CN109157662A - A kind of human serum albumins-adriamycin cross-linking agent nano particle and its application - Google Patents
A kind of human serum albumins-adriamycin cross-linking agent nano particle and its application Download PDFInfo
- Publication number
- CN109157662A CN109157662A CN201810575098.1A CN201810575098A CN109157662A CN 109157662 A CN109157662 A CN 109157662A CN 201810575098 A CN201810575098 A CN 201810575098A CN 109157662 A CN109157662 A CN 109157662A
- Authority
- CN
- China
- Prior art keywords
- adriamycin
- human serum
- cross
- serum albumins
- linking agent
- 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
- 229940009456 adriamycin Drugs 0.000 title claims abstract description 51
- 210000002966 serum Anatomy 0.000 title claims abstract description 40
- 239000003431 cross linking reagent Substances 0.000 title claims abstract description 32
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 20
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 claims abstract description 73
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 53
- 102000009027 Albumins Human genes 0.000 claims abstract description 40
- 108010088751 Albumins Proteins 0.000 claims abstract description 40
- 238000004132 cross linking Methods 0.000 claims abstract description 14
- 230000002147 killing effect Effects 0.000 claims abstract description 9
- 150000003384 small molecules Chemical class 0.000 claims abstract description 7
- 206010006187 Breast cancer Diseases 0.000 claims description 18
- 208000026310 Breast neoplasm Diseases 0.000 claims description 18
- 238000002360 preparation method Methods 0.000 claims description 18
- 239000003814 drug Substances 0.000 claims description 15
- 229940079593 drug Drugs 0.000 claims description 13
- 229960004679 doxorubicin Drugs 0.000 claims description 8
- IFYDWYVPVAMGRO-UHFFFAOYSA-N n-[3-(dimethylamino)propyl]tetradecanamide Chemical compound CCCCCCCCCCCCCC(=O)NCCCN(C)C IFYDWYVPVAMGRO-UHFFFAOYSA-N 0.000 claims description 8
- JWDFQMWEFLOOED-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) 3-(pyridin-2-yldisulfanyl)propanoate Chemical compound O=C1CCC(=O)N1OC(=O)CCSSC1=CC=CC=N1 JWDFQMWEFLOOED-UHFFFAOYSA-N 0.000 claims description 6
- 239000003795 chemical substances by application Substances 0.000 claims description 6
- 238000001514 detection method Methods 0.000 claims description 6
- 230000003902 lesion Effects 0.000 claims description 6
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 4
- 230000009467 reduction Effects 0.000 claims description 4
- 125000003396 thiol group Chemical group [H]S* 0.000 claims description 4
- 210000004369 blood Anatomy 0.000 claims description 2
- 239000008280 blood Substances 0.000 claims description 2
- 239000012216 imaging agent Substances 0.000 claims description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 2
- 238000002386 leaching Methods 0.000 claims 1
- 210000004881 tumor cell Anatomy 0.000 abstract description 14
- 238000003384 imaging method Methods 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 8
- 208000007433 Lymphatic Metastasis Diseases 0.000 abstract description 7
- 230000005764 inhibitory process Effects 0.000 abstract 1
- 238000002626 targeted therapy Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 16
- 210000004027 cell Anatomy 0.000 description 15
- 108060001084 Luciferase Proteins 0.000 description 12
- 239000005089 Luciferase Substances 0.000 description 10
- 241000699666 Mus <mouse, genus> Species 0.000 description 7
- 210000001519 tissue Anatomy 0.000 description 7
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 210000001165 lymph node Anatomy 0.000 description 5
- 238000000108 ultra-filtration Methods 0.000 description 5
- 210000003462 vein Anatomy 0.000 description 5
- 238000002835 absorbance Methods 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 230000008685 targeting Effects 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 241000699660 Mus musculus Species 0.000 description 3
- 230000002378 acidificating effect Effects 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 235000013339 cereals Nutrition 0.000 description 3
- 210000004940 nucleus Anatomy 0.000 description 3
- 238000011580 nude mouse model Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 238000011729 BALB/c nude mouse Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000002512 chemotherapy Methods 0.000 description 2
- 230000009514 concussion Effects 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 230000001665 lethal effect Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000012460 protein solution Substances 0.000 description 2
- PRDFBSVERLRRMY-UHFFFAOYSA-N 2'-(4-ethoxyphenyl)-5-(4-methylpiperazin-1-yl)-2,5'-bibenzimidazole Chemical compound C1=CC(OCC)=CC=C1C1=NC2=CC=C(C=3NC4=CC(=CC=C4N=3)N3CCN(C)CC3)C=C2N1 PRDFBSVERLRRMY-UHFFFAOYSA-N 0.000 description 1
- 208000001894 Nasopharyngeal Neoplasms Diseases 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- HPZOOQSXPMEJBV-ODCFVKFUSA-N Tirilazad mesylate Chemical compound CS(O)(=O)=O.O=C([C@@H]1[C@@]2(C)CC=C3[C@@]4(C)C=CC(=O)C=C4CC[C@H]3[C@@H]2C[C@H]1C)CN(CC1)CCN1C(N=1)=CC(N2CCCC2)=NC=1N1CCCC1 HPZOOQSXPMEJBV-ODCFVKFUSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 230000000973 chemotherapeutic effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 229940127089 cytotoxic agent Drugs 0.000 description 1
- 239000002254 cytotoxic agent Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000036267 drug metabolism Effects 0.000 description 1
- 238000002296 dynamic light scattering Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000005170 neoplastic cell Anatomy 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000005740 tumor formation Effects 0.000 description 1
- 210000001364 upper extremity Anatomy 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 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
- A61K47/51—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
- A61K47/643—Albumins, e.g. HSA, BSA, ovalbumin or a Keyhole Limpet Hemocyanin [KHL]
-
- 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0032—Methine dyes, e.g. cyanine dyes
- A61K49/0034—Indocyanine green, i.e. ICG, cardiogreen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
- A61K49/0056—Peptides, proteins, polyamino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nanotechnology (AREA)
- Oncology (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Crystallography & Structural Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to a kind of human serum albumins-adriamycin cross-linking agent and its applications, the human serum albumins synthesized by small molecule crosslinking technological-adriamycin cross-linking agent HSA-DOX is capable of forming nano particle, the HSA-DOX nano particle is capable of the lymphatic metastasis of selective killing tumour cell, inhibition tumour, can be used for the targeted imaging and targeted therapy of tumour.Compared with uncrosslinked adriamycin, HSA-DOX of the invention can significantly increase oncotherapy effect, reduce gross tumor volume.
Description
Technical field
The invention belongs to field of medicaments and field of biotechnology, and in particular to a kind of nano anti-cancer drug, in particular to one
Kind human serum albumins-adriamycin cross-linking agent and its purposes for treating tumour.
Background technique
Tumour is still a kind of common disease and frequently-occurring disease for seriously threatening the mankind for health now, is to cause dead main original
One of because.Chemotherapy is to kill tumour cell using cytotoxic drug, inhibits the growth and breeding of tumour cell and promotes tumour thin
A kind of therapeutic modality of born of the same parents' differentiation, it is a kind of systemic treatment means.But in clinical treatment, chemotherapeutic treatment is killing
The colleague of tumour cell also kills normal cell and immunocyte together.Wherein, adriamycin is the most frequently used, most important medicine
One of object.
The adriamycin of clinical use is also Doxorubicin, is a kind of important chemotherapeutics for the treatment of tumour, but due to its production
Raw bio-toxicity limits its extensive use clinically.It, can be simultaneously in chemotherapy process since it does not have tumor-targeting
Tumor tissues and normal tissue are killed, adriamycin drug, which is used for a long time, can generate serious side effect, lead to heart, liver, big
The damage of brain and kidney.
Summary of the invention
Based on the above-mentioned problems in the prior art, the present invention is crosslinked adriamycin with human serum albumins, passes through people's blood
Pure albumen enhancing reduces the toxic side effect of adriamycin to the targeting of tumour.Present inventors have unexpectedly found that human serum albumins
The adriamycin of crosslinking is capable of the targeting breast cancer cell of selectivity, can be used in breast cancer treatment, and may be used also after label
It is detected for breast cancer living imaging.In addition, for therapeutic effect, the adriamycin of human serum albumins crosslinking is and same
The adriamycin (uncrosslinked) of active dose is compared, and oncotherapy effect is higher, and safety is also more preferable.
Specifically, on the one hand, the present invention provides human serum albumins-adriamycin cross-linking agent in preparation lesion detection agent
Purposes, the human serum albumins-adriamycin cross-linking agent is Doxorubicin molecules to be crosslinked white based on small molecule crosslinking technological
The nano particle formed on albumen.
