CN110279868A - A kind of transporter or excretion body are preparing the application in targeted drug - Google Patents

A kind of transporter or excretion body are preparing the application in targeted drug Download PDF

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CN110279868A
CN110279868A CN201910594049.7A CN201910594049A CN110279868A CN 110279868 A CN110279868 A CN 110279868A CN 201910594049 A CN201910594049 A CN 201910594049A CN 110279868 A CN110279868 A CN 110279868A
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drug
excretion body
transporter
cell
tissue
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谢秋玲
熊盛
刘晨雪璇
王凯
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Jinan University
University of Jinan
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/42Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/46Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50Medicinal 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
    • A61K47/51Medicinal 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
    • A61K47/62Medicinal 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
    • A61K47/64Drug-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
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents

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Abstract

The present invention provides a kind of transporters or excretion body to prepare the application in targeted drug.The transporter can efficient targets neoplastic cells, be especially targeted α v β 3 and/or the highly expressed tumour cell of 5 integrin of α v β or tissue;And the excretion body containing the transporter also can efficiently target 3/5 integrin of α v β, the excretion body can contain small-molecule drug, the nucleic acid drug of oncotherapy as targeting vector, polypeptide drugs, pharmaceutical grade protein can also be carried by way of amalgamation and expression, kill to targeting the 3/5 highly expressed tumour of integrin of α v β, drug is significantly increased to the fragmentation effect of tumour cell, it is expected to be used for preparing the drug of targets neoplastic cells or tissue, have a good application prospect.

