The tripterine derivate the present invention relates to is in addition because cytotoxicity is weaker than Tripterine, thereby will have better security before metabolism.In a word, the tripterine derivate the present invention relates to is compared with former medicine, at stability, water-soluble, activity and security tool, improve significantly, and be more preferably antitumor drug of a class.

Claims

1. A tripterine derivative as shown in formula (I),

Wherein, R ₁ is benzyl; R ₂ , R ₃ and N form a ring, and the ring is N-substituted piperazinyl;

The structure of tripterine derivatives shown in formula (I) is as follows:

2. a preparation method of tripterine derivatives shown in formula (I) in claim 1, is characterized in that described tripterine derivatives are prepared as follows: triphosgene is dissolved in two Chloromethane is formulated into a triphosgene solution with a concentration of 0.1-0.5 mol/L, and the methylpiperazine dichloromethane solution and triethylamine with a concentration of 0.1-0.5 mol/L are added dropwise in the triphosgene solution under ice bath, Stir at room temperature for 1 to 2 hours, then add dropwise the dichloromethane solution of tripteryglide compound shown in formula (II) with a concentration of 0.1 to 0.5 mol/L, continue to stir for 20 to 48 hours, and spot the plate for detection. ) stop the reaction after the reaction of the tripteryglide compound, add 3 to 6 times the volume of deionized water in the reaction solution A to the reaction solution A, then extract with ethyl acetate, take the organic layer, wash with a saturated NaCl solution, and use Dry over anhydrous Na ₂ SO ₄ , filter with suction, and concentrate to obtain a dark red oily crude product, and then the crude product is eluted by flash column chromatography with a mixed solvent of n-hexane:acetone volume ratio of 3:1 as the eluent , point plate detection, collect the eluate that shows strong ultraviolet light at 254nm and has a polarity greater than that of the corresponding compound such as formula (II), combine the eluent, and evaporate the eluent solvent under reduced pressure to obtain the product after drying to obtain Leigong Luminol derivatives; the tripteryne compound: triphosgene: methylpiperazine: triethylamine has a material ratio of 1:1.2～1.5:1.4～2.0:3～5;

Wherein, when _R is benzyl, the tripterine compound is tripterine benzyl ester.

3. the method for claim 2 is characterized in that described tripterine benzyl ester is prepared as follows: tripterine shown in formula (Ⅲ) is dissolved in N,N-dimethylformamide Prepare a tripterine solution with a concentration of 0.1 to 0.3 mol/L, use sodium bicarbonate as a catalyst, add R ₁ X, said X is Br or I, and said R ₁ is benzyl, and stir at room temperature for 12 ~ 24 hours, add 3 ~ 6 times the volume of deionized water in the reaction solution B to the reaction solution B, extract with ethyl acetate, take the organic layer, wash with saturated sodium thiosulfate, then wash with saturated NaCl solution, and then use Dry over _Na2SO4 , filter with suction, and concentrate the organic phase to obtain a red oil, which is eluted by flash column chromatography with a volume ratio of n _- hexane:ethyl acetate of 3:1 as the eluent, detected by spot plate detection, and collected containing The eluate of tripterine ester, the combined eluent, and the red solid obtained by evaporating the elution solvent under reduced pressure is tripterine ester; described tripterine: R ₁ X: sodium bicarbonate The amount ratio of substances is 1:1.2～2.0:3.0～5.0;