WO2018102973A1 - 一种peg化维生素e-杠柳次苷偶联物纳米粒及其制法和用途 - Google Patents

一种peg化维生素e-杠柳次苷偶联物纳米粒及其制法和用途 Download PDF

Info

Publication number
WO2018102973A1
WO2018102973A1 PCT/CN2016/108628 CN2016108628W WO2018102973A1 WO 2018102973 A1 WO2018102973 A1 WO 2018102973A1 CN 2016108628 W CN2016108628 W CN 2016108628W WO 2018102973 A1 WO2018102973 A1 WO 2018102973A1
Authority
WO
WIPO (PCT)
Prior art keywords
vitamin
nanoparticles
barocin
pegylated
conjugated
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2016/108628
Other languages
English (en)
French (fr)
Inventor
徐希明
张辉云
余江南
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jiangsu University
Original Assignee
Jiangsu University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Jiangsu University filed Critical Jiangsu University
Priority to PCT/CN2016/108628 priority Critical patent/WO2018102973A1/zh
Priority to CN201680001826.7A priority patent/CN107735108A/zh
Publication of WO2018102973A1 publication Critical patent/WO2018102973A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
    • A61K31/355—Tocopherols, e.g. vitamin E
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
    • A61K31/585—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin containing lactone rings, e.g. oxandrolone, bufalin

