CN108671237A - It is a kind of to enhance the carrier and its preparation method and application that drug targeting delivers - Google Patents
It is a kind of to enhance the carrier and its preparation method and application that drug targeting delivers Download PDFInfo
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- CN108671237A CN108671237A CN201810552574.8A CN201810552574A CN108671237A CN 108671237 A CN108671237 A CN 108671237A CN 201810552574 A CN201810552574 A CN 201810552574A CN 108671237 A CN108671237 A CN 108671237A
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- 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
- A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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Abstract
The present invention relates to a kind of carrier of enhancing drug targeting delivering and its preparation method and applications, the carrier for enhancing drug targeting delivering is the leucocyte for being derived from the homologous allosome through pre- immunity inoculation, and the preparation method of the carrier of the enhancing drug targeting delivering includes the following steps：(1) pre- immune formulation is prepared, pre- immunity inoculation is carried out 1~3 time to homologous allosome；(2) leucocyte in the homologous allosome body of separation and Extraction；(3) target cell in leucocyte is detached, the carrier of drug targeting delivering is as enhanced.Compared with prior art, the present invention provides a kind of completely new thinking for enhancing cancer target, a kind of potential cell carrier platform not only can be provided to solve cancer target bottleneck, moreover it is possible to potential application of the further exploration immunocyte platform on this basis in antineoplastic target treatment, targeting mechanism etc..
Description
Technical field
The invention belongs to biotechnology, the carrier more particularly, to a kind of enhancing drug targeting delivering and its acquisition side
Method and application.
Background technology
Conventional nano drug has met with challenge in the targeting feasibility of human body.The effect of Nano medication system only office at present
It is limited to improve the stability of drug, reduce toxic side effect, extend circulation time etc., targeting is little.Clinical research is aobvious
ShowNano medication is enriched with advantage compared with naked adriamycin drug without apparent tumour.Taxanes Nano medication in 2016
NK105 and BIND-014, CRLX-101 [6] clinical three phases are announced to fail, and being sent to great expectations becomes the first targeted nano in the whole world
Drug Bind Therapeutics company's the same years also announcement of bankruptcy.Targeting efficiency lowly has been considered as numerous Nano medications
The major reason of clinic conversion failure.It is more and more statistics indicate that, even if Nano medication system surfaces connect it is upper can be actively
It identifies the targeting ligand of tumor tissues or cell, such as antibody, polypeptide, folic acid, can not also change drug in tumor tissues
Abundance.Even it is reported that, the introducing of targeting ligand may cause reaction instead, for example cause nano-particles size
Become larger, to be easier, by RES system acquisitions, or to change nanoparticle surface property so that nano-particle be not easy into
Enter tumor tissues, cell.
Enhance strategy about cancer target, at present in addition to improving tumor microenvironment, such as tumor vessel normalization and cell
Outside film engineering, living cells carrier (Live cell carrier) receives favor due to non-EPR dependences.Leucocyte has
Immune response, cell adherence, passes through the specific functions such as physiologic barrier arrival non-vascular region at cell interaction, in cell
It is considered as important new force in the application of carrier cancer targets.In the past few years, researchers are using immunocyte intrinsic
Tumour chemotaxis carries out target tumor delivering research as " Trojan Horse " drug delivery system, however, although theoretical
Upper leucocyte such as monocyte has accurate targeting ability (directional migration) to inflammation part, but Targeting Effect is poor in practical applications
Strong man anticipates.It is particularly important for oncotherapy how leucocyte tumor-targeting is enhanced.
Invention content
The purpose of the present invention is exactly to provide a kind of enhancing drug targeting delivering to solve above-mentioned targeting bottleneck problem
Carrier and its preparation method and application.
The purpose of the present invention can be achieved through the following technical solutions：
A kind of carrier of enhancing drug targeting delivering, is the leucocyte for being derived from the homologous allosome through pre- immunity inoculation.
The leucocyte includes combination one or more in neutrophil leucocyte, Monocytes/Macrophages or NK cells etc..
The preparation method of the carrier of the enhancing drug targeting delivering, includes the following steps：
(1) pre- immune formulation is prepared, pre- immunity inoculation is carried out 1~3 time to homologous allosome；
(2) leucocyte in the homologous allosome body of separation and Extraction；
(3) target cell in leucocyte is detached, the carrier of drug targeting delivering is as enhanced.