Description

A kind of transporter or excretion body are preparing the application in targeted drug
Technical field
The invention belongs to field of biotechnology, in particular to a kind of transporter or excretion body are in preparing targeted drug Using.
Background technique
Excretion body (Exosomes) is a kind of endogenous membrane vesicle that the cell that size is 30~150nm is secreted, with cell Biological function and intercellular signal transmitting have close relationship, in recent years excretion body as pharmaceutical carrier research increasingly Paid attention to.Because of its special structure, artificial synthesized pharmaceutical carrier is compared, excretion body carries out medicament transport as pharmaceutical carrier There is unique advantage, the membrane structure for being mainly reflected in excretion body is identical as cell membrane, the efficiency that drug enters cell can be improved, Blood-brain barrier can be even penetrated, while the reaction of adverse immune caused by excretion body is extremely low, there is infiltration to be well detained (EPR) effect It should be to slow release effect etc..
Although excretion body has many good qualities as pharmaceutical carrier, but lack targeting.On the other hand, it is with antibody drug The targeted drug of representative is a Main way of current drug research, however these drugs be all often with cell surface by Body combines, and needs to enter the drug having an effect into the cell and be difficult to targeting.
So if researching and developing a kind of excretion body with targeting, the drug acted on into the cell, targeting ground enhancing medicine are contained The delivering of object, not only can be improved drug availability, can also reduce drug side-effect.
Summary of the invention
The primary purpose of the present invention is that the shortcomings that overcoming the prior art and deficiency, it is swollen in preparation to provide a kind of transporter Application in the drug of tumor targeted drug or the highly expressed cell or tissue of targeted integration element.
Another object of the present invention is to provide a kind of excretion bodies to prepare tumor-targeting drug or the high table of targeted integration element Application in the drug of the cell or tissue reached.
A further object of the present invention provides a kind of tumor-targeting drug or the highly expressed cell or tissue of targeted integration element Drug.
A further object of the present invention is to provide the tumor-targeting drug or the targeted integration highly expressed cell of element or The preparation method of the drug of tissue.
The purpose of the invention is achieved by the following technical solution:
A kind of application of transporter in the drug for preparing the highly expressed cell or tissue of targeted integration element, the fortune The transporter that albumen designs for applicant is carried, which is proved to that excretion body can be entered, and can pass through other albumen The mode of fusion recombinant expression is brought into excretion body.The transporter is in Chinese patent application CN201811023079.4 Middle record, amino acid sequence is as shown in the SEQ ID NO.1 of Chinese patent application CN201811023079.4.Further In research, applicants have unexpectedly found that the transporter can efficient targets neoplastic cells, be especially targeted α v β 3 and/or α v β 5 be whole Close the highly expressed tumour cell of element or tissue.
The transporter is preparing the application in tumor-targeting drug.
The integrin is preferably α v β 3 and/or α v β 5;Further preferably α v β 3.
The tumor-targeting drug includes small-molecule drug, nucleic acid drug, polypeptide drugs, pharmaceutical grade protein.
A kind of excretion body is in the drug for preparing tumor-targeting drug or the highly expressed cell or tissue of targeted integration element Contain the transporter using, the excretion body or the transporter carry small-molecule drug, nucleic acid drug, At least one drug of polypeptide drugs or pharmaceutical grade protein.
The described carrying include excretion body contain the transporter and drug or the transporter and drug into Enter the excretion body after row amalgamation and expression.
The excretion body is preferably the excretion as claimed in claim 9 of Chinese patent application CN201811023079.4 Body.
The drug of a kind of tumor-targeting drug or the highly expressed cell or tissue of targeted integration element, contains the delivery egg It is white.
The drug of a kind of tumor-targeting drug or the highly expressed cell or tissue of targeted integration element is with the excretion body Carrier.
The preparation method of the drug of the tumor-targeting drug or the highly expressed cell or tissue of targeted integration element, packet It includes:
When the drug is small-molecule drug and/or nucleic acid drug, by the excretion body and the small molecule Drug is incubated for jointly;
When the drug is polypeptide drugs and/or pharmaceutical grade protein, by the transporter and the polypeptide Drug, pharmaceutical grade protein carry out amalgamation and expression.
The preparation method further comprises following steps: being centrifuged after incubation, precipitating is the tumour The drug of targeted drug or the highly expressed cell or tissue of targeted integration element.
The small-molecule drug includes adriamycin or its pharmaceutically acceptable salt.
The present invention has the following advantages and effects with respect to the prior art:
1. being expected to be used for preparing target present invention firstly discovers that transporter TP01 can efficiently target 3/5 integrin of α v β To tumour cell or the drug of tissue, the especially highly expressed tumour cell of 3/5 integrin of α v β or tissue.
2. the present invention further studies and confirms that the excretion body containing the transporter also can efficiently target α v β 3/ 5 integrins, the excretion body can contain chemical small molecule drug, the nucleic acid drug of oncotherapy as targeting vector, can also be with Polypeptide drugs, pharmaceutical grade protein are carried by way of amalgamation and expression, and it is highly expressed swollen to kill to targeting α v 3/5 integrin of β Tumor significantly increases drug to the fragmentation effect of tumour cell.
