CN109734768B - Deacetylated cedilanid glucose-based modified compound liposome and application thereof - Google Patents
Deacetylated cedilanid glucose-based modified compound liposome and application thereof Download PDFInfo
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Abstract
The invention discloses a deacetylated cedilanid glucose-based modified compound liposome and application thereof. The method is characterized in that the method is mainly characterized in that the method is modified from the glucose group of the cedilanid, partial modification of molecules is carried out by taking glucose and 2-aminoglucose as raw materials, then the modified molecules are connected with the 3-hydroxyl at the tail end of digoxin to generate a glucose group modified product of the deacetylcedilanid, and the molecules meeting the requirements are selected under the activity guidance. The anticancer effect of the compound is enhanced by synthesizing deacetylated cedilan glucose-based modified compound (2-O-benzyl-beta-D-glucopyranoside-deacetylated cedilan, etc.), the potency strength is improved, the half-life period is prolonged, and meanwhile, the modified compound is prepared into liposome, the liver targeting effect of the liposome is improved, and the cardiac toxicity is reduced.
Description
Technical Field
The invention relates to a modified compound, in particular to a new compound obtained by chemically modifying a traditional heart failure treatment drug deacetyl cedilan, and an application of the new compound in anticancer after modification of a liposome formulation, namely, the deacetyl cedilan is chemically modified into a deacetyl cedilan glucose-based modified compound (2-O-benzyl-beta-D-glucopyranoside-deacetyl cedilan) and is prepared into an application of a liver targeting liposome in an anti-liver cancer drug.
Background
In common cancers all over the world, the incidence rate of liver cancer ranks seventh, the incidence of liver cancer in China exceeds 50% of the total number of the whole world, and the mortality rate of liver cancer is not reduced even if treatment methods including tumor resection, liver transplantation, sorafenib, local treatment (radio frequency ablation) and the like are adopted, so that the significance of developing high-efficiency low-toxicity anti-liver cancer medicaments is great.
Cardiac glycosides are a class of drugs with selective cardiotonic action, the molecules of which are composed of an alcohol or alcohol-like group (ligand, aglycone or aglycone) bound to a varying number of sugar molecules. If the ligand contains steroid nucleus (steroid nucleus) with 17-carbon atoms connected to an unsaturated lactone ring and 3-carbon atoms connected to sugar molecules, the glycoside is a cardiac glycoside. It is of a wide variety including plant and vertebrate origin. Cardiac glycosides bind specifically to eukaryotic cell membrane sodium potassium atpase and have been used for over 200 years in the treatment of congestive heart failure and cardiac arrhythmias. In recent years, cardiac glycosides have been paid more and more attention to the prevention and treatment of tumors. More optimistically, many studies have now demonstrated that it has a preferential selective killing effect on malignant cells, i.e. does not affect the proliferation of normal cells. In view of this, it has become possible to make cardiac glycosides a new type of drug for the targeted treatment of tumors.
In vitro experiments found that cardiac glycosides were not effective against rodent tumor cells. Similarly, it has been difficult to demonstrate how isogenic animals act on cardiac glycosides. However, increasing research has demonstrated that cardiac glycosides are extremely sensitive to human tumor cell lines. Such as ouabain and bufalin. Han Ke-Qi and the like establish a nude mouse model of in-situ transplantation of a human hepatoma cell strain (BEL-7402), and are subjected to Bufarin abdominal cavity treatment. They found that the cardiac glycoside of this toad derivative significantly reduced tumor volume and extended animal life, and no damage to the heart, lungs, brain, and kidneys was found in preliminary pathological studies. An interesting report on the treatment of orthotopic non-small cell lung cancer (a549) animals with the semi-synthetic enolide unws-1450. UNBS-1450 is cardiac glycoside compound prepared from extract of Calotropis gigantea of African plant by semi-artificial synthesis. The Mijatovic T and the like orally take UNBS-1450 on the transplanted non-small cell lung cancer animal model for a long time, and the curative effect is obvious. In addition, in vitro experiments demonstrated that digoxin inhibits the growth of human neurofibroma cells. And also successfully reduced the mass transplanted to the mice in vivo animal experiments.