Due to the inclusive granulocyte of leucocyte, Monocytes/Macrophages or NK cells, belong to cell mixing, therefore according to need
It wants, needs the target cell of separation the inside, such as granulocyte or NK cells.
The pre- immune formulation is the nanosizing preparation containing immune agonist, or agonist function is immunized with sustained release
Other dosage forms.
The immune agonist, including liopopolysaccharides receptor stimulating agent or Toll-like receptoroid agonist, the Toll-like
Receptoroid agonist is selected from unmethylated cytidylic acid-guanylic acid (CpG) or resiquimod
(Resiquimod, R848) etc.) in one or two kinds of combination.
It is one week to be inoculated with gap periods.
The method of leucocyte in the homologous allosome body of step (2) described separation and Extraction is：It can be extracted from marrow, or can
After by abdominal cavity sterile stimulation a few hours, lavage, which is collected, obtains leucocyte.
Carrier of the carrier of the enhancing drug targeting delivering as therapeutic substance, the targeting for tumor locus are passed
It send, the therapeutic vector includes antitumor drug or tumour medicine nanometer medicament.
The antitumor drug, can be selected from Doxorubicin, daunorubicin, valrubicin, epirubicin, idarubicin,
Taxol, docetaxel, cis-platinum, carboplatin, oxaliplatin, camptothecine, vincristine, vincaleukoblastinum, 5 one fluorine urine crash smack one's lips (5 one FU),
Mitomycin, ring phosphorus phthalein amine, the cry of certain animals of first ammonia butterfly, rice support green onion is waken up, topotecan, capecitabine, deoxidation fluorine urine is general, Irinotecan, is replaced
Add fluorine, Chlorambucil, Belotecan, Ah Nagqu mile, tamoxifen, Gleevec, the general, Leuprorelin of fluorine urine, Flutamide, mile come
Bony acid, streptozotocin, vinorelbine, light Ji Mai, retinoic acid, mustargen, busulfan, prednisone, emerald green ketone, aspirin, salicylate,
The general life of brufen, tea, fenoprofen are named and pay attention to Mei Xin, bute, mustargen, dexamethasone, prednisolone, celecoxib, cut down
Examine former times, aulin, cortisone, corticosteroid, gemcitabine, cedrol or their arbitrary combination or spreading out for they in ground
At least one of biology or their Nano medication novel form.
After in the vector injection body for enhancing drug targeting delivering, there is stronger tumour concentration effect.
Compared with existing cancer target efficiency, tumour/liver, tumour/spleen and tumour/kidney enrichment ratio it is more conventional not into
2~3 times of the leucocyte of the pre- immune object acquisition of row.The present invention provides a kind of completely new thinking for enhancing cancer target, not only
A kind of potential cell carrier platform can be provided to solve cancer target bottleneck, moreover it is possible to further explore and exempt from this basis
Potential application of the epidemic disease cell platform in antineoplastic target treatment, targeting mechanism etc..
Description of the drawings
Fig. 1 is nanosizing CpG transmission electron microscopes picture and dispersion liquid pictorial diagram used in case study on implementation.
Fig. 2 is that living imaging compares picture in case：(A) nonimmune normal mouse obtains the tumour enrichment condition of granulocyte
(B) receive immune mouse and obtain granulocyte in tumor locus enrichment condition.
Specific implementation mode
A kind of carrier of enhancing drug targeting delivering, is the leucocyte for being derived from the homologous allosome through pre- immunity inoculation.
The leucocyte includes combination one or more in neutrophil leucocyte, Monocytes/Macrophages or NK cells etc..
The preparation method of the carrier of the enhancing drug targeting delivering, includes the following steps：
(1) pre- immune formulation is prepared, pre- immunity inoculation is carried out 1~3 time to homologous allosome；
(2) leucocyte in the homologous allosome body of separation and Extraction；
(3) target cell in leucocyte is detached, the carrier of drug targeting delivering is as enhanced.
The pre- immune formulation is the nanosizing preparation containing immune agonist, or agonist function is immunized with sustained release
Other dosage forms.
The immune agonist, including liopopolysaccharides receptor stimulating agent or Toll-like receptoroid agonist, the Toll-like
Receptoroid agonist is selected from unmethylated cytidylic acid-guanylic acid (CpG) or resiquimod
(Resiquimod, R848) etc.) in one or two kinds of combination.
It is one week to be inoculated with gap periods.