Purposes of the human serum albumins of the present invention-adriamycin cross-linking agent in preparation lesion detection agent, wherein described
Human serum albumins-adriamycin is by label, and the lesion detection agent is living imaging agent.
Purposes of the present invention, wherein the tumour is breast cancer, the label is label.
Second aspect, the present invention provide use of the human serum albumins-adriamycin cross-linking agent in preparation tumor
On the way, the human serum albumins-adriamycin cross-linking agent is that Doxorubicin molecules are crosslinked in albumin based on small molecule crosslinking technological
The nano particle of upper formation.
Human serum albumins of the present invention-adriamycin cross-linking agent is preparing the purposes in tumor, wherein institute
Stating tumour is breast cancer.
Human serum albumins of the present invention-adriamycin cross-linking agent is preparing the purposes in tumor, wherein institute
The drug for stating treatment tumour is the drug of selective killing breast cancer cell.
Human serum albumins of the present invention-adriamycin cross-linking agent is preparing the purposes in tumor, wherein institute
The drug for stating treatment tumour is the drug for inhibiting breast cancer cell lymphatic metastasis.
The third aspect, the present invention provide a kind of breast cancer treatment drug, including human serum albumins-adriamycin cross-linking agent is received
Rice grain is to be crosslinked Doxorubicin molecules on human serum albumins by small molecule crosslinking technological to form nano particle, institute
Stating nano particle average grain diameter is about 25nm；The ratio of HSA and DOX is 1:3.
Fourth aspect, the present invention provide a kind of preparation method of human serum albumins-adriamycin cross-linking agent, comprising:
(1) albumin is reacted albumin of the preparation with pyridine dimercapto by albumin pre-activate with SPDP；
(2) reduction of preactivated albumin is prepared sulfhydryl activated albumin by the reduction of pre-activate albumin；
(3) it is crosslinked, ALDOX is slowly added to obtain human serum albumins-in the sulfhydryl activated albumin of step (2) preparation
Adriamycin cross-linking agent.
Human serum albumins of the present invention-adriamycin cross-linking agent preparation method, it is characterised in that the human seralbumin
Albumen-adriamycin cross-linking agent is nano particle, average grain diameter 25nm；Wherein, the ratio of HSA and DOX is 1:3.
Compared with prior art, technical solution of the present invention has the advantage that
(1) present invention finds the tumor-targetings of albumin, especially breast cancer cell targeting, thus by being crosslinked
Adriamycin specificity is transported to tumor tissues by technology, is enriched with since albumin crosslinking adriamycin drug is targeted in tumor locus
Afterwards, understand the slow release Doxorubicin molecules under the acidic micro-environment of tumor tissues part, be delivered in neoplastic cell nuclei, thus specifically
Property on tumour cell generate lethal effect without influencing other normal tissues.To realize the imaging and/or treatment of tumour.
(2) human serum albumins of the invention, adriamycin, marker (such as Cy5.5) ternary complex are in addition to that can treat
Other than tumour, additionally it is possible to it realizes the targeted imaging of tumour cell, specifically demonstrates the targeted imaging to breast cancer cell, described three
The image checking result of first compound is consistent with Luciferase imaging results, shows its reliability and accuracy.In addition, for
The diagnosis of breast cancer, ternary complex of the present invention can Accurate Diagnosis breast cancer cell whether lymphatic metastasis occurs.
(3) human serum albumins of the invention-adriamycin cross-linking agent, adriamycin (uncrosslinked) phase with same active dose
Than, oncotherapy effect is higher, and due to dosage is lower, have slow release effect thus safety it is more preferable.
(4) human serum albumins-adriamycin cross-linking agent that preparation method of the present invention obtains is nano particle, not only because of crosslinking
HSA and improve drug metabolism characteristics, and since specific crosslinking agent and cross-linking method make cross-linking products show pH
Response characteristic.The human serum albumins-adriamycin nano particle keeps stablizing under the conditions of the near-neutral pH of normal body fluid, and
The human serum albumins-adriamycin nano particle can slowly discharge activity under the low ph value acid condition of tumor microenvironment
DOX molecule, to further enhance HSA-DOX tumour-specific.
