• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    The present invention relates to an anti-tumor drug with active targeting and a method of preparation thereof. Specifically, the present invention discloses a complex comprising nanocarriers and targeting molecules coupled to the surface of the nanocarrier. The targeting molecule is selected from [D-Arg25] -NPY, [D-His26] -NPY, [D-Arg25, D-His26] -NPY, and has a particle size less than 200 nm and a polydispersity index (PDI) smaller 0.5. Further, the present invention discloses a composition, a medicament, its production method and their use. The composition and the medicament of the present invention show a strong targeting effect against tumor cells and can transport anti-tumor drugs directed into tumor cells and thus effectively increase the drug concentration in tumor cells and have a strong killing effect on tumor cells and have almost no toxic side effects normal tissues and cells.
    公开号:CH710313B1
    申请号:CH00295/16
    申请日:2014-04-18
    公开日:2018-08-15
    发明作者:Li Juan；Wu Aiguo
    申请人:Ningbo Institute Of Materials Tech & Engineering Chinese Acadamy Of Sciences [Cn/Cn]；
    IPC主号:
    专利说明:

    description
    Technical Field The present invention relates to the field of pharmaceutical technology, and more particularly to an active targeting anti-tumor drug and a method of preparation thereof.
    Background [0002] A drug with active targeting is a type of drug or drug carrier whose surface is modified by a molecule actively binding to a target (eg, an antibody or ligand, etc.) to target specific antigens or receptors of certain tissues or can bind cells, thus achieving the function of targeting specific cells and tissues. Because of the high specificity, high selectivity, and high affinity between antigen and antibody, or between receptor and ligand, active targeting is more efficient than passive targeting, therefore active targeting drug targeting systems in the home and abroad are being actively investigated.
    Active-targeting drugs designed based on the principle of specific binding between an antigen and an antibody still have several problems, for example, low effective concentration of targeted drugs, strong racial-specificity, high immunogenicity and high cost of Research and Development.
    However, for drugs with active targeting, which are designed based on the principle of specific binding between ligand and receptor, high selectivity, no racial-specificity, no immunogenicity, high stability and low cost are characteristic; therefore, they have become the current focus in the design of tumor-targeting delivery systems. This includes research on tumor-targeted drugs mediated by fetal, transferrin, integrin and polypeptide receptors. In recent years, tumor-targeted drugs mediated by polypeptide receptors have attracted more and more attention.
    But so far, research on cancer drug-targeting systems has been aimed primarily at targeting tumor tissue. In recent years, the question of how more drug can be brought into tumor cells, after the drugs have reached the tumor, so as to achieve a targeted introduction of drugs in tumor cells, has become the focus of research in the field of drug-targeting systems , Many pharmaceutical delivery systems have relatively good tumor specificity and can deliver drugs to the surface of tumor cells. However, most of the drugs are released outside the target cell after the carrier has bound to the target cell because the carrier has little ability to penetrate into the target cell. Furthermore, the drugs also activate different genes of the tumor cell membrane, so that different molecular mechanisms are synergistically involved in the development of a resistant phenotype. Once drug resistance is established, it further reduces the drug's ability to penetrate into the tumor cell, resulting in an extremely low intracellular drug concentration. Thus, the growth of tumor cells can not be effectively inhibited.
    Neuropeptide Y (NPY) is a type of hormone that is widely distributed in the central and peripheral nervous system and receives homeostasis. Six types of NPY receptors have been identified and identified, NPY ΥΊ, Y2, Y3, Y4, Υδ and Y6, which are widely distributed in mammalian central and peripheral nervous systems. The function of NPY is inextricably linked to its receptors, in other words, the diversity of the receptors establishes the functional diversity of NPY. Studies have shown that the known medicines for neuropeptide receptors are primarily used to treat diseases in the range of physiological impairments, including obesity, cardiovascular diseases, high cholesterol, epilepsy, anxiety and the like. However, tumor drugs with NPY as a target are rare, in particular, no tumor drugs have been reported for the treatment of kidney cancer, gastric cancer, breast cancer and ovarian cancer.
    Agonists and inhibitors based on NPY receptors (ie ligands of the receptor) have been extensively explored, but it is still difficult to select a ligand for modification of the pharmaceutical carrier because of the diversity as well as the universality of NPY receptors , Therefore, it has become key in the design and research and development of targeted drugs to select a suitable ligand for modification of the pharmaceutical carrier so that the modified pharmaceutical carrier can bind to a receptor of the tumor cell with high specificity without the biological activity of other intracellular receptors and can bring the tumor drugs directed into the tumor cells, thus increasing the effective drug concentration in the tumor cells.
    Summary of the Invention An object of the present invention is to provide an active targeting tumor drug and the method of preparation thereof.
    The first aspect of the present invention discloses a complex comprising: a nanocarrier; and a targeting molecule coupled to the surface of the nanocarrier; wherein the targeting molecule is selected from the list: [D-Arg25] -NPY, [D-His26] -NPY, [D-Arg25, D-His26] -NPY, [Arg6, Pro34] pNPY, [Asn6, Pro34 ] pNPY, [Cys6, Pro34] pNPY, [Phe6, Pro34] pNPY, [Arg7, Pro34] pNPY, [D-His26, Pro34] NPY, [Phe7, Pro34] pNPY, [Pro30, Nie31, Bpa32, Leu34] NPY (28-36), [Pro30, Nal32, Leu34] NPY (28-36), [Pro30, Nie31, Nal32, Leu34] NPY (28-36), BIBO3304, PD160170, LY366258, J-104870, LY 357897, J -115814, and combinations thereof; and the nanocarrier has a particle size of <200 nm and a polydispersity index (PDI) less than 0.5.
    In a further preferred embodiment, the content of the targeting molecule is 1.11 to 22.2% by weight of the total weight of the complex.
    In a further preferred embodiment, the content of the targeting molecule is 5.60 to 11.1% by weight of the total weight of the complex.
    In another preferred embodiment, the complex has one or more of the following characteristics: (a) the complex binds to breast cancer cells, ovarian cancer cells, renal cancer cells or gastric cancer cells with high specificity; (b) the particle size of the nanocarrier is 10-200 nm.
    In a further preferred embodiment, the nanocarrier is selected from the following list:
    Protein-based nanoparticles, oligopeptide-based nanoparticles, phospholipid-based nanoliposome, polysaccharide-based nanoparticles, polyether-based nanoparticles, polyester-based nanoparticles, polyester-based polymeric micelles.
    In another preferred embodiment, the protein-based nanoparticle is selected from: a human serum albumin nanoparticle or a bovine serum albumin nanoparticle.
    In another preferred embodiment, the phospholipid-based nanoparticle is selected from: phosphatidylcholine nanoliposome, dipalmitoyl phosphatidylcholine nanoliposome, distearoyl phosphatidylcholine nanolipo-som, dipalmitoyl phosphatidyl ethanolamine nanoliposome, distearoyl phosphatidyl ethanolamine nanoliposome, or dipalmitoyl phosphatidylglycerol Nanoliposom.
    In a further preferred embodiment, the polyester-based nanoparticle is selected from: polyethylene glycol-polylactic acid nanoparticles, polyethylene glycol-polylactide-glycolide nanoparticles, or polyethylene glycol-poly-caprolactone nanoparticles.
    In a further preferred embodiment, the polysaccharide-based nanoparticle comprises a chitosan Na nopartikel.
    In a further preferred embodiment, the polyester-based polymeric micelle is selected from: poly-ethylene glycol-polylactic acid micelle, polyethylene glycol-polycaprolactone micelle, polyethylene glycol-distearoyl-phosphatidylethanolamine micelle, or polyethylene glycol-polyethyleneimine micelle.
    The second aspect of the present invention discloses a composition comprising: the complex of the first aspect; and an anti-tumor drug entrapped by the nanocarrier of the complex.
    In a further preferred embodiment, the anti-tumor drug is selected from the following list: doxorubicin, paclitaxel, docetaxel, cisplatin, mitoxantrone, daunorubicin, vincristine, all-trans retinoic acid, pharmorubicin, lurtotecan, irinotecan, 2-methoxyestradiol, gemcitabine , Vinorelbine, 5-fluorouracil, methotrexate, capecitabine, lomustine, etoposide, or combinations thereof.
    In a further preferred embodiment, the anti-tumor drug is embedded in the nanocarrier of the complex.
    In another preferred embodiment, in the composition, the encapsulation efficiency of the nanocarrier for the anti-tumor drug is over 80%, and preferably 90% or more.
    In a further preferred embodiment, the rate of killing of tumor cells by the composition is above 60%, and preferably above 70%, when the concentration of anti-tumor drug in the composition is 5-10 pg / ml.
    In a further preferred embodiment, tumor cells comprise the tumor cells of breast cancer, ovarian cancer, kidney cancer or stomach cancer.
    In a further preferred embodiment, the content of the anti-tumor drug is 1.0 to 3.0% by weight and preferably 1.5 to 2.7% by weight of the total weight of the composition.
    In a further preferred embodiment, the content of the targeting molecule is 1.11 to 22.2% by weight and preferably 5.60 to 11.1% by weight of the total weight of the composition.
    The third aspect of the present invention discloses a method of making the composition of the second aspect, the method comprising the steps of: (1) providing an anti-tumor drug loaded nanocarrier; and (2) coupling the nanocarrier of step (1) with a targeting molecule, thereby obtaining the composition.
    In a further preferred embodiment, the particle size of the nanocarrier is 200 nm or smaller, and preferably 10-200 nm.
    In a further preferred embodiment, the method for producing the nanocarrier comprises the following steps: (a) respectively providing an aqueous solution and an organic solution, the aqueous solution comprising an anti-tumor drug and a hydrophilic membrane material, and the organic solution comprises an emulsifier; (b) mixing the aqueous solution with the organic solution of step (a), thereby obtaining an emulsion; (c) curing the emulsion of step (b), thereby obtaining the nanocarrier.
    Or, the method of producing the nanocarrier comprises the steps of: (a) respectively providing an aqueous solution and an organic solution, wherein the aqueous solution comprises an emulsifier, and the organic solution comprises an anti-tumor drug and a hydrophobic membrane material ; (b) mixing the aqueous solution with the organic solution of step (a), thereby obtaining an emulsion; (c) curing the emulsion of step (b), thereby obtaining the nanocarrier.
    Or, the method of producing the nanocarrier comprises the steps of: (a) respectively providing an aqueous solution and an organic solution, wherein the aqueous solution comprises an anti-tumor drug, and the organic solution comprises a hydrophobic membrane material and an emulsifier ; (b) mixing the aqueous solution with the organic solution from step (a), thereby obtaining a first emulsion; (c) mixing the first emulsion of step (b) with the aqueous solution containing dissolved emulsifier, thereby obtaining a second emulsion; (d) curing the second emulsion from step (c), thereby obtaining the nanocarrier.
    Or the method of making the nanocarrier comprises the steps of: (a) providing a suspension comprising an anti-tumor drug, a nanocarrier, and an organic solvent; (b) curing the suspension of step (a), thereby obtaining the nanocarrier.
    In a further preferred embodiment, the hydrophilic membrane material is selected from: polyethylene glycol (PEG), polyoxyethylene (PEO), polyvinylpyrrolidone (PVP) or polyvinyl alcohol (PVA).
    In a further preferred embodiment, the hydrophobic membrane material is selected from: polyoxypropylene (PPO), polystyrene (PS), polyamino acid, polylactic acid (PLA), spermine or short-chain phospholipid.
    In a further preferred embodiment, the emulsifier is selected from:
    Pluronic F68, dextran 70 or sodium cholate.
    In a further preferred embodiment, the coupling reaction is selected from: (1) a condensation reaction of a carboxy group and an amino group; (2) an addition reaction of a sulfhydryl group with a maleimide group; or (3) noncovalent binding of avidin and biotin.
    The fourth aspect of the present invention discloses the use of the complex of the first aspect for the manufacture of a medicament for the treatment of cancer.
    In another preferred embodiment, cancer includes breast cancer, ovarian cancer, kidney cancer and stomach cancer. More preferably, cancer includes kidney cancer and stomach cancer.
    The fifth aspect of the present invention discloses the use of the composition of the second aspect for the manufacture of a medicament for the treatment of cancer.
    The sixth aspect of the present invention discloses a medicament comprising: the complex of the first aspect; an anti-tumor drug included in the nanocarrier of the composition; and a pharmaceutically acceptable carrier.
    In another preferred embodiment, the formulation of the medicament is selected from the following list: solid preparation, liquid preparation and injections.
    In a further preferred embodiment, the medicament is used on mammals, preferably on humans.
    In another preferred embodiment, the formulation of the medicament is an injection.
    In a further preferred embodiment, the injection method comprises intravenous, intramuscular, subcutaneous or intracavitary administration.
    The seventh aspect of the present invention discloses a method for the treatment of cancer, comprising a step of administering a safe and effective amount of the composition according to aspect two or the medicament according to aspect six.
    In another preferred embodiment, cancer includes breast cancer, ovarian cancer, kidney cancer and stomach cancer. More preferably, cancer includes kidney cancer and stomach cancer.
    It is to be understood that in the present invention, all technical features specifically described above and below (as well as in the examples) may be combined with each other to form new or preferred technical solutions that are not specifically described herein be described redundantly.
    Description of the figures [0048]
    Figure 1 shows a TEM pattern and a DLS particle size distribution distribution of composition [D-Arg25] -NPY-ANP-TXT.
    Fig. 2 shows the change of the particle size of the composition [D-Arg25] -NPY-ANP-TXT in NaCl solution, PBS solution and serum for 1-15 days.
    Fig. 3 shows a comparative profile of tumor cells MCF-7 and HEC-1B-Y5 which receive the composition [D-Arg25] -NPY-ANP-TXT.
    Detailed Description of the Invention Through extensive, intensive and long research, the inventors have unexpectedly discovered that a composition prepared by coupling certain targeting molecules to the anti-tumor drug loaded nanocarrier can be coupled to specific neuropeptide receptors Tumor cells can bind highly specific and can bring the anti-tumor drug directed into these cells, whereby the effective concentration of the drug in the tumor cells is increased, while the composition has almost no toxic side effects on normal tissues and cells. The composition of the present invention has a potent killing effect on tumor cells, particularly tumor cells of breast cancer, ovarian cancer, kidney cancer and gastric cancer, and thus it can be used for the preparation of a medicament for the treatment of the above-mentioned cancers. Based on these findings, the inventors have completed the present invention.
    As used herein, "biotin" refers to vitamin H, also called vitamin B7 or coenzyme R, with the molecular weight of 244.31 Da.
    As used herein, "avidin" refers to a glycoprotein having a molecular weight of about 60 kDa and mainly comprises the protein avidin (also called natural avidin, ovo albumin avidin or anti-biotin), streptavidin, dotteravidin, and the like.
    Targeting Molecule The targeting molecule of the present invention relates to a peptide agonist molecule or non-peptide antagonist molecule capable of binding to neuropeptide receptors with high specificity and efficiency and thereby conferring various biological functions Activity triggers. The peptide agonist molecule includes (but is not limited to): [D-Arg25] -NPY, [D-His26] -NPY, [D-Arg25, D-His26] -NPY, [Arg6, Pro34] pNPY, [Asn6, Pro34] pNPY, [Cys6, Pro34] pNPY, [Phe6, Pro34] pNPY, [Arg7, Pro34] pNPY, [D-His26, Pro34] NPY, [Phe7, Pro34] pNPY, [Pro30, Nie31, Bpa32 , Leu34] NPY (28-36), [Pro30, Nal32, Leu34lNPY (28-36) and [Pro30, Nie31, Nal32, Leu34] NPY (28-36), and the non-peptide antagonist molecule comprises (is but not limited to): BIBO3304, PD160170, LY366258, J-104870, LY357897 and J-115814.
    Through research, the inventors have unexpectedly found that the above targeting molecule of the present invention can bind highly specifically to breast cancer, ovarian cancer, kidney cancer and gastric cancer cells, but can not specifically bind to brain tumor and endometrial tumor cells.
    As used herein, "highly specific binding" refers to the fact that, under similar conditions, each time after binding to tumor cells and non-tumor cells (ie, normal cells), the rate of uptake of the complex of the present invention satisfies the following condition: ΒΊ / Β0> 2 , 5, preferably ΒΊ / Β0> 3, more preferably ΒΊ / Β0> 4; where ΒΊ is the rate of uptake of the complex by 10,000 tumor cells, and Bo is the rate of uptake of the complex by 10,000 normal cells.
    The complex and its method of preparation The complex of the present invention is a binary complex consisting of a biodegradable nanocarrier and targeting molecules with the targeting molecules coupled to the surface of the nanocarrier. Excessive targeting molecules can precipitate or agglomerate the nanoparticles of the complex, increasing the diameter of the complex nanoparticles (> 200 nm). In the present invention, the content of targeting molecules is 1.11 to 22.2% (weight%), and preferably 5.60 to 11.1% (weight%), based on the total weight of the complex; the remainder consists of the biodegradable nanocarrier.
    The complex of the present invention has a good dispersion and stability in aqueous NaCl solution, aqueous PBS solution or serum, without any precipitation or clumping phenomena.
    In the present invention, the nanoparticle may be selected from: protein-based nanoparticles, oligopeptide-based nanoparticles, phospholipid-based nanoliposome, polysaccharide-based nanoparticles, polyether-based nanoparticles, polyester-based nanoparticles, polyester-based polymeric micelles, or combinations thereof. Preference is given to protein-based nanoparticles, phospholipid-based nanoliposome and polysaccharide-based nanoparticles.
    The preferred protein-based nanoparticle comprises human serum albumin (HSA) nanoparticles or bovine serum albumin (BSA) nanoparticles, or combinations thereof.
    The preferred phospholipid-based nanoliposome includes phosphatidylcholine (PC) nanoliposome, dipalmi- toylphosphatidylcholine (DPPC) nanoliposome, distearoylphosphatidylcholine (DSPC) nanoliposome, dipalmitoylphosphatidylethanolamine (DPPE) nanoliposome, distearoylphosphatidylethanolamine (DSPE) nanoliposome, dipalmitoylphosphatidylglycerol (DPPG) nanoliposome, or combinations thereof.
    The preferred polyester-based nanoparticles include polyethylene glycol polylactic acid (PEG-PLA) nanoparticles, polyethylene glycol polylactide glycolide (PEG-PLGA) nanoparticles, polyethylene glycol polycaprolactone (PEG-PCL) nanoparticles, or combinations thereof.
    The preferred polysaccharide-based nanoparticle comprises chitosan nanoparticles.
    The preferred polyester-based polymeric micelle comprises polyethylene glycol-polylactic acid (PEG-PLA) micelles, polyethylene glycol-polycaprolactone (PEG-PCL) micelles, polyethylene glycol distearoylphosphatidylethanolamine (PEG-DSPE) micelles, polyethylene glycol polyethylenimine (PEG-PSA) micelles. cl-PEI) micelles, or combinations thereof.
    The method for preparing the complex of the invention mainly comprises the following steps: (1) preparation of the nanocarrier and (2) reaction of the nanocarriers with the targeting molecules. The methods for producing the nanocarrier may be methods known to those skilled in the art, and the reaction between the nanocarrier and the targeting molecule may be realized via chemical coupling methods.
    A preferred coupling method is as follows:
    When the nanocarriers contain carboxy groups (such as polyglutamic acid, polyaspartic acid, peptides and proteins containing glutamic acid and aspartic acid, and polysaccharides bearing a carboxy group), one may choose targeting molecules containing an amino group and having carboxy groups on the surface of the nanocarrier Activate EDAC and NHS (N-hydroxysuccinimide), then add dropwise a solution of the targeting molecule with terminal
    Adding amino groups to effect a covalent reaction between the activated carboxy groups and the amino groups of the targeting molecule to form a stable amide bond; if the nanocarriers contain amino groups (such as polylysine, histidine, polyarginine, peptides and proteins containing lysine, arginine and histidine and polysaccharides with amino groups), one can choose targeting molecules containing a carboxy group and the carboxy groups of the targeting molecule activate by the method given above to graft the activated carboxy groups onto the surface of the nanocarrier; For biodegradable nanocarriers containing neither carboxy nor amino groups (such as polyether and polyester polymers), the copolymerization method can be used to give the biodegradable nanocarriers amino or carboxy groups, and after forming into nanocarriers, the same method can be used as noted above, to graft the targeting molecules onto the surface of the nanocarrier.
    The other preferred coupling method is as follows:
    Mercapto groups are introduced at the targeting molecule and maleimide groups are introduced at the surface of the drug carrier system, and then an addition reaction is carried out in neutral or alkaline aqueous environment (e.g., in phosphate buffer) at room temperature.
    Thiolated targeting molecules are prepared primarily via the reaction between sulfhydryl reagents (such as 2-IT, SPDP, SATP, and SSDD, etc.) and amino groups of the targeting molecules.
    To introduce maleimide into the drug carrier system, lipid molecules and maleimide (MAL) modified polymeric materials (such as MAL-PEG-PLA, MAL-PEG-PLGA, MAL-PEG-PCL, MAL-PEG-DSPE) are mixed and dissolved in an organic Solvents are dissolved, and via thin-layer hydration and high-pressure homogenization, ma-lidded liposomes having hydrophobic ends (such as PLA, PLGA, PCL and DSPE) oriented in the lipid bilayer with the PEG-maleimide ends on the surface of the lipid membranes can be obtained ,
    In addition, maleimide can be introduced directly to the surface of the drug delivery system. For example, bifunctional linkers capable of forming maleimide groups (such as bifunctional propionic acid linkers in which the active esters of the molecule can react with amino groups to form maleimide groups) are useful for introducing maleimide groups into the nanocarriers.
    Another preferred coupling method is as follows:
    Biotin is introduced separately into the targeting molecule and drug delivery system, and avidin is used as a bridging reagent to make the drug targeting system. In other words, avidin and the biotin-carrying carrier system are first mixed and then unbound sites react with the biotin-bearing targeting molecules.
    For example, to prepare biotin-carrying PEG-PLGA, PLGA-COOH (molecular weight 20 kDa) is dissolved in dichloromethane. NHS and EDC in 8 times the amount of PLGA-COOH are added with stirring at room temperature to activate PLGA-COOH. The resulting active esters and NH2-PEG-biotin (molecular weight 3400 Da) are mixed and dissolved in chloroform. An adequate amount of Ν, Ν-diisopropylethylamine is added, the reaction is carried out overnight. The excess PEG molecules are washed out with methanol and the reaction product is precipitated with ether and dried in vacuo to obtain PLGA-PEG-biotin. Then, a certain amount of PLGA-PEG-COOH and PLGA-PEG-biotin are mixed and dissolved in acetone, the mixture is slowly added dropwise to deionized water and the acetone is removed by rotary evaporation at room temperature. The organic solvent is removed by ultrafiltration concentration to obtain biotin-bearing PEG-PLGA nanoparticles. The solution of biotin-carrying PEG-PLGA nanoparticles and avidin is incubated at room temperature for a period of time, then the free avidin is removed by centrifugation and washing. A certain amount of biotin-carrying targeting molecules and the resulting product are stirred at room temperature and then the free biotin-carrying targeting molecules are removed by centrifugation, thereby obtaining the drug-targeting system from PEG-PLGA nanoparticles.
    The composition and its method of preparation and its use The composition of the present invention comprises the complex of the present invention and anti-tumor drugs carried in the nanocarrier of the complex. Based on the total weight of the composition, the content of anti-tumor drugs is usually 1.5 to 3.0% by weight, preferably 2.0 to 3.0% by weight; and the content of targeting molecules is usually 1.11 to 22.2% by weight, and preferably 5.60 to 11.1% by weight.
    In order to achieve improved slow release and controlled release effect of the anti-tumor drugs and to avoid occurrence of in vivo opsonization, the particle size of the complex of the present invention is preferably 200 nm or less, and preferably 10-200 nm.
    The preferred anti-tumor drugs include (but are not limited to): doxorubicin, paclitaxel, do-cetaxel, cisplatin, mitoxantrone, daunorubicin, vincristine, all-trans retinoic acid, pharmacorubicin, lurtotecan, irinotecan, 2-methoxyestradiol , Gemcitabine, vinorelbine, 5-fluorouracil, methotrexate, capecitabine, lomustine, etoposide, or combinations thereof; and preferably include doxorubicin, paclitaxel, docetaxel, mitoxantrone, daunorubicin, irinotecan, gemcitabine, vinorelbine, capecitabine, or etoposide.
    The preparation method of the composition of the present invention mainly comprises the steps of: (1) providing an anti-tumor drug-loaded nanocarrier; and (2) coupling the nanocarrier of step (1) with targeting molecules, thereby obtaining the composition.
    In the invention, the nanocarriers can be prepared by ultrasonic emulsification. Preferably, the nanocarriers can be prepared by the following three methods: (1) Use of an aqueous solution as an aqueous phase in which hydrophilic anti-tumor drugs and hydrophilic membrane material (eg, polyethylene glycol) are dissolved, and use of an organic solvent (eg, dichloromethane) as Oil phase in which oil-soluble emulsifiers (eg sodium cholate) are dissolved; Mixing the aqueous phase with the oil phase and stirring the mixture to achieve coarse dispersion; Emulsifying the mixture with an ultrasonic cell disruptor to obtain a water-in-oil nanoemulsion; then, with magnetic stirring, adding a crosslinking agent (e.g., glutaraldehyde) to the resulting nanoemulsion to effect crosslinking and curing; Removal of the excess cross-linking agent and emulsifier, resulting in biodegradable nanocarriers in which anti-tumor drugs are embedded; (2) use of an organic solvent (eg dichloromethane) as an oil phase in which hydrophobic anti-tumor medicaments and hydrophobic membrane material (eg PACA) are dissolved, and use of an aqueous solution as aqueous phase in which water-soluble emulsifier (eg Pluronic F68, Dextran 70) is dissolved; Mixing the oil phase with the aqueous phase and stirring the mixture to achieve coarse dispersion; Emulsifying the mixture with an ultrasonic cell disruptor to obtain an oil-in-water nanoemulsion; then, with magnetic stirring, adding a crosslinking agent (e.g., glutaraldehyde) to the resulting nanoemulsion to effect crosslinking and curing; Removal of the excess cross-linking agent and emulsifier, resulting in biodegradable nanocarriers in which anti-tumor drugs are embedded; (3) use of an aqueous solution as an aqueous phase in which hydrophilic anti-tumor drugs are dissolved, and use of an organic solvent as an oil phase in which hydrophobic matter (e.g., PEG-PLA) and oil-soluble emulsifiers (e.g., sodium cholate) are dissolved; Mixing the aqueous phase with the oil phase and stirring the mixture to achieve coarse dispersion; Emulsifying the mixture with an ultrasonic cell disruptor to obtain a water-in-oil nanoemulsion; then adding the obtained water-in-oil nanoemulsion to an aqueous phase in which water-soluble emulsifiers are dissolved and performing ultrasonic emulsification to obtain a water / oil / water nanoemulsion; then, with magnetic stirring, adding a crosslinking agent (e.g., glutaraldehyde) to the resulting water / oil / water nanoemulsion to effect crosslinking and curing; Removal of the excess cross-linking agent and emulsifier, resulting in biodegradable nanocarriers, in which anti-tumor drugs are embedded.
    The above nanocarriers of the present invention can also be prepared by the solvent removal method. A preferred method comprises the steps of dissolving water-soluble antitumor drugs and oligopeptide or protein nanocarriers in aqueous NaCl solution, and adding ethanol dropwise with continuous magnetic stirring, and when the solution becomes a milky-white suspension, Adding glutaraldehyde for crosslinking and curing; Removal of excess cross-linking agent, resulting in biodegradable nanocarriers in which anti-tumor drugs are embedded.
    The reaction of the above step (2) is the same as that between nanocarriers and targeting molecules of the complex of the present invention.
    It should be understood that the above composition can also be prepared by packaging the anti-tumor drugs in the prepared complex of the present invention.
    The composition of the invention can be used for the preparation of anti-tumor drugs, especially for drugs for the treatment of breast cancer, ovarian cancer, kidney cancer and stomach cancer.
    Medicaments, composition and their method of administration The pharmaceutical or medicament of the present invention comprises an effective amount of the composition of the present invention and a pharmaceutically acceptable carrier or excipient.
    As used herein, the term "comprising" or "including" includes "comprising," "substantially consisting of ..." and "consisting of ...". As used herein, the term "pharmaceutically acceptable" component means that the component is suitable for humans and / or animals without excessive side effects (such as toxicity, stimulation, and allergies). In other words, the component is a substance with a reasonable benefit-risk ratio. As used herein, the term "effective amount" refers to an amount that exhibits activity or activity in humans and / or animals that can be tolerated by humans and / or animals.
    As used herein, the term "pharmaceutically acceptable carrier" refers to carriers used to administer the therapeutic agent, including various excipients and diluents. The term refers to such drug carriers: they themselves are not essential active ingredients and have no excessive toxicity after use. Suitable carriers are well known to those skilled in the art. A detailed discussion of the pharmaceutically acceptable excipient can be found in Remington's Pharmaceutical Sciences (Mack Pub., Co., N.J., 1991).
    The preparation of the medicament of the present invention comprises: solid preparation, liquid preparation or injection, and preferably injection.
    The medicament of the present invention is for mammals, and preferably for humans.
    In another preferred embodiment of the present invention, the drug or composition of the invention is administered one or more times a day, e.g. 1, 2, 3, 4, 5 or 6 times. The mode of administration includes, but is not limited to: oral administration, administration by injection, intracavitary administration, transdermal administration; and preferably administration by injection, comprising intravenous, intramuscular, subcutaneous and intracavitary administration. In order to determine the specific dosage in the administration of the drug or the composition of the present invention, the following factors should be considered: route of administration, state of health of the patient, etc., which are within the reach of an experienced physician. The safe and effective amount of the composition of the present invention is normally at least about 10 mg per kg of body weight per day, or at least about 85 mg per kg of body weight per day, and in most cases no more than about 200 mg per kg of body weight per day Day or at no more than about 115 mg per kg of body weight per day, and preferably at a dose of about 100 mg per kg of body weight per day. Compared to the prior art, the present invention has the following main advantages: (1) The complex and the composition of the present invention have good dispersion and stability in aqueous NaCl solution, PBS aqueous solution or serum, and no precipitation occurs or clumping phenomenon. (2) The complex and composition of the present invention can bind highly specifically to breast, ovarian, renal, and gastric cancer cells, and have a high targeting effect on tumor cells. (3) The composition and the medicament of the present invention can be directed to transport antitumor drugs into the tumor cells, thereby effectively increasing the drug concentration in the tumor cells, and have a strong killing effect against tumor cells, and have almost no toxic side effects to normal Tissues and cells.
    The above features of the present invention or the characteristics mentioned in the examples can be arbitrarily combined. All features disclosed in the description of the present invention may be arbitrarily combined, and any feature disclosed in the specification may be replaced by any replacement feature that fulfills identical, identical or similar purposes. Therefore, unless otherwise indicated, features disclosed are merely general examples of the same or similar features.
    The present invention will be further illustrated below with reference to specific examples. It should be understood that these examples are merely illustrative of the invention, but are not intended to limit the scope of the invention. The experimental methods described in the following examples without specific conditions are generally carried out under usual conditions or according to the instructions of the manufacturer. Unless otherwise indicated, parts and percentages are by weight.
    Unless otherwise defined, all terms and scientific terms used in the text have the meanings known to those skilled in the art. Furthermore, all methods and materials similar or equivalent to those mentioned in this invention may be used for this invention. The method of the preferred embodiment described herein and the materials are for illustrative purposes only.
    Example 1: Preparation of the composition [D-Arg25] -NPY-ANP-TXT
    [0090]
    (1) The preparation of BSA nanocarriers (ANP) with embedded TXT
    An aqueous 10 mM NaCl solution of pH 10.8 was prepared and the resulting solution was used to prepare an aqueous BSA solution at a concentration of 20 mg / ml. Then, 2.0 ml of ethanol in 2.0 ml of aqueous BSA solution was added, and after 10 minutes of magnetic stirring, 4.0 ml of ethanol was added dropwise at a rate of 2.0 ml / min (the volume ratio of the total ethanol added) aqueous solution of the nanocarrier was 3.0); during the addition was stirred magnetically continuously. Immediately after completion of the dropwise addition of ethanol, 8% aqueous glutaraldehyde solution was added (the mass ratio of glutaraldehyde to BSA was 0.24) and crosslinked (or cured) for 24 hours. Then 1.0 ml of glycine (40 mg / ml) was added to neutralize excess glutaraldehyde, and after 2.0 hours reaction time, the sample was centrifuged (20000xg, 20 min), the resulting sample was washed twice with 10 mM aqueous NaCl. Solution, and after freeze-drying for 48 hours, BSA nanocarriers were obtained. A solution of TXT was dispersed in an aqueous solution of 20 mg / ml BSA nanoparticles. ANP-TXT nanocarriers with embedded anti-tumor drugs were prepared by the same method as described above. (2) Coupling of [D-Arg25] -NPY targeting molecules to the surface of nanocarriers.
    Under catalyzation of EDAC (1-ethyl- (3-dimethylaminopropyl) carbodiimide) and via the chemical reaction between amino groups of the targeting molecule [D-Arg25] -NPY and carboxy groups on the surface of the nanocarrier ANP became the targeting molecule [D-Arg25] -NPY coupled to the surface of the nanocarrier. The specific preparation method was as follows: 500 pg / ml targeting molecule [D-Arg25] -NPY solution was prepared with phosphate buffer solution (PBS) as a solvent. In the ice bath, 50 mg of EDAC was dissolved in 10 ml of targeting molecule solution, then 90 ml of ANP-TXT suspension (5.0 mg / ml) in PBS was added, the mixture was stirred magnetically at room temperature and the reaction lasted 4 up to 24 hours. The sample was centrifuged (20000xg, 20 min) and the resulting sample was washed twice with PBS. Finally, the sample was freeze-dried for 48 hours and the composition [D-Arg25] -NPY-ANP-TXT, which had the targeting molecule coupled to the surface and embedded in the nanocarrier with anti-tumor medications, was obtained.
    The changes in the particle size of the composition [D-Arg25] -NPY-ANP-TXT in NaCl solution, PBS solution and serum over 1-15 days are shown in Table 1 and FIG.
    Table 1:
    Average diameter (nm)
    Day (s) -----
    NaCl PBS Serum I 116.6 115.2 118.6 3 115.2 116.8 117.5 5 113.4 114.3 116.4 7 111.6 116.9 115.2 9 111.8 115.2 111.9 II 112.6 113.4 114.6 13 110.5 111.6 113.1 15 110.4 110.3 111.9 As can be seen from Table 1 and Fig. 2, the nanoparticles had the complex uniform particle size and good dispersion in three different solvents, and the particle size was stable between 110 and 120 nm.
    According to the measuring method with a nanoparticle size analyzer, each of the nanoparticle complexes described had a PDI index of less than 0.5.
    Example 2: Activity Assay of Composition [D-Arg25] -NPY-ANP-TXT on Tumor Cells MCF-7 and HEC-1B-Y5 (1) MTT Assay (Cell Toxicity Test) 1. Prepare a single cell suspension in a medium containing Containing 10% fetal calf serum, then inoculating the suspension in 96-well plates (1.0 x 105 cells / well). The volume per well was 150 μΙ. 2. Place the plates in a cell incubator at 37 ° C and culture for 24 hours. 3. Absorb and discard the supernatant of the medium in the well, and add 200 μΙ of fresh medium with TXT or 200 μΙ solution with composition [D-Arg25] -NPY-ANP-TXT of the same concentration as TXT.
    4. Place the plates in a cell incubator at 37 ° C and culture for 4 hours, absorb and discard the supernatant of the medium in the well and replace with fresh medium containing no drug or nanoparticle composition, and continue the culture for 44 hours. 5. Add 10 μM MTT solution (5 mg / ml, in PBS, pH = 7.4) to each well, place the plates in a cell incubator at 37 ° C and continue the culture for 4 hours, stop the culture, and then Absorbing and discarding the supernatant of the medium in the well. 6. Add 150 μΙ DMSO to each well, oscillate for 10 minutes so that the crystal is completely thawed. 7. Measure the light absorbance at 550 nm wavelength of each well with enzyme-linked immunosorbent assay and record the results. The results are shown in Table 2.
    The cells used in the cell experiments include human MCF-7 breast cancer cells, human HEC-1B-Y5 endometrial tumor cells (supplied by American Type Culture Collection [ATCC] and by Sciencell [USA]).
    Table 2:
    Killing effect of composition [D-Arg25] -NPY-ANP-TXT against MCF-7 and HEC-1B-Y5 survival rate of cells (%) __ composition Composition
    Ctxt TXT 1X1 [D-Arg2S] -NPY-ANP-TXT [D-Arg25] -NPY-ANP-TXT (pg / ml) tumor cells tumor cells MCF-7 HEC-IB-Yj MCF-7 HEC-1B-Y5 0.0 ' 100 100 "100 100_ 0.5__90 91__93__98_ 1.0__80 79__86__90_ 2.0__50 55__60__85_ 5.0__20 40__32__75_ 10.0 1 20 [30 25 I 60 It can be seen from Table 2 and Fig. 3 that docetaxel (TXT) has an excellent kill effect on MCF-7 and HEC-1B-Y5 cells, and the composition [D-Arg25] -NPY-ANP-TXT has an excellent killing effect on MCF-7 cells, but the killing effect on HEC-1 B-Y5 cells is not Consequently, these results indicate that the composition [D-Arg25] -NPY-ANP-TXT can unexpectedly and significantly kill MCF-7 cells in a highly selective manner with an excellent killing effect.
    Example 3: Activity test of the composition [D-Arg25] -NPY-ANP-TXT on various tumor cells. (1) The activity test method was the same as in Example 2. The test results are shown in Tables 3-1 and 3-2 shown. In this example, the cells used in the cell experiments included human UWB1.289 ovarian tumor cells, human GIST H1 gastric cancer cells, human SW-13 kidney cancer cells, and human SMS-KAN brain tumor cells. Normal cells included human MCF-10a mammary epithelial cells, human HOSE-piC ovarian surface epithelial cells, human HRCEpiC renal cortex epithelial cells, human GES-1 magenta cells, and human HA brain astrocytes, human HUM-CELL-0111 endometrial epithelial cells (purchased from American Type Culture Collection (ATCC), Sciencell (USA) and Cell Bank of the Chinese Science Academy Type Culture Collection Committee).
    Table 3-1:
    Comparison of the killing effect of the composition [D-Arg25] -NPY-ANP-TXT on different tumor cells Survival of the cells (%) _ ™ Tumor cells ____ (Mgm) MCF-7 | UWB1.289 | SW-13 IGIST-HL | SMS-KAN | hEC-1B-Y5 0Ό_100_100_100 100 100_100_ CL5_93_94_90_91_97_95_ LO_86_88_85_82_89_86_10_60_65_63_60_82_80_ 5Ό_32_40_35_33_76_74_ 10.0 j25 | 26 OO | 24 [71 [70 ~ [0102] Table 3-2:
    Comparison of the killing effect of the composition [D-Arg25] -NPY-ANP-TXT on different normal cells Cell survival rate (%) _ ,, Normal cells_____ (gg / m) MCF-10a iHOSEpiC [HRCEpiC IgES-1 IhA Ihum-CELL- 011 1 0Ό_100 100_100 100 100 100_ 075_93_94_90_91_97_98_ LO_90_91_88_90_94_95_ 2Ό_88_84_81_84_87_85_ 5Ό_85_78_13_80_82_81_ 10.0 | 78 [75 [76 | 75 | 73 V5] It can be seen from Table 3-1 that the composition [D-Arg25] -NPY-ANP-TXT of the present invention Invention has an excellent killing effect on UWB1.289 cells, SW-13 cells and GIST-H1 cells as in MCF-7 cells. However, the killing effect of the composition [D-Arg25] -NPY-ANP-TXT is not significant in SMS-CAN cells. Thus, these results show that the composition can kill UWB1.289 cells, SW-13 cells and GIST-H1 cells highly selectively, and has an excellent killing effect.
    From the test results of Table 2, Table 3-1 and Table 3-2, it can be seen that the composition is capable of highly specific binding of [D-Arg25] -NPY-ANP-TXT to breast, ovarian, renal and gastric cancer cells It has a strong targeting effect against tumor cells and can deliver anti-tumor drugs directed into tumor cells and thus effectively increase the drug concentration in tumor cells and it has a strong killing effect on tumor cells and has virtually no toxic side effects on normal tissues and cells.
    Example 4: Activity Assay of Various Compositions with Embedded Docetaxel on MCF-7 and UWB1.289 Cells (1) Preparation of the Composition [0105]
    (a) Preparation of chitosan nanocarrier composition with embedded TXT
    (i) Preparation of embedded TXT chitosan nanocarriers
    A 0.2% (w / v) chitosan solution was prepared with 1% (w / v) acetic acid as a solvent. The docetaxel (TXT) was dispersed in the chitosan solution and the pH of the solution was adjusted to 4.7-4.8 with sodium hydroxide. An aqueous 0.3% (w / v) sodium tripolyphosphate (TPP) was prepared. With magnetic stirring, 0.1 ml of TPP solution was added to 0.5 ml of chitosan solution to give ion-crosslinked chitosan nanocarriers with embedded TXT. (ii) coupling the targeting molecule to the surface of the chitosan nanocarriers
    The coupling reaction of the targeting molecules was carried out on the surface of the obtained nanocarriers in the same manner as in step (2) of Example 1, except that the targeting molecule [D-Arg 25] -NPY was replaced with those of Table 5. (b) Preparation of a BSA nanocarrier (ANP) composition with embedded TXT. The preparation was carried out analogously to Example 1, while the targeting molecule [D-Arg25] -NPY was replaced by those of Table 5. (2) Cytotoxicity test The activity tests were carried out according to the method of Example 2, wherein the cells used in the experiments comprised MCF-7 and UWB1.289 cells.
    The test results are shown in Tables 5, wherein "+" means that the composition has a killing effect on tumor cells, "-" means that the composition has almost no or a weak killing effect on tumor cells.
    The specific meanings of the symbols are shown in Table 4 (a representative concentration was chosen and different symbols were assigned depending on the range of values): Table 4:
    C'rx (T ι values symbols (gg / ml) ___> 80 _-_ 60 <Survival rate of cells <80 -_ $ 50 <Survival rate of cells <60 -_ 40 <Survival rate of cells <50 + _ 30 <Survival rate of cells <40 ++ _ _20 <survival rate of cells <30 +++ _ [0110] Table 5:
    Comparison of the killing effect of different compositions on MCF-7 and UWB1.289 cells ___ nanocarriers BSA nanocarriers_Chitosan nanocarriers_
    Targeting molecule ~ MCF-7 UWB 1.289 MCF-7 UWB 1.289 [D-Arg25] -NPY _ ++ _ ++ _ ++ _ ++ _ [D-His26] -NPY _ ++ _ ++ _ ++ _ + + _ [D-Arg25, D-His26] -NPY _ ++ _ ++ _ ++ _ ++ _.
    [D-His26, Pro34] NPY _ ++ _ ++ _ ++ _ ++ _ [Phe7, Pro34] pNPY _ +++ +++ _ +++ +++ _ [Arg6, Pro34] pNPY _ +++ _ +++ _ +++ _ +++ _ [Asn6, Pro34] pNPY _ +++ _ +++ _ +++ _ +++ _ [Cys6, Pro3lt] pNPY _ +++ _ +++ _ ++ + _ +++ _ [Phe6, Pro34] pNPY _ +++ +++ _ +++ +++ _ [Pro30, Nie31, Bpa32, Leu34] NPY (28-36) +++ +++ _ ++ + -1 - ++ _ [Pro30, Nal32, Leu34] NPY (28-36) _ +++ +++ _ +++ ____ +++ _ [Pro30, Nie31, Nal32, Leu34] NPY (28-36 ) +++ +++ _ +++ +++ _ BIBO3304 _ +++ _ +++ _ +++ _ +++ _ • PD160170 _ ++ _ ++ _ ++ _ ++ _ LY366258 + ++ + ++ 1-104870 _ ++ _ ++ _ ++ _ ±± _ LY 357897 _ + _ + _ + _ + _ 1-115814 ++ ++ ++ ++
    Example 5: Activity Test of Various Compositions with Embedded Docetaxel on GIST-H1 and SW-13 Cells The method of preparation and the cytotoxicity assays of various compositions were performed as in Example 4. The test results are shown in Table 6.
    Table 6:
    Comparison of the killing effect of various compositions on GIST-H1 and SW-13 cells
    ~~ __________ Nanocarrier BSA Nanocarrier_Chitosan Nanocarrier_
    Targeting molecule ~~~~ -------- SW-13 GIST-HI SW-13 GIST-HI [D-Arg25] -NPY _ ++ _ ++ _ ++ _ ++ _ [D-His26 ] -NPY _ ++ _ ++ _ ++ _ ++ _ [D-Arg25, D-His26] -NPY _ ++ _ ++ _ ++ _ ++ _ [D-His26, Pro34] NPY _ ++ _ + + _ ++ _ ++ _ [Phe7, Pro34] pNPY _ +++ _ +++ _ +++ _ +++ _ [Arg6, Pro34] pNPY _ +++ _ +++ _ +++ _ ++ + _ [Asn6, Pro34] pNPY 4 - ++ 4 - ++ 4-4-4- 4-4-4- [Cys6, PrO34] pNPY 4-4-4- 4-4-4- 4-4- 4- 4-4-4- [Phe6, Pro34] pNPY_4- + 4 -_ +++ _ +++ _ +++ _ [Pro30, Nie31, Bpa32, Leu34] NPY (28-36) +++ _ +++ _ +++ _ +++ _ [Pro30, Nal32, Leu34] NPY (28-36) _ +++ _ +++ _ +++ _ +++ _ [Pro30, Nie31, Nal32, Leu34 ] NPY (28-36) +++ _ +++ _ +++ _ +++ _ BIBQ3304 _ +++ _ +++ _ +++ _ +++ _ PD160170 ++ ++ ++ ++ LY366258_ + _ ++ _ + _ ++ _ J-104870 ++ ++ ++ ++ LY 357897 _ + _ + _ + _ + _ Jl 15814 _ ++ _ ++ _ ++ _ ++ _
    Example 6: Activity test of various compositions with embedded docetaxel on SMS-ΚΑΝ and HEC-1B-Y5 cells The preparation method and the cytotoxicity tests of different compositions were carried out as in Example 4. The test results are shown in Table 7.
    Table 7:
    Comparison of the killing effect of various compositions on SMS-ΚΑΝ and HEC-1B-Y5 cells
    权利要求:
    Claims (9)
    [1]

    Nanocarrier BSA nanocarrier_Chitosan nanocarrier_ target molecule ~~~~ ~~ ----- SMS-ΚΑΝ 1HEC-1B-Y5 SMS-ΚΑΝHEC-1B-Y5 [D-Arg25] -NPY_ ~ _-_-_ -N [D-His26] -NPY _ - _2 _-_-_ [D-Arg25, D-His26] -NPY_~ _ - _ [D-His26, Pro34] NPY _ - __ - _-_ [Phe7 , Pro34] pNPY __-_ - _-_ [Arg6, Pro34jpNPY _ - _ - _ - _-_ [Asn6, Pro34] pNPY_ ~~ _ [Cys6, Pro34] pNPY _ - _ - __-_ [ Phe6, Pro34] pNPY__ [Pro30, No31, Bpa32, Leu34] NPY (28-36) __ - _ [Pro30, Nal32, Leu34] NPY (28-36) _ - _ --- __ [Pro30, Nie31, Nal32, Leu34] NPY (28-36) _-_-_ = _ BIBO3304_ ~ _-_ - __ PD160170 _ - _ - _-_ Z_ LY366258 _-_ ~ _ - _ »_ J-104870 __-_- It can be seen from the test results of Tables 5-7 that the compositions coupled to the targeting molecule of the present invention have a.. strong killing effect on MCF-7, UWB1.289, GIST-H1 and SW-13 cells. At a CTxt of 5 pg / ml, the survival rates of the MCF-7, GIST-H1 and SW-13 cells were significantly below 30%, and the survival rate of the UWB1.289 cells was 60%. However, the killing effect was not significant in the SMS-ΚΑΝ and HEC-1B-Y5 cells, the survival rate was well over 80%. Thus, the compositions of the present invention have good selectivity, they have a strong targeting effect on breast cancer, ovarian cancer, kidney cancer and gastric cancer cells, and show a strong killing effect on cancer cells and a low killing effect on brain tumors and endometrial tumor cells. The entire literature cited in the present application is to be regarded as the content of the application, as well as each individual cited document. In addition, it should be understood that those skilled in the art, after reviewing the above disclosures, may make various improvements and changes, and that these equivalents of the present invention also fall within the scope of the following claims. claims
    A complex characterized in that the complex comprises: a nanocarrier; and a targeting molecule coupled to the surface of the nanocarrier; wherein the targeting molecule is a peptide agonist molecule or a non-peptide antagonist molecule which synergizes with neuropeptide Y receptors in high specificity and efficiency, thereby providing various biological activity; wherein the peptide agonist molecule is selected from the list: [D-Arg25] -NPY, [D-His26] -NPY, [D-Arg25, D-His26] -NPY, [Arg6, Pro34] pNPY, [Asn6 , Pro34] pNPY, [Cys6, Pro34] pNPY, [Phe6, Pro34] pNPY, [Arg7, Pro34] pNPY, [D-His26, Pro34] NPY, [Phe7, Pro34] pNPY, [Pro30, Nie31, Bpa32, Leu34 NPY (28-36), [Pro30, Nal32, Leu34] NPY (28-36), [Pro30, Nie31, Nal32, Leu34] NPY (28-36), or combinations thereof; wherein the non-peptide antagonist molecule is selected from the group consisting of BIBO3304, PD160170, LY366258, J-104870, LY357897, J-115814, or combinations thereof; and the nanocarrier has a particle size of 200 nm or smaller and a polydispersity index (PDI) less than 0.5.
    [2]
    2. Complex according to claim 1, characterized in that the content of targeting molecule is 1.11 to 22.2% by weight of the total weight of the complex.
    [3]
    3. Complex according to claim 1, characterized in that the particle size of the nanocarrier is 10-200 nm.
    [4]
    4. Complex according to claim 1, characterized in that the nanocarrier is selected from the following list: protein-based nanoparticles, oligopeptide-based nanoparticles, phospholipid-based nanoliposome, polysaccharide-based nanoparticles, polyether-based nanoparticles, polyester-based nanoparticles, polyester-based polymeric micelle.
    [5]
    A composition characterized in that the composition comprises: a complex according to claim 1; and an anti-tumor drug included in the nanocarrier of the complex.
    [6]
    6. The composition of claim 5 as an agent for the treatment of cancer.
    [7]
    A preparation method of the composition according to claim 5, characterized in that it comprises the steps of: (1) providing an anti-tumor drug-loaded nanocarrier; and (2) coupling the nanocarrier of step (1) with a targeting molecule, thereby obtaining the composition.
    [8]
    8. Production method according to claim 7, characterized in that the coupling reaction is selected from: (1) a condensation reaction of a carboxy group with an amino group; (2) an addition reaction of a sulfhydryl group with a maleimide group; or (3) noncovalent binding of avidin and biotin.
    [9]
    A medicament comprising: the complex of claim 1; an anti-tumor drug entrapped in the nanocarrier of the complex; and a pharmaceutically acceptable carrier.
    类似技术:
    公开号 | 公开日 | 专利标题
    Yang et al.2015|An implantable active-targeting micelle-in-nanofiber device for efficient and safe cancer therapy
    Xu et al.2015|Versatile preparation of intracellular-acidity-sensitive oxime-linked polysaccharide-doxorubicin conjugate for malignancy therapeutic
    DE112013004278T5|2015-05-21|Curcumin-Er, a prolonged or sustained-release liposomal PLNA nanocurcum to minimize QT prolongation for cancer therapy
    CH710313B1|2018-08-15|Anti-tumor drug with active targeting and its method of production.
    WO2012153286A1|2012-11-15|Polymeric nanoparticles for drug delivery
    US11090268B2|2021-08-17|Targeted hydrophobic anti-tumor drug nanoformulation and preparation method thereof
    Gong et al.2015|Curcumin-incorporated albumin nanoparticles and its tumor image
    DE102006011507A1|2007-09-20|Active substance-loaded nanoparticles based on hydrophilic proteins
    CN101396346B|2014-01-01|Paclitaxel lipid composite
    Dhanka et al.2017|Injectable methotrexate loaded polycaprolactone microspheres: Physicochemical characterization, biocompatibility, and hemocompatibility evaluation
    WO2010106063A2|2010-09-23|Composite materials loaded with therapeutic and diagnostic agents comprising polymer nanoparticles and polymer fibers
    EP1796649A2|2007-06-20|Nanotransport system having a dendritic architecture
    Chen et al.2017|Toxicity, pharmacokinetics, and in vivo efficacy of biotinylated chitosan surface-modified PLGA nanoparticles for tumor therapy
    Bin et al.2017|Nano-carrier mediated co-delivery of methyl prednisolone and minocycline for improved post-traumatic spinal cord injury conditions in rats
    Wu et al.2020|Engineering stem cell derived biomimetic vesicles for versatility and effective targeted delivery
    DE60117583T2|2006-10-05|LIPOSOMES CAPTURING ANTICANCED AGENTS AND THEIR USE FOR THE TREATMENT OF MALIGNANT TUMORS
    CN108495619A|2018-09-04|Echinomycin preparation and preparation method thereof and application method
    Dong et al.2020|Comparative study of mucoadhesive and mucus-penetrative nanoparticles based on phospholipid complex to overcome the mucus barrier for inhaled delivery of baicalein
    CN105476975B|2019-10-08|Anti- brain tumor drug of active targeting type and preparation method thereof
    EP2741783A1|2014-06-18|Polyanion nanocomplexes for therapeutic applications
    Valente et al.2018|Evaluation of toxicity of glycerol monooleate nanoparticles on PC12 cell line
    US20130323314A1|2013-12-05|Novel Nanoparticles of Lactoferrin Useful for Preparing a Pharmaceutical Composition Facilitating Easy Delivery of the Drug and a Process for Preparing the Same
    Gao et al.2017|A rod bacterium-like magnetic polymer micelle for strongly enhancing selective accumulation and internalization of nanocarriers
    WO2002000162A2|2002-01-03|Drug-delivery systems
    KR20120046595A|2012-05-10|Method for preparing biocompatible particle with multi-core structure and biocompatible particle with multi-core structure prepared by the same
    同族专利:
    公开号 | 公开日
    US20160213788A1|2016-07-28|
    CN104415338B|2018-04-10|
    CN104415338A|2015-03-18|
    DE112014004133T5|2016-06-16|
    WO2015032207A1|2015-03-12|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    US5026685A|1988-07-15|1991-06-25|The Salk Institute For Biological Studies|NPY peptide analogs|
    CN1634591A|2004-11-11|2005-07-06|东华大学|Short peptide modified polylysine-polylactic copolymer nano particle and its preparation method and use|
    CN102048694B|2009-11-06|2013-03-13|复旦大学|Polypeptide-modified liver tumor-targeted nano medicine delivery system and preparation method thereof|
    US20140322341A1|2011-08-03|2014-10-30|Diane RUBIN|Novel hemostatic patch and uses thereof|
    CN103126993B|2011-12-05|2015-05-13|中国科学院宁波材料技术与工程研究所|Tumor cell active targeting drug delivery system based on nanometer particles and construction method thereof|CN105476975B|2015-08-27|2019-10-08|中国科学院宁波材料技术与工程研究所|Anti- brain tumor drug of active targeting type and preparation method thereof|
    CN107296963B|2016-04-15|2021-06-25|中国科学院宁波材料技术与工程研究所|Active targeting type ultrasonic/fluorescent bimodal contrast agent and preparation method and application thereof|
    CN106267185B|2016-09-19|2020-01-10|中国科学院过程工程研究所|Chitosan oligosaccharide vaccine adjuvant based on chemical coupling and application thereof|
    CN110339373A|2018-04-04|2019-10-18|中国科学院宁波材料技术与工程研究所|A kind of nano combined micella and its preparation method and application|
    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    CN201310407938.0A|CN104415338B|2013-09-09|2013-09-09|Active targeting type antineoplastic and preparation method thereof|
    PCT/CN2014/075727|WO2015032207A1|2013-09-09|2014-04-18|Active targeting antitumor drug and preparation method therefor|
    [返回顶部]