Definitions

  • the present invention relates to vitamin E-barocin conjugated nanoparticles and novel PEGylated vitamin E-barocin conjugated nanoparticles and a preparation method thereof and use as an antitumor drug.
  • cardiotonic steroid active ingredients in the treatment of cancer began in ancient China, when sputum secretions were recorded for the treatment of cancer. It has been found that the anti-tumor component of the sputum secretion is scorpion venom, a type A cardiotonic steroid component. Studies have reported that in patients with heart disease using digoxigenin, the concentration of digoxigenin in the blood is significantly negatively correlated with the incidence of tumors in the blood and urinary system, and digoxigenin-containing breast cancer is used. The mortality rate of patients was significantly lower than that of patients who did not use digoxigenin.
  • the active ingredient of cardiac steroids is mainly used as a cardiotonic agent in clinical practice, and the case of being used as a chemotherapy drug for clinical treatment of tumors has not been reported.
  • Most of the anti-tumor active ingredients of steroid steroids are in preclinical research, among which semi-synthetic drugs UNBS- 1450 and PBI-05204 have entered clinical phase I in the United States for the treatment of advanced drug-resistant malignancies.
  • the application of precursor design techniques to the development of drugs for the anti-tumor activity of cardiotonic steroids is still in its infancy.
  • scorpion venom including liposomes containing scorpion venom, chitosan nanoparticles and PEG-modified prodrugs.
  • PPM Periploclinoside
  • pharmacological activities such as cardiotonic, anti-inflammatory, anti-tumor and the like.
  • PPM is currently used less in clinical practice and is limited to research at the cellular and animal levels.
  • Han Yubo and Zhao Aiguo anti-tumor of lycopene aglycone Research on the role of [J]. Chinese Journal of Pediatric Hematology and Oncology, 2008, 13, (1): 1-5].
  • One of the objects of the present invention is to provide a vitamin E-bark glucoside conjugate and a method for synthesizing the same.
  • Another object of the present invention is to provide a novel PEGylated vitamin E-barocin conjugated nanoparticle and a preparation method thereof.
  • a third object of the present invention is to provide the use of the conjugate nanoparticles described in the treatment of liver cancer.
  • a vitamin E-barocin conjugated nanoparticle which is a self-assembled nanoparticle of a disulfide-S-S-coupled vitamin E and a scutellarin.
  • the above-mentioned vitamin E-barocin conjugated nanoparticles can be replaced by other cardiac steroid components such as perilla aglycone, scorpion venom, scallopin or leucine.
  • a method for preparing the above vitamin E-barocin conjugated nanoparticles which comprises the following steps:
  • Step 1 Weigh a certain amount of dithiol dihydroxyacetic acid into acetic anhydride, stir the reaction at 30 ° C for 2-24h, add appropriate amount of toluene after the reaction, spin dry toluene and acetic anhydride to obtain an oily liquid, add anhydrous Dissolve methyl chloride, then add vitamin E, the ratio of vitamin E to dithiol dihydroxyacetic acid is 10..1 ⁇ 5..1, and add appropriate amount of 4-dimethylaminopyridine (DMAP), DMAP and vitamins.
  • DMAP 4-dimethylaminopyridine
  • the molar ratio of E is 1..10, and the reaction is stirred at room temperature for 12 to 36 hours, and purified by a silica gel column.
  • Step 2 Weigh the product of step 1 VE-SS-COOH and periplocin, the molar ratio of VE-SS-COOH to periplocin is 1..1 ⁇ 3..1, dissolved in anhydrous dichloromethane Add DCC and DMAP, and the molar ratio of them to the lycopene is 1..1 ⁇ 2..1, stir the reaction at room temperature for 2-24h, filter, and purify by silica gel column.
  • Step 3 Prepare the step 2 product PPM-SS-VE in absolute ethanol solution at a concentration of 10 mg/mL to 40 mg/mL, slowly add dropwise to the double distilled water under stirring, spin dry to remove ethanol, and self-assemble.
  • Vitamin E-barocin conjugated nanoparticles PSSV-NPs
  • step 3 The above method for preparing vitamin E-barocin conjugated nanoparticles, wherein the organic solvent anhydrous ethanol in step 3 may be replaced by one or more mixed solvents selected from the group consisting of methanol, DMSO, acetonitrile and acetone.
  • the above preparation method of the vitamin E-barocidin conjugate nanoparticle, the purification method in the steps 1 and 2 is not limited to silica gel column purification, and can be purified by C8, C18 packed column chromatography or liquid phase preparation.
  • the preparation method of the above vitamin E-barocidin conjugate nanoparticle, the scutellarin described in step 2 may be other scorpion glycosides such as perilla aglycone, scorpion venom, squid glucoside or lycopene
  • the conjugated nanoparticles of vitamin E-barisin, scorpion venom, scallopin or leucine are prepared by replacing the steroid component.
  • a novel PEGylated vitamin E-baruin conjugated nanoparticle based on disulfide-SS-coupled vitamin E and periplocin, followed by methoxy Polyethylene glycol 2000-linoleic acid (PEG2000-LD) modified self-assembled PEGylated vitamin E-barocin conjugated nanoparticles, PEG2000-LD can pass ⁇ - ⁇ through hydrophobic end linoleic acid and vitamin E
  • the double force of the bond and the hydrophobic affinity is adsorbed on the surface of the PSSV-NPs, and the particle size of the PEGylated vitamin E-barocin conjugated nanoparticles ranges from 130 nm to 230 nm, and the zeta potential is -23.16— Between -28.42mV, the morphology of the nanoparticles was spherical and uniform, and the surface was PEGylated.
  • MPSSV-NPs novel PEGylated vitamin E-barocin conjugated nanoparticles
  • the scutellarin may be made of leucoside, scorpion venom, scallopin or leucine
  • the steroid-containing steroid component is substituted for the conjugated nanoparticles of PEGylated vitamin E-alloside, scorpion venom, scallopin or leucine.
  • Step 1 A certain amount of linoleic acid and mPEG, DCC and DMAP (the molar ratio of them are 2..1..2..1), add anhydrous dichloromethane, stir the reaction at room temperature for 12-24h, filter, use saturated chlorine After washing the methylene chloride layer with sodium chloride solution, it is dried over anhydrous sodium sulfate, and the organic layer is filtered, dried, and then washed with petroleum ether three times. The insoluble matter is filtered to obtain the desired product methoxy polyethylene glycol 2000-linoleic acid. (PEG2000-LD);
  • Step 2 Prepare the above-mentioned product PPM-SS-VE in absolute ethanol solution (10 mg/mL ⁇ 40 mg / mL), and add different amounts of the step 1 product PEG2000-LD, under stirring, will contain PEG2000-LD
  • the PPM-SS-VE anhydrous ethanol solution was slowly added dropwise to the double distilled water, and the ethanol was spin-dried to self-assemble to obtain a novel PEGylated vitamin E-baruin conjugated nanoparticles (MPSSV-NPs).
  • the vitamin E-barocin conjugated nanoparticle may be derived from vitamin E-alloside, vitamin E-quinoxin, Vitamin
  • the conjugated nanoparticles of the novel PEGylated vitamin E- lyrazine, scorpion venom, squid glucoside or leucovorin were prepared by replacing E-squid glucoside or vitamin E-barlitosine.
  • the molecular weight of mPEG in step (3) may be between 200 and 5000.