Detailed description of the invention
By reading the following detailed description of the preferred embodiment, various other advantages and benefits are common for this field
Technical staff will become clear.The drawings are only for the purpose of illustrating a preferred embodiment, and is not considered as to the present invention
Limitation.And throughout the drawings, the same reference numbers will be used to refer to the same parts.In the accompanying drawings:
Fig. 1: human serum albumins-adriamycin crosslinked nano-particles particle diameter distribution
Fig. 2: adriamycin absorbance and concentration standard curve
Fig. 3: HSA-DOX tumour cell positioning
Fig. 4: HSA-DOX selective killing breast cancer cell
Fig. 5: HSA-DOX in-vivo tumour targeting, 7h, for 24 hours, the living imaging figure of 48h, the left side is control group, and the right is
HSA-DOX experimental group
The lymphatic metastasis of Fig. 6: HSA-DOX target tumor
Fig. 7: mouse animal experiment Luciferase image
Fig. 8: transplantable tumor mouse tumor volume growth curve
Fig. 9: transplantable tumor mouse weight change curve
Specific embodiment
The illustrative embodiments of the disclosure are more fully described below with reference to accompanying drawings.Although showing this public affairs in attached drawing
The illustrative embodiments opened, it being understood, however, that may be realized in various forms the disclosure without the reality that should be illustrated here
The mode of applying is limited.It is to be able to thoroughly understand the disclosure on the contrary, providing these embodiments, and can be by this public affairs
The range opened is fully disclosed to those skilled in the art.
Embodiment according to the present invention proposes following embodiment
Albumin and adriamycin are crosslinked by embodiment 1 using protein micromolecular crosslinking technological
The preparation method of HSA-DOX
5mg albumin powder is weighed, the EDTA solution for the pH7.5 that 10mL concentration is 1mM is dissolved in, the 500mM of 8 μ L is added
SPDP solution.Room temperature concussion reaction 2h, reacts SPDP with the amino on albumin, forms the coupling of albumin and SPDP
Product.
Ultrafiltration is carried out with Millipore 10kD super filter tube after reaction, unreacted free SPDP molecule is removed, is used in combination
EDTA solution is washed three times.It is dissolved in the EDTA solution that 10mL concentration is 1mM pH7.5,6mg DTT is added, room temperature concussion is anti-
Answer 1h.
Ultrafiltration is carried out with Millipore 10kD super filter tube after reaction, removes unreacted free DTT molecule, and use EDTA
Solution is washed three times, reaction product is dissolved in the EDTA solution that 7mL concentration is 1mM pH7.5.Take the AlDOX of 15 μ L 100mM
(Aldox is bought from MedChemExpress company, CAS No.:1361644-26-9) solution, is added in the DMSO of 3mL and is mixed
It is even.3mL solution is slowly added in 7mL protein solution, and is shaken up during the addition process, shaken at room temperature reaction is overnight.After reaction
Ultrafiltration is carried out with Millipore 10kD super filter tube, removes unreacted free AlDOX molecule, is washed with deionized water and afterwards will three times
Product is placed in -20 DEG C of preservations.
The characterization of HSA-DOX
The HSA-DOX of preparation is characterized, HSA-DOX solution is placed in cuvette, is put into dynamic light scattering instrument
It is measured, as a result as shown in Figure 1, it was demonstrated that HSA-DOX is the nano particle (Fig. 1) that partial size is 25nm or so.
Adriamycin is diluted to different concentration, and its absorbance is measured by microplate reader, draws absorbance and concentration
Standard curve (Fig. 2).The absorbance for detecting HSA-DOX is 34923, and prepared HSA-DOX is calculated by regression curve
The concentration of DOX is 67 μM in solution；Meanwhile measuring protein concentration in HSA-DOX by A280 absorption is 1.9mg/mL, is calculated
Ratio to HSA and DOX is about 1:3.
HSA-DOX is placed in the PBS solution that pH is 7.5 and 5.5, after being placed at room temperature for 3h, by ultra-filtration and separation albumen and
Free DOX molecule.Protein solution after ultrafiltration is placed in measurement DOX fluorescence intensity in microplate reader, calculates protein concentration and DOX
Concentration obtains, and the ratio of HSA and DOX is still 1:3 under conditions of pH7.5, and under conditions of pH5.5 HSA and DOX ratio
Example is reduced to 1:0.5, it was demonstrated that HSA-DOX can degrade under acidic environment discharges DOX molecule.
