CN106946975A - A kind of triptolide derivative and preparation method thereof and preparation - Google Patents

Abstract

The invention discloses a kind of triptolide derivative and preparation method thereof and preparation, the chemical constitution such as formula (I) of described triptolide derivative is shown：

Description

A kind of triptolide derivative and preparation method thereof and preparation

Technical field

The present invention relates to pharmaceutical technology field, and in particular to a kind of triptolide derivative and preparation method thereof and system Agent.

Background technology

Triptolide (Triptolide, TP) is also known as Triptolide, triptolide, is from Chinese herb triperygium wilfordii A kind of Diterpenoid epoxide class lactone compound separated in (Tripterygium wilfodii Hook F) plant, with very strong Bioactivity, including (the Liu Q.Triptolide and its expanding such as anti-inflammatory, immunosupress, antitumor, antifertility multiple pharmacological functions[J].International Immunopharmacology,2011, 11(3):377-383).As tripterygium wilfordii main active, its antitumor value is widely studied, and so far at least 60 Kind of tumor cell line can be suppressed that (Luo Yongwei, Shi Chang, Liao Ming sun triptolide Anticancer Effect and Mechanisms are ground through research by TP Study carefully progress [J] CHINA JOURNAL OF CHINESE MATERIA MEDICAs, 2009,34 (16):2024-2026).TP has following four feature as antineoplastic： 1. the antitumor activity of wide spectrum is shown in mouse transplantable tumor animal model, it is former to breast cancer, carcinoma of urinary bladder, stomach cancer and melanoma The inhibiting rate of hair property tumour is not only effective to primary tumor up to 50%~90%, also effective to secondary tumors；2. can The growth and transfer for suppressing a variety of solid tumors are unrelated with the state of tumor suppressor gene (P53), and hair is remained to the tumour cell of P53 defects The effect of waving；3. to the tumour cell of high expression Mdr-p (MDR1) effectively, and these cells to Common Chemotherapy medicine such as The resistances such as taxol, therefore TP can share to increase curative effect with chemotherapeutics；4. there is stronger compared with traditional anti-tumor medicine resist Tumor promotion, produces very strong antitumor activity (Yang S, Chen J, Guo Z, et al.Triptolide during low concentration Inhibits the Growth and Metastasis of Solid Tumors[J].Molecular Cancer Therapeutics,2003,2(1):65-72).The specific mechanism of TP antitumor actions is failed to understand, is induced by many A signal pathways Apoptosis of tumor cells is probably the main mechanism of its antitumor action.In addition, TP can also be by suppressing angiogenesis, blocking carefully Born of the same parents' cycle progression, suppress the effect such as extracellular matrix degradation reach suppress growth and metastasis of tumours purpose (Meng C, Zhu H, Song H,et al.Targets and molecular mechanisms of triptolide in cancer therapy [J].Chinese Journal of Cancer Research,2014,26(5):622-626).It is showed in anti-tumor aspect The wide spectrum that goes out, the advantage such as efficiently are so that TP has wide potential applicability in clinical practice.

Although early in TP in 1972 be just proved to have leukaemia therapeutic action (Kupchan S M, Court W A, Dailey R G,et al.Tumor inhibitors.LXXIV.Triptolide and tripdiolide,novel antileukemic diterpenoid triepoxides from Tripterygium wilfordii[J].Journal of the American Chemical Society,1972,94(20):7194-7195), and in the recent decade, great Liang Ti Inside and outside research fully shows that growths of the TP to a variety of solid tumor cells and solid tumor is inhibited.But so far but still It is untapped go out it is a kind of for TP be applied to antineoplaston high potency drugs list.Wherein difficulty is mainly reflected in following side Face：

TP toxicity is big：Along with serious toxic and side effect while drug effect is played, and narrow therapeutic window is shown, caused Its clinical practice is greatly limited.Its toxic action be mainly reflected in general toxicity, target organ toxicity, genetoxic, Several aspects such as reproductive system toxicity, local excitation toxicity.And target organ toxicity is related to that body is almost all of physiological function Organ, even if the TP of therapeutic dose, which is still shown, suppression to immunologic function, excess, poisoning can then cause withering for lymphoid organ Contracting and necrosis (Li X, Jiang Z, the Zhang L.Triptolide of lymphocyte:progress on research in pharmacodynamics and toxicology[J].Journal of Ethnopharmacology,2014,155(1): 67-79)。

TP half-life shorts：Its internal supersession rate is fast, half-life short, causes largely to weaken its due anti- Tumor effect, and it enters blood volume distributed median extensively, and targeting is poor, and the generation of adverse reaction has been aggravated on the contrary.Shao etc. uses liquid Phase Chromatography/Mass Spectrometry method for combined use (LC/MS) have studied Pharmacokinetic Characteristics of the TP in rat body, as a result show that SD rats are single Secondary oral TP dosage is respectively 0.6,1.2 and 2.4mg/kg, and peak reaching time of blood concentration is in 10min or so, elimination half-life period 16.81~21.70min, the pharmacokinetic parameters for comparing three show that AUC and cmax value can't be improved with the increase of dosage, Nonlinear Dynamical Characteristics may be presented during prompting clinical practice high dose；SD rat single intravenous injection TP dosage 0.6mg/kg Its half-life period is about 15min.TP can be distributed to the change in detected tissue (blood plasma, the heart, liver, spleen, lung, kidney), and tissue rapidly Change (Shao F, Wang G, Xie H, et al.Pharmacokinetic study of parallel with the change of Plasma triptolide,a constituent of immunosuppressive chinese herb medicine,in rats [J].Biological&Pharmaceutical Bulletin,2007,30(4):702-707)。

TP preparation druggabilities are poor：Either in distilled water, 0.1mol/L HCl or pH6.8 PBS, TP solubility is equal Less than 0.3mg/mL, according to the regulation of 2015 editions description solubility terms of Chinese Pharmacopoeia, TP belongs to the atomic material for being dissolved in water and (opened Clever triptolides lipid nano particle research [D] the Central China University of Science and Technology, 2014).In addition, TP can in some other pharmacy Solubility in the solvent of receiving and oil is also no more than 0.5% (w/w), and it is conventional used for intravenous injection that this make it that it is difficult to be prepared into Solution.Even if developing into the new formulations such as some liposomes or nanoparticle, however it remains drugloading rate is low, envelop rate is poor, preparation Many drawbacks such as unstable.Lin Sui et al. attempts to prepare TP nano liposomes, and still only up to 0.016%, (China is specially for maximum drugloading rate Sharp CN105816428A)；Clever preparation TP lipid nano particles are opened, in best prescription technique lipid/medicine=50：1~100：1 condition Under, envelop rate is still no more than 60%, far below a regulation requirement (acute hearing triptolides lipid nano particle research for Chinese Pharmacopoeia [D] the Central China University of Science and Technology, 2014)；Shujuan WANG carries out study on the stability to preparation gained liposome and shown, freshly prepd TP fat Plastid room temperature, which is placed 10 days, has aggregation, is precipitated substantially after one month, even if carrying out frozen dried, freeze-dried rear rehydration weight The liposome built envelop rate compared with before freezing still has different degrees of reduction, and envelop rate is less than 40% (Shujuan WANG tripterygium wilfordii first The preparation of plain liposome and Primary Study [D] the Yangzhou Universitys of quality, 2010).Above property make it that TP is difficult to exploitation for the benefit of Ejection preparation in the active isomer of oncotherapy.

How on the premise of TP bioactivity is kept, action time in extension body, toxic and side effect is reduced, is to enable TP Safely and effectively it is applied to the key issue of clinical institute urgent need to resolve.Pro-drug (prodrug, prodrug) itself is without biological living Property or activity it is very low, by vivo metabolism after be changed into active material, this process can change the internal medicine of parent drug Phoronomics characteristic.Therefore, in the exploration to TP, Many researchers are grown tender of carries out structural modification to TP, to obtain with good The TP prodrugs of good pharmaco-kinetic properties.Its characteristic for being insoluble in water is limited to, most researchers tend to design and synthesis with good The new type water-solubility TP derivatives of good pharmaco-kinetic properties, to reduce the toxic and side effect of medicine.Such as：PG490-88Na、 WilGraf, Minnelide etc. be all TP water-soluble prodrug (Fidler J M, Li K, Chung C, et al.PG490-88, a derivative of triptolide,causes tumor regression and sensitizes tumors to chemotherapy[J].Molecular Cancer Therapeutics,2003,2(9):855-862；Zhou Z,Yang Y,Ding J,et al.Triptolide:structural modifications,structure-activity relationships,bioactivities,clinical development and mechanisms[J].Natural Product Reports,2012,29(4):457-475).Although TP is improved its preparation into salt by above prodrug well Matter, but not be envisaged to be able to reach that reduction toxicity improves the purpose of pharmacokinetics just like expected, however it remains degraded discharged Fast or conversion not exclusively waits possibility drawback.In such as dose escalation study of PG490-88Na Phase I clinical trial that What is more, by Examination object is patients with advanced solid tumors, and administering mode is rested one week every two weeks to be injected intravenously weekly once.More often go out in experiment Existing adverse reaction has anaemia, weak, nausea,vomiting,diarrhea and constipation etc., and grading is all 1~2 grade.However, pharmacokinetics Research shows that PG490-88Na pharmaco-kinetic properties have higher interindividual variation (2~3 times), and it is converted into TP degree It is difficult to predict, conversion process is slow and incomplete.Therefore, PG490-88Na is not a preferable TP derivative, the clinic Experiment has been forced to suspend (Kitzen J J, de Jonge M J, Lamers C H, et al.Phase I dose- escalation study of F60008,a novel apoptosis inducer,in patients with advanced solid tumours[J].European Journal of Cancer,2009,45(10):1764-1772)。 Hereafter, the relevant report for having no PG490-88Na clinical tests updates.It can be seen that TP is improved into whether water solubility is preferable into salt The demand that modification mode makes it possible to meet clinic need to be discussed.

