CN115120738B - Imiquimod prodrug nano-particles, and preparation method and application thereof - Google Patents
Imiquimod prodrug nano-particles, and preparation method and application thereof Download PDFInfo
- Publication number
- CN115120738B CN115120738B CN202210857719.1A CN202210857719A CN115120738B CN 115120738 B CN115120738 B CN 115120738B CN 202210857719 A CN202210857719 A CN 202210857719A CN 115120738 B CN115120738 B CN 115120738B
- Authority
- CN
- China
- Prior art keywords
- imiquimod
- imq
- polymer
- hedsma
- tumor
- 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.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
-
- 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
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/58—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly[meth]acrylate, polyacrylamide, polystyrene, polyvinylpyrrolidone, polyvinylalcohol or polystyrene sulfonic acid resin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
- A61K47/6927—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores
- A61K47/6929—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
- A61K47/6931—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer
- A61K47/6933—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer the polymer being obtained by reactions only involving carbon to carbon, e.g. poly(meth)acrylate, polystyrene, polyvinylpyrrolidone or polyvinylalcohol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
The invention belongs to the technical fields of biological medicine and molecular biology, and particularly relates to imiquimod prodrug nanoparticles, and a preparation method and application thereof. The invention uses chemical means to mask the C4 amino of imiquimod to obtain IMQ prodrug, the prodrug has GSH sensitivity, at the same time adopts RAFT polymerization technology to polymerize synthetic monomer, and makes modification of tumor targeting peptide on polymer, finally obtains nano-level nanoparticle delivery system by self-assembly technology. After intravenous injection, the nanosystems are relatively stable and will not be released in advance due to the lack of GSH environment. The nanosystems aggregate into tumor tissue through targeting of tumor targeting peptides. After endocytosis of the tumor cells, high GSH levels within the tumor cells promote disintegration of the nanosystems, and then release free IMQ. The free IMQ enhances infiltration of T lymphocytes in the tumor microenvironment by activating dendritic cells, which is beneficial for safe and effective delivery of IMQ.
Description
Technical Field
The invention belongs to the technical fields of biological medicine and molecular biology, and particularly relates to imiquimod prodrug nanoparticles, and a preparation method and application thereof.
Background
The information disclosed in the background of the invention is only for enhancement of understanding of the general background of the invention and is not necessarily to be taken as an admission or any form of suggestion that this information forms the prior art already known to a person of ordinary skill in the art.
Toll-like receptor (TLR) 7/8 small molecule agonists are potent activators of Antigen Presenting Cells (APCs) that can directly kill tumor cells and enhance infiltration of Cytotoxic T Lymphocytes (CTLs) by phagocytosis; they are widely studied and investigated as immunomodulators for cancer immunotherapy. Among them, imiquimod class drugs have been approved by the U.S. Food and Drug Administration (FDA) for localized basal cell carcinoma, and experiments directed to metastatic melanoma and localized bladder cancer are also underway. However, the uncontrolled rapid distribution of such small molecule agonists often causes dose-dependent immune-related adverse effects, including fatal severe cytokine storms, upon systemic administration, thereby limiting their clinical use. Currently, various attempts have been made to achieve site-specific immune activation and avoid unwanted systemic inflammation caused by its systemic distribution. These efforts include modification with lipid motifs, covalent conjugation to biological/nanomaterials, physical entrapment of nanoparticles by hydrophobic and electrostatic interactions, and the like. However, these strategies do not cover the amino group at position C4, which is critical to the biological activity of imidazoquinolines.
The biopharmacology of the small molecule imiquimod can be improved by a polymer-drug conjugate, which has many advantages such as enhanced solubility, controlled drug delivery, improved efficacy, and the like. Chemical polymer-drug binding not only can prolong blood circulation and increase tumor accumulation, but also can avoid systemic toxicity. One popular attachment strategy involves attaching the imidazoquinoline to the lipopolymer via a stimulus-responsive linker. The imidazoquinoline-polymer conjugate is achieved by attaching the imidazoquinoline to the backbone of the preformed polymer.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide imiquimod prodrug nanoparticles, and a preparation method and application thereof. The invention uses chemical means to mask off the C4 amino of Imiquimod (IMQ) to obtain IMQ precursor drug HEDSMA-IMQ (SS), the precursor drug has GSH sensitivity, at the same time adopts RAFT polymerization technology to polymerize synthetic monomer, and carries out modification of tumor targeting peptide (cRGD) on polymer, finally carries out self-assembly on the polymer by self-assembly technology to obtain nano-level nanoparticle delivery system. After intravenous injection, the nanosystems are relatively stable and will not be released in advance due to the lack of GSH environment. The nanosystems aggregate into tumor tissue through targeting of tumor targeting peptides. After endocytosis of the tumor cells, high GSH levels within the tumor cells promote disintegration of the nanosystems, and then release free IMQ. Free IMQ further enhances T lymphocyte infiltration in tumor microenvironment by activating Dendritic Cells (DCs), which is very beneficial for safe and effective delivery of IMQ. Based on the above results, the present invention has been completed.
In a first aspect of the invention, there is provided imiquimod prodrug nanoparticles obtained by self-assembly of an amphiphilic polymer, said amphiphilic polymer being further modified with a tumor targeting target.
Wherein the amphiphilic polymer is polymerized from poly (N, N-dimethylacrylamide) and imiquimod precursor; the imiquimod precursor is specifically obtained after the imiquimod masks the C4 amino group. The imiquimod precursor has Glutathione (GSH) sensitivity, i.e., the proto-drug is released only in high GSH environments.
In a second aspect of the present invention, there is provided a method for preparing the imiquimod prodrug nanoparticles described above, the method comprising: synthesizing a hydrophilic polymer poly (N, N-dimethylacrylamide) (pDMA), and then further polymerizing an imiquimod precursor HEDSMA-IMQ (SS) on the basis of the hydrophilic end to obtain an amphiphilic polymer pDMA-b-pIMQ (SS); and then carrying out target modification on the tail end of the polymer, and carrying out self-assembly on the polymer by a self-assembly technology to obtain the modified polymer.
In a third aspect of the invention, there is provided the use of imiquimod prodrug nanoparticles as described above in the preparation of an anti-tumor drug.
In a fourth aspect of the present invention, there is provided an antitumor drug, the active ingredient of which comprises the above nanoparticle.
According to the invention, the medicament further comprises at least one pharmaceutically inactive ingredient.
In a fifth aspect of the invention there is provided a method of treating a tumour, the method comprising administering to a subject a therapeutically effective dose of imiquimod prodrug nanoparticles or a medicament as described above.
The beneficial technical effects of one or more of the technical schemes are as follows:
the technical scheme provides imiquimod prodrug nano-particles, a preparation method and application thereof, and the invention uses a chemical means to mask off the C4 amino group of IMQ to obtain an IMQ prodrug HEDSMA-IMQ (SS), wherein the prodrug has GSH sensitivity, i.e. the proto-drug can be released only in a high GSH environment. In order to polymerize these monomers to give polymers, RAFT polymerization techniques were used to polymerize the monomers synthesized. Firstly, a hydrophilic polymer poly (N, N-dimethylacrylamide) (pDMA) is synthesized, and then HEDSMA-IMQ (SS) is further polymerized on the basis of the hydrophilic end to obtain an amphiphilic polymer pDMA-b-pIMQ (SS). Next, the polymer tail is modified with a tumor targeting peptide (cRGD) to impart tumor tissue targeting properties to the polymer. Finally, the polymers are self-assembled by a self-assembly technology to obtain a nano-level nanoparticle delivery system.
After intravenous injection, the nano system is stable and can not be released in advance due to lack of GSH environment. The nanosystems aggregate into tumor tissue through targeting of tumor targeting peptides. After endocytosis of the tumor cells, high GSH levels within the tumor cells promote disintegration of the nanosystems, and then release free IMQ. Free IMQ further enhances T lymphocyte infiltration in the tumor microenvironment by activating Dendritic Cells (DCs). The technical scheme provides a promising platform for safe and effective delivery of the IMQ, so that the method has good practical application value.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention.
FIG. 1 is a diagram of HEDSMA-IMQ and HEDCMA-IMQ synthesis in an embodiment of the invention;
FIG. 2 is a graph showing HEDSMA nuclear magnetic resonance results in an embodiment of the invention;
FIG. 3 is a graph showing HEDSMA-IMQ nuclear magnetic resonance results in an embodiment of the present invention;
FIG. 4 shows the results of HEDCMA nuclear magnetism in an embodiment of the invention;
FIG. 5 is a graph showing the results of HEDCMA-IMQ nuclear magnetism in an embodiment of the invention;
FIG. 6 is a schematic representation of the synthesis and modification of polymers in an embodiment of the invention;
FIG. 7 shows the nuclear magnetic resonance of pDMA in the example of the present invention;
FIG. 8 shows the nuclear magnetic resonance results of pDMA-b-pIMQ (SS) in the examples of the present invention;
FIG. 9 shows the nuclear magnetic resonance results of c-RGD-pDMA-b-pIMQ (SS) in the examples of the present invention;
FIG. 10 shows the nuclear magnetic resonance results of pDMA-b-pIMQ (CC) in the examples of the present invention;
FIG. 11 shows the nuclear magnetic resonance results of c-RGD-pDMA-b-pIMQ (CC) in the examples of the present invention;
FIG. 12 is the construction and characterization of nanoparticles in an embodiment of the present invention; wherein a is a construction schematic diagram of nano-scale N@SS-IMQ, b is a construction schematic diagram of cN@SS-IMQ with tumor active targeting capability, c is a particle size distribution diagram of B@SS-IMQ, d is a particle size distribution diagram of cN@SS-IMQ, e is a transmission electron microscope image of N@SS-IMQ, and f is a cN@SS-IMQd projection electron microscope image;
FIG. 13 is an in vitro release of nanoparticles in an embodiment of the invention;
FIG. 14 is a diagram showing the relationship of BMDC activation in the embodiment of the invention, wherein a is a schematic diagram of release of free IMQ by nanoparticles to activate Dendritic Cells (DCs), b is a flow chart of a DC activation experiment, c is the result of inducing DC cell surface CD40 expression by different preparation groups, d is the result of inducing DC cell surface CD80 expression by different preparation groups, e is the result of inducing DC cell surface CD86 expression by different preparation groups, and f is the result of inducing DC cell surface MHC-II expression by different preparation groups;
FIG. 15 shows the results of mouse imaging in the examples of the present invention;
FIG. 16 is a graph showing the results of tumor inhibition curves in the examples of the present invention, wherein a is a tumor growth curve of untreated mice, b is a tumor growth curve of mice treated with free IMQ, and c is a tumor growth curve of mice treated with cN@SS-IMQ.
Detailed Description
It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the present invention. As used herein, the singular is also intended to include the plural unless the context clearly indicates otherwise, and furthermore, it is to be understood that the terms "comprises" and/or "comprising" when used in this specification are taken to specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof.
The invention will now be further illustrated with reference to specific examples, which are given for the purpose of illustration only and are not intended to be limiting in any way. If experimental details are not specified in the examples, it is usually the case that the conditions are conventional or recommended by the reagent company; reagents, consumables, etc. used in the examples described below are commercially available unless otherwise specified.
Studies indicate that the concentration of Glutathione (GSH) in tumors is about 1000 times that in normal tissues, thereby providing a convenient condition for designing intelligent nano-drugs with tumor responsiveness.
In view of this, in one exemplary embodiment of the present invention, there is provided imiquimod prodrug nanoparticles obtained by self-assembly of amphiphilic polymers that are further modified with tumor-targeting targets.
Wherein the amphiphilic polymer is polymerized from poly (N, N-dimethylacrylamide) and imiquimod precursor; the imiquimod precursor is specifically obtained after the imiquimod masks the C4 amino group. The imiquimod precursor has Glutathione (GSH) sensitivity, i.e., the proto-drug is released only in high GSH environments.
The tumor targeting target may be any known substance having a tumor targeting effect, and in one embodiment of the present invention, the tumor targeting target is a tumor targeting peptide, such as cRGD. Through the guiding function of the tumor targeting peptide, the nano system can be effectively aggregated into tumor tissues.
The average grain diameter of the nano-particles is about 200nm, and the morphology is relatively round.
In still another embodiment of the present invention, there is provided a method for preparing the imiquimod prodrug nanoparticle described above, the method comprising: synthesizing a hydrophilic polymer poly (N, N-dimethylacrylamide) (pDMA), and then further polymerizing an imiquimod precursor HEDSMA-IMQ (SS) on the basis of the hydrophilic end to obtain an amphiphilic polymer pDMA-b-pIMQ (SS); and then carrying out target modification on the tail end of the polymer, and carrying out self-assembly on the polymer by a self-assembly technology to obtain the modified polymer.
In yet another embodiment of the present invention, the method for synthesizing the amphiphilic polymer pDMA-b-pIMQ (SS) comprises: the imiquimod precursor HEDSMA-IMQ (SS), poly (N, N-dimethylacrylamide) pDMA and azodiisobutyronitrile are dissolved in dioxane and subjected to freeze thawing for 2-3 times, then heating reaction is carried out overnight, and the product is obtained after precipitation and purification by cold diethyl ether.
The molar ratio of the imiquimod precursor HEDSMA-IMQ (SS), the poly (N, N-dimethylacrylamide) pDMA and the azodiisobutyronitrile is 0.1-1:0.01-0.05:0.001-0.01, preferably 0.55:0.02:0.00548.
The heating reaction can be carried out by adopting an oil bath heating mode, and the specific heating temperature is controlled to be 70-90 ℃, preferably 80 ℃.
The specific method for carrying out target modification on the tail end of the polymer comprises the following steps: EDC and NHS are adopted to activate amphiphilic polymer pDMA-b-pIMQ (SS), then the amphiphilic polymer pDMA-b-pIMQ (SS) reacts with c-RGD peptide, and the amphiphilic polymer pDMA-b-pIMQ is obtained after dialysis.
The mole ratio of the amphiphilic polymer pDMA-b-pIMQ to the c-RGD peptide is 1-5:4-10, preferably 4.33:6.63.
the specific method for self-assembling the polymer by the self-assembling technology comprises the following steps: and (3) dissolving the polymer modified by the target head in dimethyl sulfoxide, adding the polymer into water under the ultrasonic condition to obtain a reaction mixture, and purifying the reaction mixture to obtain the polymer. The colloidal solution obtained after purification can be further lyophilized to obtain lyophilized powder.
In yet another embodiment of the present invention, the poly (N, N-dimethylacrylamide) is also commercially available, and in one embodiment of the present invention, the poly (N, N-dimethylacrylamide) synthesis method comprises: dissolving N, N-dimethylacrylamide, a chain transfer agent of 4-cyano-4- [ (dodecyl sulfanyl) sulfanyl ] pentanoic acid and azodiisobutyronitrile in dioxane, removing oxygen through freeze thawing circulation, heating for reaction overnight, and adding the obtained solution into diethyl ether to precipitate a product.
The molar ratio of the poly (N, N-dimethylacrylamide), the chain transfer agent 4-cyano-4- [ (dodecylsulfanylthiocarbonyl) sulfanyl ] pentanoic acid and the azodiisobutyronitrile is 1-5:0.01-0.1:0.01-0.02, preferably 3:0.06:0.012;
the heating reaction can be carried out by adopting an oil bath heating mode, and the specific heating temperature is controlled to be 70-90 ℃, preferably 80 ℃.
The preparation method of the imiquimod precursor HEDSMA-IMQ comprises the steps of dissolving HEDSMA, 4-dimethylaminopyridine and triphosgene in anhydrous dichloromethane to obtain a mixture, adding imiquimod dissolved in N, N-dimethylformamide into the mixture, stirring overnight, washing a reaction product with water, drying an organic solvent phase, removing the organic solvent phase, and purifying to obtain the imiquimod precursor HEDSMA-IMQ.
The mol ratio of HEDSMA, 4-dimethylaminopyridine, triphosgene and imiquimod is 0.1-0.5:1-5:0.1-0.3:1-3, preferably 0.39:1.18:0.16:1.11;
wherein, the HEDSMA can be synthesized by any known method, and in a specific embodiment of the invention, the HEDSMA is prepared by adopting the following method: dissolving hydroxyethyl disulfide in anhydrous tetrahydrofuran under the protection of inert gas (such as nitrogen), adding methacryloyl chloride and triethylamine, removing tetrahydrofuran, washing with water, and drying and purifying the organic phase.
The mol ratio of the hydroxyethyl disulfide, the methacryloyl chloride and the triethylamine is 30-40:35-45:35-45, and is preferably 33.45:40:36.8.
In yet another embodiment of the present invention, there is provided the use of imiquimod prodrug nanoparticles as described above in the preparation of an antitumor drug.
Meanwhile, it should be noted that tumors are used in the present invention as known to those skilled in the art, and include benign tumors and/or malignant tumors. Benign tumors are defined as hyperproliferative cells that are unable to form aggressive, metastatic tumors in vivo. Conversely, a malignancy is defined as a cell with multiple cellular abnormalities and biochemical abnormalities that are capable of developing a systemic disease (e.g., tumor metastasis in a distant organ).
In yet another embodiment of the invention, the medicament of the invention is useful for the treatment of malignant tumors. Examples of malignant tumors that can be treated with the medicament of the invention include solid tumors and hematological tumors. Preferably, the solid tumor is selected from the group consisting of breast, bladder, bone, brain, central and peripheral nervous system, colon, endocrine gland, esophagus, endometrium, head, neck, liver, larynx and hypopharynx, mesothelioma, ovary, pancreas, prostate, rectum, kidney, small intestine, soft tissue, testis, stomach, skin, ureter, vagina and vulva, etc., without specific limitation.
In still another embodiment of the present invention, there is provided an antitumor drug whose active ingredient comprises the above nanoparticle.
According to the invention, the medicament further comprises at least one pharmaceutically inactive ingredient.
The pharmaceutically inactive ingredients may be carriers, excipients, diluents and the like which are generally used in pharmacy. Further, the composition can be formulated into various dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, sprays, etc., for oral administration, external use, suppositories, and sterile injectable solutions according to a usual method.
The non-pharmaceutically active ingredients, such as carriers, excipients and diluents, which may be included, are well known in the art and can be determined by one of ordinary skill in the art to meet clinical criteria.
In yet another embodiment of the present invention, the carriers, excipients and diluents include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil and the like.
In yet another embodiment of the invention, the medicament of the invention may be administered to the body in a known manner. For example, by intravenous systemic delivery or local injection into the tissue of interest. Alternatively via intravenous, transdermal, intranasal, mucosal or other delivery methods. Such administration may be via single or multiple doses. It will be appreciated by those skilled in the art that the actual dosage to be administered in the present invention may vary greatly depending on a variety of factors, such as the target cell, the type of organism or tissue thereof, the general condition of the subject to be treated, the route of administration, the mode of administration, and the like. In particular, experiments prove that the medicine is stable after intravenous injection, and can not be released in advance due to lack of GSH environment. The nanosystems aggregate into tumor tissue through targeting of tumor targeting peptides. After endocytosis of the tumor cells, high GSH levels within the tumor cells promote disintegration of the nanosystems, and then release free IMQ. Free IMQ further enhances T lymphocyte infiltration in the tumor microenvironment by activating Dendritic Cells (DCs).
The subject to be administered can be human and non-human mammals, such as mice, rats, guinea pigs, rabbits, dogs, monkeys, gorillas, etc.
In yet another embodiment of the invention, a method of treating a tumor is provided, the method comprising administering to a subject a therapeutically effective dose of the nanoparticle or drug described above.
The subject is an animal, preferably a mammal, most preferably a human, who has been the subject of treatment, observation or experiment. By "therapeutically effective amount" is meant that amount of active compound or pharmaceutical agent, including a compound of the present invention, which causes a biological or medical response in a tissue system, animal or human that is sought by a researcher, veterinarian, medical doctor or other medical personnel, which includes alleviation or partial alleviation of the symptoms of the disease, syndrome, condition or disorder being treated. It must be recognized that the optimal dosage and spacing of the active ingredients of the present invention is determined by its nature and external conditions such as the form, route and site of administration and the particular mammal being treated, and that such optimal dosage may be determined by conventional techniques. It must also be appreciated that the optimal course of treatment, i.e. the daily dosage of the simultaneous compounds over the nominal time period, can be determined by methods well known in the art.
The invention is further illustrated by the following examples, which are not to be construed as limiting the invention. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. The following examples are test methods in which specific conditions are noted, and are generally conducted under conventional conditions.
Examples
Test method
1 Synthesis of monomers
1.1 Synthesis of HEDSMA and HEDCMA
For HEDSMA, hydroxyethyl disulfide HEDS (4.00 mL,33.45 mmol) was dissolved in 30mL anhydrous tetrahydrofuran under nitrogen. Methacryloyl chloride (3.80 mL,40.0 mmol) and triethylamine (506. Mu.L, 36.80 mmol) were then added. Then, tetrahydrofuran was removed by rotary evaporation under reduced pressure, and after washing the product with water three times, the organic phase was dried over anhydrous sodium sulfate and then rotary evaporated. Then separating to obtain a product, and verifying the result through nuclear magnetism. Control HEDCMA was also synthesized by a similar method.
1.2 Synthesis of IMQ-HEDSMA and IMQ-HEDCMA
IMQ-HEDSMA was synthesized in two steps. First, at N 2 HEDSMA (80 mg,0.39 mmol), 4-dimethylaminopyridine (43.50 mg,1.18 mmol) and triphosgene (46.50 mg,0.16 mmol) were dissolved in anhydrous dichloromethane (DCM, 10 mL) under an atmosphere. IMQ (236.5 mg,1.11 mmol) dissolved completely in N, N-dimethylformamide (100 ml) was then slowly added to the mixture and stirred overnight. The reaction mixture was transferred to a separatory funnel and washed twice with Milli-Q water. MgSO for DCM layer 4 Dried and removed under reduced pressure. The product was purified using a silica gel column.IMQ-heddma was synthesized by a similar reaction, with only HEDSMA being replaced by heddma, and the other conditions remaining unchanged.
RAFT polymerization of 2 Polymer
2.1 Synthesis of pDMA
N, N-dimethylacrylamide DMA (3.0 mmol,297.39 mg), the chain transfer agent 4-cyano-4- [ (dodecylsulfanylthiocarbonyl) sulfanyl ] pentanoic acid (0.06 mmol,24.22 mg) and azobisisobutyronitrile (0.012 mmol,1.97 mg) were dissolved in dioxane (2M). Oxygen was removed thoroughly by three freeze-thaw cycles and reacted overnight in an oil bath at 80 ℃. The polymer was then precipitated by dropping the solution into diethyl ether to give pale yellow pDMA.
2.2 Synthesis of pDMA-b-pIMQ (SS) and pDMA-b-pIMQ (CC)
IMQ-HEDSMA (267 mg,0.55 mmol), pDMA (95.80 mg,0.02 mmol) and azobisisobutyronitrile AIBN (0.90 mg, 5.48. Mu. Mol) were dissolved in dioxane (2M) in a 25mL Schlenk tube and passed through three freeze-thaw cycles. Placed in an oil bath at 75 ℃ overnight. Purification by precipitation in ice-cold diethyl ether gave pDMA-b-pIMQ (SS).
IMQ-heddma was synthesized by the same method.
2.3 Synthesis of c-RGD-pDMA-b-pIMQ (SS) and c-RGD-pDMA-b-pIMQ (CC)
pDMA-b-pIMQ (SS) (80 mg, 4.33. Mu. Mol) was dissolved in anhydrous DCM containing 1-ethyl- (3-dimethylaminopropyl) carbodiimide (1.5 mg, 7.82. Mu. Mol) and N-hydroxysuccinimide (0.9 mg, 7.82. Mu. Mol) for 1 hour. EDC/NHS activated pDMA-b-pIMQ (SS) was allowed to react with c-RGD peptide (4 mg, 6.63. Mu. Mol) for an additional 24 hours under magnetic stirring. The c-RGD-pDMA-b-pIMQ (SS) was purified by dialysis. The c-RGD-pDMA-b-pIMQ (CC) was synthesized by the same method.
Construction of 3 nm delivery platforms cN@SS-IMQ and cN@CC-IMQ
c-RGD-pDMA-b-pIMQ (SS) or c-RGD-pDMA-b-pIMQ (CC) (4 mg mL) -1 50 μl) was dissolved in DMSO and added dropwise to Milli-Q water (1 mL) under ultrasound. The reaction mixture was purified by dialysis against Milli-Q water (frequent exchange of dialysis medium) to remove byproducts and solvents. After a few days, the colloidal solution was lyophilized to a powder.
Characterization of 4 cN@SS-IMQ
The particle size distribution of the cN@SS-IMQ was determined using an Zetasizer Nano ZS instrument. After negative staining with phosphotungstic acid (2%, wt/wt), the morphology was observed using a transmission electron microscope.
5 in vitro Release
To verify IMQ release, 1mL cn@ss-IMQ was placed in dialysis bags (3500 kDa) and incubated at 37 ℃ in 20mL PBS (with or without 10mM GSH). Then, 1mL of release medium was removed and replaced with 1mL of medium at predetermined time points (0.5, 1, 2, 4, 8, 12, 24, 48, 72, and 96 hours). The released IMQ was determined by High Performance Liquid Chromatography (HPLC): chromatographic conditions: a C18 column; mobile phase: methanol/water= (80:20, v/v); flow = 1.0mL/min; the detection wavelength is 319nm.
Activation of 6 BMDCs
First, 4T1 cells were seeded in 48-well plates (200 000 cells per well) overnight. The supernatant was then replaced with fresh RPMI 1640 medium containing PBS, free IMQ, HEDSMA-IMQ, HEDCMA-IMQ, cN@SS-IMQ or cN@CC-IMQ for 12 hours. BMDCs obtained as described above were seeded in another 48-well plate (100 000 cells per well) and supernatant was added to each well. After 36 hours, cells were washed three times with ice-cold PBS and non-specific binding sites were blocked with rat serum for 30 minutes. Cells were then stained with anti-CD 11c, anti-CD 80, anti-CD 86, anti-CD 40 and anti-MHC-II antibodies for 1 hour, fixed with 1% paraformaldehyde, and resuspended in 150 μl PBS for flow cytometry. Data was analyzed using FlowJo software.
Tumor targeting evaluation of 7 nanoparticles
The tumor targeting effect of nanoparticles with or without modified c-RGD peptides was evaluated using real-time near infrared fluorescence imaging techniques. By mixing 0.1mL of 4T1 cells (1X 10 6 Individual cells mL -1 ) Injected into mammary fat pads of mice to generate in situ 4T1 female BALB/c mice. When the tumor volume increases to approximately 300mm 3 At this time, mice were given equivalent concentrations of N@cy5.5 and cN@cy5.5 (0.2 mg/kg) intravenously. After 48 hours, the mice were temporarily anesthetized with isoflurane and the nanoparticle distribution in the mice was analyzed using in vivo imaging system for visualization in real time imaging.
8 tumor inhibiting effect
By mixing 0.1mL of 4T1 cells (1X 10 6 Individual cells mL -1 ) Injected into mammary fat pads of mice to generate in situ 4T1 female BALB/c mice. When the tumor volume increases to about 100mm 3 At this time, mice were randomly divided into three groups (n=6). Mice were intravenously injected every three days with i) PBS, ii) IMQ or iii) cN@SS-IMQ four times (IMQ, equal to 8.0mg kg) -1 ). Tumor diameters and mouse body weights were recorded every two days. Tumor growth and body weight curves were analyzed using GraphPad prism 7.0.
Test results
1 Synthesis of monomers
The monomer containing IMQ is synthesized by chemical synthesis means according to the method, namely HEDSMA-IMQ and HEDCMA-IMQ. HEDSMA-IMQ contains disulfide bonds, which can correspond to GSH in tumor microenvironment, so as to release free IMQ; and HEDCMA-IMQ contains GSH insensitive two carbon bonds, so that free IMQ cannot be released. The synthesis routes of HEDSMA-IMQ and HEDCMA-IMQ are shown in figure 1, and the related nuclear magnetism results are shown in figures 2-5.
2 Synthesis of Polymer
For the polymerization of HEDSMA-IMQ and HEDCMA-IMQ monomers, we used classical reversible addition-fragmentation chain transfer polymerization (Reversible Addition-Fragmentation Chain Transfer Polymerization), abbreviated RAFT polymerization. The specific synthesis procedure is shown in FIG. 6, in which hydrophilic pDMA is synthesized first, and then pDMA-b-pIMQ (SS) and pDMA-b-pIMQ (CC) are synthesized using pDMA as a chain transfer agent. In order to improve the tumor targeting aggregation capability of the polymer, the polymer end groups are modified by tumor targeting peptides, and cyclic RGD peptides are selected. All polymers were characterized by nuclear magnetic hydrogen spectroscopy. The results are shown in FIGS. 7-11.
Construction and characterization of nanoparticles
The polymer is prepared into nano particles by adopting a self-assembly technology, the particle size is about 200nm, and a transmission electron microscope shows that the nano particles are round in shape, and the result is shown in figure 12.
In vitro release of 4 nanoparticles
As shown in FIG. 13, the in vitro release behavior of nanoparticles was examined by dialysis, and the results showed that more than about 80% of free drug was released under GSH (10 mM), and that the IMQ release was lower in the absence of GSH. The results indicate that the synthesized nanoparticles have GSH sensitivity.
Activation of 5 BMDC
To verify whether the nanoparticles we constructed have the ability to activate DCs. We first incubated the drug with tumor cells 4T1 and then added the supernatant to bone marrow derived dendritic cells (BMDCs) and after a period of incubation examined the maturation of BMDCs using flow cytometry. The results are shown in FIG. 14, where stimulation of the drug promoted expression of CD40, CD80, CD86 and MHC-II on the cell surface of BMDC, demonstrating that the nanoparticles we prepared were able to effectively promote maturation of DC.
Tumor targeting evaluation of 6 nanoparticles
The tumor targeting of the nanoparticles is characterized by adopting a small animal imaging technology, and the result shows that the nanoparticles modified with the c-RGD peptide have stronger aggregation at the tumor part. The effect of improving the enrichment of nano particles at the tumor part after the tumor targeting peptide is modified is demonstrated, and the result is shown in figure 15.
In vivo efficacy of 7 nanoparticles
To evaluate the antitumor effect of cn@ss-IMQ, we established a magnetic BALB/c mouse model carrying breast cancer. When the volume of the primary tumor reaches more than or equal to 100mm 3 When mice were divided into 3 groups (6 mice per group): (1) PBS, (2) free IMQ and (3) cN@SS-IMQ. Mice in each group were injected intravenously every 4 days. The body weight of the mice was measured every 2 days with an electronic balance and the tumor size was measured with a vernier caliper. Tumor volumes were calculated using the following formula: v= (length x width) 2 )/2
The results show that the constructed nanoparticle has the strongest tumor inhibition effect, and the results are shown in fig. 16.
Taken together, this example reports a GSH-activatable IMQ-linked, c-RGD modified nano-immunomodulator (cn@ss-IMQ) that promotes intratumoral immune activation while limiting activation of the extratumor immune system upon systemic administration of such agents. The cN@SS-IMQ only induces immune activation in tumors with high GSH level, has no activity in non-targeted and GSH-deficient environments, and provides a safe drug delivery strategy for systemic anti-tumor reaction. This is achieved by blocking the active site (C4 amino group) in IMQ by disulfides. First, cN@SS-IMQ can significantly increase the amount of IMQ delivered to the tumor site compared to the free drug; once it enters a tumor cell with high GSH levels, cyclization of the polymeric prodrug occurs, followed by a 1, 6-elimination process to release free IMQ, thereby initiating tumor killing and promoting maturation of DCs to further enhance CTL infiltration into the tumor. After treatment with cn@ss-IMQ in female BALB/c model mice bearing 4T1, we observed effective treatment results. These observations underscore the broad use of endogenous stimuli-responsive monomer precursors in safe systemic cancer immunotherapy.
The invention is not a matter of the known technology.
The above embodiments are provided to illustrate the technical concept and features of the present invention and are intended to enable those skilled in the art to understand the content of the present invention and implement the same, and are not intended to limit the scope of the present invention. All equivalent changes or modifications made in accordance with the spirit of the present invention should be construed to be included in the scope of the present invention.
Claims (12)
1. The imiquimod prodrug nanoparticles are characterized by being obtained by self-assembly of amphiphilic polymers, wherein the amphiphilic polymers are further modified with tumor targeting targets, and the tumor targeting targets are tumor targeting peptides cRGD;
wherein the amphiphilic polymer is polymerized by poly (N, N-dimethylacrylamide) and imiquimod precursor, and the structural formula of the amphiphilic polymer is shown as follows:
the imiquimod precursor is specifically obtained after the imiquimod masks the C4 amino group.
2. A method for preparing imiquimod prodrug nanoparticles according to claim 1, comprising: synthesizing a hydrophilic polymer poly (N, N-dimethylacrylamide), and then further polymerizing an imiquimod precursor HEDSMA-IMQ on the basis of the hydrophilic end to obtain an amphiphilic polymer pDMA-b-pIMQ; and then carrying out target modification on the tail end of the polymer, and carrying out self-assembly on the polymer by a self-assembly technology to obtain the modified polymer.
3. The preparation method according to claim 2, wherein the synthesis method of the amphiphilic polymer pDMA-b-pmq comprises: dissolving imiquimod precursor HEDSMA-IMQ, poly (N, N-dimethylacrylamide) and azodiisobutyronitrile in dioxane, performing freeze thawing for 2-3 times, heating for reaction overnight, and precipitating and purifying the product by cold diethyl ether to obtain the preparation;
the mol ratio of the imiquimod precursor HEDSMA-IMQ to the poly (N, N-dimethylacrylamide) to the azodiisobutyronitrile is 0.1-1:0.01-0.05:0.001-0.01;
the heating reaction is carried out by adopting an oil bath heating mode, and the specific heating temperature is controlled to be 70-90 ℃.
4. The method of claim 3, wherein the molar ratio of the imiquimod precursor HEDSMA-IMQ, poly (N, N-dimethylacrylamide), and azobisisobutyronitrile is 0.55:0.02:0.00548;
the specific heating temperature of the heating reaction is controlled to be 80 ℃.
5. The method of claim 2, wherein the specific method for target modification of the polymer tail comprises: EDC and NHS are adopted to activate amphiphilic polymer pDMA-b-pIMQ, then the amphiphilic polymer pDMA-b-pIMQ reacts with c-RGD peptide, and the amphiphilic polymer pDMA-b-pIMQ is obtained after dialysis.
6. The method of claim 5, wherein the amphiphilic polymer pDMA-b-pIMQ to c-RGD peptide molar ratio is 1-5:4-10.
7. The method of claim 6, wherein the amphiphilic polymer pDMA-b-pIMQ to c-RGD peptide is present in a molar ratio of 4.33:6.63.
8. the method of claim 2, wherein the self-assembling of the polymer by self-assembly techniques comprises: and (3) dissolving the polymer modified by the target head in dimethyl sulfoxide, adding the polymer into water under the ultrasonic condition to obtain a reaction mixture, and purifying the reaction mixture to obtain the polymer.
9. The preparation method according to claim 2, wherein the imiquimod precursor HEDSMA-IMQ is prepared by dissolving HEDSMA, 4-dimethylaminopyridine and triphosgene in anhydrous dichloromethane to obtain a mixture, adding imiquimod dissolved in N, N-dimethylformamide into the mixture, stirring overnight, washing the reaction product with water, drying and removing the organic solvent phase, and purifying to obtain the imiquimod;
the mol ratio of HEDSMA, 4-dimethylaminopyridine, triphosgene and imiquimod is 0.1-0.5:1-5:0.1-0.3:1-3;
the HEDSMA is prepared by the following method: dissolving hydroxyethyl disulfide in anhydrous tetrahydrofuran under the protection of inert gas, adding methacryloyl chloride and triethylamine, removing tetrahydrofuran, washing with water, and drying and purifying the organic phase to obtain the catalyst;
the mol ratio of the hydroxyethyl disulfide, the methacryloyl chloride and the triethylamine is 30-40:35-45:35-45.
10. The method of claim 9, wherein the molar ratio of HEDSMA, 4-dimethylaminopyridine, triphosgene, and imiquimod is 0.39:1.18:0.16:1.11;
the molar ratio of the hydroxyethyl disulfide, the methacryloyl chloride and the triethylamine is 33.45:40:36.8.
11. Use of imiquimod prodrug nanoparticles according to claim 1 for the preparation of an antitumor drug.
12. An antitumor drug, wherein the active ingredient of the antitumor drug comprises imiquimod prodrug nanoparticles according to claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210857719.1A CN115120738B (en) | 2022-07-20 | 2022-07-20 | Imiquimod prodrug nano-particles, and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210857719.1A CN115120738B (en) | 2022-07-20 | 2022-07-20 | Imiquimod prodrug nano-particles, and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115120738A CN115120738A (en) | 2022-09-30 |
| CN115120738B true CN115120738B (en) | 2023-05-12 |
Family
ID=83384811
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210857719.1A Active CN115120738B (en) | 2022-07-20 | 2022-07-20 | Imiquimod prodrug nano-particles, and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115120738B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115120738B (en) * | 2022-07-20 | 2023-05-12 | 山东大学 | Imiquimod prodrug nano-particles, and preparation method and application thereof |
| CN116327782A (en) * | 2023-03-27 | 2023-06-27 | 山东大学 | Bone-targeted co-delivery bortezomib and pomalidomide nanocomposite as well as preparation method and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101909664A (en) * | 2008-01-15 | 2010-12-08 | 陶氏制药科学公司 | Imiquimod formulation |
| CN113081961A (en) * | 2021-04-08 | 2021-07-09 | 湘潭大学 | Immune regulator-bonded core cross-linked micelle anti-tumor prodrug with pH response and preparation method thereof |
| CN113395983A (en) * | 2019-02-08 | 2021-09-14 | 成均馆大学校产学协力团 | TOLL-like receptor 7 or 8 agonist-cholesterol complexes and uses thereof |
| WO2022087156A1 (en) * | 2020-10-20 | 2022-04-28 | The Board Of Regents Of The University Of Texas System | Proinflammatory prodrugs |
| CN115120738A (en) * | 2022-07-20 | 2022-09-30 | 山东大学 | Imiquimod prodrug nanoparticles and preparation method and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10799580B2 (en) * | 2015-09-09 | 2020-10-13 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Expression vector delivery system and use thereof for inducing an immune response |
-
2022
- 2022-07-20 CN CN202210857719.1A patent/CN115120738B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101909664A (en) * | 2008-01-15 | 2010-12-08 | 陶氏制药科学公司 | Imiquimod formulation |
| CN113395983A (en) * | 2019-02-08 | 2021-09-14 | 成均馆大学校产学协力团 | TOLL-like receptor 7 or 8 agonist-cholesterol complexes and uses thereof |
| WO2022087156A1 (en) * | 2020-10-20 | 2022-04-28 | The Board Of Regents Of The University Of Texas System | Proinflammatory prodrugs |
| CN113081961A (en) * | 2021-04-08 | 2021-07-09 | 湘潭大学 | Immune regulator-bonded core cross-linked micelle anti-tumor prodrug with pH response and preparation method thereof |
| CN115120738A (en) * | 2022-07-20 | 2022-09-30 | 山东大学 | Imiquimod prodrug nanoparticles and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115120738A (en) | 2022-09-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN115120738B (en) | Imiquimod prodrug nano-particles, and preparation method and application thereof | |
| Liu et al. | Development of a hypoxia-triggered and hypoxic radiosensitized liposome as a doxorubicin carrier to promote synergetic chemo-/radio-therapy for glioma | |
| JP6155287B2 (en) | Functional PLA-PEG copolymer, its nanoparticles, its manufacture, and its use for targeted drug delivery and imaging | |
| TWI394774B (en) | Novel block copolymers, microcell modifiers, and anticancer agents that are useful as an active ingredient | |
| EP2209496B1 (en) | Multifunctional degradable nanoparticles with control over size and functionalities | |
| CN108478531A (en) | Folate-targeted restores sensitive medicament-carried polymer nano micelle and its preparation method and application | |
| EP3313527B1 (en) | Anticancer drug-containing plant virus particles | |
| Gao et al. | Reversal of multidrug resistance by reduction-sensitive linear cationic click polymer/iMDR1-pDNA complex nanoparticles | |
| CN106237340B (en) | Application of hyaluronic acid nanoparticles in preparation of medicine for treating lymphatic system tumor | |
| CN103566379A (en) | Preparation and application of intracellular triggering reduction sensitive drug linked gene targeted co-carrier | |
| CN111514306B (en) | Fullerene nano-particles for enhancing anti-tumor immunotherapy | |
| CN108339124A (en) | A kind of preparation method and application of twin-stage Brain targeting polymer micelle delivery system | |
| Liu et al. | Mouse model to explore the therapeutic effect of nano-doxorubicin drug delivery system on bladder cancer | |
| Wu et al. | Enhanced drug delivery to hepatocellular carcinoma with a galactosylated core–shell polyphosphoester nanogel | |
| KR20090080046A (en) | Compositions and methods for ph targeted drug delivery | |
| KR101797829B1 (en) | surface charge conversion type nanoparticles for drug delivery and manufacturing method thereof | |
| WO2023005953A1 (en) | Drug loading monomolecular nano polymer, prodrug, micelle, drug delivery system, preparation method, and use | |
| WO2019096095A1 (en) | Integrin receptor-targeted anti-cancer conjugate | |
| CN115400223A (en) | Ferrocene and TLR7/8 agonist co-linked nanoparticle and preparation method and application thereof | |
| CN113967256B (en) | Nanoparticle with photothermal-chemotherapy-immune function and preparation method and application thereof | |
| TWI771652B (en) | CD44-targeted multi-arm conjugates | |
| CN110958998B (en) | Erianin derivatives and methods of using erianin derivatives | |
| CN115607680A (en) | Preparation and application of gold cluster-aptamer and derivative assembly thereof | |
| CN107184990A (en) | A kind of preparation method of antibody coupling medicine-carried nano particles | |
| CN111888333A (en) | Transferrin receptor targeted nano micelle and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |