CN114558148A - Preparation method and application of tumor switch type nano phototherapy system - Google Patents
Preparation method and application of tumor switch type nano phototherapy system Download PDFInfo
- Publication number
- CN114558148A CN114558148A CN202210271183.5A CN202210271183A CN114558148A CN 114558148 A CN114558148 A CN 114558148A CN 202210271183 A CN202210271183 A CN 202210271183A CN 114558148 A CN114558148 A CN 114558148A
- Authority
- CN
- China
- Prior art keywords
- transferrin
- nano
- stirring
- tumor
- solution
- 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.)
- Granted
Links
- 238000001126 phototherapy Methods 0.000 title claims abstract description 43
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 42
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- XXSMGPRMXLTPCZ-UHFFFAOYSA-N hydroxychloroquine Chemical compound ClC1=CC=C2C(NC(C)CCCN(CCO)CC)=CC=NC2=C1 XXSMGPRMXLTPCZ-UHFFFAOYSA-N 0.000 claims abstract description 71
- 229960004171 hydroxychloroquine Drugs 0.000 claims abstract description 55
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 claims abstract description 52
- 102000004338 Transferrin Human genes 0.000 claims abstract description 35
- 108090000901 Transferrin Proteins 0.000 claims abstract description 35
- 239000012581 transferrin Substances 0.000 claims abstract description 35
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 33
- 229960003180 glutathione Drugs 0.000 claims abstract description 26
- 238000003756 stirring Methods 0.000 claims abstract description 26
- 239000000243 solution Substances 0.000 claims abstract description 23
- 239000003814 drug Substances 0.000 claims abstract description 20
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 18
- 229940079593 drug Drugs 0.000 claims abstract description 18
- 239000007864 aqueous solution Substances 0.000 claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 13
- 229910021642 ultra pure water Inorganic materials 0.000 claims abstract description 13
- 239000012498 ultrapure water Substances 0.000 claims abstract description 13
- 108010024636 Glutathione Proteins 0.000 claims abstract description 12
- 239000012456 homogeneous solution Substances 0.000 claims abstract description 12
- 230000000259 anti-tumor effect Effects 0.000 claims abstract description 11
- 238000000502 dialysis Methods 0.000 claims abstract description 11
- 239000002245 particle Substances 0.000 claims abstract description 5
- VOADVZVYWFSHSM-UHFFFAOYSA-L sodium tellurite Chemical compound [Na+].[Na+].[O-][Te]([O-])=O VOADVZVYWFSHSM-UHFFFAOYSA-L 0.000 claims abstract description 3
- 230000002209 hydrophobic effect Effects 0.000 claims description 9
- 229910004273 TeO3 Inorganic materials 0.000 claims description 8
- 238000011161 development Methods 0.000 claims description 8
- 239000011734 sodium Substances 0.000 claims description 8
- 238000002347 injection Methods 0.000 claims description 5
- 239000007924 injection Substances 0.000 claims description 5
- 239000008194 pharmaceutical composition Substances 0.000 claims description 5
- 239000003937 drug carrier Substances 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims 2
- 239000012467 final product Substances 0.000 claims 1
- 239000008176 lyophilized powder Substances 0.000 claims 1
- 210000004881 tumor cell Anatomy 0.000 abstract description 13
- 210000004027 cell Anatomy 0.000 abstract description 10
- 239000003504 photosensitizing agent Substances 0.000 abstract description 7
- 230000004900 autophagic degradation Effects 0.000 abstract description 6
- 238000006243 chemical reaction Methods 0.000 abstract description 6
- 230000008569 process Effects 0.000 abstract description 4
- 230000002195 synergetic effect Effects 0.000 abstract description 4
- 230000008685 targeting Effects 0.000 abstract description 4
- 239000003642 reactive oxygen metabolite Substances 0.000 abstract 2
- 238000002474 experimental method Methods 0.000 description 8
- 230000005764 inhibitory process Effects 0.000 description 8
- 229910052714 tellurium Inorganic materials 0.000 description 5
- PORWMNRCUJJQNO-UHFFFAOYSA-N tellurium atom Chemical compound [Te] PORWMNRCUJJQNO-UHFFFAOYSA-N 0.000 description 5
- 238000011282 treatment Methods 0.000 description 5
- 230000008901 benefit Effects 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 241000699670 Mus sp. Species 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 102000007238 Transferrin Receptors Human genes 0.000 description 3
- 108010033576 Transferrin Receptors Proteins 0.000 description 3
- 239000012822 autophagy inhibitor Substances 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000011835 investigation Methods 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 238000002428 photodynamic therapy Methods 0.000 description 3
- 238000007626 photothermal therapy Methods 0.000 description 3
- 230000003592 biomimetic effect Effects 0.000 description 2
- 230000033558 biomineral tissue development Effects 0.000 description 2
- 230000001186 cumulative effect Effects 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000005917 in vivo anti-tumor Effects 0.000 description 2
- 210000003712 lysosome Anatomy 0.000 description 2
- 230000001868 lysosomic effect Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 208000000461 Esophageal Neoplasms Diseases 0.000 description 1
- 241001529251 Gallinago gallinago Species 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 208000002454 Nasopharyngeal Carcinoma Diseases 0.000 description 1
- 206010061306 Nasopharyngeal cancer Diseases 0.000 description 1
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 210000004957 autophagosome Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 201000004101 esophageal cancer Diseases 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000037041 intracellular level Effects 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 201000011216 nasopharynx carcinoma Diseases 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 229910000033 sodium borohydride Inorganic materials 0.000 description 1
- 239000012279 sodium borohydride Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 230000005909 tumor killing Effects 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
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
- A61K47/69—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
- A61K47/6949—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 inclusion complexes, e.g. clathrates, cavitates or fullerenes
-
- 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
- A61K31/4706—4-Aminoquinolines; 8-Aminoquinolines, e.g. chloroquine, primaquine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
-
- 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
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
- A61K47/644—Transferrin, e.g. a lactoferrin or ovotransferrin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to a preparation method and application of a tumor switch-type nano phototherapy system, which effectively solves the problem of low efficiency of the existing anti-tumor phototherapy, transferrin is dissolved in glutathione aqueous solution and stirred to open intramolecular disulfide bonds S-S, and transferrin homogeneous solution with expanded spatial structure is obtained; dripping hydroxychloroquine ethanol solution into the transferrin homogeneous solution, and stirring; dropwise adding sodium tellurite aqueous solution, then using sodium hydroxide solution to adjust pH, stirring, transferring to a dialysis bag with MWCO =100kD, and dialyzing with ultrapure water to remove free drugs, thereby obtaining the Tf-Te/HCQ nano preparation with uniform particle size, which can realize natural targeting on tumors and selective drug release in target cells, remarkably improve the photothermal conversion efficiency of photosensitizers and the generation efficiency of ROS (reactive oxygen species), and simultaneously, the loaded HCQ can selectively inhibit the autophagy of tumor cells enhanced in the phototherapy process, thereby exerting the synergistic antitumor effect.
Description
Technical Field
The invention relates to the field of pharmaceutical preparations, in particular to a preparation method and application of a tumor switch type nano phototherapy system.
Background
In recent years, phototherapy (including photothermal therapy PTT and photodynamic therapy PDT) has been developed as a potential new tumor therapy model for clinical treatment of various cancers due to its significant advantages of high efficiency, minimal invasion, low toxicity, etc. Phototherapy is the process of irradiating photosensitizer enriched in tumor with laser to generate local high temperature or active oxygen (ROS) with strong cytotoxicity to kill tumor cells. Recent studies have found that tumor cells activate autophagy under stress conditions. Initiate intracellular circulation by autophagy, perform self-detoxification, enhance resistance of tumor cells, resist PTT and PDT treatments. Therefore, the development of a novel photosensitizer and the simultaneous inhibition of the treatment resistance caused by autophagy are novel strategies for improving the phototherapy antitumor efficiency.
The tellurium nanoparticles have good photo-thermal conversion efficiency and characteristics of a photosensitizer, and have good application prospects in the field of anti-tumor phototherapy. But the potential biological toxicity and in vivo degradation problems have prevented their biological use. How to improve the biocompatibility of the nano tellurium is an important challenge to be faced.
Transferrin is the main transferrin-containing substance in blood plasma, and the expression level of transferrin receptor (TfR) on the surface of tumor cells is far higher than that of normal cells, so transferrin has natural targeting property on the tumor cells. In addition, transferrin is an excellent drug carrier, and the hydrophobic cavity of the transferrin can be loaded with various hydrophobic therapeutic drugs. Transferrin therefore has its unique advantages in antitumor therapy.
Hydroxychloroquine (HCQ) as a classical autophagy inhibitor can destroy the activity of acid hydrolase in lysosomes by inducing lysosomes to deacidify, so that damaged substances in the autophagosomes cannot be degraded, a self-protection way of tumor cells under a severe oxidative stress state is cut off, and the anti-tumor phototherapy effect is synergistically enhanced. However, HCQ has no tissue and cell selectivity, so that how to accurately snipe cancer cells, selectively inhibit autophagy of tumor cells enhanced in the phototherapy process and exert a synergistic anti-tumor effect is a problem to be solved urgently at present.
Disclosure of Invention
In view of the above situation, the present invention aims to provide a method for preparing a "switch-type" nano phototherapy system for tumor and the application thereof, which can effectively solve the problem of low efficiency of the existing anti-tumor phototherapy.
The invention has the technical scheme that the preparation method of the tumor switch type nano phototherapy system comprises the following specific steps:
(1) dissolving 10-25mg of transferrin in 2mL of glutathione aqueous solution with the concentration of 6-20mM, stirring for 30-90min at the temperature of 25-55 ℃, and opening an intramolecular disulfide bond S-S to obtain a transferrin homogeneous solution with the final concentration of 5-12.5mg/mL and a spatial structure developed; dropwise adding 0.4mL of Hydroxychloroquine (HCQ) ethanol solution of 2.5-10mg/mL into the transferrin homogeneous solution, and stirring for 4-12 h;
the preparation method of the 6-20mM Glutathione (also called GSH aqueous solution) comprises the steps of dissolving 4-12mg of Glutathione (GSH) in 2mL of ultrapure water to prepare the Glutathione aqueous solution;
(2) 2-8mg/mL sodium tellurite (Na) is added dropwise2TeO3) 1mL of the aqueous solution, then adjusting the pH value to 7-14 by using a 2M sodium hydroxide (NaOH) solution, stirring for 4-12h at 25 ℃, then transferring to a dialysis bag with MWCO =100kD, and dialyzing for 4-12h by using ultrapure water to remove free drugs, thus obtaining the tumor switch type nano phototherapy system (also known as: tumor "switch-type" nano phototherapy system Tf-Te/HCQ) based on disulfide bond reconstruction.
The invention converts intramolecular disulfide bonds of transferrin into intermolecular disulfide bonds by a disulfide bond reconstruction technology, utilizes a hydrophobic cavity to load an autophagy inhibitor Hydroxychloroquine (HCQ), and synchronously carries out in-situ biomimetic mineralization on nano tellurium (Tf-Te) to obtain a Tf-Te/HCQ nano phototherapy system, wherein the Tf-Te/HCQ nano phototherapy system is a 'switch type' nano phototherapy system for tumors.
The preparation method is simple, the prepared Tf-Te/HCQ nano preparation has uniform grain diameter of 50-500nm, can realize natural targeting on tumors and selective drug release in target cells, and obviously improves the photo-thermal conversion and ROS generation efficiency of the photosensitizer. Meanwhile, the loaded HCQ can selectively inhibit autophagy enhanced by tumor cells in the phototherapy process, and plays a role in synergistic antitumor effect.
Detailed Description
The following examples are provided to explain the present invention in detail.
Example 1
(1) Dissolving 15mg transferrin in 2mL of 12mM glutathione aqueous solution, stirring for 60min at 37 ℃, and opening intramolecular disulfide bonds S-S to obtain a transferrin homogeneous solution with a final concentration of 7.5mg/mL and a spatial structure development; dropwise adding 0.4mL of hydroxychloroquine ethanol solution of 6mg/mL, and stirring for 8h to enable HCQ load to enter a hydrophobic cavity of transferrin;
(2) 5mg/mL of Na was added dropwise2TeO3Adjusting the pH value to 10 by using 2M NaOH solution, stirring for 8h at 25 ℃, transferring to a dialysis bag with MWCO =100kD, and dialyzing for 8h by using ultrapure water to remove free drugs to obtain the Tf-Te/HCQ based on the disulfide bond reconstruction tumor switch type nano phototherapy system.
Example 2
(1) Dissolving 10mg of transferrin in 2mL of 6mM glutathione aqueous solution, stirring for 30min at 25 ℃, and opening intramolecular disulfide bonds S-S to obtain transferrin homogeneous solution with final concentration of 5mg/mL and spatial structure development; dropwise adding 0.4mL of hydroxychloroquine ethanol solution of 2.5mg/mL, and stirring for 4h to make HCQ load enter a hydrophobic cavity of transferrin;
(2) 2mg/mL of Na was added dropwise2TeO3Adjusting the pH value to 7 by using 2M NaOH solution, stirring for 4h at 25 ℃, transferring to a dialysis bag with MWCO =100kD, and dialyzing for 4h by using ultrapure water to remove free drugs to obtain the Tf-Te/HCQ based on the disulfide bond reconstruction tumor switch type nano phototherapy system.
Example 3
(1) Dissolving 25mg transferrin in 2mL of glutathione aqueous solution with the concentration of 20mM, stirring for 90min at 55 ℃, and opening intramolecular disulfide bonds S-S to obtain transferrin homogeneous solution with the final concentration of 12.5mg/mL and spatial structure development; dropwise adding 0.4mL of hydroxychloroquine ethanol solution of 10mg/mL, and stirring for 12h to enable HCQ load to enter a hydrophobic cavity of transferrin;
(2) 8mg/mL of Na was added dropwise2TeO3Adjusting the pH value to 14 by using 2M NaOH solution, stirring for 12h at 25 ℃, transferring to a dialysis bag with MWCO =100kD, and dialyzing for 12h by using ultrapure water to remove free drugs to obtain the Tf-Te/HCQ based on the disulfide bond reconstruction tumor switch type nano phototherapy system.
The particle size of the tumor switch type nano phototherapy system is 50-500 nm.
Another objective of the present invention is to provide an application of the "switch-type" nano phototherapy system (Tf-Te/HCQ for short) in preparing anti-tumor drugs, such as breast cancer, lung cancer, melanoma, nasopharyngeal carcinoma, esophageal cancer, etc.
A pharmaceutical composition of a tumor 'switch type' nano phototherapy system comprises the tumor 'switch type' nano phototherapy system and a pharmaceutically acceptable carrier; the pharmaceutical composition is an injection or a freeze-dried powder injection.
The invention obtains consistent results through repeated experiments, and the related experimental data are as follows:
experiment 1: characterization of Tf-Te/HCQ
The transmission electron microscope result of the tumor 'switch type' nano phototherapy system prepared by the method shows that the nano preparation (namely the tumor 'switch type' nano phototherapy system prepared by the method) has uniform particle size, and the average particle size is 50-500 nm.
Experiment 2: photothermal conversion Performance evaluation of Tf-Te/HCQ
Investigation of the formulation concentration: the Tf-Te/HCQ nano preparation with the concentration of 0-80 mug/mL is prepared by adding ultrapure water into the Tf-Te/HCQ nano preparation, the Tf-Te/HCQ nano preparation with different concentrations (0-80 mug/mL) is irradiated for 0-5min under 808nm laser, and the change of temperature along with the concentration is observed and recorded every 1 min.
Examination of laser power: placing 40 μ g/mL Tf-Te/HCQ nanometer preparation under 808nm laser with power of 1-5W/cm2Irradiating for 0-5min, and observing and recording the change of temperature with power every 1 min.
The results show that the temperature change of the Tf-Te/HCQ has obvious concentration dependence and power dependence. The temperature rise is faster the greater the concentration and laser power are for the same time of irradiation. The photothermal conversion efficiency can reach 70 percent, thereby realizing the purpose of tumor photothermal treatment.
Experiment 3: GSH response drug release characteristic investigation of Tf-Te/HCQ drug delivery system
The Tf-Te/HCQ nanopreparations (i.e. the tumor "switch-type" nano phototherapy system prepared in the present application) were placed in dialysis bags (MWCO =3500 Da), immersed in phosphate PBS buffer of different GSH concentrations [ ph7.4 ] (no GSH was added to this group); GSH at pH7.4+ 20. mu.M; pH7.4+5mM GSH, then placed in a constant temperature shaker for release (100 r/min, 37 ℃). Taking out a small amount of release medium at a preset time point, measuring HCQ absorbance at 343nm by adopting an ultraviolet spectrophotometry, and calculating the cumulative drug release percentage.
The result shows that HCQ release has obvious GSH dependence, and the release speed is as follows: pH7.4+5mM GSH > pH7.4+ 20. mu.M GSH > pH7.4. Wherein, the cumulative drug release percentages of the pH7.4+5mM GSH, pH7.4+20 μ M GSH and pH7.4 groups at 72h are 85%, 45% and 30% respectively, which shows that the Tf-Te/HCQ nano preparation can release drug at fixed points in the microenvironment with high GSH of tumors.
Experiment 4: experimental investigation of in vitro cell inhibition
4T1 tumor cells were selected at 2X 10 in logarithmic growth phase 4cell/mL density, placed in 96 wellsIn the plate, the plate was cultured for 24 hours. After the cells were completely adhered, the different preparations in the following experimental groups were added, incubated for 4 hours and irradiated with 808nm laser for 1min (2W/cm)2) Then continuously culturing for 24 hours, and finally determining the cell inhibition rate by adopting an SRB method;
the experimental groups were as follows: 1) blank group: 1640 medium without any formulation. 2) Tf-Te group: dissolving Tf-Te in 1640 culture medium to obtain Tf-Te dispersion solution with concentration of 10 μ g/mL (the Tf-Te is prepared by dissolving 10-25mg transferrin in 1mL ultrapure water, adjusting pH to 7-14 with 2M NaOH solution, and adding 2-8mg/mL Na dropwise2TeO31mL of aqueous solution, reacting for 2h, and adding 0.5-2mg of sodium borohydride (NaBH)4) Reacting for 4-12h, transferring to a dialysis bag with MWCO =100kD, and dialyzing with ultrapure water for 4-12h to remove free drug to obtain Tf-Te). 3) HCQ group: HCQ was dissolved in 1640 medium to prepare a free HCQ solution having a concentration of 10. mu.g/mL. 4): Tf-Te/HCQ group: the Tf-Te/HCQ nano preparation prepared by the application is added into a 1640 culture medium to be dissolved, and the Tf-Te/HCQ preparation with the concentration of 10 mug/mL is prepared.
The results show that the cell inhibition rates of the Tf-Te group, the HCQ group and the Tf-Te/HCQ group are respectively 52%, 48% and 89%. Compared with other groups, the Tf-Te/HCQ group has the highest cell inhibition rate, and shows that the Tf-Te/HCQ prepared by the method can realize high-efficiency killing of tumor cells under laser irradiation.
Experiment 5: evaluation of in vivo antitumor Effect
4T1 tumor cells were inoculated subcutaneously into Bablc mice when tumor volume reached-100 mm3At the time, tumor-bearing mice were randomly divided into 8 groups: 1) a normal saline control group, 2) an HCQ group, 3) a Tf-Te group, 4) a Tf-Te/HCQ group, 5) a normal saline control + laser group, 6) an HCQ + laser group, 7) a Tf-Te + laser group, 8) a Tf-Te/HCQ + laser group; wherein all laser groups irradiated the tumor part with 808nm laser for 1min (2W/cm) 4 hours after intravenous administration2). The single doses of HCQ group and HCQ + laser group in the above administration groups were 10mg/kg, the single doses of Tf-Te group and Tf-Te + laser group were 8mg/kg, and the single doses of HCQ and Tf-Te in the Tf-Te/HCQ group and Tf-Te/HCQ + laser group were 8mg/kgThe amounts were 10mg/kg and 8mg/kg, respectively. The drug is administrated by tail vein injection every other day for 7 times. At the end of the pharmacodynamic experiment, all mice were sacrificed, tumors were removed, weighed, and tumor inhibition rate was calculated.
Results show that the tumor inhibition rates of the HCQ group, the Tf-Te/HCQ group, the normal saline contrast + laser group, the HCQ + laser group, the Tf-Te + laser group and the Tf-Te/HCQ + laser group are 33%, 23%, 46%, 5.2%, 39%, 58% and 87% respectively. Compared with other administration groups, the Tf-Te/HCQ + laser group has the highest tumor inhibition rate in vivo, which shows that the Tf-Te/HCQ can obviously inhibit the growth of 4T1 tumor under laser irradiation, and has good in vivo anti-tumor effect.
Experiments show that the Tf-Te/HCQ prepared by the preparation method has stable and reliable drug effect and good product quality. Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) the characteristics of high expression of transferrin receptor on the surface of tumor cells and high intracellular level of GSH are utilized to realize natural targeting of tumor and micro-environment response drug release;
(2) the synthesis of the nano tellurium photosensitizer is improved through a biomimetic mineralization technology, and the photo-thermal conversion efficiency of the nano tellurium photosensitizer is obviously improved.
(3) And the compound has synergistic effect with the hydroxychloroquine which is an autophagy inhibitor, so that the phototherapy efficiency of the tumor is improved.
The characteristics show that the preparation method is simple, easy to operate, low in cost, good in anti-tumor effect, and huge in economic and social benefits, and develops new application of the tumor treatment medicine.
Claims (8)
1. A preparation method of a tumor switch type nano phototherapy system is characterized by comprising the following specific steps:
(1) dissolving 10-25mg transferrin in 2mL of glutathione aqueous solution with the concentration of 6-20mM, and stirring for 30-90min at the temperature of 25-55 ℃ to obtain transferrin homogeneous solution with the final concentration of 5-12.5mg/mL and spatial structure development; dripping 0.4mL of hydroxychloroquine ethanol solution of 2.5-10mg/mL into the transferrin homogeneous solution, and stirring for 4-12 h;
(2) dropwise adding 1mL of 2-8mg/mL sodium tellurite aqueous solution, adjusting pH to 7-14 with 2M sodium hydroxide solution, stirring at 25 deg.C for 4-12h, transferring to dialysis bag with MWCO =100kD, and dialyzing with ultrapure water for 4-12h to remove free drug.
2. The method of claim 1, wherein the nano phototherapy system is a "switch type" nano phototherapy system,
(1) dissolving 15mg transferrin in 2mL of 12mM glutathione aqueous solution, stirring for 60min at 37 ℃, and opening intramolecular disulfide bonds S-S to obtain a transferrin homogeneous solution with a final concentration of 7.5mg/mL and a spatial structure development; dropwise adding 0.4mL of hydroxychloroquine ethanol solution of 6mg/mL, and stirring for 8h to enable HCQ load to enter a hydrophobic cavity of transferrin;
(2) 5mg/mL of Na was added dropwise2TeO3Adjusting pH to 10 with 2M NaOH solution in 1mL of water solution, stirring at 25 deg.C for 8h, transferring to MWCO =100kD dialysis bag, and dialyzing with ultrapure water for 8h to remove free drug.
3. The method of claim 1, wherein the nano phototherapy system is a "switch type" nano phototherapy system,
(1) dissolving 10mg transferrin in 2mL of glutathione aqueous solution with the concentration of 6mM, stirring for 30min at 25 ℃, and opening intramolecular disulfide bonds S-S to obtain transferrin homogeneous solution with the final concentration of 5mg/mL and spatial structure development; dropwise adding 0.4mL of hydroxychloroquine ethanol solution of 2.5mg/mL, and stirring for 4h to make HCQ load enter a hydrophobic cavity of transferrin;
(2) 2mg/mL of Na was added dropwise2TeO3Adjusting pH to 7 with 2M NaOH solution in 1mL of water solution, stirring at 25 deg.C for 4h, transferring to MWCO =100kD dialysis bag, and dialyzing with ultrapure water for 4h to remove free drug.
4. The method of claim 1, wherein the nano phototherapy system is a "switch type" nano phototherapy system,
(1) dissolving 25mg of transferrin in 2mL of glutathione aqueous solution with the concentration of 20mM, stirring for 90min at 55 ℃, and opening intramolecular disulfide bonds S-S to obtain transferrin homogeneous solution with the final concentration of 12.5mg/mL and a spatial structure development; dropwise adding 0.4mL of hydroxychloroquine ethanol solution of 10mg/mL, and stirring for 12h to enable HCQ load to enter a hydrophobic cavity of transferrin;
(2) 8mg/mL of Na is added dropwise2TeO3Adjusting pH to 14 with 2M NaOH solution, stirring at 25 deg.C for 12h, transferring to MWCO =100kD dialysis bag, and dialyzing with ultrapure water for 12h to remove free drug to obtain the final product.
5. The method of claim 1, wherein the particle size of the tumor "on-off" nano phototherapy system is 50-500 nm.
6. Use of the tumor "switched" nano phototherapy system of any one of claims 1 to 5 in the preparation of an anti-tumor medicament.
7. A pharmaceutical composition of a tumor switch type nano phototherapy system is characterized by comprising the tumor switch type nano phototherapy system and a pharmaceutically acceptable carrier.
8. The pharmaceutical composition of claim 7, wherein the pharmaceutical composition is an injection or lyophilized powder for injection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210271183.5A CN114558148B (en) | 2022-03-18 | 2022-03-18 | Preparation method and application of tumor switch-type nano phototherapy system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210271183.5A CN114558148B (en) | 2022-03-18 | 2022-03-18 | Preparation method and application of tumor switch-type nano phototherapy system |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114558148A true CN114558148A (en) | 2022-05-31 |
| CN114558148B CN114558148B (en) | 2024-03-08 |
Family
ID=81720267
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210271183.5A Active CN114558148B (en) | 2022-03-18 | 2022-03-18 | Preparation method and application of tumor switch-type nano phototherapy system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114558148B (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002094271A1 (en) * | 2001-05-15 | 2002-11-28 | Faulk Pharmaceuticals, Inc. | Targeted delivery of bioaffecting compounds for the treatment of cancer |
| US20160310594A1 (en) * | 2013-10-16 | 2016-10-27 | Jinis Co., Ltd. | Sensitizing composition using electromagnetic waves for thermal therapy of cancers, and cancer therapy using same |
| CN107427579A (en) * | 2015-01-19 | 2017-12-01 | 锡拉莱斯科技有限公司 | Metal sugar-protein compound and its purposes as chemotherapy compound |
| CN108126199A (en) * | 2017-12-29 | 2018-06-08 | 青岛大学 | A kind of degradable double-bang firecracker answers the preparation method of intelligent macromolecule/molybdenum sulfide pharmaceutical carrier |
| CN108379228A (en) * | 2018-02-28 | 2018-08-10 | 湖南大学 | A kind of albumin nanoparticle and its preparation method and application of package pharmacological active substance |
| CN112451675A (en) * | 2019-09-06 | 2021-03-09 | 复旦大学 | Synergistic combination medicine for treating breast cancer |
| CN113304264A (en) * | 2021-06-13 | 2021-08-27 | 重庆医科大学 | Quercetin tellurium nanoparticles and preparation method thereof |
| CN114163547A (en) * | 2021-12-14 | 2022-03-11 | 中国药科大学 | Pharmaceutical composition for mild photothermal treatment of tumors and preparation method and application thereof |
-
2022
- 2022-03-18 CN CN202210271183.5A patent/CN114558148B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002094271A1 (en) * | 2001-05-15 | 2002-11-28 | Faulk Pharmaceuticals, Inc. | Targeted delivery of bioaffecting compounds for the treatment of cancer |
| US20040157767A1 (en) * | 2001-05-15 | 2004-08-12 | Faulk W. Page | Targeted delivery of bioaffecting compounds for the treatment of cancer |
| US20160310594A1 (en) * | 2013-10-16 | 2016-10-27 | Jinis Co., Ltd. | Sensitizing composition using electromagnetic waves for thermal therapy of cancers, and cancer therapy using same |
| CN107427579A (en) * | 2015-01-19 | 2017-12-01 | 锡拉莱斯科技有限公司 | Metal sugar-protein compound and its purposes as chemotherapy compound |
| CN108126199A (en) * | 2017-12-29 | 2018-06-08 | 青岛大学 | A kind of degradable double-bang firecracker answers the preparation method of intelligent macromolecule/molybdenum sulfide pharmaceutical carrier |
| CN108379228A (en) * | 2018-02-28 | 2018-08-10 | 湖南大学 | A kind of albumin nanoparticle and its preparation method and application of package pharmacological active substance |
| CN112451675A (en) * | 2019-09-06 | 2021-03-09 | 复旦大学 | Synergistic combination medicine for treating breast cancer |
| CN113304264A (en) * | 2021-06-13 | 2021-08-27 | 重庆医科大学 | Quercetin tellurium nanoparticles and preparation method thereof |
| CN114163547A (en) * | 2021-12-14 | 2022-03-11 | 中国药科大学 | Pharmaceutical composition for mild photothermal treatment of tumors and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 汪巧莉: "双功能碲纳米点用于肿瘤光治疗的研究", pages 2 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114558148B (en) | 2024-03-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Xu et al. | Smart polydopamine-based nanoplatforms for biomedical applications: state-of-art and further perspectives | |
| CN113521098B (en) | Platinum (IV) and cRGD modified GA/Fe nano-particle carried doxorubicin and method for targeted therapy of tumors by using same | |
| CN110665003A (en) | Double-drug-loading carrier-free nanoparticle and preparation method thereof | |
| Wang et al. | Chitosan-and hyaluronic acid-based nanoarchitectures in phototherapy: Combination cancer chemotherapy, immunotherapy and gene therapy | |
| CN113018267B (en) | Unsaturated fatty acid-photosensitizer co-assembled nanoparticles and construction method and application thereof | |
| CN111632144B (en) | High-drug-loading-rate photo-chemotherapy bifunctional nanoparticles and preparation method thereof | |
| Lin et al. | A review on reactive oxygen species (ROS)-inducing nanoparticles activated by uni-or multi-modal dynamic treatment for oncotherapy | |
| CN112546025B (en) | Preparation method of Ce6@CMCS-DSP-IPI549 anti-tumor nano-delivery system | |
| CN110251672B (en) | Nano diagnosis and treatment agent and preparation method and application thereof | |
| CN114558148B (en) | Preparation method and application of tumor switch-type nano phototherapy system | |
| CN107812189B (en) | Hypocrellin nano preparation for actively targeting specific tumor cells and preparation method and application thereof | |
| CN117338925A (en) | Application of albumin drug-loaded nano-particles for double light treatment in preparation of drugs for treating cervical cancer | |
| CN113384698B (en) | Self-assembled nano-medicament for synergetic chemotherapy/acousto-photodynamic therapy and application thereof | |
| CN112870356B (en) | Series of medicines for photodynamic therapy of tumor and application thereof | |
| Zhang et al. | A NIR-driven green affording-oxygen microrobot for targeted photodynamic therapy of tumors | |
| CN116036289A (en) | Nanometer preparation for enhancing iron death treatment effect, and preparation method and application thereof | |
| KR102513560B1 (en) | Pharmaceutical composition for the treatment of cancer comprising phycocyanin-anticancer agent conjugate, composition for photodynamic therapy comprising the same and manufacturing method thereof | |
| CN115252788A (en) | Multi-mode anti-tumor nano-drug carrier, drug delivery system, and preparation method and application thereof | |
| CN113908276A (en) | Light-controlled drug release nano particle and preparation method and application thereof | |
| CN113827724A (en) | Drug-loaded Prussian blue @ manganese fibrin composite gel, and preparation method and application thereof | |
| CN113679836A (en) | Targeted ZIF-8-polydopamine prodrug nanoparticle and preparation method and application thereof | |
| CN114028565B (en) | 3D-COF drug carrying system for treating breast cancer and preparation method thereof | |
| CN114917191B (en) | Preparation method and application of vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS | |
| CN117257760A (en) | Preparation method and application of brain glioma-targeted copper ion hybrid polydopamine nanoparticle | |
| WO2022087865A1 (en) | Application of hydrogen peroxide in solid tumor treatment |
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 |