And on the other hand, the focus that nanometer formulation is developed as current research has shown to be widely applied prospect, its is excellent Gesture is：While Nano medication can reduce toxicity by targeting, further improve and treat on the basis of validity is ensured Effect；Medicine is wrapped in nanoparticle, particularly long-circulating nanoparticles, with obvious slow-releasing and controlled-releasing action, medicine is slow down significantly The supersession rate of thing in vivo, extends half-life period, makes it have time enough by drug targeting in tumor locus, reduces normal Distribution in tissue, so as to improve the security of clinical application.Above feature exactly TP is in the urgent need to the ideal effect reached. And with the development of preparation technique, existing many nanometer formulation sequential uses are in clinic, such as Evacet at presentInjection taxol (albumin combination type), trade nameTaxol polymer micelleDeng (Barenholz Y.—The first FDA-approved nano-drug: Lessons learned[J].Journal of Controlled Release,2012,160(160):117-134；Gong Ji Virtue, Lu Ming, Li Jie wait injections taxol (albumin combination type) treatment of advanced gastric cancer [J] Peking University journal medical science Version, 2014,46 (1):144-148；Lee K S,Chung H C,Im S A,et al.Multicenter phase II trial of Genexol-PM,a Cremophor-free,polymeric micelle formulation of paclitaxel,in patients with metastatic breast cancer[J].Breast Cancer Research and Treatment,2008,108(2):241-250).Above-mentioned preparation has absolutely proved that nanometer formulation is used to resist The feasibility of tumour medicine listing, can provide important references for TP exploitation, or it will be more just to be developed into nanometer formulation True research direction.

But as TP preparation druggability defects are determined, attempt that TP directly is prepared into liposome, micella, nanoparticle etc. The exploitativeness for wrapping up nanometer formulation is very poor, main reason is that TP is fat-soluble too low, is mismatched with the compatibility of lapping It is caused.How to improve TP it is fat-soluble by be nanometer formulation exploitation emphasis.In view of prodrug can significantly improve the change of parent drug Learn property, if seem significant using the means to meet preparation druggability.Chinese patent CN105263475A In disclose a kind of TP prodrugs " MRx102 ", but MRx102 does not have a druggability, or druggability is poor, is mainly reflected in：1. it is real Apply oil phase amount used in MRx102 emulsion formulations in example big, most emulsion oil phase consumptions are 40%, the erect image patent institute State, " classical emulsion formulations contain 10~30% triglycerides ", and the emulsion that 40% oil phase will result directly in preparation is thick Thick emulsifiable paste shape (Fidler J M, An J, Carter B Z, et al.Preclinical antileukemic activity,toxicology,toxicokinetics and formulation development of triptolide derivative MRx102[J].Cancer Chemotherapy and Pharmacology,2014,73(5):961- 974), this bacteria removing for necessarily affecting preparation, it is impossible to cross 0.22 μm of filter membrane or carry out autoclaving, in addition potential syringeability Deficiency, more likely causes smoothly to inject, causes patient's poor compliance；2. solubility is too poor in oil, aside from three octanoic acids The emulsion that tricaprylin in the security of glyceride, the patent table 4 even with 29.4% is prepared as oil phase, MRx102 solubility 0.681 μ g/mL of, directly result in drugloading rate too low；3. stability is too poor, in the patent table 5, even with 40% tricaprylin prepares the emulsion of 2mg/mL drugloading rates as emulsion made from oil phase (by taking E-3 as an example), and its medicine contains It is respectively 1529,1514,1353,1176 μ g/mL that (solubility), which is measured, from 0 hour, 1 hour, 24 hours, 8 days, that is, prepare Drugloading rate reduces 41.2% after emulsion is placed 8 days, without druggability；4. containing high content in MRx102 emulsion formulations In tricaprylin, the patent clearly, tricaprylin is partly or entirely replaced to reduce MRx102 in breast with soybean oil Solubility in agent, illustrate tricaprylin can not shortage property, and the composition is currently used primarily in skin care item and cosmetics, There is hidden danger for its security in intravenous administration formulation；5. dissolve, added in formula in emulsion in order to increase MRx102 Ionic surfactant sodium taurocholate carries out solubilising, this also illustrates the prodrug and without significantly improving fat-soluble spy Point.And not only toxicity is larger for ionic surfactant, while there is stronger haemocylolysis (Cui Fude pharmacies [M] north Capital:People's Health Publisher, 2011:42).In addition the nanometer formulations such as other liposomes, micella are also not directed in patent Preparation, or carried out the exploitation of Fat Emulsion and also belong to helpless choosing.

To sum up, although the problem of having a large amount of researchs for being directed to triptolide, triptolide preparation druggability Still protrude, be badly in need of further research and development to improve above mentioned problem, exploitation is applied to the high potency drugs of antineoplaston for TP.

The content of the invention

The present invention has carried out substantial amounts of structural modification to solve the defect of TP druggabilities, for TP and worked, targetedly Start with from fat-soluble prodrug angle, discovery is fat-soluble after TP is bonded saturated fatty acid in research significantly improves, by further It is triptolide fat obtained by the modification of different chain length saturated fatty acid in the range of 2n (n is 2~9) that screening, which has synthesized carbon number, Fat acid esters, as a result shows, above triptolide fatty acid ester preparation druggability is superior to TP.And attempt be prepared into liposome, Micella, nanoparticle, Fat Emulsion, which are respectively provided with, good contains effect.Further in vivo and in vitro shows, prepares gained nanometer system Agent also has simultaneously significantly reduces toxicity, the advantage such as extension half-life period.

The first object of the present invention is to provide a kind of triptolide derivative, specifically a kind of triptolide fat Acid esters, shown in its chemical constitution such as formula (I)：

In formula (I), R represents the straight chained alkyl acyl group that carbon number is 2n, and n is 2~9, preferably 4~9, more preferably 7~9.

The second object of the present invention is the preparation method of the triptolide fatty acid ester described in offer, described Thunder God Rattan A prime fatty acid ester can be obtained by triptolide with saturated fatty acid by esterification, and synthetic route is：

Wherein, ROH is saturated fatty acid, and R is defined as described in any one of claims 1 to 3；

Specific steps include：

(a) saturated fatty acid, condensing agent and catalyst are dissolved in organic solvent, 0~10 DEG C is cooled under stirring to mix Close liquid；

(b) by triptolide dissolving instill in organic solvent in above-mentioned mixed liquor, under the conditions of 0~10 DEG C react 15~ 45 minutes, then continue to react at room temperature, esterification terminates rear isolated triptolide fatty acid ester.

In the preparation method of triptolide fatty acid ester,

Described condensing agent is paranitrobenzoyl chloride, N, N'- dicyclohexylcarbodiimides (DCC), N, N'- diisopropyls Carbodiimide (DIC), 1- ethyls-(3- dimethylaminopropyls) phosphinylidyne diimmonium salt hydrochlorate (EDCHCl), the chloro- 1- methyl of 2- Pyridine iodide (CMPI), hexafluorophosphoric acid BTA -1- bases-epoxide tripyrrole alkyl phosphorus (PyBOP), 2- (7- azobenzenes And triazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid esters (HATU), O- (5- Chloro-Benzotriazole -1- bases)-two (diformazans Amido) carbon hexafluorophosphate (HCTU), O- (BTA -1- bases epoxide)-two piperidines carbon hexafluorophosphates (HBPipU), the one or two in N-hydroxy-succinamide (NHS) and N- hydroxy thiosuccinimides (sulfo-NHS) More than, preferably DCC or EDCHCl；

Described catalyst is I-hydroxybenzotriazole (HOBT), DMAP (DMAP), triethylamine and N, N- One or more in diisopropylethylamine (DIPEA), preferably DMAP or DIPEA；

Described organic solvent is dichloromethane, chloroform, N,N-dimethylformamide (DMF) and N, N- dimethyl second One or more in acid amides (DMA), organic solvent is preferably anhydrous.

Described saturated fatty acid is 1.2~4 with triptolide mol ratio:1, preferably 2~3:1；Described condensing agent Mol ratio with triptolide is 1.2~4:1, preferably 2~3:1.

The described triptolide fatty acid ester of the present invention can be used for preparing antitumor or anti-inflammatory drug.

The third object of the present invention is the nanometer formulation of the triptolide fatty acid ester described in offer, the nanometer system Agent is liposome, polymer micelle, albumin nano granular or Fat Emulsion, preferred liposome.

In some preferred embodiments of the present invention, described liposome prepared by following ingredients according to w/v and Into：

Above-mentioned formula is preferably：

Wherein,

Described phosphatide is yolk phospholipid, the phosphatide of soybean lecithin and various animal origins, hydrogenation yolk phospholipid, hydrogen Change soybean lecithin, DPPC, dimyristoyl phosphatidyl choline, DSPC, phosphatidyl Monoethanolamine, DSPG, DPPG, DOPC, cuorin and sphingomyelins In one or more；It is preferred that yolk phospholipid or soybean lecithin.

Described PEGylation phosphatide be PEGylation DSPE, PEGylation DPPE, One or more in the nutmeg phosphatidyl-ethanolamine of PEGylation two, preferably PEGylation DSPE；Institute The PEG mean molecule quantities for stating PEGylation phosphatide are 1000~8000, preferably 1500~3500, more preferably 2000.

Described freeze drying protectant is the one or two in trehalose, sucrose, maltose, lactose, mannitol, glucose More than；It is preferred that trehalose or sucrose.

Described pH adjusting agent is in NaOH, sodium acetate, acetic acid, phosphate, carbonate, hydrochloric acid, citric acid etc. It is one or more kinds of；It is preferred that NaOH, hydrochloric acid or citric acid.

Described liposome can be prepared by following steps：

(A) configuration of liposome crude product, weighs formula ratio triptolide fatty acid ester, phosphatide, PEGylation phosphatide and courage solid Alcohol, adds appropriate organic solvent I and obtains organic phase in heating for dissolving at 25~75 DEG C, then organic phase is slowly injected into 25~75 DEG C In appropriate water for injection, marginal not enters side and stirred and evenly mixed, and produces liposome crude product；

(B) preparation of liposome solutions, is placed in high pressure homogenizer emulsifying by the liposome crude product or is placed in crowded Go out in device to pass sequentially through the extruded film extrusion of different pore size or be placed in high pressure homogenizer after homogeneous row extrusion again, obtain liposome Solution；

(C) formula ratio freeze drying protectant is weighed, the liposome solutions are dissolved in；Plus water for injection constant volume, regulation pH to rule Definite value；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced；

Or,

Step (A) is replaced with：(A ') weighs formula ratio triptolide fatty acid ester, phosphatide, PEGylation phosphatide, cholesterol It is dissolved in organic solvent II, vacuum revolving removes solvent and obtains lipid membrane under the conditions of 40~60 DEG C, is carried out with appropriate water for injection Aquation obtains liposome crude product.

