CN110200920B - Reduction-sensitive pharmaceutical composition and preparation and application thereof - Google Patents
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Abstract
The invention discloses a reduction-sensitive pharmaceutical composition, and a preparation method and an application thereof, wherein the pharmaceutical composition is nanoparticles (according to the situation, the pharmaceutical composition also comprises an auxiliary agent acceptable in pharmacodynamics) consisting of bis-n-alkyl dithio-ortho-alkanoic acid glycerophosphorylcholine and a medicament, has reduction-sensitive rapid degradability, good stability, long circulation and targeted disintegration and rapid medicament release functions. The reduction-sensitive pharmaceutical composition is prepared by a film dispersion method, a reverse phase evaporation method, a freeze drying method, an ultrasonic dispersion method, a spray drying method, a film extrusion method or a high-pressure homogenization method, and the process is simple. The reduction sensitive pharmaceutical composition is used for preparing antitumor drugs, antiviral drugs and antibacterial drugs.
Description
Technical Field
The invention discloses a reduction-sensitive pharmaceutical composition and preparation and application thereof, and relates to the technical field of medicines.
Background
The nano-particles are important drug carriers, comprise liposomes, solid lipid nanoparticles, nano-micelles and the like, and can load and control various drugs. The hydrophobic drug is typically in the lipid of the nanoparticle. In the in vivo circulation process of the drug-loaded nanoparticles, the drug is easy to leak out of the nanoparticles, and the exertion of the drug effect is influenced.
The hydrophilic macromolecule such as polyethylene glycol is covered on the surface of the liposome, so that the circulation time of the liposome in blood can be prolonged, and the invisible nanoparticles are also called to slow the leakage of the medicine. The invisible nanoparticles not only have long circulation time, but also can enhance the permeation effect by utilizing tumor capillaries, improve the tumor targeting effect of the medicament and improve the anti-tumor effect. However, due to the long circulation time, the nanoparticles are easy to deposit on the hand and foot parts, for example, the doxorubicin is administered by nanoparticle loading injection in the most common administration mode, but the doxorubicin long-circulation nanoparticle deposition on the hand and foot parts often causes hand and foot diseases. Therefore, the drug-loaded nanoparticles, particularly the long-circulating nanoparticles, circulate in blood to be beneficial to reaching tumor sites, but need to be rapidly disintegrated and released at the tumor sites, namely, the drug-loaded nanoparticles are required to have a certain tumor targeted degradation effect.
The glutathione concentration in inflammatory tissues, tumor tissues, particularly tumor cells is far higher than that in normal tissues, and the microenvironment of the tissues can promote the rupture of the disulfide bond-containing prodrug molecules or long-circulating phospholipid molecules, so that the release of the drugs and the improvement of the effective drug concentration are realized. However, the previously reported reduction-sensitive nanoparticles generally use normal long-chain fatty acid phospholipids, and the breakage of disulfide bonds does not easily lead to the complete disintegration of the nanoparticles.
Therefore, the invention is especially needed to invent a functional phospholipid which has the assembly capability and the reduction-sensitive targeted breakage, and the assembled nano-particles of the phospholipid release quickly due to the complete disintegration of the reduction sensitivity, thereby overcoming the defect that the release of the long-circulating nano-particles is too slow.
Anthracyclines such as doxorubicin are an important class of antitumor drugs, but generally have cardiotoxicity, myelotoxicity and the like. Loaded in liposome can reduce toxicity to some extent, but is easy to leak and releases drug too quickly. The nano-particle carrier constructed by adopting the polyethylene glycol macromolecular phospholipid can prolong the drug release time, but the long circulation time causes the nano-particles to be easily deposited on the hand and foot parts to cause hand and foot diseases. Therefore, it is particularly desirable to invent a drug delivery system that can quickly release drugs at the tumor site while preventing leakage through rapid degradation of the microenvironment response at the tumor site.
Disclosure of Invention
The technical problem is as follows: the invention aims to provide a reduction-sensitive pharmaceutical composition, and a preparation method and an application thereof, wherein the pharmaceutical composition is nanoparticles (including pharmaceutically acceptable auxiliaries according to the situation) composed of bis-n-alkyl dithio-n-alkanoic acid glycerophosphorylcholine and a medicament, and has reduction-sensitive rapid degradability; simultaneously provides a preparation method based on the pharmaceutical composition and application thereof in preparing antitumor drugs, antiviral drugs and antibacterial drugs.
The technical scheme is as follows: the invention provides a reduction-sensitive pharmaceutical composition which is a nanoparticle and comprises di-n-alkyl dithio-n-alkanoic acid glycerophosphorylcholine and a drug.
Wherein:
the reduction-sensitive pharmaceutical composition further comprises a pharmaceutically acceptable adjuvant.
The reduction-sensitive pharmaceutical composition is in the form of nanoparticles, the nanoparticles are liposomes, solid lipid nanoparticles or micelles, and the particle size of the particles is 10-1000 nm.
The structural general formula of the di-n-alkyl dithio-n-alkanoic acid glycerophosphorylcholine is as follows:
wherein m is a positive integer of 2 to 10, and n is a positive integer of 7 to 15.
The medicine is an anti-tumor medicine, an anti-virus medicine or an antibacterial medicine.
The anti-tumor drug is one of paclitaxel, docetaxel, cabazitaxel, camptothecin, hydroxycamptothecin, 7-ethyl-10-hydroxycamptothecin, irinotecan, topotecan, lurtotecan, belotecan, 7-hydroxymethyl camptothecin, vincristine, vinorelbine, ansamycin, rapamycin, cytarabine, etoposide, mitoxantrone, doxorubicin, zorubicin, epirubicin, idarubicin, sambicin, tamoxifen, triptolide, mivaquoid, ursolic acid or platinum drugs.
The platinum drug comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, leplatin or cis-bisneodecanoic acid-trans-R, R-1, 2-cyclohexanediamine platinum (II).
The antibacterial drug is one of amphotericin B, gentamicin, nystatin, fluconazole, amikacin, levofloxacin or econazole nitrate.
The antiviral drug is ribavirin or adefovir dipivoxil.
The pharmaceutically acceptable auxiliary agent is one or more of phospholipid, glyceride, fatty acid, tween, poloxamer or cholesterol.
The phospholipid comprises lecithin and synthetic phospholipid.
The phospholipid comprises a targeting phospholipid.
The phospholipid comprises distearoyl glycerol phosphatidyl ethanolamine-polyethylene glycol and distearoyl glycerol phosphatidyl ethanolamine-polyethylene glycol-folic acid.
The invention also provides a preparation method of the reduction-sensitive pharmaceutical composition, and the reduction-sensitive pharmaceutical composition is prepared by a film dispersion method, a reverse phase evaporation method, a freeze-drying method, an ultrasonic dispersion method, a spray-drying method, a film extrusion method or a high-pressure homogenization method.
The invention also provides an application of the reduction sensitive pharmaceutical composition, and the reduction sensitive pharmaceutical composition is applied to preparation of anti-tumor agents, antiviral agents and antibacterial agents.
Wherein:
the medicament is a liquid preparation, a solid preparation, a semi-solid preparation, a capsule, a granule or a gel.
The medicament can be administered in unit dosage form, and the administration route can be intestinal or parenteral, such as oral, muscle, subcutaneous, nasal, oral mucosa, skin, peritoneum or rectum.
In view of the in vitro degradation rate, the pharmaceutical composition of the present invention exhibits reduction-sensitive rapid degradability.
Has the advantages that: compared with the prior art, the invention has the following advantages:
the reduction-sensitive pharmaceutical composition is nanoparticles, has reduction-sensitive rapid degradability, and can rapidly break the disulfide bond of the bis-n-alkyl dithio-ortho-alkanoic acid glycerophosphorylcholine in the presence of high-concentration glutathione in inflammation, tumor parts or tumor cells, so that the liposome is disintegrated, the drug is rapidly released, and the drug effect is exerted;
the reduction-sensitive pharmaceutical composition has good stability and long circulation, and has the functions of targeted disintegration and rapid drug release at the tumor part due to the reduction microenvironment response;
the reduction-sensitive pharmaceutical composition can be prepared very easily by simple processes such as a film method, a high-pressure homogenization method and the like, and the particle diameter is 10-1000 nm;
the reduction-sensitive pharmaceutical composition is liposome, solid lipid nanoparticle or micelle;
the anti-tumor agent prepared from the reduction sensitive pharmaceutical composition has excellent anti-tumor activity;
the reduction-sensitive pharmaceutical composition of the present invention can be used as a liquid preparation, a solid preparation, a semisolid preparation, a sterile preparation and a sterile preparation.
Drawings
FIG. 1 is a transmission electron micrograph of Doxorubicin (DOX) reduction-responsive liposomes;
FIG. 2 is the structural diagram of the dithiophospholipid liposome DOX/12-SS3-LLP liposome in different states, wherein, the diagram a is the transmission electron microscope diagram of the dithiophospholipid liposome 12-SS3-LLP (the insertion diagram is the frozen transmission electron microscope diagram), and the diagram b is the transmission electron microscope diagram of the dithiophospholipid liposome 12-SS3-LLP after being treated by dithiothreitol;
FIG. 3 is a graph of the release profile of DOX-loaded reduction-sensitive liposomes in DTT media with (or without) dithiothreitol;
FIG. 4 is a schematic representation of the evaluation of the in vitro anti-tumor effect of reduction-sensitive liposomes using an A549 cell model and an MCF-7 cell model, wherein panel a is a schematic representation of the evaluation of the in vitro anti-tumor effect of reduction-sensitive liposomes using the A549 cell model and panel b is a schematic representation of the evaluation of the in vitro anti-tumor effect of reduction-sensitive liposomes using the MCF-7 cell model;
fig. 5 is a graph showing the in vivo anti-tumor effect and toxicity of doxorubicin-loaded reduction-sensitive liposomes, wherein graph a is the change in tumor volume, graph b is the change in mouse body weight, and graph c is the tumor weight.
Detailed Description
The technical scheme of the invention is further explained in detail by combining the drawings and the embodiments in the specification.
The reduction-sensitive pharmaceutical composition is a nanoparticle, and comprises bis-n-alkyl dithio-ortho-alkanoic acid glycerophosphorylcholine and a drug.
Wherein:
the reduction-sensitive pharmaceutical composition further comprises a pharmaceutically acceptable adjuvant.
The reduction-sensitive pharmaceutical composition is nanoparticles, the particle size of the reduction-sensitive pharmaceutical composition is 10-1000 nm, and the nanoparticles can be liposomes, solid lipid nanoparticles or micelles;
the di-n-alkyl dithio-ortho-alkanoic acid glycerophosphorylcholine has the following structure:
wherein m is a positive integer of 2 to 10, and n is a positive integer of 7 to 15.
The medicine is antitumor medicine, antiviral medicine, and antibacterial medicine.
The anti-tumor drug is one of the following drugs: paclitaxel, docetaxel, cabazitaxel, camptothecin, hydroxycamptothecin, 7-ethyl-10-hydroxycamptothecin, irinotecan, topotecan, lurtotecan, belotecan, 7-hydroxymethyl camptothecin, vincristine, vinorelbine, ansamycin, rapamycin, cytarabine, etoposide, mitoxantrone, doxorubicin, zorubicin, epirubicin, idarubicin, sambafloxacin, tamoxifen, triptolide, mivaquoin, ursolic acid, platinum drug.
The platinum drug comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, leplatin or cis-bisneodecanoic acid-trans-R, R-1, 2-cyclohexanediamine platinum (II).
The antibacterial drug is one of the following: amphotericin B, gentamicin, nystatin, fluconazole, amikacin, levofloxacin or econazole nitrate.
The antiviral drug is ribavirin or adefovir dipivoxil.
The pharmaceutically acceptable auxiliary agent is one or more of phospholipid, glyceride, fatty acid, tween, poloxamer or cholesterol.
The phospholipid comprises lecithin and synthetic phospholipid.
The phospholipid comprises a targeting phospholipid.
The phospholipid comprises distearoyl glycerol phosphatidyl ethanolamine-polyethylene glycol and distearoyl glycerol phosphatidyl ethanolamine-polyethylene glycol-folic acid.
A method for preparing reduction-sensitive pharmaceutical composition comprises film dispersion method, reverse phase evaporation method, freeze drying method, ultrasonic dispersion method, spray drying method, film extrusion method or high pressure homogenization method.
An application of a reduction-sensitive pharmaceutical composition, which is applied to the preparation of anti-tumor drugs, antiviral drugs and antibacterial drugs.
Wherein:
the medicament is a liquid preparation, a solid preparation, a semi-solid preparation, a capsule, a granule or a gel.
The pharmaceutical composition can be prepared into a medicament which can be administered in a unit dosage form, and the administration route can be intestinal or parenteral, such as oral, muscle, subcutaneous, nasal cavity, oral mucosa, skin, peritoneum or rectum.