Embodiment 2, albumin are coupled adriamycin drug selective killing tumour cell
HSA-DOX can enter tumour cell: HSA-DOX is added to the MDA-MB-231 cell culture medium of adhere-wall culture
In, and to be individually added into AlDOX as control, 4h is incubated in 37 DEG C of incubators, and carried out to nucleus with Hoechst33342
Dye 5min.It can be observed that the fluorescence (blue) of the fluorescence (red) of DOX and Hoechst generate altogether under Laser Scanning Confocal Microscope
It positions (Fig. 3).Similar to AlDOX control group result, HSA-DOX proves that HSA-DOX discharges in the acidic environment of tumour cell
DOX, so that DOX can enter nucleus.
HSA-DOX selective killing tumour cell: in tumour cell (MDA-MB-231) and normal cell (HEK293)
The HSA-DOX of various concentration is added, makes wherein 0.4,0.8,1.6 μM of DOX concentration.After 37 DEG C are incubated for for 24 hours, with MTT to thin
Cytoactive is measured (Fig. 4).The result shows that as HSA-DOX concentration increases, for the killing ability of MDA-MB-231 cell
It is significantly improved, and there is no apparent lethal effect for HEK293 cell, therefore can prove that HSA-DOX has selectivity
Effect of the killing tumor cell without influencing normal cell.
Embodiment 3, animal model compliance test result
1, the tumor-targeting of HSA-DOX in vivo:
In the subcutaneous injection 3x10 of Balb/c nude mice5It is transferred to the MDA-MD-231 cell of Luciferase luciferase.To
Gross tumor volume in 5-10mm3,100 μ L Cy5.5 of tail vein injection label HSA-DOX, for 24 hours after it is right in living imaging instrument
(Fig. 5) is imaged in Cy5.5 and Luciferase fluorescence.It can be seen that the fluorescence of Cy5.5 and Luciferase has significantly altogether
Positioning, it was demonstrated that HSA-DOX can be enriched in tumor tissues, have good tumor-targeting.
2, the lymphatic metastasis of HSA-DOX target tumor:
The MDA-MD-231 cell of Luciferase luciferase is transferred in the tail vein injection 5x105 of Balb/c nude mice.
Culture after two weeks tumour cell generate lymphatic metastasis, 100 μ L Cy5.5 of tail vein injection label HSA-DOX, for 24 hours after in work
It carries out Cy5.5 and Luciferase fluorescence to the thigh lymph node of nude mice in body imager to be imaged (Fig. 6), and by nude mice
Upper limb and lymph node of lower extremity, which take out, carries out fluorescence imaging.Wherein, Cy5.5 and Luciferase generates obvious common location, and solution is cut
Four lymph nodes in two have occurred MDA-MB-231 Nasopharyngeal neoplasms, that is, in the lymph node for having Luciferase fluorescence
There is the fluorescence of Cy5.5 simultaneously, and without obvious Cy5.5 fluorescence in the lymph node not shifted.Prove that HSA-DOX can not only target
To tumor tissues, moreover it is possible to the lymphatic metastasis of target tumor.
3, validity and safety of the HSA-DOX for oncotherapy:
4-6 weeks nude mice is chosen, 5x10 is subcutaneously injected5The luciferase containing Luciferase MDA-MB-231 cell.?
Tumor model tumor formation is stablized, and after gross tumor volume is 5-10mm3, is equally divided into three groups (control group, DOX group, HSA-DOX groups).It is right
HSA-DOX group every 100 μ L PBS solution of tail vein injection, Doxorubicin solution and HSA-DOX solution respectively are combined according to group, DOX,
So that DOX concentration is 2mg/kg.Tail vein is administered once every three days, carries out Luciferase imaging to tumour after two weeks
(Fig. 7).Imaging results illustrate that the fluorescence intensity for injecting the mouse tumor of HSA-DOX is lower, i.e., have significantly to the growth of tumour
Inhibiting effect.Meanwhile the volume of mouse tumor is measured over the course for the treatment of and is calculated (the long x wide 2/2 of tumour), also record
Mouse weight data, as a result as shown in Figure 8,9.The above results illustrate to be administered alone AlDOX it is unobvious to the therapeutic effect of tumour,
And HSA-DOX obviously can be such that gross tumor volume reduces.Also, during the administration, mouse weight has no significant change, it was demonstrated that its
With good safety.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
In the technical scope disclosed by the present invention, any changes or substitutions that can be easily thought of by anyone skilled in the art,
It should be covered by the protection scope of the present invention.Therefore, protection scope of the present invention should be with the protection model of the claim
Subject to enclosing.