  • the invention further provides for the characterization of novel PEGylated vitamin E-barocyanine conjugate nanoparticles.
  • the particle size range of the nanoparticles was determined by dynamic light scattering method to be between 130 nm and 230 nm, and the zeta potential was between -23.16 and 28.42 mV.
  • the morphology of the nanoparticles was observed by projection electron microscopy. The shape of the nanoparticles was uniform and uniform, and the surface was PEGylated.
  • the present invention further provides studies on the stability and in vitro release characteristics of novel PEGylated vitamin E-barisin conjugate nanoparticles.
  • the invention further provides pharmacokinetic parameters of novel PEGylated vitamin E-barocin conjugated nanoparticles.
  • the circulation time of the nanoparticles in rats was significantly longer than that of the prototype drug.
  • the present invention further provides tissue distribution characteristics of novel PEGylated vitamin E-barocyanine conjugate nanoparticles.
  • the tissue distribution experiment was carried out with H22 solid tumor of liver cancer as a model. The results showed that the nanoparticles had obvious liver and tumor targeting.
  • the invention further provides the use of novel PEGylated vitamin E-barocin conjugated nanoparticles in the treatment of liver cancer.
  • In vivo anti-tumor activity experiments were carried out using H22 liver cancer solid tumors as a model. The experimental results show that the nanoparticles can significantly inhibit the growth of H22 solid tumors and can be used to treat liver cancer.
  • the present invention synthesizes a vitamin E-baruin conjugate, and it can self-assemble into nanoparticles (PSSV-NPs) in water, and the first modified PEGylated vitamin E- is modified by PEG2000-LD using PESV-NPs. Periplocin conjugated nanoparticles.
  • novel PEGylated vitamin E-barocin conjugated nanoparticles prepared by the invention have reduced particle size, improved stability, prolonged circulation time, increased tumor tissue distribution, and enhanced antitumor effect in vivo.
  • High-efficiency, low-toxicity and nano-targeted drug delivery of steroidal active ingredients provides new carriers.
  • Figure 1 is a 1 H-NMR spectrum of the target product (PPM-SS-VE);
  • Figure 2 is a 1H-NMR spectrum of the target product (mPEG2000-LD);
  • Figure 3 is a projection electron micrograph of PSSV-NPs and MPSSV-NPs
  • Figure 5 is an in vitro release experiment: cumulative release rate (A) and PPM-SS-VE of MPSSV-NP in different MPSSV-NPs in four media containing GSH (10 mM, 1 mM, 10 ⁇ M, and 1 ⁇ M GSH) Degradation rate of the complex (B);
  • Figure 6 is a plot of the in vivo drug dosage of PSSV-NPs and MPSSV-NPs after injection;
  • Figure 8 is a pharmacodynamic diagram of PSSV-NPs and MPSSV-NPs against H22 solid tumors of liver cancer: (A) a graph showing the change in volume growth of solid tumors after administration; (B) a graph of changes in body weight of mice after administration; (C) Solid tumor after 11 days of administration; mass of solid tumor after 11 days of administration (D).
  • VE-SS-COOH 120 mg
  • periplocin 100 mg
  • DCC dicyclohexylcarbodiimide
  • DMAP 4-dimethylaminopyridine
  • the nanoparticle was prepared by one-step nanoprecipitation method. The procedure was as follows: Prepare 20 mg/mL of PPM-SS-VE anhydrous ethanol solution, and slowly add 200 uL to 2 mL of double distilled water under stirring, and spin dry to remove ethanol to obtain PSSV. -NPs.
  • the novel PEGylated vitamin E-barocidin conjugate nanoparticles (MPSSV-NPs) are as above, except that different amounts of mPEG2000-LD are added to the PPM-SS-VE anhydrous ethanol solution (mass ratio is 1.. 0.1, 1..0.2, 1..0.4, 1..0.8).
  • the particle size and zeta potential of the nanoparticles were determined by dynamic light scattering using Bruker particle size and Zeta potential analyzer. Dilute the sample to the appropriate concentration with deionized water before assay. Take the appropriate amount of the nanoparticle solution, drop it on the copper grid, negatively stain the phosphotungstic acid for 1 min, blot the filter paper, dry it, and observe the appearance of the nanoparticle under the transmission electron microscope and take a picture, as shown in Figure 3, PSSV-NPs and MPSSV.
  • the morphology of the NPs nanoparticles is regular spherical and uniform, while the surface of MPSSV-NPs has a PEGylated shell.
  • the degradation rate of MPSSV-NPs at low concentrations of GSH (1 ⁇ M and 10 ⁇ M) was significantly lower than that at high concentrations (1 mM and 10 mM), and the release rate of PPM was significantly lower than that at high concentrations at low concentrations.
  • GSH concentration in the plasma environment is between 1 ⁇ M and 10 ⁇ M
  • the GSH concentration in the tumor environment is between 1 mM and 10 mM
  • MPSSV-NPs slowly release PPM in the plasma environment, allowing the nanoparticles to have sufficient time in the plasma. It accumulates in tumor tissue by EPR effect, and can rapidly release PPM after reaching the tumor environment, indicating that MPSSV-NPs have certain tumor targeting properties.
  • mice One week after the inoculation, the volume of the right subcutaneous subcutaneous tumor of the mouse was about 1 cm3, and the tumor formation rate was 100%, and the body weight of the mice was weighed one by one.
  • Thirty female H22 solid tumor ICR mice were randomly divided into groups. Five mice in each group were subjected to tissue distribution experiments. During the environmental adaptation period, they were fed normally with water. They were fasted for 12 hours before the experiment and were given free access to water.
  • PPM with a concentration of 0.8 mg/mL and PSPV-NPs and MPSSV-NPs of 1.66 mg/mL were prepared separately and administered intravenously at a dose of 4 mg/kg and 8.3 mg/kg, respectively.
  • the injection volume of the mouse weighing 20 g was 0.1 mL, and the injection volume was adjusted according to the actual body weight of the mouse.
  • the rats were sacrificed at 1, 2, and 4 hours after administration. Blood, heart, liver, spleen, lung, kidney, and tumor were taken. The mice were harvested from the eyeballs and allowed to stand for half an hour. After centrifugation at 6000 rpm for 10 minutes, the plasma was taken and placed at -20. °C refrigerator; accurately weighed amount of mouse organs, according to 0.2g ⁇ mL -1 0.9% NaCl solution was added, high-speed shearing machine crushed tissue sample, placed in -20 °C refrigerator. As a result, as shown in Fig. 7, MPSSV-NPs have certain liver and tumor targeting properties.
  • mice One week after inoculation, the diameter of the right subcutaneous subcutaneous tumor of the mouse was about 0.5 cm in diameter, and the tumor formation rate was 100%.
  • the body weight of the mice was weighed one by one, and they were divided into 4 groups by random number table, each group 6 only.
  • PPM group 4 mg/kg was injected into the tail vein of mice, and administered once every 2 days.
  • PSSV-NP group The mice were injected with 8.3 mg/kg in the tail vein and administered once every 2 days.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Epidemiology (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Molecular Biology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicinal Preparation (AREA)

Abstract

一种维生素E-杠柳次苷偶联物纳米粒，它是通过二硫醇二羟基乙酸作为连接剂，将杠柳次苷和维生素E进行偶联形成前体药物，该前体药物能在水介质中可自组装为纳米粒。还公开了一种新型PEG化维生素E-杠柳次苷偶联物纳米粒(MPSSV-NPs)及其制备方法，它是在二硫键-S-S-偶联维生素E和杠柳次苷的基础上，再以甲氧基聚乙二醇2000-亚油酸(PEG2000-LD)修饰的自组装PEG化维生素E-杠柳次苷偶联物纳米粒，该纳米粒有如下功效：1、该纳米前药能提高原型药杠柳次苷的体内外稳定性，延长体内循环时间；2、该纳米前药的释放和降解对谷胱甘肽(GSH)敏感；3具有显著的肝/肝癌靶向性；4，提高了原型药杠柳次苷对肝癌的治疗效果。

Description

一种PEG化维生素E-杠柳次苷偶联物纳米粒及其制法和用途 技术领域
本发明涉及维生素E-杠柳次苷偶联物纳米粒和新型PEG化维生素E-杠柳次苷偶联物纳米粒及其制备方法和作为抗肿瘤药物的用途。
背景技术
强心甾类固醇活性成分应用于肿瘤治疗始于中国的古代，当时记载着蟾蜍分泌物可用于治疗癌症。目前已发现蟾蜍分泌物中起抗肿瘤作用的成分为蟾毒灵，一种甲型强心甾类固醇成分。有研究报道，在因心脏病而使用洋地黄毒苷的患者中发现血液中洋地黄毒苷的浓度高低和血液及泌尿系统肿瘤发生率呈显著负相关性，且使用洋地黄毒苷的乳腺癌患者的死亡率明显低于未使用洋地黄毒苷的患者。目前强心甾类固醇活性成分主要作为强心剂应用于临床，而作为化疗药物用于肿瘤临床治疗的案例还没有报道，大部分强心甾类固醇抗肿瘤活性成分处于临床前研究，其中半合成药物UNBS-1450和PBI-05204已经在美国进入临床Ⅰ期用于治疗晚期耐药恶性肿瘤的研究。而将前体设计技术应用于具有抗肿瘤活性的强心甾类固醇成分的药物开发研究尚处于起步阶段。目前大部分研究工作者将目光关注于对蟾毒灵的研究，其中包括包载蟾毒灵的脂质体、壳聚糖纳米粒和PEG修饰的前体药物。
杠柳次苷(PPM)具有多种药理活性，如强心、抗炎、抗肿瘤等。目前PPM在临床应用的较少，局限于在细胞和动物水平的研究。有研究表明PPM的苷元具有明显体内外抑瘤效果，其效果与5-Fu相当，但有一定的不良反应如食欲减退、脱毛、抗拒等[参见：韩宇博和赵爱国，杠柳苷元的抗肿瘤作用研究[J].中国小儿血液与肿瘤杂志，2008，13，(1)：1-5]。对PPM的研究较少，体外细胞毒性评价显示其明显抑制PC3、U937、HCT-8、Bel-7402、BGC823、A549和A2780等肿瘤细胞的增殖(IC50值在0.02-0.29μM，显著高于杠柳苷元)(参见：Spera D,Siciliano T,De T N,et al.Planta Med.2007,73:384-387)。药动学研究表明PPM大鼠口服给药后在4h左右达到最大浓度，组织分布研究显示其在肝脏分布最多， 其次为结肠和心脏(参见：Yan K,Wang X,Jia Y,et al..Biomed Chromatogr.2015,30:1195-1201)。对于杠柳次苷的剂型研究还无见报道。
中国专利申请CN201010583339.0《一种杠柳次苷的制备方法》，是关于杠柳次苷的制备方法，未涉及到该药在抗肿瘤活性方面的应用。
发明内容
本发明的目的之一是，提供维生素E-杠柳次苷偶联物及其合成方法。
本发明的目的之二是，提供一种新型PEG化维生素E-杠柳次苷偶联物纳米粒及其制备方法。
本发明的目的之三是，提供所述的偶联物纳米粒在治疗肝癌中的应用。
本发明的技术方案如下：
一种维生素E-杠柳次苷偶联物纳米粒，它是以二硫键-S-S-偶联维生素E和杠柳次苷的自组装纳米粒。
上述的维生素E-杠柳次苷偶联物纳米粒，所述的杠柳次苷可以由杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷等其他强心甾类固醇成分所替代，得到的维生素E-杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷的偶联物纳米粒。
一种制备上述的维生素E-杠柳次苷偶联物纳米粒的方法，它包括下列步骤：
步骤1：称取一定量二硫醇二羟基乙酸加入到乙酸酐中，在30℃搅拌反应2-24h，反应后加入适量甲苯，加压旋干甲苯和乙酸酐得油状液体，加入无水二氯甲烷溶解，随后加入维生素E，维生素E与二硫醇二羟基乙酸的物质的量之比为10︰1～5︰1，以及加入适量的4-二甲氨基吡啶(DMAP)，DMAP与维生素E的摩尔比为1︰10，室温下搅拌反应12～36h，采用硅胶柱纯化，洗脱剂为正己烷/乙酸乙酯/乙酸＝10/1/1，得产物VE-S-S-COOH；
步骤2：称取步骤1的产物VE-S-S-COOH和杠柳次苷，VE-S-S-COOH和杠柳次苷的摩尔比为1︰1～3︰1，加入无水二氯甲烷溶解后，加入DCC和DMAP，它们与和杠柳次苷的摩尔比为1︰1～2︰1，室温搅拌反应2-24h，过滤，采用硅胶柱纯化，洗脱剂为石油醚/乙酸乙酯＝2/3，收集目标产物PPM-S-S-VE，即为维生素E-杠柳次苷偶联物；
步骤3：配制步骤2产物PPM-S-S-VE的无水乙醇溶液，浓度为10mg/mL～40mg/mL，在搅拌条件下，慢慢滴加入到双蒸水中，旋干除去乙醇，自组装得 维生素E-杠柳次苷偶联物纳米粒(PSSV-NPs)。
上述制备维生素E-杠柳次苷偶联物纳米粒的方法，步骤3中有机溶剂无水乙醇可以选自甲醇、DMSO、乙腈、丙酮中的一种或者多种混合溶剂所替代。
上述的维生素E-杠柳次苷偶联物纳米粒的制备方法，步骤1和2中的纯化方法不限于硅胶柱纯化，可以采用C8、C18填料柱层析或液相制备纯化。
上述的维生素E-杠柳次苷偶联物纳米粒的制备方法，步骤2所述的杠柳次苷可以由杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷等其他强心甾类固醇成分所替代制得维生素E-杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷的偶联物纳米粒。
一种新型PEG化维生素E-杠柳次苷偶联物纳米粒(MPSSV-NPs)，它是在二硫键-S-S-偶联维生素E和杠柳次苷的基础上，再以甲氧基聚乙二醇2000-亚油酸(PEG2000-LD)修饰的自组装PEG化维生素E-杠柳次苷偶联物纳米粒，PEG2000-LD能通过疏水端亚油酸与维生素E通过π-π键和疏水性相亲的双重作用力吸附在PSSV-NPs表面，所述的PEG化维生素E-杠柳次苷偶联物纳米粒的粒径范围为130nm-230nm之间，zeta电位为-23.16—-28.42mV之间，投射电镜观察纳米粒形态呈规则球形，分散均匀，表面有PEG化外壳。
上述的新型PEG化维生素E-杠柳次苷偶联物纳米粒(MPSSV-NPs)，所述的杠柳次苷可以由杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷等其他强心甾类固醇成分所替代得到PEG化维生素E-杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷的偶联物纳米粒。
一种制备上述的新型PEG化维生素E-杠柳次苷偶联物纳米粒的方法，
步骤1：将一定量亚油酸与mPEG、DCC和DMAP(它们的摩尔比为2︰1︰2︰1)，加入无水二氯甲烷，常温下搅拌反应12-24h，过滤，用饱和氯化钠溶液洗二氯甲烷层后，加入无水硫酸钠干燥，过滤有机层，旋干后加入石油醚洗三次，过滤得不溶物即为目标产物甲氧基聚乙二醇2000-亚油酸(PEG2000-LD)；
步骤2：配制上述的产物PPM-S-S-VE的无水乙醇溶液(10mg/mL～40mg/mL)，并加入不同量的步骤1产物PEG2000-LD，在搅拌条件下，将含有PEG2000-LD的PPM-S-S-VE的无水乙醇溶液慢慢滴加入到双蒸水中，旋干除去乙醇，自组装得新型PEG化维生素E-杠柳次苷偶联物纳米粒(MPSSV-NPs)。
上述制备新型PEG化维生素E-杠柳次苷偶联物纳米粒的方法，步骤(2)中所述的维生素E-杠柳次苷可以由维生素E-杠柳苷元、维生素E-蟾毒灵、维生素 E-海葱次苷或维生素E-杠柳毒苷等所替代，制得新型PEG化维生素E-杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷的偶联物纳米粒。
上述制备新型PEG化维生素E-杠柳次苷偶联物纳米粒的方法，步骤(3)中mPEG的分子量可以是200-5000之间。
本发明进一步提供了新型PEG化维生素E-杠柳次苷偶联物纳米粒的表征。动态光散射法测定纳米粒的粒径范围为130nm-230nm之间，zeta电位为-23.16—-28.42mV之间。投射电镜观察纳米粒形态呈规则球形，分散均匀，表面有PEG化外壳。
本发明进一步提供了新型PEG化维生素E-杠柳次苷偶联物纳米粒的稳定性和体外释放特性研究。一个月内该纳米粒在水和PBS(pH＝7.4)介质中的粒径无明显变化。该纳米粒在PBS(pH＝7.4)介质中药物释放和降解速率随着谷胱甘肽浓度的升高而提高，具有一定的氧化还原感应性。
本发明进一步提供了新型PEG化维生素E-杠柳次苷偶联物纳米粒的药动学参数。该纳米粒在大鼠体内的循环时间明显长于原型药。
本发明进一步提供了新型PEG化维生素E-杠柳次苷偶联物纳米粒的组织分布特性。以H22肝癌实体瘤为模型进行组织分布实验，结果表明，该纳米粒明显具有肝脏和肿瘤靶向性。
本发明进一步提供了新型PEG化维生素E-杠柳次苷偶联物纳米粒在治疗肝癌中的应用。以H22肝癌实体瘤为模型进行体内抗肿瘤活性实验。实验结果表明，该纳米粒可以明显抑制H22肝癌实体瘤的生长，可用于治疗肝癌。
有益效果
(1)本发明合成维生素E-杠柳次苷偶联物，且其在水中能自组装成纳米粒(PSSV-NPs)，并首次采用PEG2000-LD修饰PSSV-NPs得到新型PEG化维生素E-杠柳次苷偶联物纳米粒。
(2)本发明制备得到新型PEG化维生素E-杠柳次苷偶联物纳米粒的粒径降低、稳定性提高、体内循环时间延长、肿瘤组织分布提高、体内抑瘤效果增强，为强心甾类固醇活性成分高效低毒和纳米化靶向给药提供了新载体。
附图说明
图1为目标产物(PPM-S-S-VE)的1H-NMR图谱；
图2为目标产物(mPEG2000-LD)的1H-NMR图谱；
图3为PSSV-NPs和MPSSV-NPs的投射电镜图；
图4为PSSV-NPs和不同比例(质量比为1︰0、1︰0.1、1︰0.2、1︰0.4、1︰0.8)的MPSSV-NPs在水介质(A)和pH＝7.4的PBS(B)介质中的稳定性考察；
图5为体外释放实验：MPSSV-NP在不同MPSSV-NP在含GSH的四种介质(10mM、1mM、10μM、和1μM GSH)中的PPM的累计释放率(A)和PPM-S-S-VE偶联物的降解率(B)；
图6为PSSV-NPs和MPSSV-NPs大鼠注射给药后体内药时曲线；
图7为PSSV-NPs和MPSSV-NPs尾静脉注射给药后各个时间点后血浆及各个脏器中药物含量(μg/g)(n＝5)；
图8为PSSV-NPs和MPSSV-NPs对H22肝癌实体瘤的药效图：(A)给药后实体瘤体积生长变化曲线图；(B)给药后小鼠的体重变化图；(C)给药11天后的实体瘤；给药11天后的实体瘤的质量(D)。
具体实施方式
以下所列实施例有助于本领域技术人员更好地理解本发明，但不以任何方式限制本发明。
实施例1.维生素E-杠柳次苷偶联物的制备
称取二硫醇二羟基乙酸(0.2g,1.10mmol)加入到3mL乙酸酐中，在30℃搅拌反应2h，反应后加入适量甲苯，加压旋干甲苯和乙酸酐得油状液体，加入2mL无水二氯甲烷溶解，随后加入VE(0.1g,0.23mmol)和适量DMAP，室温下搅拌反应5min，采用硅胶柱纯化，洗脱剂为正己烷/乙酸乙酯/乙酸＝10/1/1，得产物VE-S-S-COOH(114mg)。
称取VE-S-S-COOH(120mg)和杠柳次苷(100mg)，加入6mL无水二氯甲烷溶解后，加入二环己基碳二亚胺(DCC)(48mg)和4-二甲氨基吡啶(DMAP)(30mg)，室温搅拌反应2h，过滤除去二环己基脲(DCU)，采用硅胶柱纯化，洗脱剂为石油醚/乙酸乙酯＝2/3，收集目标产物PPM-S-S-VE(80mg，得率为37.6％)。
上述目标产物采用质谱和1H-NMR进行表征，如图1所示。
实施例2.mPEG2000-LD的合成、纯化和表征
称取亚油酸(500mg)、mPEG(100mg)、DCC(400mg)和DMAP(240mg)，加入20mL无水二氯甲烷，搅拌反应12h，过滤除去DCU，用饱和氯化钠溶液洗二氯甲烷层后，加入无水硫酸钠干燥，过滤有机层，旋干后加入石油醚洗三次，过滤得不溶物即为目标产物PEG2000-LD，采用1H-NMR进行表征，如图2所示。
实施例3.PPM-S-S-VE纳米粒(PSSV-NPs)和新型PEG化维生素E-杠柳次苷偶联物纳米粒(MPSSV-NPs)的制备
采用一步纳米沉淀法制备纳米粒，步骤如下：配制PPM-S-S-VE无水乙醇溶液20mg/mL，在搅拌条件下，吸取200uL慢慢滴加入到2mL双蒸水中，旋干除去乙醇，得PSSV-NPs。新型PEG化维生素E-杠柳次苷偶联物纳米粒(MPSSV-NPs)步骤如上，不同的是在PPM-S-S-VE无水乙醇溶液中加入不同含量的mPEG2000-LD(质量比为1︰0.1、1︰0.2、1︰0.4、1︰0.8)。
实施例4.PSSV-NPs和新型PEG化维生素E-杠柳次苷偶联物纳米粒(MPSSV-NPs)的表征
采用bruke粒径与Zeta电位分析仪，以动态光散射法测定纳米粒的粒径与Zeta电位。测定前用去离子水稀释样品至适当浓度。取纳米粒溶液适量，滴加在铜网上，磷钨酸负染1min，滤纸吸干，干燥后置于透射电镜下观察纳米粒的外观形态并拍照，如图3所示，PSSV-NPs和MPSSV-NPs纳米粒形态均呈规则球形，分散均匀，而MPSSV-NPs表面有PEG化外壳。
实施例5稳定性试验
制备不同质量比例PEG化的MPSSV-NPs(1︰0.1、1：0.2、1︰0.4、1︰0.8)，分别分散在水、PBS(pH＝7.4)的介质中，常温放置，在时间点第1、2、3、5、10、15、20、25、30天时测量粒径，评价其稳定性。如图4所示，mPEG-LD修饰的MPSSV-NPs不仅能提高其在PBS(pH＝7.4)中的稳定性，而且PEG化后 能提高其在体内的循环时间。
实施例6体外释放
将100uL的浓度为1mg/mL的MPSSV-NPs放入到介质2.5mL的不同浓度GSH(1μM、10μM、1mM、10mM)的PBS(pH＝7.4)当中，放置在37℃中水浴震荡，转速为100rpm，在时间点0.5h、1h、2h、4h、8h、12h、24h时取100ul，加300uL甲醇溶解，液相测定PPM和PPM-S-S-VE的含量，计算释放率。如图5所示，MPSSV-NPs在GSH低浓度时(1μM和10μM)的降解速率明显低于高浓度(1mM和10mM))，且在低浓度时PPM的释放率明显低于高浓度时，而体内血浆环境的GSH浓度处于1μM-10μM之间，而肿瘤环境的GSH浓度处于1mM-10mM之间，因此MPSSV-NPs在体内血浆环境缓慢释放PPM，使纳米粒在体内血浆中有足够的时间通过EPR效应而聚集在肿瘤组织中，而到达肿瘤环境后能快速释放PPM，表明MPSSV-NPs具有一定的肿瘤靶向性。
实施例7药动学实验
15只健康雄性SD大鼠在适应环境三天后进行实验。随机分为3组，每组5只大鼠，给药前禁食12小时，自由饮水。按4mg·kg-1的剂量分别尾静脉注射杠柳次苷、PSSV-NPs和MPSSV-NPs溶液。于给药后0.25、0.5、1、1.5、2、4、6、8、12、24h大鼠眼球后丛静脉采血约0.5mL，置肝素抗凝管中，4000r·min-1离心10min分离血浆，-20℃下冷冻存储直至分析。如图6所示，mPEG-LD修饰的PSSV-NPs能显著提高其在体内血浆中的滞留时间
实施例8在H22实体瘤小鼠中体内组织分布实验
接种1周后，小鼠右侧腋下皮下瘤体积长到约1cm3左右大小，成瘤率为100％，逐个称取小鼠体重。30只雌性H22实体瘤ICR小鼠随机分组，每组5只小鼠，进行组织分布实验，环境适应期间正常进水进食，于实验前12小时禁食，自由饮水。分别制备浓度为0.8mg/mL的PPM及1.66mg/mL的纳米前药PSSV-NPs和MPSSV-NPs，尾静脉注射给药，给药剂量分别为4mg/kg和8.3mg/kg。体重为20g的小鼠注射体积为0.1mL，按小鼠实际体重调整注射体积。 于给药后1、2和4小时处死，取血、心、肝、脾、肺、肾、瘤，小鼠摘眼球取血，静置半小时以后，6000rpm离心10min取血浆，置于-20℃冰箱保存；精密称取一定量小鼠各脏器，按0.2g·mL-1加入0.9％NaCl溶液，高速剪切机捣碎组织样品，置于-20℃冰箱保存。结果如图7所示，MPSSV-NPs具有一定肝和肿瘤靶向性。
实施例9纳米前药在H22实体瘤小鼠中药效学研究
接种1周后，小鼠右侧腋下皮下瘤直径长到约0.5cm左右大小，成瘤率为100％，逐个称取小鼠体重，用随机数字表法将其分成4组，每组6只。
(1)空白模型组:小鼠尾静脉注射PBS(pH＝7.4)5mL/kg，每2天给药1次。
(2)PPM组:小鼠尾静脉注射4mg/kg，每2天给药1次。
(3)PSSV-NP组:小鼠尾静脉注射8.3mg/kg，每2天给药1次。
(4)MPSSV-NP组:小鼠尾静脉注射8.3mg/kg，每2天给药1次。
结果如图8所示，表明PPM、PSSV-NPs和MPSSV-NPs能显著抑制肿瘤的生长，而MPSSV-NPs的抑制作用最强。

Claims (11)

  1. 一种维生素E-杠柳次苷偶联物纳米粒，其特征是：维生素E-杠柳次苷偶联物是通过二硫醇二羟基乙酸作为连接剂，将杠柳次苷和维生素E进行偶联形成前体药物，该前体药物能在水介质中可自组装为纳米粒。
  2. 根据权利要求1所述的维生素E-杠柳次苷偶联物纳米粒，其特征是：所述的杠柳次苷由杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷所替代，得到的维生素E-杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷的偶联物纳米粒。
  3. 一种制备权利要求1所述的维生素E-杠柳次苷偶联物纳米粒的方法，其特征是它包括下列步骤：
    步骤1：称取一定量二硫醇二羟基乙酸加入到乙酸酐中，在30℃搅拌反应2-24h，反应后加入适量甲苯，加压旋干甲苯和乙酸酐得油状液体，加入无水二氯甲烷溶解，随后加入维生素E，维生素E与二硫醇二羟基乙酸的物质的量之比为10︰1～5︰1，以及加入适量的4-二甲氨基吡啶(DMAP)，DMAP与维生素E的摩尔比为1︰10，室温下搅拌反应12～36h，采用硅胶柱纯化，洗脱剂为正己烷/乙酸乙酯/乙酸＝10/1/1，得产物VE-S-S-COOH；
    步骤2：称取步骤1的产物VE-S-S-COOH和杠柳次苷，VE-S-S-COOH和杠柳次苷的摩尔比为1︰1～3︰1，加入无水二氯甲烷溶解后，加入二环己基碳二亚胺(DCC)和DMAP，它们与和杠柳次苷的摩尔比为1︰1～2︰1，室温搅拌反应2-24h，过滤，采用硅胶柱纯化，洗脱剂为石油醚/乙酸乙酯＝2/3，收集目标产物PPM-S-S-VE，即为维生素E-杠柳次苷偶联物；
    步骤3：配制步骤2产物PPM-S-S-VE的无水乙醇溶液，浓度为10mg/mL～40mg/mL，在搅拌条件下，慢慢滴加入到双蒸水中，旋干除去乙醇，自组装得维生素E-杠柳次苷偶联物纳米粒(PSSV-NPs)。
  4. 根据权利要求3所述制备维生素E-杠柳次苷偶联物纳米粒的方法，其特征是：步骤3中所述的有机溶剂无水乙醇以甲醇、DMSO、乙腈或丙酮中的一种或者多种的混合溶剂所替代。
  5. 根据权利要求3所述的维生素E-杠柳次苷偶联物纳米粒的制备方法，其特征是：步骤1和2中的纯化方法采用C8或C18填料柱层析或液相制备纯化。
  6. 根据权利要求3所述的维生素E-杠柳次苷偶联物纳米粒的制备方法，其特征是：步骤2所述的杠柳次苷由杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷所替代制得维生素E-杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷的偶联物纳米粒。
  7. 一种新型PEG化维生素E-杠柳次苷偶联物纳米粒，其特征是：它是在二硫键-S-S-偶联维生素E和杠柳次苷的基础上，再以甲氧基聚乙二醇2000-亚油酸(PEG2000-LD)修饰的自组装PEG化维生素E-杠柳次苷偶联物纳米粒，投射电镜观察纳米粒形态呈规则球形，分散均匀，表面有PEG化外壳，所述的PEG化维生素E-杠柳次苷偶联物纳米粒的粒径范围为130nm-230nm之间，zeta电位为-23.16—-28.42mV之间。
  8. 根据权利要求7所述的新型PEG化维生素E-杠柳次苷偶联物纳米粒，其特征是：所述的杠柳次苷由杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷所替代得到PEG化维生素E-杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷的偶联物纳米粒。
  9. 一种制备权利要求7上述的新型PEG化维生素E-杠柳次苷偶联物纳米粒的方法，其特征是它包括如下步骤：
    步骤1：将一定量亚油酸与聚乙二醇(mPEG)、DCC和DMAP，加入无水二氯甲烷，(mPEG)、DCC和DMAP的摩尔比为2︰1︰2︰1，常温下搅拌反应12-24h，过滤，用饱和氯化钠溶液洗二氯甲烷层后，加入无水硫酸钠干燥，过滤有机层，旋干后加入石油醚洗三次，过滤得不溶物即为目标产物PEG2000-LD，所述的mPEG的数均分子量是200-5000之间；
    步骤2：配制上述的产物PPM-S-S-VE的无水乙醇溶液，浓度为10mg/mL～40mg/mL，并加入不同量的步骤1产物PEG2000-LD，在搅拌条件下，将含有PEG2000-LD的PPM-S-S-VE的无水乙醇溶液慢慢滴加入到双蒸水中，旋干除去乙醇，自组装得新型PEG化维生素E-杠柳次苷偶联物纳米粒(MPSSV-NPs)。
  10. 根据权利要求9所述的制备新型PEG化维生素E-杠柳次苷偶联物纳米粒的方法，其特征是：步骤(2)中所述的维生素E-杠柳次苷由维生素E-杠柳苷元、维生素E-蟾毒灵、维生素E-海葱次苷或维生素E-杠柳毒苷所替代，制得新型PEG化维生素E-杠柳苷元、蟾毒灵、海葱次苷或杠柳毒苷的偶联物纳米粒。
  11. 权利要求1所述的维生素E-杠柳次苷偶联物纳米粒或权利要求7所述的新型PEG化维生素E-杠柳次苷偶联物纳米粒在制备抗肿瘤药物中的应用。
PCT/CN2016/108628 2016-12-06 2016-12-06 一种peg化维生素e-杠柳次苷偶联物纳米粒及其制法和用途 Ceased WO2018102973A1 (zh)

Priority Applications (2)

Application Number Priority Date Filing Date Title
PCT/CN2016/108628 WO2018102973A1 (zh) 2016-12-06 2016-12-06 一种peg化维生素e-杠柳次苷偶联物纳米粒及其制法和用途
CN201680001826.7A CN107735108A (zh) 2016-12-06 2016-12-06 一种peg化维生素e‑杠柳次苷偶联物纳米粒及其制法和用途

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2016/108628 WO2018102973A1 (zh) 2016-12-06 2016-12-06 一种peg化维生素e-杠柳次苷偶联物纳米粒及其制法和用途

Publications (1)

Publication Number Publication Date
WO2018102973A1 true WO2018102973A1 (zh) 2018-06-14

Family

ID=61201131

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2016/108628 Ceased WO2018102973A1 (zh) 2016-12-06 2016-12-06 一种peg化维生素e-杠柳次苷偶联物纳米粒及其制法和用途

Country Status (2)

Country Link
CN (1) CN107735108A (zh)
WO (1) WO2018102973A1 (zh)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116650443A (zh) * 2023-06-01 2023-08-29 常州大学 一种还原响应型载药纳米粒及其制备方法

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111494641B (zh) * 2020-04-22 2021-08-03 南开大学 肿瘤微环境响应性的表面电荷可反转纳米药物递送载体
CN114904013A (zh) * 2021-02-08 2022-08-16 中国科学院上海药物研究所 硫酸乙酰肝素-骨化三醇偶联物，包含其的乙酰肝素酶响应性纳米粒，其制备方法及用途
CN115624631B (zh) * 2021-12-14 2024-10-01 盐城工学院 一种蟾毒灵前药自组装纳米粒及其制备方法与应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1850847A (zh) * 2006-05-10 2006-10-25 天津中医药大学 一种杠柳苷元的制备方法
CN102154417A (zh) * 2010-12-13 2011-08-17 天津中医药大学 一种杠柳次苷的制备方法
CN103288911A (zh) * 2013-06-04 2013-09-11 暨南大学 蟾毒灵糖基化衍生物及其制法和在制备抗肿瘤药物中的用途
CN105687251A (zh) * 2016-01-21 2016-06-22 中国人民解放军第四军医大学 蟾酥提取物在制备治疗人脑胶质瘤药物中的应用

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104667292B (zh) * 2015-02-02 2017-09-05 北京大学 一种还原响应型药物偶联物纳米粒的制备及其应用

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1850847A (zh) * 2006-05-10 2006-10-25 天津中医药大学 一种杠柳苷元的制备方法
CN102154417A (zh) * 2010-12-13 2011-08-17 天津中医药大学 一种杠柳次苷的制备方法
CN103288911A (zh) * 2013-06-04 2013-09-11 暨南大学 蟾毒灵糖基化衍生物及其制法和在制备抗肿瘤药物中的用途
CN105687251A (zh) * 2016-01-21 2016-06-22 中国人民解放军第四军医大学 蟾酥提取物在制备治疗人脑胶质瘤药物中的应用

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116650443A (zh) * 2023-06-01 2023-08-29 常州大学 一种还原响应型载药纳米粒及其制备方法

Also Published As

Publication number Publication date
CN107735108A (zh) 2018-02-23

Similar Documents

Publication Publication Date Title
Yi et al. Cytotoxic effect of novel Flammulina velutipes sterols and its oral bioavailability via mixed micellar nanoformulation
CN108478803A (zh) 氧化还原超敏感二硫键桥连前药自组装纳米粒的构建
JP2002535242A (ja) 新規な粒子状剤型
CN111632032A (zh) 一种天然小分子共组装纳米药物传输系统及其制备方法与应用
CN111484501A (zh) 羟基喜树碱亚油酸酯小分子前药及其自组装纳米粒的构建
CN113264906B (zh) 多西他赛二聚体小分子前药及其自组装纳米粒的构建
Xu et al. Cabazitaxel prodrug nanoassemblies with branched chain modifications: Narrowing the gap between efficacy and safety
Zhou et al. A facile and universal method to achieve liposomal remote loading of non-ionizable drugs with outstanding safety profiles and therapeutic effect
WO2018102973A1 (zh) 一种peg化维生素e-杠柳次苷偶联物纳米粒及其制法和用途
Cheng et al. Improve bile duct-targeted drug delivery and therapeutic efficacy for cholangiocarcinoma by cucurbitacin B loaded phospholipid complex modified with berberine hydrochloride
KR101429668B1 (ko) 양친성 저분자량 히알루론산 복합체를 포함하는 나노 입자 및 그의 제조 방법
CN111097052A (zh) 用于肿瘤主动靶向治疗的两亲性前药及其纳米颗粒的制备方法、应用
CN104826122A (zh) 绿原酸及其衍生物的脂质修饰物及其制备和纯化方法
CN104045823B (zh) 一种甘草次酸衍生物及其制备方法和应用
Xu et al. The design and synthesis of redox-responsive oridonin polymeric prodrug micelle formulation for effective gastric cancer therapy
KR101332001B1 (ko) 양친성 저분자량 히알루론산 복합체를 포함하는 나노 입자 및 그의 제조 방법
JP2023526707A (ja) カバジタキセル弱塩基性誘導体およびその製剤
CN104826118A (zh) 绿原酸及其衍生物脂质修饰物的应用
CN102716080B (zh) 含异穿心莲内酯固体脂质纳米粒的混悬液及其制备方法和应用
CN119528925B (zh) 喜树碱类前药及其载药脂质体和应用
CN115212314A (zh) 一种白蛋白结合型抗肿瘤药及其纳米复合物、制备方法和应用
CN110772644B (zh) 聚乙二醇修饰的强心苷类化合物前药及其抗肿瘤用途
CN103127522B (zh) 一种脂肪酸-姜黄素衍生物的缀合物及其应用
CN105777769B (zh) 一种7-乙基-10-羟基喜树碱的衍生物及其制备方法与应用
WO2018102974A1 (zh) 一种天然活性成分强心甾类固醇与奥曲肽的偶联物及其制法和用途

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 16923236

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 16923236

Country of ref document: EP

Kind code of ref document: A1