Detailed description of the invention
Fig. 1 is the PCR products electrophoresis map of transporter TP01, and swimming lane 1 is Marker, and the PCR that swimming lane 2~3 is TP01 is produced Object.
Fig. 2 is the PCR products electrophoresis map of fusion protein TP01-EGFP, wherein swimming lane 1 is Marker, and swimming lane 2~3 is attached most importance to Histone TP01-EGFP.
Fig. 3 is the fluorescent microscopy images figure (100 ×) for having transfected the cell of TP01-EGFP recombinant plasmid.
Fig. 4 is transmission electron microscope (TEM) photo figure of excretion body.
Fig. 5 is the NTA Analysis of test results figure of excretion body.
Fig. 6 is containing the Western blotting result figure after the cracking of TP01-EGFP excretion body, wherein swimming lane 1,2 is Contain TP01-EGFP excretion body;Swimming lane 3 is blank excretion body.
Fig. 7 is the Western blotting testing result figure of 3 expression quantity of α v β in different cells.
Fig. 8 is the transfection of the flow cytomery after EX-TP01-EGFP and EX-EGFP transfection A549 and Raji cell Result figure.
Fig. 9 is the Western of intracellular EGFP after EX-TP01-EGFP and EX-EGFP transfection A549 and Raji cell Blotting testing result figure.
Figure 10 is the canonical plotting of small-molecule drug DOX.
Figure 11 is cell toxicity test of the excretion body EX-TP01-EGFP and EX-EGFP to A549 cell for having contained DOX Result analysis chart.
Specific embodiment
Present invention will now be described in further detail with reference to the embodiments and the accompanying drawings, but embodiments of the present invention are unlimited In this.
The building of recombinant vector of the embodiment 1 containing transporter TP01
It constructs to obtain containing transporter TP01 referring to the method for Chinese patent application CN201811023079.4 embodiment 1 Recombinant vector.
Fig. 1 is the PCR product electrophoresis result of transporter TP01.
The preparation of 2 TP01-EGFP fusion protein excretion body of embodiment
TP01-EGFP fusion egg is prepared referring to the method for Chinese patent application CN201811023079.4 embodiment 2 It is ultrawhite to secrete body.
Fig. 2 is the PCR product electrophoresis result of fusion protein TP01-EGFP.
TP01-EGFP Transfected Recombinant Plasmid HEK 293F cell (invitrogen company) takes cell drop on glass slide, Under the microscope in fluorescence microscopy, as a result as shown in figure 3, most cells show green fluorescence (Fig. 3 B) under the conditions of fluorescence, Illustrate that the plasmid Successful transfection with EGFP enters cell, and expresses green fluorescent protein.Fig. 3 A is (i.e. visible under bright-field Under the conditions of light) observation result figure.
Referring to the side of 2 step 3 of Chinese patent application CN201811023079.4 embodiment " extraction and detection of excretion body " Method, step, by the form of transmission electron microscope observing excretion body (EX-TP01-EGFP), as a result as shown in Figure 4;Using NanoSight NS300 instrument (being purchased from Malvern) is measured by partial size of the NTA to excretion body with concentration, as a result such as Fig. 5 Shown, NTA detects particle diameter distribution between 60~180nm, and peak-peak belongs to the particle size range of excretion body in 120nm.
" inspection of recombinant protein is carried in excretion body referring to 2 step 4 of Chinese patent application CN201811023079.4 embodiment Method, the step of survey ", examine the albumen containing TP01-EGFP excretion body after cracking by Western blotting It surveys, two repetitions is set.As a result as shown in fig. 6, can detecte TP01 albumen and EGFP albumen in excretion body, simultaneously CD63, CD9 and CD81 are equally existed as excretion body Marker albumen, illustrate that TP01-EGFP fusion protein is present in excretion body In (EXO-TP01-1 of swimming lane 1 and the EXO-TP01-2 of swimming lane 2).
The detection of 3 difference tumour cell α v β of embodiment, 3 expression quantity
Quantitative analysis is carried out to the content of α v β 3 in various kinds of cell:
1. cell to be measured
HEK293T/F cell: people's renal epithelial cell (is purchased from Shanghai Jiang Lin Biotechnology Co., Ltd)
HepG2 cell: Hepatoblastoma (is purchased from Shanghai Yu Bo Biotechnology Co., Ltd)
A549 cell: human lung carcinoma cell (is purchased from Shanghai Jiang Lin Biotechnology Co., Ltd)
3T3 cell: mouse fibroblast (is purchased from Shanghai Jiang Lin Biotechnology Co., Ltd)
Raji cell: human lymphoma cell (is purchased from Shanghai Fu Sheng Industrial Co., Ltd.)
Daudi cell: people's Hematologic Malignancy Cell (grinds Biotechnology Co., Ltd purchased from Shanghai one)
2. specific detection method
Above several cells are cultivated respectively, and calculate cell quantity with Countstar cell counter, take 1 × 10 respectively5 A cell is cracked.By cell RIPA lysate (Beyotime;P0013B half an hour, cracking process) are cracked on ice Middle PMSF (the Beyotime that final concentration of 1mM is added;ST506), protein degradation is prevented.After half an hour, lysate is centrifuged, Take supernatant BCA kit (Thermo;23227) total protein concentration is detected.Loading total protein concentration is adjusted with PBS, so that phase With under loading volume, total protein concentration equivalent.Then the 5 of addition total volume 20% × non-reduced loading buffer (is purchased from Shanghai Yan Jin Biotechnology Co., Ltd, YJ0053A), sample 15min is heated after mixing under the conditions of 70 DEG C, is centrifuged after cooling, As sample.
Western Blot method utilizes 3 antibody (abcam of Anti- α v β with embodiment 2;Ab179475 it) carries out Western Blot verifying, it is A549 cell that this receptor, which expresses highest cell, as the result is shown.α on HepG2 cell and 3T3 cell 3 integrin protein receptor of v β also has expression, and the content of the α v β 3 of HEK293 cell is less.Raji cell and Daudi cell In 3 integrin receptor albumen (Fig. 7) of α v β is not detected.
Therefore, the A549 human lung carcinoma cell rich in α v β 3 is chosen as positive cell;Raji people of the selection without α v β 3 is drenched Bar oncocyte carries out follow-up study as negative cells.
Excretion body targeting transfection of the embodiment 4 containing TP01-EGFP is rich in 3 tumour cell of α v β
1. the acquisition of the excretion body without TP01
Hong Xun Biotechnology Co., Ltd in Suzhou is entrusted to synthesize EGFP gene, gene order is with embodiment 2, referring to embodiment EGFP gene segment is building up on expression vector pcDNA3.4 (purchased from Invitrogen company) by 2 method.
After Transfected Recombinant Plasmid HEK 293F cell (invitrogen company), excretion body EX-EGFP is obtained, excretion body It obtains and detection method is the same as embodiment 2.
2. the targeting transfection of excretion body is rich in 3 tumour cell of α v β
2 step of embodiment " extractions and detection of 3. excretion bodies " are extracted obtained excretion body EX-TP01-EGFP (to contain TP01-EGFP) and the present embodiment step 1 excretion body EX-EGFP (containing only EGFP) obtained without TP01 transfects sky respectively White A549 cell and Raji cell is observed and is detected with flow cytometer and WB method to be compared.
(1) flow cytomery: the cell of blank is added in 6 orifice plates, cultivates the excretion of 1% (v/v) of addition after 48h Body is removed culture solution, cell is washed three times with PBS, cell dissociation is got off using pancreatin, takes 200 μ L cells after cultivating 48h Liquid.It is set as control group with the cell of corresponding untransfected excretion body, for adjusting FSC voltage, SSC voltage and its respective streams Speed.The green channel fluorescent value of cellular control unit is detected, and Marker is set;The fluorescent value of test experience group cell and analysis Its result.
As shown in figure 8, transfection efficiency of the excretion body (EX-TP01-EGFP) in A549 cell containing TP01 is 16.83%, and the transfection efficiency of the excretion body (EX-EGFP) without TP01 only has 1.19%;And the Raji negative for α v β 3 Cell, two kinds of excretion bodies wink transfer efficient all close to 0.
WB detection method is the same as embodiment 2, Anti-EGFP antibody (Proteintech Group, USA simultaneously;66002-1- Ig the content of EGFP in recipient cell) is detected, the amount of EGFP is significantly larger than EX- in the A549 cell after EX-TP01-EGFP transfection EGFP transfects the EGFP amount after cell, and EGFP is not detected in Raji cell.Illustrate excretion body tool of the present invention containing TP01 There is the targeting for α v β 3.
Embodiment 5 targets excretion body EX-TP01-EGFP and contains small molecule anti-neoplastic drug doxorubicin DOX
By small molecule chemotherapeutic drug doxorubicin hydrochloride (DOX) (be purchased from Sigma) with various concentration (5mg/mL, 1mg/mL, 0.5mg/mL, 0.1mg/mL) it is dissolved in DMEM serum free medium (purchased from Sigma), it is measured using ultraviolet specrophotometer Light absorption value under its 480nm draws standard curve, as shown in Figure 10.
Drug crosses 0.22 μm of filter membrane after dilution, excretion the body EX-TP01-EGFP and EX- that will be obtained in embodiment 2 and 4 EGFP(107A/mL) respectively with drug (concentration 1mg/mL) by volume 1:1 in 37 DEG C of incubators jointly apply educate 2h; 10000g is centrifuged 70min, separates supernatant drug, is precipitated as excretion body and continues to employ, namely contain the excretion body EX-TP01- of drug EGFP and EX-EGFP.Supernatant measures the light absorption value under its 480nm with ultraviolet spectrometry degree meter, brings the bent formula y=of mark into 0.0029x+0.016 calculates drug concentration after carrying medicine, thus estimates excretion body drugloading rate roughly.It is found through detection, two kinds of excretions Drugloading rate in body is respectively 88ng/107A excretion body and 83ng/107A excretion body, the two are not significantly different.
Embodiment 6 has contained lethal effect of the targeting excretion body for tumour cell of drug
The excretion body for containing drug uses the DMEM culture medium containing 0.4% North America fetal calf serum (purchased from gibco) (to be purchased from Gibco it) dilutes.Take A549 cell according to 5 × 105Cell is simultaneously cultivated with the basis DMEM that 10mL contains 10% North America fetal calf serum Base dilution, is added 96 orifice plates, and every 100 μ L of hole reserves several holes as the culture medium blank control that cell is not added.37 DEG C of cultures It will be sucked out afterwards containing the culture medium of 10% North America fetal calf serum DMEM for 24 hours, the continuous plasma-free DMEM medium that is added of Cell relay is put Culture medium is sucked out after entering incubator culture 2h;By with containing 0.4% North America fetal calf serum dilution after load medicine excretion body according to institute DOX dose is needed to be added in cell, 37 DEG C of culture 48h.
By CCK-8 solution (be purchased from Pu Zhi Biotechnology Co., Ltd), it is added 96 orifice plates in the case of being protected from light, every 10 μ L of hole, 37 DEG C are protected from light culture 1h;Microplate reader detects 450nm OD value, calculates Cell viability according to following formula, formula is as follows:
Cell inhibitory rate (%)=100- (A dosing-A blank)/(A not dosing-A blank) × 100
Wherein:
A dosing: cell is added, carries medicine excretion body (carrying the amount of medicine excretion body according to used in DOX medicine calculation), CCK- 8 solution mix aperture light absorption values;
A blank: cell-free medium and CCK-8 solution mix aperture light absorption value is added;
A not dosing: cell, CCK-8 solution, no medicine feeding hole light absorption value is added.
As a result such as Figure 11, while the corresponding IC50 value of abscissa concentration calculation according to corresponding to figure upper 50% inhibiting rate, Two kinds of load medicine excretion bodies are compared, its IC50 value (1.8ng/mL) of EX-TP01-EGFP is lower than EX-EGFP (2.6ng/mL), are said The tumor cytotoxicity effect of the excretion body EX-TP01-EGFP of bright targeting is better than EX-EGFP, can prove that the presence of TP01 makes Excretion body after carrying medicine has potential targeting effect.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention, It should be equivalent substitute mode, be included within the scope of the present invention.