Cardiac glycosides have been found to be useful in the prevention or treatment of malignant diseases through complex cellular signal transduction mechanisms. Mijatovic T and the like carry out chemical structure modification on cardiac glycoside, successfully remove the effect of the cardiac glycoside on cardiac muscle, and find that the compounds without cardiac activity cause T lymphoblast to show time-dependent apoptosis. The activity of antitumor cells is enhanced. But has little effect on Peripheral Blood Mononuclear Cells (PBMC). The cardiac glycoside is expected to be transformed into a specific anti-tumor medicament without cardiotoxicity. The cardiac glycoside used as a traditional cardiac medicine and known chemotherapeutic medicines such as paclitaxel, carboplatin, adriamycin, vincristine, cyclophosphamide and the like have different anticancer mechanisms, and if the cardiac glycoside and the known chemotherapeutic medicines can be used together with the chemotherapeutic medicines, the effects of synergy, reduction of adverse reactions and even resistance to cell drug resistance can be produced. Even though it is still in the in vitro or animal experimental stage, the future therapeutic prospects for tumors are undoubtedly optimistic. Recent research finds that the combination of the cedilanid and tumor necrosis factor has the effect of resisting glioblastoma, the liver cancer in vitro has the effect of resisting liver cancer, the state of a cancer suppressor gene PTEN is related to apoptosis caused by protein kinase C delta, and in early research work, the early research finds that the cardiac glycoside drug deacetylated cedilanid (British name: Deslanoside, also known as deacetylated hairy flower glycoside, deacetylated hairy flower glycoside C) has strong selective liver cancer resisting effect in vitro and has the IC (half inhibitory concentration) IC (IC) effect on SMMC-7721 human liver cancer cells500.18. mu.g/ml, IC of human normal human liver cell line LO25075.70. mu.g/ml, differing by a factor of 420. However, deacetyl cedilan (cedilan) can lose glucose and acetic acid in vivo by hydrolysis to form digoxin with almost no anti-liver cancer effect, so that the potency of the in vivo anti-cancer activity of deacetyl cedilan is reduced and the time (time) for maintaining the effect is shortened. In addition, cardiac glycosides are at greater risk for anticancer use due to their ubiquitous cardiotoxicity.
Disclosure of Invention
The invention aims to provide deacetylated cedilanid glucose-based modified compound liposome and application thereof, aiming at the defect that deacetylated cedilanid can lose glucose and acetic acid in vivo through hydrolysis and becomes digoxin with almost no anti-liver cancer effect. The anticancer effect of the compound is enhanced by synthesizing deacetylated cedilan glucose-based modified compound (2-O-benzyl-beta-D-glucopyranoside-deacetylated cedilan, etc.), the potency strength is improved, the half-life period is prolonged, and meanwhile, the modified compound is prepared into liposome, the liver targeting effect of the liposome is improved, and the cardiac toxicity is reduced.
One of the objects of the present invention is: discloses a synthesis method of deacetylated cilaria glucosyl modified compounds (2-O-benzyl-beta-D-glucopyranoside-deacetylated cilaria, etc.).
The second purpose of the invention is that: discloses a preparation method of a deacetylated cilaria glucose-based modified compound liposome preparation.
The third purpose of the invention is: discloses application of a deacetylated cilaria glucose-based modified compound liposome preparation in an anti-liver cancer medicament.
The structural formula of the deacetyl cedilanid is as follows:
since the glucose group of deacetyl cedilan is easily hydrolyzed by enzyme, hydrolyzed to digoxin to lose the activity thereof, and digoxin is highly toxic, it becomes important to maintain the stability of glucose group. Because the activity of deacetyl cedilanid is closely related to the molecular structure, the modification of partial structure is supposed to be carried out under the condition of keeping the main structure of the molecule unchanged, thereby achieving the aim of maintaining the stable existence of glucosyl to keep or improve the molecular activity. The method is mainly characterized in that glucose group is modified, glucose and 2-aminoglucose are used as raw materials to perform partial modification of molecules, and then the modified molecules are connected with terminal 3-hydroxyl of digoxin to generate a glucose group modified product of deacetyl cedilan, and the molecules meeting the requirements are selected under the activity guidance.
The synthesis of the deacetylated cedilanid glucosyl modified compound is specifically illustrated by the synthesis example of 2-O-benzyl-beta-D-glucopyranoside-deacetylated cedilanid:
(1) the structural formula of the 2-O-benzyl-beta-D-glucopyranoside-deacetyl cedilanid is as follows:
(2) the synthetic route is as follows:
(3) the specific synthesis steps are as follows:
synthesis of compound 1: under ice bath, 140ml of methanol and 2.4 ml of acetyl chloride are added into a dry reaction bottle provided with a drying tube, stirred for 5 minutes, naturally stirred to room temperature and stirred for 20 minutes, 10 g (54.6mmol) of glucose is added, and the mixture is heated to reflux for reaction for 2 hours. Cooling the reaction solution to room temperature, adding anhydrous potassium carbonate to neutralize to be neutral, filtering, and concentrating to obtain a crude product of the compound 1 (1-O-methyl glucoside);
synthesis of compound 2: dissolving the obtained crude product of the compound 1 in 34 ml of DMF, dropwise adding 16 ml (105.2mmol) of benzaldehyde, quickly adding 1.72 g (9mmol) of p-toluenesulfonic acid, reacting overnight at 50 ℃, wherein TLC shows that the reaction is complete, pouring the reactant into a mixed solution of 100 ml of ice water and 140ml of petroleum ether, violently shaking, separating out a milky solid, performing suction filtration, and recrystallizing with absolute ethyl alcohol to obtain a compound 2;
synthesis of compound 3: the obtained compound 2 and Bu2SnO 14 g (44.8mmol) and toluene 180 ml are added into a round bottom flask provided with a water separator, reflux reaction is carried out for 3-4 hours until no water is separated out, the reaction solution is concentrated to 60 ml, anhydrous DMF30 ml is added, 7 ml (58.4mmol) benzyl bromide is added dropwise, reaction is carried out for 5-6 hours at the temperature of 100 ℃ and 110 ℃, TLC detection is carried out after the reaction is finished, reactants are diluted by ethyl acetate 100 ml, proper amount of water and saturated saline are sequentially used for washing for 2-3 times, and an organic layer is dried by anhydrous sodium sulfateDrying, filtering, concentrating under reduced pressure, and purifying by silica gel column chromatography (petroleum ether and ethyl acetate mixed elution) to obtain compound 3;
synthesis of compound 5: stirring the obtained compound 3, 0.3 g of concentrated sulfuric acid, 90 ml of acetic acid and 30 ml of acetic anhydride at room temperature for reaction for 30 minutes to 1 hour, and monitoring by TLC to obtain a compound 4 after the reaction is finished; adding 11 ml of bromoacetyl and 7 ml of methanol, and reacting for 2 hours in a dark place; diluting the reaction mixture with 200 ml of dichloromethane, washing the reaction mixture with proper amount of water, saturated sodium bicarbonate solution and saturated brine for 2-3 times respectively, drying an organic layer with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure to obtain a crude compound 5;
synthesis of compound 7: dissolving 0.97 g (5mmol) of digoxin compound 6 and 0.17 g (5mmol) of diphenyl tin chloride in 100 ml of acetonitrile, and stirring at room temperature in the dark for reaction for 10 minutes; adding 1.74 g (7.5mmol) of silver oxide, 0.7 g (3.75mmol) of 55 '-dimethyl-2, 2' -bipyridine, weighing 3 g (7.5mmol) of the compound 5 obtained in the step (iv), adding the mixture into the reaction system, stirring vigorously at 35 ℃ for 24 hours, cooling the reaction mixture to room temperature, quenching the reaction system with 1ml of saturated aqueous ammonium chloride solution, diluting with 1:1 chloroform, filtering off insoluble salt residues, concentrating the solution under reduced pressure, subjecting the residue to column chromatography to obtain acetyl of the compound 7, dissolving the acetyl in 140ml of methanol, adding 40g of sodium methoxide, and treating at room temperature to obtain the compound 7.