Claims (10)
1. purposes of the human serum albumins-adriamycin cross-linking agent in preparation lesion detection agent, human serum albumins-Ah mould
Plain cross-linking agent is that Doxorubicin molecules are crosslinked to the nano particle formed on albumin based on small molecule crosslinking technological.
2. purposes of the human serum albumins as described in claim 1-adriamycin cross-linking agent in preparation lesion detection agent, feature
It is that the human serum albumins-adriamycin is by label, the lesion detection agent is living imaging agent.
3. purposes as claimed in claim 1 or 2, it is characterised in that the tumour is breast cancer, and the label is
Label.
4. purposes of the human serum albumins-adriamycin cross-linking agent in preparation tumor, the human serum albumins-Ah
Mycin cross-linking agent is that Doxorubicin molecules are crosslinked to the nano particle formed on albumin based on small molecule crosslinking technological.
5. purposes of the human serum albumins as claimed in claim 4-adriamycin cross-linking agent in preparation tumor, special
Sign is that the tumour is breast cancer.
6. purposes as described in claim 4 or 5, it is characterised in that the drug of the treatment tumour is selective killing breast cancer
The drug of cell.
7. purposes as described in claim 4 or 5, it is characterised in that the drug of the treatment tumour is to inhibit breast cancer cell leaching
Fawn on the drug of transfer.
8. a kind of breast cancer treatment drug, including human serum albumins-adriamycin cross-linking agent nano particle are by adriamycin point
Son is crosslinked on human serum albumins by small molecule crosslinking technological forms nano particle, and the nano particle average grain diameter is about
25nm；The ratio of HSA and DOX is 1:3.
9. a kind of human serum albumins-adriamycin cross-linking agent preparation method, comprising:
(1) albumin is reacted albumin of the preparation with pyridine dimercapto by albumin pre-activate with SPDP；
(2) reduction of preactivated albumin is prepared sulfhydryl activated albumin by the reduction of pre-activate albumin；
(3) it is crosslinked, ALDOX is slowly added to obtain human serum albumins-Ah mould in the sulfhydryl activated albumin of step (2) preparation
Plain cross-linking agent.
10. human serum albumins as claimed in claim 9-adriamycin cross-linking agent preparation method, it is characterised in that people's blood
Clear albumin-adriamycin cross-linking agent is nano particle, average grain diameter 25nm；Wherein, the ratio of HSA and DOX is 1:3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810575098.1A CN109157662B (en) | 2018-06-06 | 2018-06-06 | Human serum albumin-adriamycin cross-linked substance nano-particles and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810575098.1A CN109157662B (en) | 2018-06-06 | 2018-06-06 | Human serum albumin-adriamycin cross-linked substance nano-particles and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109157662A true CN109157662A (en) | 2019-01-08 |
| CN109157662B CN109157662B (en) | 2021-07-20 |
Family
ID=64897117
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810575098.1A Active CN109157662B (en) | 2018-06-06 | 2018-06-06 | Human serum albumin-adriamycin cross-linked substance nano-particles and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109157662B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114306356A (en) * | 2021-12-17 | 2022-04-12 | 兰州大学 | Folic acid/cis-aconitic anhydride bond adriamycin/human serum albumin (FA/CAD/HSA) composition and preparation method and application thereof |
| CN114569736A (en) * | 2021-12-17 | 2022-06-03 | 兰州大学 | Cis-aconitic anhydride bond adriamycin/human serum albumin (CAD/HSA) composition and preparation method and application thereof |
| CN114917238A (en) * | 2022-05-30 | 2022-08-19 | 郑州大学 | Preparation method and application of pancreatic cancer chemotherapy-immunotherapy drug delivery system |
| WO2023104171A1 (en) * | 2021-12-09 | 2023-06-15 | 深圳先进技术研究院 | Chimeric antigen receptor t cell secreting bispecific antibodies, preparation method therefor and use thereof |