In view of the in vitro degradation rate, the pharmaceutical composition of the present invention exhibits reduction-sensitive rapid degradability.
From the in vitro drug effect screening, the reduction sensitive drug composition of the invention has good anti-inflammatory and anti-tumor activity.
The preparation method of the reduction-sensitive pharmaceutical composition is characterized in that the reduction-sensitive pharmaceutical composition is prepared from di-n-alkyl dithio-n-alkanoic acid glycerophosphorylcholine, a medicament and a pharmacodynamically acceptable auxiliary agent by a film dispersion method, a reverse phase evaporation method, a freeze drying method, an ultrasonic dispersion method, a spray drying method, a film extrusion method, a high-pressure homogenization method or the like.
The present invention is further illustrated by the following examples, but is not limited to the following examples.
In the examples, the phospholipid compounds represented by the following symbols and structural formulae are as follows:
8-SS3-PC di-n-octyl dithiopropionic acid glycerophosphocholine,
10-SS3-PC di-n-decyl dithiopropionic acid glycerophosphocholine,
12-SS3-PC di-n-dodecyl dithiopropionic acid glycerophosphocholine,
14-SS3-PC di-n-tetradecyl dithiopropionic acid glycerophosphocholine,
16-SS3-PC Glycerol phosphocholine bis-n-hexadecyldithiophosphate.
10-SS6-PC bis-n-decyldithio-n-hexanoic acid glycerophosphorylcholine
8-SS11-PC bis-n-octyldithion-undecyl glycerophosphocholine
14-SS6-PC Di-n-tetradecyl dithiohexanoic acid glycerophosphocholine
Example 1
A reduction-sensitive pharmaceutical composition is Doxorubicin (DOX) reduction-responsive liposome, is granular and has the particle size of 110 nanometers, and comprises di-n-dodecyl dithiopropionic acid glycerophosphocholine, doxorubicin, cholesterol and distearoyl phosphatidylcholine DSPC.
The preparation method comprises the following steps:
adding 0.5mmol of di-n-dodecyl dithiopropionic acid glycerophosphorylcholine 12-SS3-PC, 0.5mmol of distearoyl phosphatidylcholine DSPC, 0.3mmol of cholesterol and 20ml of trichloromethane into an eggplant-shaped bottle for dissolving, slowly evaporating to dryness by spinning at 60 ℃, purging with nitrogen for 10 minutes to form a film on the inner wall of the eggplant-shaped bottle; adding 20ml ammonium sulfate solution, forming membrane at 60 deg.C, and filtering with 200nm filter membrane to obtain liposome solution; dialyzing the liposome solution with semipermeable membrane, removing ammonium sulfate from liposome, adding doxorubicin hydrochloride 0.1mmol into the liposome solution, and incubating at 50 deg.C; the doxorubicin enters the liposome to form sulfate gel, and the sulfate gel is deposited in the liposome to obtain the doxorubicin reduction response liposome.
The average particle size of the liposome is measured to be 110nm by adopting a laser particle sizer, and the form of the nano particles is measured by a transmission electron microscope, so that a spherical structure is shown (figure 1); and (3) freeze-drying the liposome nanoparticle solution to obtain powdery nanoparticles.
The doxorubicin reduction response liposome prepared in the embodiment is applied to preparation of an antitumor agent.
Example 2
A reduction-sensitive pharmaceutical composition is paclitaxel reduction-responsive liposome, is granular, has particle size of 108nm, and comprises di-n-tetradecyl dithiopropionic acid glycerophosphorylcholine, paclitaxel, dipalmitoyl phosphatidylcholine, and cholesterol.
The preparation method comprises the following steps:
adding 0.1mmol of paclitaxel, 14-SS3-PC 1.0mmol of di-n-tetradecyl dithiopropionic acid glycerophosphocholine, 1.0mmol of diphosphonocholine dipalmitate, 0.3mmol of cholesterol and 20ml of chloroform into a solanaceous bottle for dissolving, slowly evaporating to dryness at 60 ℃, purging with nitrogen for 10 minutes to form a film on the inner wall of the solanaceous bottle, adding 20ml of PBS (pH 7.4) solution, forming the film at 60 ℃, and filtering with a 200nm filter membrane to obtain the liposome nanoparticles.
Measuring the average particle diameter by a laser particle sizer to be 108nm, and measuring the form of the nano particles by a transmission electron microscope to show a spherical structure; and (3) freeze-drying the paclitaxel reduction response liposome nanoparticle solution to obtain powdery nanoparticles.
The paclitaxel reduction response liposome prepared in the embodiment is applied to preparing an anti-tumor agent.
Example 3
A reduction-sensitive pharmaceutical composition is a docetaxel reduction-responsive liposome, is granular and has a particle size of 112 nanometers, and comprises di-n-tetradecyl dithiopropionic acid glycerophosphocholine, docetaxel and an auxiliary agent lecithin and cholesterol.
The preparation method comprises the following steps:
docetaxel 0.1mmol, di-n-tetradecyl dithiopropionic acid glycerophosphocholine 14-SS3-PC 0.2
Adding mmol, lecithin 0.7mmol, cholesterol 0.3mmol, and chloroform 20ml into a eggplant-shaped bottle, dissolving, slowly evaporating to dryness at 60 deg.C, purging with nitrogen for 10min to form a film on the inner wall of the eggplant-shaped bottle; adding 20ml PBS (pH 7.4) solution, filming at 60 deg.C, and filtering with 200nm filter membrane to obtain liposome nanoparticle solution.
The docetaxel reduction response liposome prepared by the embodiment is applied to preparing an antitumor agent.
Example 4
A reduction-sensitive pharmaceutical composition is paclitaxel reduction-response stealth liposome, is granular and has a particle size of 120 nanometers, and comprises di-n-tetradecyl dithiopropionic acid glycerophosphorylcholine, paclitaxel and distearoyl phosphatidylcholine DSPC, distearoyl phosphatidylethanolamine-polyethylene glycol and cholesterol.
The preparation method comprises the following steps:
0.1mmol of paclitaxel, 0.5mmol of di-n-tetradecyl dithiopropionic acid glycerophosphorylcholine 14-SS3-PC, 0.5mmol of distearoyl phosphatidylcholine (DSPC), 0.1mmol of distearoyl phosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000, PEG molecular weight 2000), 0.3mmol of cholesterol and 20ml of trichloromethane are added into a solanaceous bottle for dissolving, slowly and spirally evaporated to dryness at 60 ℃, nitrogen is used for purging for 10 minutes, and a layer of film is formed on the inner wall of the solanaceous bottle; adding 20ml PBS (pH 7.4) solution, filming at 60 deg.C, and filtering with 200nm filter membrane to obtain liposome nanoparticle solution; and (3) freeze-drying the liposome nanoparticle solution to obtain powdery nanoparticles.