The preparation of the deacetyl cilaria dextrose modified compound liposome injection comprises
(1) Preparation of deacetylated cilaria glucose-based modified compound liposome microcapsule
Preparing capsules by a reverse titration method, weighing 100mg of modified compound 7, mixing and homogenizing with 20mL of 18mg/mL sodium alginate solution, extracting with a container, dropwise adding into a 40mL beaker containing 37mg/mL CaCl2 solution, performing suction filtration after 20 minutes, washing, and freeze-drying to obtain the complement antibody micro-capsules, wherein the maximum encapsulation rate of the capsules reaches 70%. The scale-up is carried out in pilot plant test and industrial production.
(2) Preparation of modified compound liposome microcapsule injection
Dissolving modified liposome microcapsule in 10% ethanol to obtain a solution with a concentration of 0.1mg/1ml, filtering for sterilization, manually or mechanically filling 1ml per capsule, sealing, shading, and sealing for storage.
The invention has the advantages that:
1. in vitro experiments, the deacetylated cedilan glucose-based modified compound has toxicity on liver cancer cells equivalent to cedilan, but has the potency over 100 times higher than that of the common drug 5 fluorouracil, and has lower toxicity on normal cells;
2. in an in vivo animal experiment, compared with the cedilanid, the deacetylated cedilanid glucosyl modified compound has obviously enhanced inhibition rate on liver cancer; the deacetylated glucose-based modified compound of the cedilanid is not easy to be hydrolyzed into digoxin without anticancer effect by enzyme in the liver, so that the anticancer time is obviously prolonged.
3. In an in vivo animal experiment for evaluating cardiotoxicity, compared with a deacetylated cedilan glucose-based modified compound, the deacetylated cedilan glucose-based modified compound prepared into liposome improves the targeting property to the liver, reduces the cardiotoxicity of cardiac glycoside compounds, reduces the occurrence of arrhythmia and obviously improves the therapeutic index of the medicament. Experiments prove that the deacetylated cedilanid glucosyl modified compound is an ideal, efficient and low-toxicity anti-liver cancer drug.
Drawings
FIG. 1 shows the cardiotoxicity test of liposome of example 2-O-benzyl-beta-D-glucopyranoside-deacetylated cilantro,
electrocardiogram for heart rate change and arrhythmia occurrence in unit time after administration of the mouse;
in the figure, A: group A electrocardiogram; b: b group electrocardiogram; c: and C group electrocardiograms.
Detailed Description
The in vitro and in vivo anticancer experiment of the medicine is carried out by establishing in vitro cell culture and in vivo animal models, and the experimental method comprises the following steps:
culturing cancer cells and normal cells;
secondly, performing deacetylation of the glucose-based modified compound of the cedilanid and comparative experiments on the in-vitro anticancer and normal cell toxicity of the cedilanid;
establishing a liver cancer nude mouse animal model, and developing an in vivo anticancer comparative experiment of deacetylated cedilan glucose-based modified compound liposome and cedilan liposome;
and fourthly, evaluating the toxicity of the acetylated cediland glucose-based modified compound liposome and cediland heart.
1. In vitro anticancer experiments
Human tumor cell lines were cultured in DMEM medium (containing 10% inactivated fetal calf serum, 100U/ml penicillin and 100. mu.g/ml streptomycin), and human normal liver cell line HL-7702 was cultured in RPMI1640 medium (containing 20% inactivated fetal calf serum, 100U/ml penicillin and 100. mu.g/ml streptomycin). The six cell strains are all cultured in an incubator with 37 ℃ and 5% CO2 saturated humidity, the culture solution is changed and passaged once for two to three days, and cells in the logarithmic growth phase are taken for experiment.
MTT method detects the effect of anticancer medicine on the proliferation activity of tumor cells. Taking cells in logarithmic growth phase at 1 × 104The density of the/k wells was seeded in 96 well plates at 180. mu.l per well. The test was carried out with a negative control group (PBS) and a positive control group (5-FU) (final concentration: 10)-1mmol/L), different concentrations (2X 10)-2，4×10-3，8×10-4，1.6×10-4，3.2×10-5mmol/L) of the drug groups, each group is provided with 5 multiple wells, cultured overnight in an incubator with 5% CO2 saturated humidity at 37 ℃, and 20 μ L of the drug solution (final concentration is 2X 10)-2，4×10-3，8×10-4，1.6×10-4，3.2×10-5mmol/L), adding the medicine, culturing for 48h, adding 20 mul of MTT solution (5mg/ml) into each hole, after culturing for 4h, removing the culture solution by suction, adding 150 mul of DMSO into each hole, slightly shaking for 10min, dissolving crystals, measuring OD value at 490nm wavelength by a microplate reader, and calculating the cell growth inhibition rate according to the following formula:
cell growth inhibition (%) [1- (drug-treated group/negative control group) ] × 100%.
Drug treatment group: cell-containing medium, MTT, drug;
negative control group: cell-containing medium, MTT, PBS;
blank group: cell and drug free media, MTT;
the experiment was repeated 3 times.
The results of 2-O-benzyl- β -D-glucopyranoside-deacetylated cedilan and deacetylated cedilan in vitro anti-human hepatoma cells (HepG2) and on normal hepatocytes are shown in table 1, table 2:
TABLE 1 Effect of 2-O-benzyl- β -D-glucopyranoside-deacetylated cilaria (A) and deacetylated cilaria (B) against human hepatoma cells in vitro (HepG2) (A)
n＝5)
Note: p<0.05, compared to the PBS control group;#P>0.05, group A was compared to group B.
TABLE 2 in vitro Effect of 2-O-benzyl- β -D-glucopyranoside-deacetylated cilaria (A) and deacetylated cilaria (B) on human Normal hepatocytes (HL-7702) (results)
n＝5)
Note: p >0.05, group a, group B compared to PBS control group; # P >0.05, group A compared to group B.
2. In vivo antitumor assay
Human liver cancer cells (HCCLM3) were routinely cultured for 48 hours and made up to 5X 10 with PBS6And (5) standby. 18-20g of nude mice (provided by BALB/c-nu Guangdong laboratory animal center), breeding for one week under aseptic condition by adopting 5 multiplied by 106HCCLM3 cell nude mouse back 0.1 ml/one subcutaneous inoculation molding, until the tumor mass is about 0.5cm long, the divided administration is started, the animals are divided into 3 groups; control group(PBS group), 2-O-benzyl-beta-D-glucopyranoside-deacetyl cedilan group (A) (1.0mg/kg), and cedilan group (B) (1.0mg/kg) were administered intraperitoneally 1 time per day, 9 times, animals were killed by inebriation at experiment 10, and the masses were taken out and weighed by electronic balance. The statistical method comprises the following steps: the mice were sacrificed the day after the last administration, the body weights were weighed, tumors were removed and weighed, and the tumor Inhibition Rate (IR) was calculated as (average tumor weight of control group-average tumor weight of administration group)/average tumor weight of control group × 100%. Statistical processing is carried out on all data by using SPSS 17.0 software, results are expressed by means of mean +/-standard deviation, mean comparison between two groups is carried out by using t test, single-factor variance analysis is adopted for mean comparison between multiple groups, and significant difference is found when P is less than 0.05. And (3) data analysis: mean tumor volumes are expressed in ± s, and the results are shown in table 3:
TABLE 3 Effect of 2-O-benzyl- β -D-glucopyranoside-deacetylated cilantrum (A) and deacetylated cilantrum (B) on mouse subcutaneous implanted hepatoma cell tumors
| Group of | Dosage (mg/kg) | Number of animals (n) | Tumor weight (g) | Tumor inhibition Rate (%) |
| PBS group | -- | 10 | 0.34±0.23 | -- |
| A | 1.0 | 10 | 0.11±0,02 | 67.64*# |
| B | 1.0 | 10 | 0.19±0.05 | 44.10* |
Note: p <0.05 compared to PBS group; # P <0.05 compared to group B.
Description of the drawings: in vivo animal experiments, the deacetylated cedilanid glucose-based modified compound (A) and the deacetylated cedilanid (B) have inhibition effects on tumors, and the inhibition rate of (A) on liver cancer is obviously higher than that of (B). The reason is that: in vivo, the glucose group of deacetylated cedilanid is easily hydrolyzed by enzyme to generate digoxin, thereby losing related anticancer activity, and the deacetylated cedilanid glucose group modified compound (A) is modified to maintain the stability of glucose group and keep the stability of anticancer activity.
3.2-O-benzyl-beta-D-glucopyranoside-deacetylated cilansetron liposome cardiotoxicity test
Selecting 30 Kunming mice with SPF grade of 18-20g, and dividing the mice into three groups, namely a normal control group (A), wherein the female and male mice can be used simultaneously (provided by SPF animal room of Guilin medical institute); 2-O-benzyl-beta-D-glucopyranoside-deacetylated cedilanid liposome administration group (group B) and 2-O-benzyl-beta-D-glucopyranoside-deacetylated cedilanid liposome administration group (group C), wherein 10 of each group is prepared by carrying out intraperitoneal injection anesthesia on 45mg/kg pentobarbital sodium under the laboratory condition of quiet room temperature of 22-26 ℃, fixing a mouse in a standing position, carrying out abdominal injection administration on 2mg/kg after the mouse is settled, recording the electrocardiogram of the mouse for 10 minutes by using an electrocardiogram instrument (single-lead ECC-11A type, frequency response range of 0-90 Hz, paper speed of 50mmPs and standard II lead) after the administration is carried out for 10 minutes, calculating the occurrence frequency of arrhythmia, expressing the result by mean +/-standard deviation, comparing the mean between two groups, and carrying out t test, and obtaining the result shown in Table 4, FIG. 1;
TABLE 4 variation of heart rate and occurrence of arrhythmia per unit time (10 min) after administration to mice
Note: p <0.05, group B compared with group C, showed that the liver targeting property was improved and the cardiotoxicity was reduced by making liposomes from 2-O-benzyl- β -D-glucopyranoside-deacetylated cedilanid.
In fig. 1, diagram a: group A electrocardiogram; and B: b group electrocardiogram, the heart rate is slightly slowed down, the rhythm is normal, and no arrhythmia is found; and (C) figure: the heart rate of the C group of electrocardiograms is obviously slowed down, atrioventricular conduction block is seen in the electrocardiograms, the P-Q interval is prolonged, QRS-T waves disappear, and atrioventricular conduction block arrhythmia is obvious.
Claims (4)
1. The synthesis of the deacetylated glucose-based modified compound of the cedilanid is characterized in that: the deacetylated cedilanid glucosyl modified compound is 2-O-benzyl-beta-D-glucopyranoside-deacetylated cedilanid, and the structural formula is as follows:
the synthetic route is as follows:
2. the synthesis of deacetylated glucose-modified cilaria lemaneiformis compound according to claim 1, wherein the specific synthesis steps are as follows:
synthesis of compound 1: under ice bath, adding 140ml of methanol and 2.4 ml of acetyl chloride into a dry reaction bottle provided with a drying tube, stirring for 5 minutes, naturally stirring, heating to room temperature, stirring for 20 minutes, adding 10 g of glucose, heating to reflux for reaction for 2 hours, cooling the reaction solution to room temperature, adding anhydrous potassium carbonate to neutralize to neutrality, filtering, and concentrating to obtain a crude product of the 1-O-methyl glucoside compound 1;
synthesis of compound 2: dissolving the obtained crude product of the compound 1 in 34 ml of DMF, dropwise adding 16 ml of benzaldehyde, quickly adding 1.72 g of p-toluenesulfonic acid, reacting overnight at 50 ℃, displaying complete reaction by TLC, pouring the reactant into a mixed solution of 100 ml of ice water and 140ml of petroleum ether, violently shaking, separating out milky white solid, performing suction filtration, and recrystallizing with absolute ethyl alcohol to obtain a compound 2;
synthesis of compound 3: the obtained compound 2 and Bu2SnO 14 g and toluene 180 ml are added into a round-bottom flask provided with a water separator, reflux reaction is carried out for 3-4 hours until no water is separated out, the reaction solution is concentrated to 60 ml, anhydrous DMF30 ml is added, benzyl bromide 7 ml is dropwise added, reaction is carried out for 5-6 hours at the temperature of 110 ℃, TLC detection reaction is finished, reactants are diluted by ethyl acetate 100 ml, appropriate amount of water and saturated saline are sequentially used, washing is carried out for 2-3 times, an organic layer is dried by anhydrous sodium sulfate, filtering and reduced pressure concentration are carried out, and the organic layer is purified by silica gel column chromatography to obtain a compound 3;
synthesis of compound 5: stirring the obtained compound 3, 0.3 g of concentrated sulfuric acid, 90 ml of acetic acid and 30 ml of acetic anhydride at room temperature for reaction for 30 minutes to 1 hour, and monitoring by TLC to obtain a compound 4 after the reaction is finished;
adding 11 ml of bromoacetyl and 7 ml of methanol, and reacting for 2 hours in a dark place; diluting the reaction mixture with 200 ml of dichloromethane, washing the reaction mixture with proper amount of water, saturated sodium bicarbonate solution and saturated brine for 2-3 times respectively, drying an organic layer with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure to obtain a crude compound 5;
synthesis of compound 7: dissolving 0.97 g of digoxin compound 6 and 0.17 g of diphenyl tin chloride in 100 ml of acetonitrile, and stirring for reaction for 10 minutes at room temperature in a dark place; adding 1.74 g of silver oxide and 0.7 g of 5,5 '-dimethyl-2, 2' -bipyridine, weighing 3 g of the compound 5 obtained in the step (iv), adding the mixture into a reaction system, stirring vigorously at 35 ℃ for 24 hours, cooling the reaction mixture to room temperature, quenching the reaction system with 1ml of saturated aqueous ammonium chloride solution, diluting the reaction system with 1:1 parts of chloroform and acetone, filtering out insoluble salt residues, concentrating the solution under reduced pressure, carrying out column chromatography on the residue to obtain an acetyl compound of the compound 7, dissolving the acetyl compound in 140ml of methanol, adding 40g of sodium methoxide, and treating at room temperature to obtain the compound 7.
3. The preparation of the deacetyl cedilanid glucosyl-modified compound liposome injection prepared in claim 2, characterized in that: comprises that
(1) Preparation of deacetylated cilaria glucose-based modified compound liposome microcapsule
Preparing capsule by reverse titration, weighing 100mg of modified compound 7, mixing with 20mL of sodium alginate solution with concentration of 18mg/mL, homogenizing, extracting with container, and dripping 40mL of CaCl containing 37mg/mL dropwise2In a beaker of the solution, after 20 minutes, the complement antibody micro-capsules are obtained by suction filtration, washing and freeze drying, and the maximum encapsulation rate reaches 70 percent;
(2) preparation of modified compound liposome microcapsule injection
Dissolving modified compound liposome microcapsule in 10% ethanol to obtain a solution with concentration of 0.1mg/1ml, filtering for sterilization, manually or mechanically filling 1ml per capsule, sealing, shading, and sealing for storage.
4. Use of the glucose-modified desciclovir liposome injection prepared by the method of claim 3 in preparing anti-liver cancer drugs.
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| WO2012074547A2 (en) * | 2010-11-29 | 2012-06-07 | New York University | STEROID COMPOUNDS AS RORγt MODULATORS AND USES THEREOF |
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| CN102416019A (en) * | 2011-11-18 | 2012-04-18 | 桂林医学院 | Application of cedilanid in preparation of anti-liver cancer and anti-bladder cancer medicament |
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| Regioselective Diversification of a Cardiac Glycoside, Lanatoside C,by Organocatalysis;Yoshihiro Ueda et al.;《J. Org. Chem.》;20120807;第77卷;第7850-7857页 * |
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