Claims (10)

1. a kind of application of transporter in the drug for preparing the highly expressed cell or tissue of targeted integration element, feature exist In:
The SEQ ID NO.1 institute of the amino acid sequence of the transporter such as Chinese patent application CN201811023079.4 Show.
2. transporter according to claim 1 is in the drug for preparing the highly expressed cell or tissue of targeted integration element Using, it is characterised in that:
The integrin is α v β 3 and/or α v β 5.
3. transporter described in claim 1 is preparing the application in tumor-targeting drug.
4. transporter according to claim 3 is preparing the application in tumor-targeting drug, it is characterised in that:
The tumor-targeting drug includes small-molecule drug, nucleic acid drug, polypeptide drugs, pharmaceutical grade protein.
5. a kind of excretion body answering in the drug for preparing tumor-targeting drug or the highly expressed cell or tissue of targeted integration element With, it is characterised in that:
The excretion body contains transporter described in claim 1 or transporter described in claim 1 carry it is small At least one of molecular drug, nucleic acid drug, polypeptide drugs or pharmaceutical grade protein.
6. excretion body according to claim 5 is preparing tumor-targeting drug or the highly expressed cell of targeted integration element or group The application in drug knitted, it is characterised in that:
The integrin is α v β 3 and/or α v β 5;
The carrying includes that the excretion body contains the transporter and drug or the transporter and drug Enter the excretion body after carrying out amalgamation and expression;
The excretion body is the excretion body as claimed in claim 9 of Chinese patent application CN201811023079.4.
7. the drug of a kind of tumor-targeting drug or the highly expressed cell or tissue of targeted integration element, it is characterised in that:
Contain transporter described in claim 1.
8. the drug of a kind of tumor-targeting drug or the highly expressed cell or tissue of targeted integration element, it is characterised in that:
Using the described in any item excretion bodies of claim 5 or 6 as carrier.
9. the preparation side of the drug of tumor-targeting drug according to any one of claims 8 or the highly expressed cell or tissue of targeted integration element Method, which comprises the steps of:
When the drug is small-molecule drug and/or nucleic acid drug, by the excretion body and the small-molecule drug It is common to be incubated for;
When the drug is polypeptide drugs and/or pharmaceutical grade protein, by the transporter and the polypeptide drugs, Pharmaceutical grade protein carries out amalgamation and expression.
10. the drug of tumor-targeting drug according to claim 9 or the highly expressed cell or tissue of targeted integration element Preparation method, it is characterised in that:
The preparation method further comprises following steps: being centrifuged after incubation, precipitating is the cancer target The drug of drug or the highly expressed cell or tissue of targeted integration element;
The small-molecule drug includes adriamycin or its pharmaceutically acceptable salt.
CN201910594049.7A 2019-07-03 2019-07-03 A kind of transporter or excretion body are preparing the application in targeted drug Pending CN110279868A (en)

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CN111424017A (en) * 2020-03-27 2020-07-17 暨南大学 Exosome loading shRNA (short hairpin ribonucleic acid) and construction method and application thereof
CN111840513A (en) * 2020-06-12 2020-10-30 广东工业大学 Composite exosome loaded with tumor apoptosis promoting protein and anticancer small molecules and preparation method and application thereof
CN114540307A (en) * 2022-02-16 2022-05-27 多莱泌生物科技(武汉)有限公司 Preparation method of liver-targeting engineered exosome

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111424017A (en) * 2020-03-27 2020-07-17 暨南大学 Exosome loading shRNA (short hairpin ribonucleic acid) and construction method and application thereof
CN111424017B (en) * 2020-03-27 2022-08-09 暨南大学 Exosome loading shRNA (short hairpin ribonucleic acid) and construction method and application thereof
CN111840513A (en) * 2020-06-12 2020-10-30 广东工业大学 Composite exosome loaded with tumor apoptosis promoting protein and anticancer small molecules and preparation method and application thereof
CN114540307A (en) * 2022-02-16 2022-05-27 多莱泌生物科技(武汉)有限公司 Preparation method of liver-targeting engineered exosome

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