One or more of the described organic solvent I in absolute ethyl alcohol, propane diols, the tert-butyl alcohol；Consumption be 1~ 10% grams per milliliter, described organic solvent I is retained in liposome, or after the emulsification of liposome crude product again by ultrafiltration or Scrapper thin film evaporator is removed.

Described organic solvent II is the one or more in dichloromethane, chloroform, absolute ethyl alcohol, and consumption is 1~10% grams per milliliter.

Described freeze drying protectant can be directly added into liposome solutions made from step (B), can also be first dissolved in note Penetrate with being mixed again with liposome solutions in water (in being soluble in the aqueous phase plus).

The extrusion membrane aperture in step (B) is selected from 2.0 μm, 1.0 μm, 0.8 μm, 0.6 μm, 0.4 μm, 0.2 μm, 0.1 μ One or more in m and 0.05 μm, pass sequentially through large aperture to small-bore during extrusion.

The beneficial effects of the present invention are：

The present invention has carried out substantial amounts of structural modification for TP and worked, and starts with from fat-soluble angle is improved, in TP C14 Position OH bonding saturated fatty acids, introducing aliphatic acid while hydroxyl is reduced into polarity into ester can effectively strengthen fat-soluble, and It is stable in properties, it is not oxidizable, TP nanometer formulation druggabilities can be significantly improved.Carbon number has been synthesized for 2n by further screening Triptolide fatty acid ester preparation druggability obtained by the modification of different chain length saturated fatty acid is superior in the range of (n is 2~9) TP, is respectively provided with good by nanometer formulations such as liposome, micella, nanoparticle, the Fat Emulsions of triptolide fatty acid ester preparation Good contains effect, the nanometer formulation being made has that drugloading rate is big, envelop rate is high, it is stable in properties the features such as.It is further internal Outer research shows, prepares gained nanometer formulation simultaneously also with toxicity is significantly reduced, extends the advantages such as half-life period.The present invention is into Work(exploitation triptolide nanometer formulation can establish solid foundation for the research of triptolide there is provided possible with application.

Embodiment

Below in conjunction with specific embodiment, the invention will be further described.It should be understood that following examples are merely to illustrate this Invention is not for restriction the scope of the present invention.

TP of the present invention reaches Pharmaceutical Technology Co., Ltd again purchased from Jiangsu；Saturated fatty acid has purchased from Chinese medicines group chemical reagent Limit company.

The experimental method of unreceipted actual conditions in the following example, generally according to normal condition, or according to manufacturer Proposed condition.

The preparation of the different triptolide fatty acid esters of embodiment 1

1.1.1 the preparation of triptolide stearate (TP-SA)

Throwing amount 3mmol stearic acid, 3mmol DCC, 3mmol DMAP in reaction vessel, add the anhydrous dichloromethanes of 20mL Alkane dissolves, and is stirred 30 minutes under condition of ice bath；1mmol TP is dissolved in appropriate anhydrous methylene chloride and is slowly added dropwise into anti- Answer and reacted under system, condition of ice bath 30 minutes, continue reaction at room temperature and stay overnight, reactant is obtained into Thunder God through silica gel column separating purification Rattan A prime stearate 532.2mg.Yield 84.9%.

¹H NMR(DMSO-_d6, 600MHz) and δ 4.98 (1H, s, 14-CH), 4.86 (1H, d, J=18.88Hz, 19-CH), 4.77 (1H, d, J=18.88Hz, 19-CH), 3.95 (1H, d, 11-CH), 3.69 (1H, d, 12-CH), 3.56 (1H, d, 7- CH),2.28-2.39(2H,m,2ˊ-CH₂),1.73-2.00(1H,m,15-CH),1.80-1.85(2H,m,3ˊ-CH₂),1.57- 1.59(2H,m,2-CH₂),1.24-1.34(28H,m,14×CH₂),0.92(3H,s,20-CH₂),0.85-0.87(6H,m,16、 17-CH₂),0.76(3H,t,18ˊ-CH₂).

¹³C NMR(DMSO,150MHz)δ175.52,170.59,71.08,70.66,63.74,63.08,61.30, 59.84,55.29,54.93,35.51,34.19,31.78,29.57,29.51,29.37,29.23,29.19,28.71, 28.11,25.06,22.85,22.57,17.90,17.05,17.00,14.39,14.13.

ESI-MS(m/z)：627.8[M+H]⁺.

Chemical structural formula is as follows：

1.1.2 the preparation of triptolide stearate (TP-SA)

Throwing amount 2.5mmol stearic acid, 2.5mmol EDCHCl, 2.5mmol DIPEA in reaction vessel, are added 20mL anhydrous methylene chlorides dissolve, and are stirred 30 minutes under condition of ice bath；1mmol TP is dissolved in appropriate anhydrous methylene chloride And be slowly added dropwise into reaction system, reacted under condition of ice bath 30 minutes, continue reaction at room temperature and stay overnight, by reactant through silicagel column Isolate and purify to obtain triptolide stearate 524.0mg.Yield 83.6%.

1.2.1 the preparation of triptolide palmitate (TP-PA)

Throwing amount 3mmol palmitic acids, 3mmol DCC, 3mmol DMAP in reaction vessel, add the anhydrous dichloromethanes of 20mL Alkane dissolves, and is stirred 30 minutes under condition of ice bath；1mmol TP is dissolved in appropriate anhydrous methylene chloride and is slowly added dropwise into anti- Answer and reacted under system, condition of ice bath 45 minutes, continue reaction at room temperature and stay overnight, reactant must be purified through silica gel column separating purification Triptolide palmitate 504.7mg.Yield 84.3%.

¹H NMR(DMSO-_d6, 600MHz) and δ 4.96 (1H, s, 14-CH), 4.86 (1H, d, J=18.88Hz, 19-CH), 4.73 (1H, d, J=18.86Hz, 19-CH), 3.98 (1H, d, 11-CH), 3.69 (1H, d, 12-CH), 3.57 (1H, d, 7- CH),2.28-2.42(2H,m,2ˊ-CH₂),1.73-2.00(1H,m,15-CH),1.80-1.85(2H,m,3ˊ-CH₂),1.57- 1.61(2H,m,2-CH₂),1.24-1.35(24H,m,12×CH₂),0.93(3H,s,20-CH₂),0.85-0.88(6H,m,16、 17-CH₂),0.78(3H,t,18ˊ-CH₂).

¹³C NMR(DMSO,150MHz)δ178.52,173.59,70.88,70.26,63.64,63.18,61.30, 59.34,55.29,55.03,35.91,34.21,31.78,29.77,29.23,29.17,28.71,28.05,25.13, 22.80,22.57,17.90,17.35,17.09,14.42,14.11.

ESI-MS(m/z)：599.8[M+H]⁺.

Chemical structural formula is as follows：

1.2.2 the preparation of triptolide palmitate (TP-PA)

Throwing amount 2mmol palmitic acids, 2mmol EDCHCl, 2mmol DIPEA in reaction vessel, add 20mL anhydrous Dichloromethane dissolves, and is stirred 30 minutes under condition of ice bath；1mmol TP is dissolved in appropriate anhydrous methylene chloride and slowly dripped Add and reacted under reaction system, condition of ice bath 30 minutes, continue reaction at room temperature and stay overnight, by reactant through silica gel column separating purification Triptolide palmitate 495.2mg must be purified.Yield 82.7%.

1.3.1 the preparation of triptolide myristinate (TP-MA)

Throwing amount 3mmol myristic acids, 3mmol DCC, 3mmol DMAP in reaction vessel, add the anhydrous trichlorines of 20mL Methane dissolves, and is stirred 15 minutes under condition of ice bath；By 1mmol TP be dissolved in appropriate anhydrous chloroform and be slowly added dropwise into Reacted under reaction system, condition of ice bath 30 minutes, continue reaction at room temperature and stay overnight, reactant is obtained into thunder through silica gel column separating purification Public rattan A prime myristinate 464.5mg.Yield 81.4%.

¹H NMR(DMSO-_d6, 600MHz) and δ 4.97 (1H, s, 14-CH), 4.84 (1H, d, J=18.88Hz, 19-CH), 4.73 (1H, d, J=18.83Hz, 19-CH), 3.97 (1H, d, 11-CH), 3.66 (1H, d, 12-CH), 3.61 (1H, d, 7- CH),2.27-2.41(2H,m,2ˊ-CH₂),1.73-2.00(1H,m,15-CH),1.80-1.85(2H,m,3ˊ-CH₂),1.57- 1.61(2H,m,2-CH₂),1.22-1.35(20H,m,10×CH₂),0.90(3H,s,20-CH₂),0.83-0.87(6H,m,16、 17-CH₂),0.74(3H,t,18ˊ-CH₂).

¹³C NMR(DMSO,150MHz)δ178.92,173.10,70.90,62.73,61.64,59.34,58.43, 55.29,55.03,49.97,46.13,46.05,35.91,34.21,34.19,32.22,31.88,31.52,29.57, 29.23,29.17,28.71,28.05,25.13,22.80,22.57,17.90,17.35,17.09,14.42,14.08.

ESI-MS(m/z)：571.7[M+H]⁺.

Chemical structural formula is as follows：

1.3.2 the preparation of triptolide myristinate (TP-MA)

Throwing amount 2mmol myristic acids, 2mmol EDCHCl, 2mmol DIPEA in reaction vessel, add 20mL without Water dichloromethane dissolves, and is stirred 15 minutes under condition of ice bath；1mmol TP is dissolved in appropriate anhydrous chloroform and slow It is added dropwise under reaction system, condition of ice bath and reacts 30 minutes, continues reaction at room temperature and stay overnight, reactant is pure through silica gel post separation Change to obtain triptolide myristinate 476.5mg.Yield 83.5%.

1.4.1 the preparation of triptolide laurate (TP-LA)

Throwing amount 1.2mmol laurate, 1.2mmol DIC, 1.2mmol HOBT in reaction vessel, add 20mL anhydrous DMF dissolves, and is stirred 20 minutes under condition of ice bath；By 1mmol TP be dissolved in appropriate anhydrous methylene chloride and be slowly added dropwise into Reacted under reaction system, condition of ice bath 30 minutes, continue reaction at room temperature and stay overnight, reactant is obtained into thunder through silica gel column separating purification Public rattan A prime laurate 397.3mg.Yield 73.2%.

¹H NMR(DMSO-_d6,600MHz)δ:4.78 (1H, s, 14-CH), 4.50 (1H, d, J=18.77Hz, 19-CH), 4.43 (1H, d, J=18.77Hz, 19-CH), 3.35 (2H, s, 11,12-CH), 3.21 (1H, t, 7-CH), 2.56-2.60 (1H, m,4-CH),2.32-2.37(2H,m,2ˊ-CH₂),2.16-2.21(1H,m,3-CH),1.80-1.85(2H,m,3ˊ-CH₂), 1.73-1.77(1H,m,15-CH),1.57-1.61(4H,m,1、2-CH₂),1.44-1.50(2H,m,6-CH₂),1.25-1.30 (16H,m,8×CH₂),1.04-1.10(1H,m,5-CH),0.93(3H,s,20-CH₂),0.85-0.88(6H,m,16、17- CH₂),0.78(3H,t,12ˊ-CH₂).

¹³C NMR(150MHz,DMSO)δ：178.9,173.3,71.0,62.6,61.5,59.3,58.7,52.4,49.7, 46.0,34.9,34.2,31.6,29.6,29.3,22.7,21.6,19.3,18.5,14.3.

ESI-MS(m/z)：543.7[M+H]⁺.

Chemical structural formula is as follows：

1.5.1 the preparation of triptolide decylate (TP-DA)

Throwing amount 1.5mmol capric acid, 1.5mmol paranitrobenzoyl chloride, 1.5mmol triethylamine in reaction vessel, Add and stirred 5 minutes under the dissolving of 20mL anhydrous methylene chlorides, condition of ice bath；1mmol TP is dissolved in appropriate anhydrous dichloromethane In alkane and it is slowly added dropwise into reaction system, is reacted under condition of ice bath 30 minutes, continue reaction at room temperature and stay overnight, by reactant through silicon Glue column separating purification obtains triptolide decylate 390.6mg.Yield 75.9%.

¹H NMR(DMSO-_d6,600MHz)δ:4.88 (1H, s, 14-CH), 4.54 (1H, d, J=18.71Hz, 19-CH), 4.33 (1H, d, J=18.71Hz, 19-CH), 3.35 (2H, s, 11,12-CH), 3.21 (1H, t, 7-CH), 2.56-2.60 (1H, m,4-CH),2.32-2.37(2H,m,2ˊ-CH₂),2.16-2.25(1H,m,3-CH),1.82-1.85(2H,m,3ˊ-CH₂), 1.73-1.77(1H,m,15-CH),1.57-1.61(4H,m,1、2-CH₂),1.44-1.50(2H,m,6-CH₂),1.25-1.30 (12H,m,6×CH₂),1.04-1.10(1H,m,5-CH),0.93(3H,s,20-CH₂),0.85-0.88(6H,m,16、17- CH₂),0.78(3H,t,10ˊ-CH₂).

¹³C NMR(150MHz,DMSO)δ：178.9,173.5,71.7,62.6,61.6,59.3,58.4,52.3,49.9, 46.0,34.8,34.2,31.5,29.5,26.3,25.0,22.7,21.6,19.1,14.0.

ESI-MS(m/z)：515.6[M+H]⁺.

Chemical structural formula is as follows：

1.6.1 the preparation of triptolide caprylate (TP-CA)

Throwing amount 4mmol octanoic acids, 4mmol DCC, 4mmol DMAP in reaction vessel, add the anhydrous DMA dissolvings of 20mL, Stirred 30 minutes under condition of ice bath；1mmol TP is dissolved in appropriate anhydrous methylene chloride and is slowly added dropwise into reaction system, Reacted under condition of ice bath 45 minutes, continue reaction at room temperature and stay overnight, reactant is obtained into triptolide through silica gel column separating purification Caprylate 411.2mg.Yield 84.5%.

¹H NMR(DMSO-_d6,600MHz)δ:4.81 (1H, s, 14-CH), 4.47 (1H, d, J=18.68Hz, 19-CH), 4.22 (1H, d, J=18.68Hz, 19-CH), 3.37 (2H, s, 11,12-CH), 3.17 (1H, t, 7-CH), 2.56-2.60 (1H, m,4-CH),2.33-2.37(2H,m,2ˊ-CH₂),2.16-2.21(1H,m,3-CH),1.80-1.86(2H,m,3ˊ-CH₂), 1.73-1.78(1H,m,15-CH),1.57-1.61(4H,m,1、2-CH₂),1.39-1.50(2H,m,6-CH₂),1.25-1.30 (8H,m,4ˊ、5ˊ、6ˊ、7ˊ-CH₂),1.04-1.10(1H,m,5-CH),0.93(3H,s,20-CH₂),0.85-0.88(6H,m, 16、17-CH₂),0.78(3H,t,18ˊ-CH₂).

¹³C NMR(150MHz,DMSO)δ：178.8,173.2,71.1,62.6,61.3,59.6,58.2,52.3,49.9, 46.0,34.9,34.2,31.5,29.0,26.3,25.0,22.7,21.6,19.0,13.9.

ESI-MS(m/z)：487.6[M+H]⁺.

Chemical structural formula is as follows：

1.7.1 the preparation of triptolide capronate (TP-HA)

Throwing amount 2.5mmol caproic acids, 2.5mmol HATU, 2.5mmol DMAP in reaction vessel, add 20mL anhydrous DMF dissolves, and is stirred 15 minutes under condition of ice bath；By 1mmol TP be dissolved in appropriate anhydrous chloroform and be slowly added dropwise into Reacted under reaction system, condition of ice bath 30 minutes, continue reaction at room temperature and stay overnight, reactant is obtained into thunder through silica gel column separating purification Public rattan A prime capronate 352.6mg.Yield 76.9%.

¹H NMR(DMSO-_d6,600MHz)δ:4.88 (1H, s, 14-CH), 4.45 (1H, d, J=18.78Hz, 19-CH), 4.15 (1H, d, J=18.78Hz, 19-CH), 3.43 (2H, s, 11,12-CH), 3.20 (1H, t, 7-CH), 2.66-2.76 (1H, m,4-CH),2.28-2.35(2H,m,2ˊ-CH₂),2.17-2.20(1H,m,3-CH),1.80-1.85(2H,m,3ˊ-CH₂), 1.73-1.79(1H,m,15-CH),1.57-1.61(4H,m,1、2-CH₂),1.39-1.50(2H,m,6-CH₂),1.25-1.30 (4H,m,4ˊ、5ˊ-CH₂),1.04-1.10(1H,m,5-CH),0.93(3H,s,20-CH₂),0.85-0.88(9H,m,16、17、6 ˊ-CH₂).

¹³C NMR(150MHz,DMSO)δ：178.5,172.0,71.3,63.1,61.8,59.8,58.6,53.0,49.6, 46.3,34.9,34.2,32.2,31.8,26.5,22.4,21.7,19.3,14.1.

ESI-MS(m/z)：459.5[M+H]⁺.

Chemical structural formula is as follows：

1.8.1 the preparation of triptolide butyrate (TP-BA)

Throwing amount 3mmol butyric acid, 3mmol EDCHCl, 3mmol DIPEA in reaction vessel, add 20mL anhydrous three Chloromethanes dissolves, and is stirred 30 minutes under condition of ice bath；1mmol TP is dissolved in appropriate anhydrous chloroform and is slowly added dropwise Enter to react under reaction system, condition of ice bath 45 minutes, continue reaction at room temperature and stay overnight, reactant is obtained through silica gel column separating purification Triptolide butyrate 341.8mg.Yield 79.4%.

¹H NMR(DMSO-_d6,600MHz)δ:4.96 (1H, s, 14-CH), 4.86 (1H, d, J=18.88Hz, 19-CH), 4.73 (1H, d, J=18.86Hz, 19-CH), 3.98 (1H, d, 11-CH), 3.93 (1H, d, 12-CH), 3.70 (1H, d, 7- CH),2.66-2.76(1H,m,4-CH),2.28-2.42(2H,m,2ˊ-CH₂),2.17-2.20(1H,m,3-CH),1.80- 1.85(2H,m,3ˊ-CH₂),1.73-1.78(1H,m,15-CH),1.57-1.61(4H,m,1、2-CH₂),1.39-1.50(2H, m,6-CH₂),1.04-1.10(1H,m,5-CH),0.99(3H,t,4ˊ-CH₃),0.93(3H,s,20-CH₂),0.85-0.88 (6H,m,16、17-CH₂).

¹³C NMR(150MHz,DMSO)δ：178.9,173.1,70.9,62.7,61.7,59.3,58.4,52.4,49.9, 46.1,36.4,34.1,32.2,31.9,26.3,21.6,19.2,18.9,13.5.

ESI-MS(m/z)：431.5[M+H]⁺.

Chemical structural formula is as follows：

Embodiment 2：The preparation of triptolide fatty acid ester difference nanometer formulation

The different triptolide fatty acid esters of gained are prepared with embodiment 1 and carry out preparation druggability investigation, preparation bag is investigated Include liposome, polymer micelle, albumin nano granular, Fat Emulsion.