The method of leucocyte in the homologous allosome body of step (2) described separation and Extraction is：It can be extracted from marrow, or can
After by abdominal cavity sterile stimulation a few hours, lavage, which is collected, obtains leucocyte.
Carrier of the carrier of the enhancing drug targeting delivering as therapeutic substance, the targeting for tumor locus are passed
It send, the therapeutic vector includes antitumor drug or tumour medicine nanometer medicament.
The antitumor drug, can be selected from Doxorubicin, daunorubicin, valrubicin, epirubicin, idarubicin,
Taxol, docetaxel, cis-platinum, carboplatin, oxaliplatin, camptothecine, vincristine, vincaleukoblastinum, 5 one fluorine urine crash smack one's lips (5 one FU),
Mitomycin, ring phosphorus phthalein amine, the cry of certain animals of first ammonia butterfly, rice support green onion is waken up, topotecan, capecitabine, deoxidation fluorine urine is general, Irinotecan, is replaced
Add fluorine, Chlorambucil, Belotecan, Ah Nagqu mile, tamoxifen, Gleevec, the general, Leuprorelin of fluorine urine, Flutamide, mile come
Bony acid, streptozotocin, vinorelbine, light Ji Mai, retinoic acid, mustargen, busulfan, prednisone, emerald green ketone, aspirin, salicylate,
The general life of brufen, tea, fenoprofen are named and pay attention to Mei Xin, bute, mustargen, dexamethasone, prednisolone, celecoxib, cut down
Examine former times, aulin, cortisone, corticosteroid, gemcitabine, cedrol or their arbitrary combination or spreading out for they in ground
At least one of biology or their Nano medication novel form.
After in the vector injection body for enhancing drug targeting delivering, there is stronger tumour concentration effect.
By taking mouse as an example, Enhancement test murine interleukin cancer target new method is provided, this method includes following step
Suddenly：
(1) suitable pre- immune formulation is prepared, pre- immunity inoculation was carried out 1~3 time to 4~13 weeks between twenty and fifty phase mouse, is connect
Kind gap periods are one week, wherein pre- immune formulation is the nanosizing preparation containing immune agonist or has sustained release agonist work(
Can other dosage forms, agonist, including liopopolysaccharides, Toll-like receptoroid agonist (such as unmethylated cytidine
Acid-guanylic acid (CpG), resiquimod (Resiquimod, R848) etc.) one or two kinds of combination；
(2) after by step (1) mouse peritoneal by sterile stimulation a few hours, lavation, which is collected, obtains leucocyte.It is wherein sterile
Stimulant includes various sterile preparations, such as common a certain concentration thioglycolate.
(3) target cell (neutrophil leucocyte, Monocytes/Macrophages or NK cells etc.) in leucocyte is detached, injection is homologous
In allosome tumor-bearing mice body, there is stronger tumour concentration effect.
By taking mouse as an example, the mechanism for the targeting enhancing being related to：The reality used in cell carrier area researches
It is generally also to live in gnotobasis to test room mouse, and the immune system of experiment mice is without cause of disease stimulating exercise, immune system
It is difficult to reach maturity, the immunocyte chemotaxis and targeting detached in vivo from it is undesirable to be will be understood by.And to mouse
After carrying out immunostimulation, response of the immune system to inflammatory signals of mouse can be significantly improved, thus is substantially increased pair
The targeting ability of tumour.
The present invention is described in detail with specific embodiment below in conjunction with the accompanying drawings.
Embodiment
1, by the bable/c mouse subcutaneous injection nanosizings CpG of 7~8w, used nanosizing CpG transmission electron microscope pictures
And dispersion liquid pictorial diagram is as shown in Figure 1, injecting 1 time or 1 times a week, adding up after injecting 3 times.It is collected by peritoneal lavage method
Internal leucocyte, its neutrophil leucocyte of separation and Extraction.
2, fluorescent marker (Dir labels) is carried out to neutrophil leucocyte
3, the mouse of lotus 4T1 tumours, when 5~8mm of tumor size or so, injection 5 × 106~107A/200uL's or so is glimmering
The neutrophil leucocyte of signal.
4, neutrophil leucocyte is injected in homologous allosome tumor-bearing mice body, and interval different time carries out living imaging, and comparison is non-
The granulocyte that immune group mouse obtains.Targeting Effect refers to Fig. 2, and the tumour that the nonimmune normal mouses of Fig. 2 (A) obtain granulocyte is rich
Collect situation, Fig. 2 (B) receives immune mouse and obtains granulocyte in tumor locus enrichment condition.
The above description of the embodiments is intended to facilitate ordinary skill in the art to understand and use the invention.
Person skilled in the art obviously easily can make various modifications to these embodiments, and described herein general
Principle is applied in other embodiment without having to go through creative labor.Therefore, the present invention is not limited to the above embodiments, ability
Field technique personnel announcement according to the present invention, improvement and modification made without departing from the scope of the present invention all should be the present invention's
Within protection domain.
Claims (9)
1. a kind of carrier of enhancing drug targeting delivering, which is characterized in that it is to be derived from the homologous allosome through pre- immunity inoculation
Leucocyte.
2. a kind of carrier of enhancing drug targeting delivering according to claim 1, which is characterized in that the leucocyte includes
One or more combination in neutrophil leucocyte, Monocytes/Macrophages or NK cells.
3. the preparation method of the carrier of enhancing drug targeting delivering as claimed in claim 1 or 2, which is characterized in that including following
Step：
(1) pre- immune formulation is prepared, pre- immunity inoculation is carried out 1~3 time to homologous allosome；
(2) leucocyte in the homologous allosome body of separation and Extraction；
(3) target cell in leucocyte is detached, the carrier of drug targeting delivering is as enhanced.
4. the preparation method of the carrier of enhancing drug targeting delivering according to claim 3, which is characterized in that described pre- immune
Preparation is the nanosizing preparation containing immune agonist, or other dosage forms of agonist function are immunized with sustained release.
5. the preparation method of the carrier of enhancing drug targeting delivering according to claim 4, which is characterized in that described immune sharp
Dynamic agent, including liopopolysaccharides receptor stimulating agent or Toll-like receptoroid agonist, the Toll-like receptoroid agonist are selected from not
One or two kinds of combination in the cytidylic acid-guanylic acid or resiquimod that methylate.
6. the preparation method of the carrier of enhancing drug targeting delivering according to claim 3, which is characterized in that between inoculation every other week
Phase is one week.
7. the preparation method of the carrier of enhancing drug targeting delivering according to claim 3, which is characterized in that step (2) institute
The method for stating the leucocyte in the homologous allosome body of separation and Extraction is：Extracted from marrow, or by the sterile stimulation in abdominal cavity after, lavation
Method, which is collected, obtains leucocyte.
8. the application of the carrier of enhancing drug targeting delivering as described in claim 1, which is characterized in that the enhancing drug targeting
Carrier of the carrier of delivering as therapeutic substance, the therapeutic substance include antitumor drug or tumour medicine nanometer medicine
Agent.
9. the application of the carrier of enhancing drug targeting delivering according to claim 8, which is characterized in that the antineoplastic
Object, selected from Doxorubicin, daunorubicin, valrubicin, epirubicin, idarubicin, taxol, docetaxel, cis-platinum, card
Platinum, oxaliplatin, camptothecine, vincristine, vincaleukoblastinum, 5 one fluorine urine crash smack one's lips, mitomycin, ring phosphorus phthalein amine, the cry of certain animals of first ammonia butterfly, rice
It asks green onion to wake up, topotecan, capecitabine, deoxygenate general fluorine urine, Irinotecan, Tegafur, Chlorambucil, Belotecan, Ah that
Bent mile, tamoxifen, Gleevec, fluorine urine is general, Leuprorelin, Flutamide, mile come bony acid, streptozotocin, vinorelbine, light Ji Mai,
Retinoic acid, busulfan, prednisone, emerald green ketone, aspirin, salicylate, brufen, the general life of tea, fenoprofen, is named and pays attention at mustargen
Mei Xin, bute, mustargen, dexamethasone, prednisolone, celecoxib, valdecoxib, aulin, cortisone, cortex
Steroids, gemcitabine, cedrol or at least one of their arbitrary combination or their derivative or they receive
Rice pharmaceutical dosage form.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112138157A (en) * | 2020-09-29 | 2020-12-29 | 同济大学 | Optically controlled granulocyte biological agent and preparation and application thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112138157A (en) * | 2020-09-29 | 2020-12-29 | 同济大学 | Optically controlled granulocyte biological agent and preparation and application thereof |
| CN112138157B (en) * | 2020-09-29 | 2022-02-18 | 同济大学 | Optically controlled granulocyte biological agent and preparation and application thereof |
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