| WO2023104137A1 (en) * | 2021-12-07 | 2023-06-15 | 深圳先进技术研究院 | Method for preparing tumor antigen-responsive living cellular nano backpacks and use thereof |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1737010A (en) * | 2004-08-19 | 2006-02-22 | 张阳德 | Galactose albumin preparation method |
| CN101267842A (en) * | 2005-09-20 | 2008-09-17 | 博洛尼亚大学病理学研究学院 | Use of conjugates of doxorubicin with lactosaminated albumin |
| US7445764B1 (en) * | 1999-06-10 | 2008-11-04 | Ktb Tumorforschungsgesellsschaft Mbh | Carrier-drug conjugate |
| CN101433723A (en) * | 2008-12-23 | 2009-05-20 | 重庆莱美药业股份有限公司 | PH sensitivity type anticancer prodrug as well as preparation method and application thereof |
| CN101569609A (en) * | 2009-02-24 | 2009-11-04 | 高其品 | Method for preparing adriamycin-albumin microsphere with targeting effect |
| CN102688499A (en) * | 2012-06-05 | 2012-09-26 | 中国科学院过程工程研究所 | Albumin-polyethylene glycol-medicinal molecular conjugate |
| CN103864921A (en) * | 2014-01-16 | 2014-06-18 | 苏州大学 | Folic acid-adriamycin immunological preparation for treating cancer through dual targets and preparation method thereof |
| CN104162172A (en) * | 2013-09-27 | 2014-11-26 | 深圳先进技术研究院 | Paclitaxel-containing polymer albumin nanosphere and preparation method and application thereof |
| CN105288639A (en) * | 2015-11-23 | 2016-02-03 | 中国药科大学 | Preparation and applications of Doxorubicin-loading active targeting albumin nanometer carrier |
-
2018
- 2018-06-06 CN CN201810575098.1A patent/CN109157662B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7445764B1 (en) * | 1999-06-10 | 2008-11-04 | Ktb Tumorforschungsgesellsschaft Mbh | Carrier-drug conjugate |
| CN1737010A (en) * | 2004-08-19 | 2006-02-22 | 张阳德 | Galactose albumin preparation method |
| CN101267842A (en) * | 2005-09-20 | 2008-09-17 | 博洛尼亚大学病理学研究学院 | Use of conjugates of doxorubicin with lactosaminated albumin |
| CN101433723A (en) * | 2008-12-23 | 2009-05-20 | 重庆莱美药业股份有限公司 | PH sensitivity type anticancer prodrug as well as preparation method and application thereof |
| CN101569609A (en) * | 2009-02-24 | 2009-11-04 | 高其品 | Method for preparing adriamycin-albumin microsphere with targeting effect |
| CN102688499A (en) * | 2012-06-05 | 2012-09-26 | 中国科学院过程工程研究所 | Albumin-polyethylene glycol-medicinal molecular conjugate |
| CN104162172A (en) * | 2013-09-27 | 2014-11-26 | 深圳先进技术研究院 | Paclitaxel-containing polymer albumin nanosphere and preparation method and application thereof |
| CN103864921A (en) * | 2014-01-16 | 2014-06-18 | 苏州大学 | Folic acid-adriamycin immunological preparation for treating cancer through dual targets and preparation method thereof |
| CN105288639A (en) * | 2015-11-23 | 2016-02-03 | 中国药科大学 | Preparation and applications of Doxorubicin-loading active targeting albumin nanometer carrier |
Non-Patent Citations (13)
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023104137A1 (en) * | 2021-12-07 | 2023-06-15 | 深圳先进技术研究院 | Method for preparing tumor antigen-responsive living cellular nano backpacks and use thereof |
| WO2023104171A1 (en) * | 2021-12-09 | 2023-06-15 | 深圳先进技术研究院 | Chimeric antigen receptor t cell secreting bispecific antibodies, preparation method therefor and use thereof |
| CN114306356A (en) * | 2021-12-17 | 2022-04-12 | 兰州大学 | Folic acid/cis-aconitic anhydride bond adriamycin/human serum albumin (FA/CAD/HSA) composition and preparation method and application thereof |
| CN114569736A (en) * | 2021-12-17 | 2022-06-03 | 兰州大学 | Cis-aconitic anhydride bond adriamycin/human serum albumin (CAD/HSA) composition and preparation method and application thereof |
| CN114917238A (en) * | 2022-05-30 | 2022-08-19 | 郑州大学 | Preparation method and application of pancreatic cancer chemotherapy-immunotherapy drug delivery system |
| CN114917238B (en) * | 2022-05-30 | 2024-02-02 | 郑州大学 | Preparation method and application of pancreatic cancer chemotherapy-immunotherapy drug delivery system |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109157662B (en) | 2021-07-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109157662A (en) | A kind of human serum albumins-adriamycin cross-linking agent nano particle and its application | |