The preparation of 2.1 different triptolide fatty acid ester liposomes

2.1.1a the preparation of triptolide stearate (TP-SA) liposome

Weigh triptolide stearate 0.2g, yolk phospholipid (PC-98T) 2g and cholesterol 0.2g and add the anhydrous second of 5g In alcohol, in heating for dissolving at 60 DEG C, organic phase is obtained；Organic phase is slowly injected into 80mL 60 DEG C of waters for injection, marginal not enters side Stir and evenly mix, produce liposome crude product；Liposome crude product is placed in extruder, pass sequentially through 0.2 μm of aperture, 0.1 μm, 0.05 μm extruded film extrusion, obtain liposome solutions；Ultrafiltration removes ethanol in liposome；25g sucrose is weighed, above-mentioned liposome is dissolved in molten Liquid；Plus water for injection is settled to 100mL, with NaOH, salt acid for adjusting pH to 4.0；0.22 μm of membrane filtration, is dispensed, freezing Dry, seal gland, produce.

2.1.1b the preparation of triptolide stearate (TP-SA) liposome

Weigh triptolide stearate 0.2g, yolk phospholipid (EPCS) 2g, PEGylation DSPE (DSPE-PEG2000) 0.2g and cholesterol 0.2g are dissolved in q. s. methylene chloride, and vacuum revolving removes solvent and obtained under the conditions of 45 DEG C Lipid membrane, carries out aquation with 80mL waters for injection and obtains liposome crude product；Liposome crude product is placed in homogeneous in high pressure homogenizer Emulsification, obtains liposome solutions；25g trehaloses are weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection is settled to 100mL, hydrogen is used Sodium oxide molybdena, salt acid for adjusting pH to 6.0；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.1c the preparation of triptolide stearate (TP-SA) liposome

Weigh triptolide stearate 0.3g, hydrogenated phospholipid (HSPC) 3g, PEGylation DSPE (DSPE-PEG2000) 0.3g and cholesterol 0.3g are dissolved in appropriate chloroform, and vacuum revolving removes solvent and obtained under the conditions of 45 DEG C Lipid membrane, carries out aquation with 75mL waters for injection and obtains liposome crude product；Liposome crude product is placed in homogeneous in high pressure homogenizer Emulsification, obtains liposome solutions；30g sucrose is weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection is settled to 100mL, hydrogen-oxygen is used Change sodium, lemon acid for adjusting pH to 8.0；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.1d the preparation of triptolide stearate (TP-SA) liposome

Weigh triptolide stearate 0.2g, yolk phospholipid (PC-98T) 2g, PEGylation distearyl monoethanolamine (DSPE-PEG2000) 0.2g and cholesterol 0.2g are dissolved in q. s. methylene chloride, and vacuum revolving removes solvent and obtained under the conditions of 45 DEG C Lipid membrane, weighs 25g sucrose and is dissolved in 80mL water for injection hydrated films and obtain liposome crude product；Liposome crude product is placed in high pressure Emulsifying in homogenizer, obtains liposome solutions；25g sucrose is weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection is settled to 100mL, with NaOH, salt acid for adjusting pH to 6.5；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.2a the preparation of triptolide palmitate (TP-PA) liposome

Weigh triptolide palmitate 0.2g, yolk phospholipid (PC-98T) 2g and cholesterol 0.2g and add the anhydrous second of 5g Alcohol obtains organic phase in heating for dissolving at 60 DEG C；Organic phase is slowly injected into 80mL 60 DEG C of waters for injection, marginal not enters side stirring Mix, produce liposome crude product；Liposome crude product is placed in extruder, 0.2 μm, 0.1 μm, 0.05 μm of aperture is passed sequentially through Extruded film is extruded, and obtains liposome solutions；Ultrafiltration removes ethanol in liposome；25g sucrose is weighed, above-mentioned liposome solutions are dissolved in； Plus water for injection is settled to 100mL, with NaOH, salt acid for adjusting pH to 4.0；0.22 μm of membrane filtration, packing, freezing is dry It is dry, gland is sealed, is produced.

2.1.2b the preparation of triptolide palmitate (TP-PA) liposome

Weigh triptolide palmitate 0.2g, yolk phospholipid (EPCS) 2g, PEGylation DSPE (DSPE-PEG2000) 0.2g and cholesterol 0.2g are dissolved in q. s. methylene chloride, and vacuum revolving removes solvent and obtained under the conditions of 45 DEG C Lipid membrane, carries out aquation with 80mL waters for injection and obtains liposome crude product；Liposome crude product is placed in homogeneous in high pressure homogenizer Emulsification, obtains liposome solutions；25g trehaloses are weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection is settled to 100mL, hydrogen is used Sodium oxide molybdena, salt acid for adjusting pH to 6.0；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.2c the preparation of triptolide palmitate (TP-PA) liposome

Weigh triptolide palmitate 0.3g, hydrogenated phospholipid (HSPC) 3g, PEGylation DSPE (DSPE-PEG2000) 0.3g and cholesterol 0.3g are dissolved in appropriate chloroform, and vacuum revolving removes solvent and obtained under the conditions of 45 DEG C Lipid membrane, carries out aquation with 75mL waters for injection and obtains liposome crude product；Liposome crude product is placed in homogeneous in high pressure homogenizer Emulsification, obtains liposome solutions；30g sucrose is weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection is settled to 100mL, hydrogen-oxygen is used Change sodium, lemon acid for adjusting pH to 8.0；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.2d the preparation of triptolide palmitate (TP-PA) liposome

Weigh triptolide palmitate 0.2g, yolk phospholipid (PC-98T) 2g, PEGylation distearyl monoethanolamine (DSPE-PEG2000) 0.2g and cholesterol 0.2g are dissolved in q. s. methylene chloride, and vacuum revolving removes solvent and obtained under the conditions of 45 DEG C Lipid membrane, weighs 25g sucrose and is dissolved in 80mL water for injection hydrated films and obtain liposome crude product；Liposome crude product is placed in high pressure Emulsifying in homogenizer, obtains liposome solutions；25g sucrose is weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection is settled to 100mL, with NaOH, salt acid for adjusting pH to 6.5；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.3a the preparation of triptolide myristinate (TP-MA) liposome

Weigh triptolide myristinate 0.2g, yolk phospholipid (EPCS) 2g, PEGylation distearoylphosphatidyl ethanol Amine (DSPE-PEG2000) 0.2g and cholesterol 0.2g are dissolved in q. s. methylene chloride, and vacuum revolving removes solvent under the conditions of 45 DEG C Lipid membrane is obtained, aquation is carried out with 80mL waters for injection and obtains liposome crude product；Liposome crude product is placed in high pressure homogenizer Matter is emulsified, and obtains liposome solutions；25g trehaloses are weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection is settled to 100mL, use NaOH, salt acid for adjusting pH to 6.0；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.3b the preparation of triptolide myristinate (TP-MA) liposome

Weigh triptolide myristinate 0.3g, hydrogenated phospholipid (HSPC) 3g, PEGylation distearoylphosphatidyl ethanol Amine (DSPE-PEG2000) 0.3g and cholesterol 0.3g are dissolved in appropriate chloroform, and vacuum revolving removes solvent under the conditions of 45 DEG C Lipid membrane is obtained, aquation is carried out with 75mL waters for injection and obtains liposome crude product；Liposome crude product is placed in high pressure homogenizer Matter is emulsified, and obtains liposome solutions；30g sucrose is weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection is settled to 100mL, hydrogen is used Sodium oxide molybdena, lemon acid for adjusting pH to 8.0；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.4a the preparation of triptolide laurate (TP-LA) liposome

Weigh triptolide laurate 0.5g, yolk phospholipid (E80) 5g, PEGylation DSPE (DSPE-PEG5000) 0.7g and cholesterol 0.5g adds 4g propane diols in heating for dissolving at 75 DEG C, obtains organic phase；By organic phase It is slowly injected into 80mL 75 DEG C of waters for injection, marginal not enters side and stirred and evenly mixed, produces liposome crude product；Liposome crude product is placed in In extruder, the extruded film extrusion in 1.0 μm, 0.6 μm, 0.4 μm, 0.2 μm, 0.1 μm, 0.05 μm of aperture is passed sequentially through, lipid is obtained Liquid solution；Ultrafiltration removes propane diols in liposome；25g lactose is weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection is settled to 100mL, pH to 7.0 is adjusted with hydrochloric acid, phosphate；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.4b the preparation of triptolide laurate (TP-LA) liposome

Weigh triptolide laurate 0.2g, soybean lecithin (PL-100M) 4g, the myristoyl phosphatidyl of PEGylation two Monoethanolamine (DMPE-PEG3500) 0.4g and cholesterol 0.4g are dissolved in appropriate chloroform, and vacuum revolving is removed under the conditions of 45 DEG C Solvent obtains lipid membrane, and carrying out aquation with 80mL waters for injection obtains liposome crude product；Liposome crude product is placed in extruder, according to The secondary extruded film by 0.2 μm, 0.1 μm, 0.05 μm of aperture is extruded, and obtains liposome solutions；25g sucrose is weighed, above-mentioned fat is dissolved in Plastid solution；Plus water for injection is settled to 100mL, pH to 7.0 is adjusted with phosphate；0.22 μm of membrane filtration, packing, freezing is dry It is dry, gland is sealed, is produced.

2.1.5a the preparation of triptolide decylate (TP-DA) liposome

Weigh triptolide decylate 0.7g, soybean lecithin (S100) 7g, PEGylation DSPE (DSPE-PEG2000) 1g and cholesterol 1g adds 10g absolute ethyl alcohols in heating for dissolving at 30 DEG C, obtains organic phase；Organic phase is delayed In slow injection 75mL 30 DEG C of waters for injection, marginal not enters side and stirred and evenly mixed, and produces liposome crude product；Liposome crude product is placed in crowded Go out in device, pass sequentially through the extruded film extrusion in 0.2 μm, 0.1 μm, 0.05 μm of aperture, obtain liposome solutions；Wipe film evaporates Remove ethanol in liposome；30g mannitol is weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection is settled to 100mL, vinegar is used Acid, phosphate regulation pH to 6.0；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.5b the preparation of triptolide decylate (TP-DA) liposome

Weigh triptolide decylate 0.2g, yolk phospholipid (EPCS) 2g, PEGylation DSPE (DSPE-PEG8000) 0.2g and cholesterol 0.2g adds the 8g tert-butyl alcohols in heating for dissolving at 50 DEG C, obtains organic phase；By organic phase It is slowly injected into 80mL 50 DEG C of waters for injection, marginal not enters side and stirred and evenly mixed, produces liposome crude product；Liposome crude product is placed in Emulsifying in high pressure homogenizer, obtains liposome solutions；Wipe film evaporation removes the tert-butyl alcohol in liposome；Weigh 20g seas Algae sugar, is dissolved in above-mentioned liposome solutions；Plus water for injection is settled to 100mL, with NaOH, salt acid for adjusting pH to 6.0；0.22 μm membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.6a the preparation of triptolide caprylate (TP-CA) liposome

Weigh triptolide caprylate 0.05g, yolk phospholipid (EPCS) 0.5g, the palmityl phosphatidyl ethanol of PEGylation two Amine (DPPE-PEG2000) 0.4g is dissolved in q. s. methylene chloride, and vacuum revolving removes solvent and obtains lipid membrane under the conditions of 50 DEG C, Aquation, which is carried out, with 90mL waters for injection obtains liposome crude product；Liposome crude product is placed in extruder, the μ of aperture 0.4 is passed sequentially through M, 0.2 μm, 0.1 μm, 0.05 μm of extruded film extrusion, obtain liposome solutions；4g maltose is weighed, above-mentioned liposome is dissolved in molten Liquid；Plus water for injection is settled to 100mL, with NaOH, salt acid for adjusting pH to 6.0；0.22 μm of membrane filtration, is dispensed, freezing Dry, seal gland, produce.

2.1.6b the preparation of triptolide caprylate (TP-CA) liposome

Weigh triptolide caprylate 0.1g, soybean lecithin (DOPC) 1g, PEGylation DSPE (DSPE-PEG1500) 1g and cholesterol 0.1g adds the 3g tert-butyl alcohols in heating for dissolving at 25 DEG C, obtains organic phase；Organic phase is delayed In slow injection 95mL 25 DEG C of waters for injection, marginal not enters side and stirred and evenly mixed, and produces liposome crude product；Liposome crude product is placed in crowded Go out in device, pass sequentially through the extruded film extrusion in 0.2 μm, 0.1 μm, 0.05 μm of aperture, obtain liposome solutions；Ultrafiltration removes liposome The middle tert-butyl alcohol；10g lactose is weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection is settled to 100mL, with NaOH, hydrochloric acid Adjust pH to 4.0；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.6c the preparation of triptolide caprylate (TP-CA) liposome

Weigh triptolide caprylate 0.4g, yolk phospholipid (EPCS) 4g, PEGylation DSPE (DSPE-PEG2000) 0.4g and cholesterol 0.4g are dissolved in appropriate absolute ethyl alcohol, and vacuum revolving removes solvent and obtained under the conditions of 45 DEG C Lipid membrane, carries out aquation with 85mL waters for injection and obtains liposome crude product；Liposome crude product is placed in homogeneous in high pressure homogenizer Emulsification, obtains liposome solutions；15g trehaloses are weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection is settled to 100mL, hydrogen is used Sodium oxide molybdena, salt acid for adjusting pH to 7.0；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.7a the preparation of triptolide capronate (TP-HA) liposome

Weigh triptolide capronate 1g, yolk phospholipid (PC-98T) 10g, PEGylation DSPE (DSPE-PEG2000) 0.5g and cholesterol 0.5g are dissolved in q. s. methylene chloride, and vacuum revolving removes solvent and obtained under the conditions of 45 DEG C Lipid membrane, weighs 30g sucrose and is dissolved in 80mL water for injection hydrated films and obtain liposome crude product；Liposome crude product is placed in high pressure Emulsifying in homogenizer, obtains liposome solutions；30g trehaloses are weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection constant volume To 100mL, with NaOH, salt acid for adjusting pH to 8.0；0.22 μm of membrane filtration, is dispensed, and freeze-drying seals gland, i.e., .

2.1.7b the preparation of triptolide capronate (TP-HA) liposome

Weigh triptolide capronate 0.2g, soybean lecithin (SPC-3) 2g, PEGylation DSPE (DPPE-PEG2000) 0.2g is dissolved in appropriate absolute ethyl alcohol, and vacuum revolving removes solvent and obtains lipid membrane under the conditions of 50 DEG C, uses 75mL waters for injection carry out aquation and obtain liposome crude product；Liposome crude product is placed in extruder, pass sequentially through 2.0 μm of aperture, 1.0 μm, 0.4 μm, 0.1 μm, 0.05 μm of extruded film extrusion, obtains liposome solutions；40g sucrose is weighed, above-mentioned liposome is dissolved in Solution；Plus water for injection is settled to 100mL, pH to 5.0 is adjusted with acetic acid, phosphate；0.22 μm of membrane filtration, is dispensed, freezing Dry, seal gland, produce.

2.1.8a the preparation of triptolide butyrate (TP-BA) liposome

Weigh triptolide butyrate 0.2g, hydrogenated phospholipid (HSPC) 3g, PEGylation DSPE (DSPE-PEG6000) 0.3g and cholesterol 0.3g adds 6g propane diols in heating for dissolving at 55 DEG C, obtains organic phase；By organic phase It is slowly injected into 85mL 55 DEG C of waters for injection, marginal not enters side and stirred and evenly mixed, produces liposome crude product；Liposome crude product is placed in Emulsifying in high pressure homogenizer, then be placed in extruder, the extruded film extrusion in 0.05 μm of aperture is crossed, liposome solutions are obtained；It is super Filter propane diols in liposome；9g glucose, 6g mannitol are weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection constant volume To 100mL, pH to 10.0 is adjusted with NaOH；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

2.1.8b the preparation of triptolide butyrate (TP-BA) liposome

Weigh triptolide butyrate 0.2g, phosphatide (DSPC) 2g, PEGylation DSPE (DSPE- PEG2000) 0.2g and cholesterol 0.8g adds 1g absolute ethyl alcohols in heating for dissolving at 50 DEG C, obtains organic phase；Organic phase is slow In 50 DEG C of waters for injection for injecting 80mL, marginal not enters side and stirred and evenly mixed, and produces liposome crude product；Liposome crude product is placed in extrusion In device, the extruded film extrusion in 2.0 μm, 1.0 μm, 0.4 μm, 0.05 μm of aperture is passed sequentially through, liposome solutions are obtained；Ultrafiltration removes degreasing Ethanol in plastid；25g trehaloses are weighed, above-mentioned liposome solutions are dissolved in；Plus water for injection is settled to 100mL, with NaOH, Salt acid for adjusting pH is to 6.0；0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced.

The preparation of 2.2 different triptolide fatty acid ester polymer micelles

2.2.1a the preparation of triptolide stearate (TP-SA) polymer micelle

Triptolide stearate 0.2g, MPEG-PLA (mPEG-PDLLA) 1.2g is weighed to be dissolved in In appropriate acetonitrile；Vacuum revolving removes solvent film forming under the conditions of 60 DEG C；60 DEG C of water for injection shaking aquations are preheated to 90mL；Plus Enter the dissolving of 20g trehaloses, be settled to 100mL, 0.22 μm of membrane filtration is dispensed, and freeze-drying is sealed gland, produced.

2.2.1b the preparation of triptolide palmitate (TP-PA) polymer micelle

Triptolide palmitate 0.3g, MPEG-PLA (mPEG-PDLLA) 1.5g is weighed to be dissolved in In appropriate acetonitrile；Vacuum revolving removes solvent film forming under the conditions of 60 DEG C；60 DEG C of water for injection shaking aquations are preheated to 90mL；Plus Enter the dissolving of 20g sucrose, be settled to 100mL, 0.22 μm of membrane filtration is dispensed, and freeze-drying is sealed gland, produced.

2.2.2 the preparation of triptolide myristinate (TP-MA) polymer micelle

Weigh triptolide myristinate 0.2g, MPEG-PLA (mPEG-PDLLA) 1.4g molten In appropriate acetonitrile；Vacuum revolving removes solvent film forming under the conditions of 60 DEG C；60 DEG C of water for injection shaking aquations are preheated to 85mL； 12g glucose, the dissolving of 8g mannitol are added, 100mL is settled to, 0.22 μm of membrane filtration is dispensed, and gland is sealed in freeze-drying, Produce.

2.2.3 the preparation of triptolide caprylate (TP-CA) polymer micelle

Triptolide caprylate 0.4g, MPEG-PLA (mPEG-PDLLA) 5g is weighed to be dissolved in right amount In acetonitrile；Vacuum revolving removes solvent film forming under the conditions of 55 DEG C；55 DEG C of water for injection shaking aquations are preheated to 85mL；It is settled to 100mL, 0.22 μm of membrane filtration is dispensed, and freeze-drying is sealed gland, produced.

2.2.4 the preparation of triptolide butyrate (TP-BA) polymer micelle

Weigh triptolide butyrate 0.5g, MPEG-PLA (mPEG-PDLLA) 2.5g be dissolved in it is suitable Measure in acetonitrile；Vacuum revolving removes solvent film forming under the conditions of 50 DEG C；60 DEG C of water for injection shaking aquations are preheated to 85mL；Add 20g maltose is dissolved, and is settled to 100mL, and 0.22 μm of membrane filtration is dispensed, and freeze-drying is sealed gland, produced.

The preparation of 2.3 triptolide fatty acid ester albumin nano granulars

2.3.1 the preparation of triptolide stearate (TP-SA) albumin nano granular

Weighing human serum albumins 4.5g and adding 100mL waters for injection low temperature and stir makes mixing, treats that temperature is down to 2 DEG C, drop 1mL chloroforms are added, it is for aqueous phase that 10000rpm, which shears 5min, under the conditions of 0 DEG C；Weigh triptolide stearate 0.5g molten In absolute ethyl alcohol-chloroform mixed solvent 3mL (1：2) it is for organic phase；In high speed shear disperse aqueous phase under the conditions of be slowly added dropwise into Organic phase, 10000rpm shearings 5min obtains colostrum；Colostrum is transferred in high pressure homogenizer, with homogenization pressure 5000psi, 10000psi, 15000psi, 20000psi are respectively circulated 2 times；Homogeneous is diluted after terminating with 5 times of amounts, 2 DEG C of waters for injection, using scraper plate Formula thin film evaporation removes solvent, and concentration is settled to 100mL；0.22 μm of membrane filtration is degerming, dispenses, and gland is sealed in freeze-drying, Produce.

2.3.2 the preparation of triptolide palmitate (TP-PA) albumin nano granular

Weighing human serum albumins 4.5g and adding 100mL waters for injection low temperature and stir makes mixing, treats that temperature is down to 2 DEG C, drop 1mL chloroforms are added, it is for aqueous phase that 10000rpm, which shears 3min, under the conditions of 0 DEG C；Weigh triptolide palmitate 0.4g molten In absolute ethyl alcohol-chloroform mixed solvent 3mL (1：4) it is for organic phase；In high speed shear disperse aqueous phase under the conditions of be slowly added dropwise into Organic phase, 10000rpm shearings 5min obtains colostrum；Colostrum is transferred in high pressure homogenizer, with homogenization pressure 5000psi, 10000psi, 15000psi, 20000psi are respectively circulated 2 times；Homogeneous is diluted after terminating with 5 times of amounts, 2 DEG C of waters for injection, using scraper plate Formula thin film evaporation removes solvent, and concentration is settled to 100mL；0.22 μm of membrane filtration is degerming, dispenses, and gland is sealed in freeze-drying, Produce.

2.3.3 the preparation of triptolide myristinate (TP-MA) albumin nano granular

Weighing human serum albumins 4.5g and adding 100mL waters for injection low temperature and stir makes mixing, treats that temperature is down to 2 DEG C, drop 1mL chloroforms are added, it is for aqueous phase that 10000rpm, which shears 3min, under the conditions of 0 DEG C；Weigh triptolide myristinate 0.45g It is dissolved in absolute ethyl alcohol-chloroform mixed solvent 2.5mL (1：4) it is for organic phase；Disperse slowly to drip under the conditions of aqueous phase in high speed shear Organic phase is added, 12000rpm shearings 8min obtains colostrum；Colostrum is transferred in high pressure homogenizer, with homogenization pressure 10000psi, 15000psi, 20000psi are respectively circulated 2 times；Homogeneous is diluted after terminating with 5 times of amounts, 2 DEG C of waters for injection, using scraper plate Formula thin film evaporation removes solvent, and concentration is settled to 100mL；0.22 μm of membrane filtration is degerming, dispenses, and gland is sealed in freeze-drying, Produce.

2.3.4 the preparation of triptolide caprylate (TP-CA) albumin nano granular

Weighing human serum albumins 4.5g and adding 100mL waters for injection low temperature and stir makes mixing, treats that temperature is down to 2 DEG C, drop 1mL chloroforms are added, it is for aqueous phase that 10000rpm, which shears 5min, under the conditions of 0 DEG C；Triptolide caprylate 0.5g is weighed to be dissolved in Absolute ethyl alcohol-chloroform mixed solvent 4mL (1：3) it is for organic phase；It is slowly added dropwise under the conditions of high speed shear disperses aqueous phase into having Machine phase, 10000rpm shearings 5min obtains colostrum；Colostrum is transferred in high pressure homogenizer, 2 are circulated with homogenization pressure 15000psi Secondary, 20000psi is circulated 4 times；Homogeneous is diluted after terminating with 5 times of amounts, 2 DEG C of waters for injection, is removed using wipe film evaporation molten Agent, concentration is settled to 100mL；0.22 μm of membrane filtration is degerming, dispenses, and freeze-drying is sealed gland, produced.

2.3.5 the preparation of triptolide butyrate (TP-BA) albumin nano granular

Weighing human serum albumins 4.5g and adding 100mL waters for injection low temperature and stir makes mixing, treats that temperature is down to 2 DEG C, drop 1mL chloroforms are added, it is for aqueous phase that 10000rpm, which shears 3min, under the conditions of 0 DEG C；Triptolide butyrate 0.4g is weighed to be dissolved in Absolute ethyl alcohol-chloroform mixed solvent 2mL (1：5) it is for organic phase；It is slowly added dropwise under the conditions of high speed shear disperses aqueous phase into having Machine phase, 10000rpm shearings 3min obtains colostrum；Colostrum is transferred in high pressure homogenizer, with homogenization pressure 5000psi, 10000psi, 15000psi, 20000psi are respectively circulated 2 times；Homogeneous is diluted after terminating with 5 times of amounts, 2 DEG C of waters for injection, using scraper plate Formula thin film evaporation removes solvent, and concentration is settled to 100mL；0.22 μm of membrane filtration is degerming, dispenses, and gland is sealed in freeze-drying, Produce.

The preparation of 2.4 triptolide fatty acid ester Fat Emulsions

2.4.1 the preparation of triptolide stearate (TP-SA) Fat Emulsion

Weigh injection medium chain triglyceride 5g, triptolide stearate 0.2g, heating water bath is under the conditions of 70 DEG C It is stirred to dissolve, obtains oil phase；Weigh phosphatide (PL-100M) 3g to add in 90mL waters for injection, shearing makes to disperse, and obtains aqueous phase；Will Oil phase and aqueous phase are mixed at 70 DEG C, while emulsifying 5min with emulsification pretreatment machine, obtain colostrum, 100mL is settled to water for injection； Colostrum is placed in high pressure homogenizer further emulsifying, with salt acid for adjusting pH to 5.0,0.22 μm of membrane filtration is crossed, dispenses, Seal, sterilize 15min at 121 DEG C, produces.

2.4.2 the preparation of triptolide palmitate (TP-PA) Fat Emulsion

Weigh injection medium chain triglyceride 2.5g, soybean oil 2.5g, triptolide palmitate 0.2g, heating water bath It is stirred to dissolve under the conditions of to 60 DEG C, obtains oil phase；Weigh phosphatide (PL-100M) 3g to add in 90mL waters for injection, shearing makes point Dissipate, obtain aqueous phase；Oil phase and aqueous phase are mixed at 60 DEG C, while emulsifying 5min with emulsification pretreatment machine, colostrum is obtained, uses water for injection It is settled to 100mL；Colostrum is placed in high pressure homogenizer further emulsifying, pH to 7.0 is adjusted with phosphate, 0.22 μm is crossed Membrane filtration, is dispensed, sealing, and sterilize 30min at 100 DEG C, produces.

2.4.3 the preparation of triptolide myristinate (TP-MA) Fat Emulsion

Weigh injection soybean oil 6g, triptolide myristinate 0.3g, heating water bath to stirring under the conditions of 70 DEG C Make dissolving, obtain oil phase；Weigh phosphatide (PL-100M) 3g to add in 90mL waters for injection, shearing makes to disperse, and obtains aqueous phase；By oil phase Mixed with aqueous phase at 70 DEG C, while emulsifying 8min with emulsification pretreatment machine, obtain colostrum, 100mL is settled to water for injection；Will be just Breast is placed in high pressure homogenizer further emulsifying, pH to 6.5 is adjusted with phosphate, respectively through 0.45 μm, 0.22 μm of filter membrane Filtration sterilization, is dispensed, and sealing is produced.

2.4.4 the preparation of triptolide caprylate (TP-CA) Fat Emulsion

Injection medium chain triglyceride 4g, soybean oil 4g, triptolide caprylate 0.4g are weighed, heating water bath is to 80 DEG C Under the conditions of be stirred to dissolve, obtain oil phase；Weigh phosphatide (PC-98T) 4g, glycerine 1g to add in 85mL waters for injection, shearing makes point Dissipate, obtain aqueous phase；Oil phase and aqueous phase are mixed at 80 DEG C, while emulsifying 5min with emulsification pretreatment machine, colostrum is obtained, uses water for injection It is settled to 100mL；Colostrum is placed in high pressure homogenizer further emulsifying, with salt acid for adjusting pH to 6.0,0.45 μm of filter membrane Filtering, is dispensed, sealing, and sterilize 15min at 121 DEG C, produces.

2.4.5 the preparation of triptolide butyrate (TP-BA) Fat Emulsion

Weigh injection medium chain triglyceride 4g, oleic acid 0.01g, triptolide butyrate 0.2g, heating water bath to 80 It is stirred to dissolve under the conditions of DEG C, obtains oil phase；Weigh phosphatide (SPC-3) 3g to add in 90mL waters for injection, shearing makes to disperse, and obtains water Phase；Oil phase and aqueous phase are mixed at 65 DEG C, while emulsifying 6min with emulsification pretreatment machine, colostrum is obtained, is settled to water for injection 100mL；Colostrum is placed in high pressure homogenizer further emulsifying, with newborn acid for adjusting pH to 5.0,0.22 μm of filter membrane mistake is crossed Filter, is dispensed, sealing, and sterilize 15min at 121 DEG C, produces.

As a result show：Triptolide fatty acid ester can effectively prepare liposome, polymer micelle, albumin nano The nanometer formulations such as grain, Fat Emulsion, druggability is good.

Embodiment 3：The measure of liposomal particle size and envelop rate

Different triptolide fatty acid esters and thunder are prepared respectively using 2.1.1b prescribe medicine fat ratios of embodiment as unified prescription Public rattan A prime liposome, determines liposomal particle size and envelop rate.It the results are shown in Table 1,

The liposomal particle size of table 1 and entrapment efficiency determination result

Test result indicate that, triptolide fatty acid ester can be prepared into liposome, particle diameter distribution under the conditions of identical prescription 100~130nm, envelop rate is more than 90%；And because medicine is separated out in triptolide preparation process, liposome can not effectively be made Standby, envelop rate is less than 50%.Integrated comparative is found simultaneously, and the saturated fatty acid of carbon number >=8 is modified and obtains triptolide Fatty acid ester has smaller particle diameter, and is more evenly distributed, therefore it is preferred that carbon number is the saturated fat of 2n (n is 4~9) Acid.

Embodiment 4：Cytotoxicity experiment

Based on embodiment 3 preferably on the basis of, prepare carbon original respectively by uniformly prescription of the prescribe medicine fat ratios of embodiment 2.1.1b Subnumber is the triptolide fatty acid ester liposome of 2n (n is 4~9) different chain length saturated fatty acid modification, with TP (DMSO) antitumor cytotoxicity that solution carries out MCF-7 and A549 cells as control is studied, and concrete operation step is as follows：

1) the good cell of growth conditions is diluted to suitable concentration with certain culture medium, then by every 5000 cells in hole Density be inoculated in 96 well culture plates, per the μ L of hole 100.By culture plate in incubator preculture 24h (37 DEG C, 5%CO₂)。

2) 10 μ L various concentrations medicines are separately added into culture plate (equimolar is administered, each 3 multiple holes of concentration).Will training Support plate and be incubated 48h in incubator.

3) the CCK-8 μ L of culture medium 10 are added into every hole.Culture plate is incubated 4h in incubator.

4) absorbance at 450nm is determined with ELIASA.Average, calculate inhibiting rate.

Computing formula：Inhibiting rate=[(Ac-As)/(Ac-Ab)] × 100%

As：Experimental port (culture medium, CCK-8 containing cell, medicine)

Ac：Control wells (culture medium and CCK-8, non-drug containing containing cell)

Ab：Blank well (culture medium, CCK-8 without cell and medicine)

As a result：Carbazole alkaloid IC under each drug concentration is calculated respectively₅₀Value is shown in Table 2.

The cytotoxic effect IC of table 2₅₀It is worth result

Test result indicate that, triptolide fatty acid ester preparation gained liposome is compared for TP, cytotoxic effect It is lower.In terms of being embodied in the inhibitory action for cell growth, triptolide fatty acid ester liposome reaches that half suppresses institute The drug concentration IC needed₅₀Value is higher, and the saturated fatty acid modification gained triptolide fatty acid ester of carbon number >=14 Liposome has higher IC than other again₅₀Concentration, therefore further preferably carbon number 2n (n is 7~9) saturated fatty acid.

Embodiment 5：Rat Internal pharmacokinetics are studied

Based on embodiment 4 preferably on the basis of, prepare C respectively by uniformly prescription of the prescribe medicine fat ratios of embodiment 2.1.1b_2n(n For 7~9) in the range of triptolide fatty acid ester (TP-MA, TP-PA, TP-SA) fat for modifying of different chain length saturated fatty acid Plastid, using TP (DMSO) solution as with reference to progress rat Internal pharmacokinetics research.Rat tail vein is administered, and dosage is equivalent TP 3mg/kg, takes blood time point setting 2,5,10,15,30,45,60,90,120,180min, each 5 SD rats (♂) of every group of medicine.

Sample treatment and analysis method：Blood plasma plus 4 times of amount methanol extraction albumen are taken, 12000r/min centrifugation 10min take Methanol is flung to clearly, remaining raffinate adds 400 μ L methyl tertiary butyl ether(MTBE)s to be extracted twice, 12000r/min centrifugations 10min.Collect organic extraction Phase is taken, concentration is volatilized, plus the μ L of methanol 100 redissolve, centrifuging and taking supernatant enters liquid phase measurement cubage blood concentration, using DAS 2.0 softwares carry out pharmacokinetics fitting, and half-life period the results are shown in Table 3.

The medicine Half-life in vivo result of table 3

Test result indicate that, triptolide fatty acid ester prepares gained liposome can effectively extend medicine effect compared to TP Half-life period, significantly improve pharmacokinetic property.

The preferred embodiment to the invention is illustrated above, but the invention be not limited to it is described Embodiment, those of ordinary skill in the art can also make a variety of equivalent on the premise of without prejudice to the invention spirit Modification or replacement, these equivalent modifications or replacement are all contained in the application claim limited range.

Claims (17)

1. a kind of triptolide fatty acid ester, shown in its chemical constitution such as formula (I)：

In formula (I), R represents the straight chained alkyl acyl group that carbon number is 2n, and n is 2~9.

2. triptolide fatty acid ester as claimed in claim 1, it is characterised in that n is 4~9.

3. triptolide fatty acid ester as claimed in claim 1, it is characterised in that n is 7~9.

4. a kind of method of the triptolide fatty acid ester prepared described in any one of claims 1 to 3, it is characterised in that institute The triptolide fatty acid ester stated is obtained by triptolide with saturated fatty acid by esterification, and synthetic route is：

Wherein, ROH is saturated fatty acid, and R is defined as described in any one of claims 1 to 3；

Specific steps include：

(a) saturated fatty acid, condensing agent and catalyst are dissolved in organic solvent, 0~10 DEG C is cooled under stirring and obtains mixed liquor；

(b) triptolide dissolving is instilled in above-mentioned mixed liquor in organic solvent, 15~45 points is reacted under the conditions of 0~10 DEG C Clock, then continues to react, esterification terminates rear isolated triptolide fatty acid ester at room temperature.

5. method as claimed in claim 4, it is characterised in that

Described condensing agent be paranitrobenzoyl chloride, N, N'- dicyclohexylcarbodiimides, N, N'- DICs, 1- ethyls-(3- dimethylaminopropyls) phosphinylidyne diimmonium salt hydrochlorate, the chloro- 1- methyl pyridinium iodides of 2-, hexafluorophosphoric acid benzene And triazol-1-yl-epoxide tripyrrole alkyl phosphorus, 2- (7- azos BTA)-N, N, N', N'- tetramethylurea hexafluorophosphoric acids Ester, O- (5- Chloro-Benzotriazole -1- bases)-two (dimethylamino) carbon hexafluorophosphates, O- (BTA -1- base oxygen Base) one or two in-two piperidines carbon hexafluorophosphates, N-hydroxy-succinamide and N- hydroxy thiosuccinimides More than；

Described catalyst is in I-hydroxybenzotriazole, DMAP, triethylamine and N, N- diisopropylethylamine It is one or more kinds of；

Described organic solvent is in dichloromethane, chloroform, N,N-dimethylformamide and DMAC N,N' dimethyl acetamide It is one or more kinds of.

6. method as claimed in claim 4, it is characterised in that described saturated fatty acid is with triptolide mol ratio 1.2~4:1, described condensing agent and the mol ratio of triptolide are 1.2~4:1.

7. the answering in antitumor or anti-inflammatory drug is prepared of the triptolide fatty acid ester described in any one of claims 1 to 3 With.

8. a kind of nanometer formulation of the triptolide fatty acid ester described in any one of claims 1 to 3, the nanometer formulation is Liposome, polymer micelle, albumin nano granular or Fat Emulsion.

9. nanometer formulation as claimed in claim 8, it is characterised in that described liposome is by following ingredients according to bulking value Than being formulated：

10. nanometer formulation as claimed in claim 9, it is characterised in that described liposome is by following ingredients according to weighing body Product ratio is formulated：

11. the nanometer formulation as described in claim 9 or 10, it is characterised in that

Described phosphatide is yolk phospholipid, soybean lecithin, phosphatide, hydrogenation yolk phospholipid, hydrogenated soya phosphatide, two palmityl phosphatide Phatidylcholine, dimyristoyl phosphatidyl choline, DSPC, phosphatidyl-ethanolamine, distearoylphosphatidyl are sweet One or more in oil, DPPG, DOPC, cuorin and sphingomyelins；

Described PEGylation phosphatide is PEGylation DSPE, PEGylation DPPE, PEG Change the one or more in two nutmeg phosphatidyl-ethanolamines, the PEG mean molecule quantities of the PEGylation phosphatide for 1000~ 8000；

Described freeze drying protectant be in trehalose, sucrose, maltose, lactose, mannitol, glucose it is one or two kinds of with On；

Described pH adjusting agent is one kind in NaOH, sodium acetate, acetic acid, phosphate, carbonate, hydrochloric acid, citric acid etc. Or it is two or more.

12. the nanometer formulation as described in claim 9 or 10, it is characterised in that

Described phosphatide is yolk phospholipid or soybean lecithin；

Described PEGylation phosphatide is the PEGylation DSPE that mean molecule quantity is 2000；

Described freeze drying protectant is trehalose or sucrose；

Described pH adjusting agent is NaOH, hydrochloric acid or citric acid.

13. a kind of preparation method of liposome, the formula of the liposome is as described in any one of claim 9~12, the system Preparation Method includes step：

(A) preparation of liposome crude product, weighs formula ratio triptolide fatty acid ester, phosphatide, PEGylation phosphatide and cholesterol, Add appropriate organic solvent I and obtain organic phase in heating for dissolving at 25~75 DEG C, then organic phase is slowly injected into 25~75 DEG C suitable Measure in water for injection, marginal not enters side and stirred and evenly mixed, and produces liposome crude product；

(B) preparation of liposome solutions, is placed in high pressure homogenizer emulsifying by the liposome crude product or is placed in extruder In pass sequentially through different pore size extruded film extrusion or be placed in high pressure homogenizer after homogeneous again row extrusion, obtain liposome solutions；

(C) formula ratio freeze drying protectant is weighed, the liposome solutions are dissolved in；Plus water for injection constant volume, regulation pH to setting； 0.22 μm of membrane filtration, is dispensed, and freeze-drying is sealed gland, produced；

Or,

Step (A) is replaced with：(A ') weighs formula ratio triptolide fatty acid ester, phosphatide, PEGylation phosphatide, cholesterol and is dissolved in In organic solvent II, vacuum revolving removes solvent and obtains lipid membrane under the conditions of 40~60 DEG C, and aquation is carried out with appropriate water for injection Obtain liposome crude product.

14. the preparation method of liposome as claimed in claim 13, it is characterised in that described organic solvent I is selected from anhydrous One or more in ethanol, propane diols, the tert-butyl alcohol；Described organic solvent I is retained in liposome, or in lipid Removed again by ultrafiltration or scrapper thin film evaporator after the emulsification of body crude product.

15. the preparation method of liposome as claimed in claim 13, it is characterised in that described organic solvent II is dichloromethane One or more in alkane, chloroform, absolute ethyl alcohol.

16. the preparation method of liposome as claimed in claim 13, it is characterised in that described freeze drying protectant is in liposome In solution it is additional or in being soluble in the aqueous phase in plus.

17. the preparation method of liposome as claimed in claim 13, it is characterised in that the extrusion fenestra in step (B) One or more of the footpath in 2.0 μm, 1.0 μm, 0.8 μm, 0.6 μm, 0.4 μm, 0.2 μm, 0.1 μm and 0.05 μm, are squeezed Large aperture is passed sequentially through when going out to small-bore.