The paclitaxel reduction response liposome prepared in the embodiment is applied to preparing an anti-tumor agent.
For ease of control, paclitaxel was prepared as a common stealth liposome, using the same procedure as above, except that 14-SS3-PC was replaced with equal moles of DSPC.
Example 5
A reduction sensitive pharmaceutical composition is a doxorubicin reduction response invisible liposome, is granular and has the particle size of 116 nanometers, and comprises di-n-tetradecyl dithiopropionic acid glycerophosphocholine, doxorubicin, auxiliary agents cholesterol and distearoyl phosphatidylcholine DSPC.
The preparation method comprises the following steps:
adding 0.5mmol of di-n-tetradecyl dithiopropionic acid glycerophosphorylcholine 14-SS3-PC, 0.5mmol of distearoyl phosphatidylcholine (DSPC), 0.3mmol of cholesterol and 20ml of chloroform into an eggplant-shaped bottle for dissolving, slowly evaporating to dryness by spinning at 60 ℃, purging with nitrogen for 10 minutes, and forming a film on the inner wall of the eggplant-shaped bottle; adding 20ml ammonium sulfate solution, forming membrane at 60 deg.C, and filtering with 200nm filter membrane. The resulting liposome solution was dialyzed with a semipermeable membrane to remove ammonium sulfate from the outer surface of the liposome. Doxorubicin hydrochloride 0.1mmol was added to the liposome solution and incubated at 50 ℃. The doxorubicin enters the liposome to form sulfate gel which is deposited inside the liposome; then, adding 0.1mmol of distearoyl phosphatidyl ethanolamine-polyethylene glycol (DSPE-PEG2000) and the liposome solution for incubation for 30 minutes to obtain doxorubicin reduction response invisible liposomes, and freeze-drying the liposome nanoparticle solution to obtain powdery nanoparticles.
The doxorubicin reduction-response stealth liposome prepared in the embodiment is applied to preparation of an antitumor agent.
Example 6
A reduction-sensitive pharmaceutical composition is cytarabine liposome, is granular and has a particle size of 130 nm, and comprises di-n-tetradecyl dithiopropionic acid glycerol phosphocholine and cytarabine.
The preparation method comprises the following steps:
0.5mmol of di-n-tetradecyl dithiopropionic acid glycerophosphorylcholine 14-SS3-PC, 1.0mmol of dipalmitoyl phosphatidylglycerol, 0.2mmol of cholesterol and 0.3mmol of triolein are put into 3mL of chloroform-diethyl ether (volume ratio is 1: 1) mixed solvent, and stirred and mixed to obtain suspension. Cytarabine hydrochloride aqueous solution 2mL (concentration 0.1M) was added to the above suspension, and stirred at 10000rpm at high speed to give a colostrum. And (3) quickly injecting 1mL of colostrum into 10mL of mixed solution containing 4% of glucose and 40mM of lysine by mass fraction, performing vortex oscillation to obtain multiple emulsion, shaking in a water bath at 37 ℃, and diluting to 10mL by using normal saline to obtain liposome suspension. 10mL of physiological saline was added, centrifugation was carried out at 3000rpm for 5min, the supernatant was discarded, and the precipitate was washed with physiological saline 3 times. The precipitate was diluted with physiological saline to prepare cytarabine multivesicular liposome with a concentration of 5 mM.
The cytarabine lipidosome particle prepared in the embodiment is applied to preparing an anti-tumor medicament.
Example 7
A reduction sensitive medicinal composition is irinotecan liposome, is granular and has the particle size of 120 nanometers, and comprises di-n-tetradecyl dithiopropionic acid glycerol phosphocholine, cytarabine cholesterol, distearoyl glycerol phosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000) and distearoyl phosphatidylcholine (DSPC).
The preparation method comprises the following steps:
dissolving 0.5mmol of di-n-tetradecyl dithiopropionic acid glycerophosphorylcholine 14-SS3-PC, 1.0mmol of distearoyl phosphatidylcholine (DSPC), 0.2mmol of cholesterol and 0.1mmol of distearoyl phosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000) in a proper amount of 2mL of anhydrous ethanol, dropwise adding the solution into a sucrose octasulfate triethylamine solution at the temperature of 60 ℃, and stirring for 10 min; the ethanol was removed by rotary evaporation under reduced pressure and homogenized through a 0.2 μm filter. Dialyzing the blank liposome suspension with PBS, and removing sucrose octasulfate triethylamine in the external water phase to obtain blank liposome; and respectively sucking blank liposome and irinotecan drug solution, and incubating and carrying the drug for 30min under the water bath condition of 60 ℃ to obtain the irinotecan liposome.
The irinotecan liposome particles prepared in the embodiment are applied to preparation of antitumor agents.
Example 8
A reduction sensitive medicinal composition is vincristine liposome, is granular, has particle size of 132nm, and comprises di-n-hexadecyl dithiopropionic acid glycerol phosphocholine, vincristine, cholesterol, distearoyl glycerol phosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000, PEG molecular weight 2000) and distearoyl phosphatidylcholine (DSPC).
The preparation method comprises the following steps:
0.1mmol of vincristine, 0.5mmol of di-n-hexadecyl dithiopropionic acid glycerophosphorylcholine 16-SS3-PC, 2.0mmol of distearoyl phosphatidylcholine (DSPC), 0.1mmol of distearoyl phosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000), 0.3mmol of cholesterol and 20ml of chloroform are added into a solanform bottle to be dissolved, slowly and spirally evaporated at 60 ℃ for 10 minutes, and nitrogen is used for purging to form a layer of film on the inner wall of the solanform bottle. Adding 20ml PBS (pH 7.4) solution, filming at 60 deg.C, and filtering with 200nm filter membrane to obtain liposome nanoparticle solution.
The vincristine liposome particles prepared in the embodiment are applied to preparation of antitumor agents.
Example 9
A reduction sensitive medicinal composition is mitoxantrone liposome, is granular and has a particle size of 120nm, and comprises di-n-hexadecyl dithiopropionic acid glycerol phosphocholine, mitoxantrone, pharmaceutically acceptable auxiliary agents cholesterol, distearoyl glycerol phosphatidyl ethanolamine-polyethylene glycol (DSPE-PEG2000) and distearoyl phosphatidylcholine (DSPC).
The preparation method comprises the following steps:
0.1mmol of mitoxantrone, 0.5mmol of di-n-hexadecyl dithiopropionic acid glycerophosphorylcholine 16-SS3-PC, 2.5mmol of distearoyl phosphatidylcholine (DSPC), 0.1mmol of distearoyl phosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000), 0.3mmol of cholesterol and 20ml of chloroform are added into a solanform bottle to be dissolved, slowly and spirally evaporated to dryness at 60 ℃, nitrogen is used for purging for 10 minutes, and a layer of film is formed on the inner wall of the solanform bottle. Adding 20ml PBS (pH 7.4) solution, filming at 60 deg.C, and filtering with 200nm filter membrane to obtain liposome nanoparticle solution.
The mitoxantrone liposome particles prepared in this example are used for preparing antitumor agents.
Example 10
A reduction sensitive medicinal composition is amphotericin B liposome, is granular, has a particle size of 136 nm, and comprises di-n-decyl dithio-n-hexanoate glycerophosphorylcholine, amphotericin B, cholesterol, distearoyl glycerophosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000) and distearoyl phosphatidylcholine (DSPC).
The preparation method comprises the following steps:
0.5mmol of amphotericin B, 0.5mmol of bis-n-decyldithiophosphoric acid n-hexanoic acid glycerophosphorylcholine 10-SS6-PC, 5.0mmol of distearoylphosphatidyl choline (DSPC), 0.3mmol of distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000), 1.0mmol of cholesterol and 20ml of chloroform are added into a solanaceous bottle for dissolving, slowly and spirally evaporated at 60 ℃ for 10 minutes, and nitrogen is used for purging to form a layer of film on the inner wall of the solanaceous bottle. Adding 20ml PBS (pH 7.4) solution, filming at 60 deg.C, and filtering with 200nm filter membrane to obtain liposome nanoparticle solution.
The amphotericin B liposome particles prepared in this example are used for preparing antibacterial agents.
Example 11
A reduction sensitive medicinal composition is an amikacin liposome, is granular, has a particle size of 142 nanometers, and comprises di-n-octyl dithio-n-undecanoic acid glycerol phosphocholine, amikacin, cholesterol, distearoyl glycerol phosphatidyl ethanolamine-polyethylene glycol (DSPE-PEG4000, PEG molecular weight 4000) and distearoyl phosphatidylcholine (DSPC).
The preparation method comprises the following steps:
0.1mmol of amikacin, 0.5mmol of bis-n-octyl dithio-n-undecyl glycerophosphorylcholine 8-SS11-PC, 0.5mmol of distearoyl phosphatidylcholine DSPC, 0.1mmol of distearoyl phosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000), 0.3mmol of cholesterol and 20ml of chloroform are added into a solanform bottle to be dissolved, slowly and spirally evaporated to dryness at 60 ℃, nitrogen is used for purging for 10 minutes, and a layer of film is formed on the inner wall of the solanform bottle. Adding 20ml PBS (pH 7.4) solution, filming at 60 deg.C, and filtering with 200nm filter membrane to obtain liposome nanoparticle solution.
The amikacin liposome particles prepared in the embodiment are applied to preparation of antibacterial agents.
Example 12
A reduction-sensitive medicinal composition is a cisplatin liposome, is granular, has a particle size of 110 nanometers, and comprises di-n-tetradecyl dithiocaproic acid glycerophosphocholine, cisplatin, lecithin and cholesterol.
The preparation method comprises the following steps:
putting di-n-tetradecyl dithiocaproic acid glycerophosphorylcholine 14-SS6-PC, lecithin, cholesterol and DSPE-PEG2000 (molar ratio is 2:2:2:0.3) into an eggplant-shaped bottle, adding 15m L chloroform for dissolving, removing chloroform in a water bath at 45 ℃ by rotary evaporation, and forming a layer of phospholipid film on the wall of the bottle; the residual solvent was removed in vacuo and purged with nitrogen. After the phospholipid membrane was formed, 9m L physiological saline solution containing 9.0mg of cisplatin was added, and the mixture was subjected to shake hydration in a constant temperature oscillator at 37 ℃ for 1 hour; after hydration is finished, vortex oscillation is carried out for 5min, and ultrasonic treatment is carried out for 10min, so as to obtain the multilayer liposome suspension. Sequentially extruding the liposome suspension through 450nm and 220nm polycarbonate filter membranes for 5 times respectively; the cisplatin stealth liposome with the average particle size of about 110nm and uniform distribution is obtained.
The cisplatin liposome particles prepared in the embodiment are applied to preparation of antitumor drugs.
Example 13
A reduction-sensitive medicinal composition is topotecan liposome, is granular, has a particle size of 120nm, and comprises di-n-tetradecyl dithiopropionic acid glycerol phosphocholine, irinotecan, cholesterol, distearoyl glycerol phosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000) and distearoyl phosphatidylcholine (DSPC).
The preparation method comprises the following steps:
dissolving 0.5mmol of di-n-tetradecyl dithiopropionic acid glycerophosphorylcholine 14-SS3-PC, 0.5mmol of DSPC, 0.2mmol of cholesterol and 0.1mmol of DSPE-PEG20000.1mmol in a proper amount of 2mL of absolute ethyl alcohol, dropwise adding the solution into a sucrose octasulfate triethylamine solution with the temperature of 60 ℃, and stirring for 10 min; removing ethanol by rotary evaporation under reduced pressure, and homogenizing with 0.2 μm filter membrane; dialyzing the blank liposome suspension with PBS, and removing sucrose octasulfate triethylamine in the external water phase to obtain blank liposome; respectively sucking blank liposome and topotecan drug solution, and incubating and carrying the drug for 30min under the water bath condition of 60 ℃ to obtain the irinotecan liposome.
The topotecan liposome particles prepared in the embodiment are applied to preparation of antitumor drugs.
Example 14
A reduction-sensitive pharmaceutical composition is doxorubicin reduction-sensitive stealth liposome series particles, and comprises a bis-n-alkyl dithio-n-alkanoic acid glycerophosphocholine series (bis-n-octyl dithio-propionic acid glycerophosphocholine 8-SS3-PC, bis-n-decyl dithio-propionic acid glycerophosphocholine 10-SS3-PC, bis-n-dodecyl dithio-propionic acid glycerophosphocholine 12-SS3-PC, bis-n-tetradecyl dithio-propionic acid glycerophosphocholine 14-SS3-PC, bis-n-hexadecyl dithio-propionic acid glycerophosphocholine 16-SS3-PC), doxorubicin, cholesterol, distearoyl glycerol phosphatidylethanolamine-polyethylene glycol (DSPE-PEG 2000).
Preparation of doxorubicin reduction-sensitive stealth liposome series:
0.9mmol of di-n-alkyl dithio-n-alkanoic acid glycerophosphocholine series (di-n-octyl dithio propionic acid glycerophosphocholine 8-SS3-PC, di-n-decyl dithio propionic acid glycerophosphocholine 10-SS3-PC, di-n-dodecyl dithio propionic acid glycerophosphocholine 12-SS3-PC, di-n-tetradecyl dithio propionic acid glycerophosphocholine 14-SS3-PC, di-n-hexadecyl dithio propionic acid glycerophosphocholine 16-SS3-PC), and is mixed with distearoyl phosphatidyl ethanolamine polyethylene glycol DSPE-PEG20000.1mmol, cholesterol 0.3mmol and chloroform 20ml, the solvent is evaporated by spinning at 60 ℃, 20ml of ammonium sulfate solution is added, the membrane is formed at 60 ℃, and the membrane is filtered by an ultrasonic homogenization and 200nm filter membrane to obtain the reduction sensitive stealth liposome series nanoparticle solution, respectively marked as 8-SS3-LLP, 10-SS3-LLP, 12-SS3-LLP, 14-SS3-LLP and 16-SS 3-LLP.
Dialyzing the liposome solution with semipermeable membrane to remove ammonium sulfate from liposome; adding 0.1mmol of doxorubicin hydrochloride into the liposome solution, and incubating at 50 ℃; the doxorubicin enters the liposome to form sulfate gel to be deposited in the liposome to obtain doxorubicin reduction response invisible liposome series which are respectively marked as DOX/8-SS3-LLP, DOX/10-SS3-LLP, DOX/12-SS3-LLP, DOX/14-SS3-LLP and DOX/16-SS 3-LLP.
The doxorubicin common stealth liposome (denoted DOX/LP) was prepared as above except that 14-SS3-PC was replaced with equimolar DSPC.
The content of doxorubicin in the liposome solution was measured by uv spectroscopy, and the doxorubicin loading efficiency (EE%) and the drug loading amount (DL%) were calculated, and the results are shown in table 1.
The mean diameter of liposome particles and surface potential were measured by dynamic laser light scattering (DLS) method, and the results are shown in table 1. It can be seen that the size of the dithio phospholipid long-circulating liposome is about 100 nanometers, and the surface of the dithio phospholipid long-circulating liposome is negatively charged, which indicates that the liposome has stability.
Transmission electron microscopy (TEM, JEOL) and cryo-transmission electron microscopy (Tecnai G2F 20200 kV, FEI) the structure of DOX/12-SS3-LLP liposomes was observed and the results are shown in FIG. 2 a. It can be seen that the liposomes are of regular spherical structure with a diameter smaller than the liposome hydration kinetics diameter measured by DLS. Cryo-transmission electron microscopy showed that the dithiophospholipid liposomes 12-SS3-LLP are in the typical unilamellar liposome structure with a bilayer thickness of about 5 nm (inset panel in FIG. 2 a).
TABLE 1 characterization of drug-loaded dithiophospholipid stealth liposomes
Example 15
Lysis of DOX/12-SS3-LLP reduction-sensitive stealth liposomes
The degradation experiments simulated an in vivo reducing environment using DTT solution. Dithiophospholipid stealth liposomes DOX/12-SS3-LLP (10. mu.g/mL) of example 14 were incubated with 0.5mL of aqueous dithiothreitol (DTT, concentration 10mmol/L) for 1 hour at room temperature. The structural changes of the liposomes were observed by transmission electron microscopy and the results are shown in FIG. 2 b.
As can be seen in fig. 2b, the liposomes disintegrated completely into fragments under the effect of DTT. It is clear that the dithiophospholipids are cleaved into non-amphiphilic molecules under reducing conditions, resulting in complete disintegration of the liposomes. The dithio phospholipid cracking process is beneficial to the sensitive complete disintegration and rapid and full drug release of long-circulating liposome.
Example 16
Paclitaxel reduction response stealth liposome in-vitro degradation drug release test
Paclitaxel reduction-responsive stealth liposome nanoparticles prepared in example 4 were dissolved in 10mL of PBS solution and divided into 2 portions, one of which was 0.5mL of Glutathione (GSH) in PBS buffer (GSH concentration 50mmol) and the other was 0.5mL of PBS buffer. The control was a normal long circulating liposome (DOX/LP) loaded with the same molar content of paclitaxel (except 14-SS3-PC was replaced with glyceryl phosphorylcholine distearate DSPC, the 2-component ratios and methods were the same as in example 4). Placing the mixture in an incubator to incubate at 37 ℃, and detecting the content of paclitaxel by using high performance liquid chromatography (Agilent 1100LC, Zorbax reversed phase C18 column, 150X 4.6mm, 5 μm, sample size of 20 μ L, column temperature of 25 ℃, detection wavelength of 254nm, gradient elution: 2-90% buffer B/A, flow rate of 1.0mL/min, buffer A: deionized water of 0.1% TFA, buffer B: acetonitrile of 0.1% TFA), and calculating the release amount of paclitaxel.
The result shows that the paclitaxel released by the GSH-treated paclitaxel reduction response stealth liposome solution after 3 hours reaches 82 percent of the total amount; paclitaxel reduction-responsive stealth liposome solution without GSH treatment released only 11% of the total amount of paclitaxel after 3 hours. Paclitaxel was released after 3 hours from the paclitaxel loaded normal long circulating liposomes to 9% of the total amount. Therefore, the paclitaxel reduction response stealth liposome has reduction sensitivity, can quickly release paclitaxel original drug under the action of GSH, and has the release speed far higher than that of the common long-circulating liposome.
Example 17
Doxorubicin reduction response stealth liposome in vitro degradation drug release test
The doxorubicin reduction-responsive stealth liposome solution series prepared in example 14, DOX/8-SS3-LLP, DOX/10-SS3-LLP, DOX/12-SS3-LLP, DOX/14-SS3-LLP, DOX/16-SS3-LLP, each 2mL (DOX concentration 0.1mg/mL), were added to a dialysis bag (MWCO 8000). The release medium is PBS buffer solution containing (or not containing) 10mM dithiothreitol DTT. The dialysis bag and the liposome are immersed into a release medium, and the system is maintained at a constant temperature of 37 ℃; at different time points, 1mL of degradation medium was sampled while 1mL of fresh release medium was replenished, the DOX concentration of each sample was measured using an ultraviolet spectrophotometer, and the final release rate was calculated.
As shown in FIG. 3, compared with the DOX-loaded common stealth liposome (DOX/LP, example 14), the DOX-loaded reduction-sensitive long-circulating liposome has high release rate in the release medium containing 10mM DTT, and the cumulative release amount within 24 hours can reach about 80%; in contrast, in a release medium without DTT, the cumulative release amount of the DOX-loaded reduction-sensitive stealth liposome can only reach 10% -20%, so that the DOX-loaded reduction-sensitive stealth liposome has obvious reduction responsiveness and can quickly release the loaded DOX drug.
Example 18
In vitro tumor cell killing experiment
In example 14 evaluation of antitumor Effect in vitro of DOX-loaded reduction-sensitive stealth liposomes (DOX/8-SS3-LLP, DOX/10-SS3-LLP, DOX/12-SS3-LLP, DOX/14-SS3-LLP, DOX/16-SS3-LLP) two cell models, A549 and MCF-7, were used. A549 and MCF-7 cells were seeded at a density of 10000 cells/well in 96-well culture plates; setting five concentration gradients of 0.001, 0.01, 0.1, 1 and 10 mug/mL, and the incubation time is 24 hours; after the incubation was completed, the medium in each well was aspirated, 20. mu.L of MTT solution was added, and the mixture was incubated at 37 ℃ for 4 hours. Then, the MTT solution in each well was aspirated off, and 150. mu.L of DMSO solution was added; finally, the absorbance (OD) of each well was read at 490nm wavelength using a microplate reader. Cell viability was calculated according to the formula.
ODdrugOD value, OD of well representing incubation drugcontrolThe OD value, OD of the wells incubated with the medium without the drug aloneblankOD of wells with DMSO only.
The results are shown in FIG. 4, and compared with DOX/LP (example 14), DOX/8-SS3-LLP, DOX/10-SS3-LLP, DOX/12-SS3-LLP, DOX/14-SS3-LLP have a relatively stronger effect of killing cells. DOX is a free drug, has the fastest cellular uptake speed, and therefore, often shows a stronger effect of killing cells.
Experimental example 19
Antitumor test in animal body
Preparation of a nude mouse model: 2-month-old, 15-20 g female BALB/c mice were selected. Collecting the cultured MCF-7 cell suspension at a concentration of 1X 107Each dose of each dose was inoculated subcutaneously into the right axilla of a nude mouse at 0.1ml until the average tumor size was 0.1 cc.
Grouping and administration: measuring the diameter of the transplanted tumor of the nude mouse by using a vernier caliper, and enabling the tumor to grow to 75mm3The animals were randomly divided into 20 experimental animals and 5 experimental animals. Meanwhile, each group of nude mice starts tail vein injection administration, the administration scheme is shown in the group and the administration scheme, and the antitumor effect of the tested sample is dynamically observed by using the method for measuring the tumor diameter. The formula for Tumor Volume (TV) is: TV 1/2 × a × b2(formula 3-2), wherein a and b represent the length and width, respectively.
Groups and dosing regimens: the dosage is 5 mg/kg.
Blank group: saline, i.v. once every three days, 0.2ml in volume for 3 consecutive weeks.
Control group: doxorubicin was dissolved in saline and injected intravenously every three days in a volume of 0.2ml for 3 consecutive weeks. The group of common Dox stealth liposomes loaded with doxorubicin (example 14) was injected intravenously every three days in a volume of 0.2ml for 3 consecutive weeks.
Drug group: DOX/12-SS 3-group LLP (example 14); DOX/16-SS 3-group LLP (example 14). Intravenous injection, 0.2ml in volume, every three days for 3 weeks.
During the experiment, the body weight and tumor size of the mice were recorded every two days. After 21 days of dosing, all groups of mice were sacrificed and tumors were isolated and recorded by weighing, and the results are shown in fig. 5.
Both DOX/12-SS3-LLP and DOX/16-SS3-LLP showed significant reduction in tumor volume and weight compared to the control group and the blank group, indicating a better tumor growth inhibition effect (FIGS. 5a and 5 c). Wherein DOX/16-SS3-LLP has the best effect, and has better effect of inhibiting tumor growth than DOX/12-SS 3-LLP. As can be seen from the body weight change curve of the mice (fig. 5b), the doxorubicin saline group had the lowest body weight and showed significant toxic and side effects; the body weight of the reduction-sensitive long-circulating liposome group was high, indicating insignificant toxicity.
Example 20
A reduction-sensitive pharmaceutical composition is paclitaxel reduction-response solid lipid nanoparticles with particle size of 132nm, and comprises di-n-tetradecyl dithiopropionic acid glycerophosphocholine, paclitaxel, stearic acid, and Poloxamer Poloxamer 188.
The preparation method comprises the following steps:
paclitaxel 0.1g, palmitic acid 5.0g, lecithin 2.0g are mixed, heated to 80 ℃ for slow dissolution, added into 20ml PBS (pH 7.4) containing di-n-tetradecyl dithiopropionic acid glycerophosphocholine 14-SS3-PC 0.5g and poloxamer Poloxamer 1880.5 g of water solution, stirred at high speed, homogenized under high pressure for 3 times, cooled to obtain solid lipid nanoparticles.
The average particle size was measured by a laser particle sizer to 132nm, and the morphology of the nanoparticles was measured by a transmission electron microscope to show a spherical structure.
Example 21
A reduction-sensitive pharmaceutical composition is a paclitaxel reduction-response nano micelle, has a particle size of 120nm, and comprises di-n-tetradecyl dithiopropionic acid glycerophosphocholine, paclitaxel, tricaprylin and tween 80.
The preparation method comprises the following steps:
0.1mmol of docetaxel, 14-SS3-PC 1.0mmol of di-n-tetradecyl dithiopropionic acid glycerophosphocholine, 1.0mmol of diphosphonocholine dipalmitate, 2.0mmol of glycerol monocaprylate, 1.0mmol of glycerol tricaprylate and 800.3 mmol of Tween are heated to 60 ℃ for slow dissolution, 20ml of PBS (pH 7.4) solution is added, high-speed stirring is carried out, and the average value of a high-pressure homogenizer is carried out for 3 times, so that the nano micelle is obtained.
The average particle size was measured by a laser particle sizer to be 120nm, and the morphology of the nanoparticles was measured by a transmission electron microscope to show a spherical structure.
Example 22
A reduction-sensitive liposome pharmaceutical composition is a mivawood peptide liposome, is granular and has a particle size of 132nm, and comprises di-n-tetradecyl dithiopropionic acid glycerol phosphocholine, mivawood peptide, cholesterol, distearoyl glycerol phosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000) and distearoyl phosphatidylcholine (DSPC).
The preparation method comprises the following steps:
dissolving 0.5mmol of di-n-tetradecyl dithiopropionic acid glycerophosphorylcholine 14-SS3-PC, 0.5mmol of DSPC, 0.2mmol of cholesterol, DSPE-PEG20000.1mmol and 0.1mmol of mivawood peptide in 5mL of dichloromethane, adding into a 250mL eggplant-shaped bottle, removing the solvent by rotary evaporation, and adding 5mL of physiological saline for hydration; performing ultrasonic treatment for 5 minutes, homogenizing through a 0.2 mu m filter membrane to obtain the mivabradine lipidosome nano particles, wherein the average diameter of the nano particles measured by a laser particle size analyzer is 132 nm.
The mivajatide liposome particles prepared in the embodiment are applied to preparation of antitumor drugs.
Example 23
A reduction-sensitive medicinal composition is triptolide liposome, is in the form of granule with particle diameter of 150nm, and comprises di-n-tetradecyl dithiopropionic acid glycerophosphocholine, triptolide, cholesterol, DSPE-PEG2000, distearoyl glycerol phosphatidylethanolamine-polyethylene glycol-folic acid (DSPE-PEG2000-FA), and distearoyl phosphatidylcholine (DSPC).
The preparation method comprises the following steps:
dissolving 0.5mmol of di-n-tetradecyl dithiopropionic acid glycerophosphorylcholine 14-SS3-PC, 0.5mmol of DSPC, 0.2mmol of cholesterol, 0.1mmol of DSPE-PEG20000.1 mmol, 0.02mmol of DSPE-PEG2000-FA and 0.05mmol of triptolide in 5mL of dichloromethane, adding into a 250mL eggplant-shaped bottle, removing the solvent by rotary evaporation, and adding 5mL of physiological saline for hydration; carrying out ultrasonic treatment for 5 minutes, and homogenizing through a 0.2 mu m filter membrane to obtain the triptolide liposome nanoparticles with targeted folate receptors, wherein the average diameter of the nanoparticles measured by a laser particle sizer is 150 nm.
The triptolide liposome nanoparticles prepared in the embodiment are applied to preparation of antitumor drugs.
Example 24
A reduction sensitive medicinal composition is a tamoxifen liposome, is granular and has a particle size of 142 nanometers, and comprises di-n-tetradecyl dithiopropionic acid glycerol phosphocholine, tamoxifen, cholesterol, distearoyl glycerol phosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000) and distearoyl phosphatidylcholine (DSPC).
The preparation method comprises the following steps:
dissolving 0.5mmol of di-n-tetradecyl dithiopropionic acid glycerophosphorylcholine 14-SS3-PC, 0.5mmol of DSPC, 0.2mmol of cholesterol, DSPE-PEG20000.1mmol and 0.05mmol of tamoxifen in 10mL of diethyl ether, adding into a 250mL eggplant-shaped bottle, removing the solvent by rotary evaporation, and adding 5mL of physiological saline for hydration; and (3) carrying out ultrasonic treatment for 5 minutes, homogenizing through a 0.2-micron filter membrane to obtain tamoxifen liposome nanoparticles, wherein the average diameter of the nanoparticles measured by a laser particle sizer is 142 nanometers.
The tamoxifen liposome particles prepared in the embodiment are applied to preparing antitumor agents.
The above examples are only preferred embodiments of the present invention, it should be noted that: it will be apparent to those skilled in the art that various modifications and equivalents can be made without departing from the spirit of the invention, and it is intended that all such modifications and equivalents fall within the scope of the invention as defined in the claims.
Claims (7)
1. A reduction-sensitive pharmaceutical composition, comprising: the medicine composition is nanoparticles and comprises di-n-alkyl dithio-n-alkanoic acid glycerophosphocholine and a medicine;
wherein the structural general formula of the di-n-alkyl dithio-n-alkanoic acid glycerophosphorylcholine is as follows:
wherein m is a positive integer of 2 to 10, and n is a positive integer of 7 to 15;
the nano-particles are liposomes, solid lipid nanoparticles or micelles, and the particle size of the particles is 10-1000 nm.
2. A reduction-sensitive pharmaceutical composition according to claim 1, wherein: the reduction-sensitive pharmaceutical composition further comprises a pharmaceutically acceptable adjuvant.
3. A reduction-sensitive pharmaceutical composition according to claim 1, wherein: the medicine is an anti-tumor medicine, an anti-virus medicine or an antibacterial medicine.
4. A reduction-sensitive pharmaceutical composition according to claim 3, wherein: the anti-tumor drug is one of paclitaxel, docetaxel, cabazitaxel, camptothecin, hydroxycamptothecin, 7-ethyl-10-hydroxycamptothecin, irinotecan, topotecan, lurtotecan, belotecan, 7-hydroxymethyl camptothecin, vincristine, vinorelbine, ansamycin, rapamycin, cytarabine, etoposide, mitoxantrone, doxorubicin, zorubicin, epirubicin, idarubicin, sambicin, tamoxifen, triptolide, mivaquoid, ursolic acid or platinum drugs; wherein the platinum drug comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, leplatin or cis-bisneodecanoic acid-trans-R, R-1, 2-cyclohexanediamine platinum (II).
5. A reduction-sensitive pharmaceutical composition according to claim 3, wherein: the antibacterial drug is one of amphotericin B, gentamicin, nystatin, fluconazole, amikacin, levofloxacin or econazole nitrate; the antiviral drug is ribavirin or adefovir dipivoxil.
6. A reduction-sensitive pharmaceutical composition according to claim 2, wherein: the pharmaceutically acceptable auxiliary agent is one or more of phospholipid, glyceride, fatty acid, tween, poloxamer or cholesterol.
7. A process for preparing a reduction-sensitive pharmaceutical composition according to any one of claims 1 to 6, wherein: the reduction sensitive medicine composition is prepared by a film dispersion method, a reverse phase evaporation method, a freeze drying method, an ultrasonic dispersion method, a spray drying method, a film extrusion method or a high-pressure homogenization method;
the reduction-sensitive pharmaceutical composition is applied to preparation of an anti-tumor medicament, an anti-virus medicament or an antibacterial medicament, and the anti-tumor medicament, the anti-virus medicament or the antibacterial medicament is a liquid preparation, a solid preparation or a semisolid preparation.
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