| Song et al. | Ferritin: a multifunctional nanoplatform for biological detection, imaging diagnosis, and drug delivery | |
| Song et al. | Bioconjugated manganese dioxide nanoparticles enhance chemotherapy response by priming tumor-associated macrophages toward M1-like phenotype and attenuating tumor hypoxia | |
| Tian et al. | JQ1-loaded polydopamine nanoplatform inhibits c-MYC/programmed cell death ligand 1 to enhance photothermal therapy for triple-negative breast cancer | |
| Mu et al. | Anti-HER2/neu peptide-conjugated iron oxide nanoparticles for targeted delivery of paclitaxel to breast cancer cells | |
| Son et al. | Folate-modified PLGA nanoparticles for tumor-targeted delivery of pheophorbide a in vivo | |
| Yang et al. | NIR-activated self-sensitized polymeric micelles for enhanced cancer chemo-photothermal therapy | |
| Ma et al. | Indocyanine green-based theranostic nanoplatform for NIR fluorescence image-guided chemo/photothermal therapy of cervical cancer | |
| Jeon et al. | The intratumoral administration of ferucarbotran conjugated with doxorubicin improved therapeutic effect by magnetic hyperthermia combined with pharmacotherapy in a hepatocellular carcinoma model | |
| CN111888336B (en) | Calcium carbonate poly (lactic acid-glycolic acid) composite particles and preparation and application thereof | |
| Lu et al. | Liposomal IR-780 as a highly stable nanotheranostic agent for improved photothermal/photodynamic therapy of brain tumors by convection-enhanced delivery | |
| CN105307499B (en) | For tumor toxicity and the targeting corroles of MRI | |
| Wu et al. | Biocompatible AIEgen/p-glycoprotein siRNA@ reduction-sensitive paclitaxel polymeric prodrug nanoparticles for overcoming chemotherapy resistance in ovarian cancer | |
| Wu et al. | PDA-based drug delivery nanosystems: a potential approach for glioma treatment | |
| Liu et al. | Systemic Co-delivery of drugs by a pH-and photosensitive smart nanocarrier to treat cancer by chemo-photothermal-starvation combination therapy | |
| Albatany et al. | Brain tumor acidification using drugs simultaneously targeting multiple pH regulatory mechanisms | |
| Dai et al. | Erythrocyte membrane-camouflaged nanoparticles as effective and biocompatible platform: Either autologous or allogeneic erythrocyte-derived | |
| Zhang et al. | Arsenite-loaded albumin nanoparticles for targeted synergistic chemo-photothermal therapy of HCC | |
| Fang et al. | Therapeutic efficacy and cardioprotection of nucleolin-targeted doxorubicin-loaded ultrasound nanobubbles in treating triple-negative breast cancer | |
| Fan et al. | Biomimetic PLGA-based nanocomplexes for improved tumor penetration to enhance chemo-photodynamic therapy against metastasis of TNBC | |
| Kang et al. | Multifunctional theranostic nanoparticles for enhanced tumor targeted imaging and synergistic FUS/chemotherapy on murine 4T1 breast cancer cell | |
| Zeng et al. | Highly efficient Chemo/Photothermal therapy alleviating tumor hypoxia against cancer and attenuate liver metastasis in vivo | |
| Kourani et al. | Inulin coated Mn3O4 nanocuboids coupled with RNA interference reverse intestinal tumorigenesis in Apc knockout murine colon cancer models | |
| Dai et al. | Erythrocyte Membrane-Camouflaged Aggregation-Induced Emission Nanoparticles for Fetal Intestinal Maturation Assessment | |
| Liu et al. | An excipient-free “sugar-coated bullet” for the targeted treatment of orthotopic hepatocellular carcinoma |
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 | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20190426 Address after: 100871 No. 5, the Summer Palace Road, Beijing, Haidian District Applicant after: Peking University Address before: No. 46, Jiaojiang District, Taizhou, Zhejiang Province Applicant before: Zhejiang Hisun Pharmaceutical Co., Ltd. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |