CN109439665A - A kind of targeting combines the aptamer drug conjugates and application thereof of CD133 albumen - Google Patents
A kind of targeting combines the aptamer drug conjugates and application thereof of CD133 albumen Download PDFInfo
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- CN109439665A CN109439665A CN201811484364.6A CN201811484364A CN109439665A CN 109439665 A CN109439665 A CN 109439665A CN 201811484364 A CN201811484364 A CN 201811484364A CN 109439665 A CN109439665 A CN 109439665A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/115—Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
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- 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
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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
- A61K47/50—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
- A61K47/54—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 an organic compound
- A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
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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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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/16—Aptamers
Abstract
The present invention provides the aptamer drug conjugates and application thereof that a kind of targeting combines CD133 albumen, nucleic acid aptamer sequence such as SEQ ID NO.1, the present invention filters out the aptamers that targeting combines CD133 albumen using the method for cell screening from the library ssDNA, picks out aptamers AP-1-M of the invention.In one of scheme of the invention finally by physics inlay in the way of classical chemotherapeutic drugs Doxorubicin (DOX) connected with AP-1-M, aptamer-cell toxicant conjugate (AP-1-M-Dox) in conjunction with ATC can be targeted and probe into its cytotoxic effect by obtaining, preferable target is provided clinically to screen and studying novel targeted drug, new method and thinking are also provided for clinical treatment ATC.
Description
Technical field
The invention belongs to field of biological pharmacy, in particular to a kind of targeting combines aptamer-cell of CD133 albumen
Malicious conjugate.
Background technique
CD133 albumen is epicyte protein superfamily member, the sugar for being about 120kDa containing 865 amino acid, molecular weight
Albumen, molecular structure include: the extracellular end N-, 2 extracellular Loop rings, 5 transmembrane domains, 59 amino acid
The end cell C- and 8 N- glycosylate end.The prediction size of CD133 albumen be 97kDa, but it is actual glycosylation CD133 molecule
Amount is 120kDa.In recent years, researcher has found that CD133 albumen is present in the tumor cell surface of a variety of stem-like cells, wherein wrapping
Liver cancer, colon cancer and oophoroma are included, this provides preferable specific target spot clinically to screen and studying newtype drug, from
And to research and develop the thinking that more safely and effectively molecular targeted agents provide.In addition, also studies have reported that CD133 albumen is not
There is expression in differentiated thyroid carcinoma (Anaplastic thyroid carcinoma, ATC), this has prompted memebrane protein CD133
It is expected to targeted therapy of one of the combination target spot as aptamers for ATC.
Aptamer, also known as chemical antibody, has accurately targeting, by single-stranded ribonucleotide or deoxyribose
Nucleotide is constituted.Although most aptamers itself are without therapeutic effect, aptamer can be connected with drug toxicity
Composition targeting aptamers conjugate is connect, drug " accurate " can be transported to target cell, so that aptamers coupling drug is by target
Cell endocytic, portion discharges drug-induced apoptosis in the cell, has not only effectively improved the drug concentration of tumor by local, but also can be very big
Ground reduces the toxicity of other tissues, organ in vivo, to have the function that really to target Synergy and attenuation.In addition, aptamer
It also has the advantage that 1. in-vitro screenings, rapid artificial can synthesize；2. target type is extensive, protein or intact cell etc. are equal
It can be used as Screening target；3. molecular weight is little, it is not immunoreacted, is easily specifically bound with target molecule, affinity is strong, and solves
It can achieve pmol to nmol from constant；4. chemical stability is good, denaturation is reversible with renaturation, is easy to room temperature transport and long-term preservation
Deng.
Thyroid cancer can be divided into papillary carcinoma according to pathological, filter blocking cancer, Hurthle Cell cancer, cephaloma and not
Break up cancer, wherein undifferentiated type thyroid cancer (Anaplastic thyroid cancer, ATC) is a kind of high malignancy tumour,
Poor prognosis, it is median survival interval about 3-7 months, annual to deposit rate about 10%.Although ATC only accounts for the 2% of thyroid malignancy,
But due to the thyroid cancer patients substantial amounts in China, the absolute quantity of ATC patient is not low.In addition, because ATC causes extremely
The patient died accounts for about the 14%-39% of thyroid tumors total toll, is most fatal one of the malignant tumour of the mankind, and hand
The traditional treatments means such as art, radiotherapy, 131I are nearly unavailable to ATC.Therefore the therapeutic effect of such patient how is improved and improves,
The median survival interval for extending patient, is the difficult point and hot spot in ATC research all the time.
Currently, main direction of studying one of of the aptamer coupling drug as macromolecular targeted therapy malignant tumour,
It is increasingly becoming the hot spot of research in recent years.However the correlation of aptamer and its cell toxicant conjugate for thyroid cancer is ground
Study carefully less, especially there is not been reported for the research of relevant aptamer and cell toxicant conjugate with ATC.
Summary of the invention
The present invention utilizes RT-PCR, Western blot and laser confocal microscope (Confocal Microscopy) etc.
Method, which confirms CD133 albumen, to be had expression in undifferentiated type thyroid carcinoma cell and is expressed on cell membrane, and normal thyroid
Cell does not express CD133 albumen, this prompt memebrane protein CD133 can be used as one of combination target spot of ATC.
Subsequent inventor filters out targeting using the method for cell screening from the library ssDNA and combines the suitable of CD133 albumen
Ligand, and the ssDNA filtered out is detected by the method for high-flux sequence, eventually by the methods of flow cytometry pair
Aptamers AP-1-M aptamers i.e. of the invention are picked out in the analysis of the stability and compatibility of gained sequence.Then pass through
Immunofluorescence technique proves that aptamers AP-1-M can specifically be targeted in conjunction with CD133 albumen, and and expression CD133 albumen
ATC cell combination is to play the role of targeting conveying.
In one of scheme of the invention finally by physics inlay in the way of by classical chemotherapeutic drugs Doxorubicin
(DOX) it is connected with AP-1-M, aptamer-cell toxicant conjugate (AP-1-M-Dox) in conjunction with ATC can be targeted and visit by obtaining
Study carefully its cytotoxic effect, provides preferable target clinically to screen and studying novel targeted drug, be also clinical treatment
ATC provides new method and thinking.
Wherein one side according to the present invention provides a kind of aptamer for targeting and combining CD133 albumen, targeting knot
Close SEQ ID NO.1 in the nucleic acid aptamer sequence such as sequence table of CD133 albumen, sequence are as follows: 5 '-TACCAGCCGTTTCCC
CGGAGGGTCACCCCTGACGCATTCGGTTGAC-3’。
Wherein one side according to the present invention, provides a kind of application for targeting and combining the aptamer of CD133 albumen,
Application of the above-mentioned aptamers as the chemotherapy of tumors target medicine carrier of the CD133 positive.
It is even in conjunction with the aptamer drug of CD133 albumen to provide a kind of targeting for wherein one side according to the present invention
Join object, targeting combine CD133 albumen aptamer drug conjugates be the sequence such as aptamer of SEQ ID NO.1 and
The conjugate of cytotoxic drug.
It is even in conjunction with the aptamer drug of CD133 albumen to provide a kind of targeting for wherein one side according to the present invention
The coupling mode of connection object, above-mentioned aptamer and cytotoxic drug is by being rich in GC in the connection of the end of aptamer 5 '
Base-pair is to cytotoxic drug.
It is even in conjunction with the aptamer drug of CD133 albumen to provide a kind of targeting for wherein one side according to the present invention
Join object, the end of conjugate aptamer 5 ' connects 8 pairs of GC base sequences.
It is even in conjunction with the aptamer drug of CD133 albumen to provide a kind of targeting for wherein one side according to the present invention
Join object, wherein cytotoxic drug is adriamycin.
It is even in conjunction with the aptamer drug of CD133 albumen to provide a kind of targeting for wherein one side according to the present invention
Join the purposes of object, any one above-mentioned targeting combines the aptamer drug conjugates of CD133 albumen in preparation for controlling
Treat the application in the antineoplastic agents of the CD133 positive.
Wherein one side according to the present invention, provides a kind of tumor drug formulation of targeted therapy CD133 positive, the system
Agent contains any one of the above targeting and combines acceptable carrier on the aptamer drug conjugates and pharmacy of CD133 albumen
Or excipient.
It is even in conjunction with the aptamer drug of CD133 albumen to provide a kind of targeting for wherein one side according to the present invention
Join the preparation method of object, GC base sequence long in 5 ' end connection the preceding paragraphs of aptamer first, then again by above-mentioned core
Sour aptamers and cytotoxic drug are incubated for, and molar concentration rate 1:10 is best concentration ratio, and obtains aptamers with this
Drug conjugates (drug couples body) --- AP-1-M-Dox.
The aptamers that targeting combines CD133 albumen are filtered out from the library ssDNA using the method for cell screening, and to sieve
The ssDNA selected is detected by the method for high-flux sequence, eventually by the methods of flow cytometry to gained sequence
The analysis of stability and compatibility, picks out aptamers.Then prove that aptamers can be specifically by immunofluorescence technique
Targeting combines CD133 albumen, and and expresses the ATC cell combination of CD133 albumen to play the role of targeting conveying.This is
The targeted therapy of ATC provides new idea and method.
Present invention targeting combines the aptamer of CD133 albumen to also have the advantage that 1. be in-vitro screening, can be quick
It is artificial synthesized；2. molecular weight is little, it is not immunoreacted, is easily specifically bound with target molecule, affinity is strong, and dissociation constant
Pmol be can achieve to nmol；3. chemical stability is good, denaturation is reversible with renaturation, is easy to room temperature transport and long-term preservation etc..
One of purpose of the invention is to provide a kind of aptamer-cell that CD133 albumen is combined by targeting
Malicious conjugate AP-1-M-Dox.DNA sequence dna rich in GC base-pair can be polymerize with itself, bag structure be formed, thus object
Reason inlays adriamycin.
Based on this, AP-1-M-Dox provided by the invention is to connect upper 8 pairs of GC base sequences by the 5 ' ends in AP-1-M,
AP-1-M can be carried directly cytotoxic drug (such as adriamycin), the ATC cell of the selective killing CD133 positive.Therefore,
Another aspect of the invention provides AP-1-M-Dox of the invention and is preparing the tumour (such as ATC) for treating the CD133 positive
Application in preparation.
One of beneficial effect of the invention is: CD133 aptamer of the invention includes (1) can selectivity
The combination CD133 positive tumour cell, and it is weaker with the cell combination of CD133 feminine gender；(2) it preferably can directly carry thin
Cytotoxic drugs adriamycin, selective combination and the tumour cell for killing the CD133 positive.
In the present invention wherein embodiment, aptamer AP-1-M is coupled with classical chemotherapeutic drugs Doxorubicin, is obtained
AP-1-M-Dox simultaneously confirms it and truly has preferable cytotoxic effect, and due to the aptamer AP-1-M property of can choose
In conjunction with the tumour cell of the CD133 positive, and it is weaker with the cell combination of CD133 feminine gender, therefore another aspect of the present invention, it provides
Aptamers as the CD133 positive chemotherapy of tumors target medicine carrier application, chemotherapeutics is incorporated into aptamer
On, when carrying out tumor chemoradiotherapy, the tumour cell of the CD133 positive is killed to the property of can choose, and there is no toxicity to normal cell,
Above-mentioned chemotherapeutics (cytotoxic drug), can be selected from metabolism class drug for example methotrexate (MTX), fluorouracil, floxuridine,
Gemcitabine, Raltitrexed, antitumor antibiotic class drug such as mitomycin C, bleomycin, Doxorubicin, epirubicin, pyrrole are soft
Than star, plant bases such as vincaleukoblastinum, taxol, hydroxycamptothecin, antitumor steroids such as tamoxifen, Letrozole, prednisone,
Miscellany such as cis-platinum, carboplatin, mitoxantrone, anti-tumor small molecular targeted drug such as Gefitinib, Imatinib or Lapatinib, more
Preferably, it is mould to be selected from fluorouracil, floxuridine, methotrexate (MTX), cis-platinum, gemcitabine, Raltitrexed, mitogen for above-mentioned chemotherapeutics
Element, bleomycin, vincaleukoblastinum, taxol, hydroxycamptothecin, carboplatin, tamoxifen, Letrozole, prednisone, Gefitinib, she
Imatinib or Lapatinib.
Detailed description of the invention
Fig. 1 is the present invention wherein expression of CD133 albumen and positioning scenarios figure in different cell strains in an embodiment；
Fig. 2 is the specific detection figure of a wherein embodiment aptamers AP-1-M of the invention；
Fig. 3 is the combination situation detection of wherein embodiment aptamers AP-1-M and FRO a cell of the invention.
Fig. 4 is the coupling synthesis mode schematic diagram of wherein embodiment aptamers an AP-1-M and Dox of the invention；
Fig. 5 is the specific detection figure of a wherein embodiment aptamers AP-1-M of the invention.
Specific embodiment
The present invention will be further described below with reference to the drawings.
Embodiment 1: the expression of CD133 albumen and positioning scenarios in different cell strains
As shown in Figure 1, Caco-2: colon adenocarcinoma cell；FRO: undifferentiated type thyroid carcinoma cell；NTH: normal thyroid
Cell；293T: human embryonic kidney cell line.
The expression of CD133 albumen in different cell strains is observed using Laser Scanning Confocal Microscope (Confocal Microscopy)
Situation (Fig. 1), red fluorescence (i.e. the bright spot of membranaceous distribution in microphoto is opened in the lower left corner four in Fig. 1) represent memebrane protein CD133
The expression and distribution of albumen, blue-fluorescence (i.e. each microphoto center shape speck of Fig. 1) represent DAPI, indicator cells core.As a result it shows
Show, undifferentiated type thyroid cancer FRO cell and as CD133 albumen in the colon adenocarcinoma cell Caco-2 cell of positive controls
It is expressed on cell membrane, and the HEK-293T cell and normal thyroid cell Nthy-ori3-1 (NTH) as negative control are no
Express CD133 albumen.
Embodiment 2: aptamers AP-1-M is in conjunction with CD133 positive cell and negative cells.
As shown in Fig. 2, by marking AP-1-M aptamer using FITC, to be overexpressed the HEK- of CD133 albumen
293T cell is negative cells as the HEK-293T cell of positive cell, wild type, fits AP-1-M nucleic acid under 4 DEG C of environment
Ligand and positive cell and negative cells are incubated for, then observation discovery AP-1-M and positive cell under laser confocal microscope
In conjunction with and be positioned on cell membrane (Fig. 2-A), and (Fig. 2-B) is not combined with negative cells, this also illustrates AP-1-M is to be directed to
The targeting aptamer of CD133 albumen.
As a result it indicates, AP-1-M is the aptamer for specifically binding CD133 albumen, and surveys nucleic acid with flow cytometry
The affine linearity curve of aptamers show that the Kd value of AP-1-M is 101.4nM.
Embodiment 3: the combination of aptamers CD133 and undifferentiated type thyroid carcinoma cell FRO.
As shown in figure 3, AP-1-M and FRO cell is incubated for 30 minutes in 4 DEG C and 37 DEG C of environment respectively, find at 4 DEG C
When under environment, since the endocytosis of cell is suppressed, AP-1-M and FRO cell combination is simultaneously located on cell membrane (Fig. 3-A)；And
Due to the endocytosis of cell under 37 DEG C of environment, AP-1-M is swallowed (Fig. 3-B) intracellular by FRO into the cell.
As a result illustrate AP-1-M can with FRO cell combination and be located on cell membrane and can by it is interior swallow it is intracellular to reaching
Target the purpose of conveying.
Embodiment 4: the method that aptamers AP-1-M couples drug
As shown in figure 4, present invention research is preferably rich in the DNA sequence dna of GC base-pair, due to being rich in the DNA sequence of GC base-pair
Column can be polymerize with itself, form bag structure, so that physics is inlayed containing the drug for having Megumi ring, such as adriamycin, flesh-coloured mycin
Deng having the fluorescence spectrum of itself, and the characteristic being quenched in GC base-pair using adriamycin, detection and calculate Aptamer's
Drugloading rate.
Upper 8 pairs of GC base sequences are connected at the 5 ' ends of AP-1-M first, then again match AP-1-M and Dox with various concentration
Than being incubated for, after being incubated for 2 hours, compare absorbance height at 590nM using spectrophotometer, it is final to determine that 1:10 is most
Good concentration proportioning, and aptamers drug is obtained with this and couples body --- AP-1-M-Dox.
Based on this, AP-1-M-Dox of the invention can prepare tumour (such as ATC) preparation for treating the CD133 positive.
Embodiment 5:AP-1-M-Dox makees the drug toxicity of normal thyroid cell and undifferentiated type thyroid carcinoma cell
With
The specific detection figure (A: normal thyroid cell Nthy-ori3-1 of aptamers AP-1-M as shown in Figure 5；B:
Undifferentiated type thyroid carcinoma cell: FRO)
Normal thyroid cell strain Nthy-ori3-1 (NTH) and undifferentiated type thyroid gland (ATC) cell strain FRO are taken respectively
In 96 orifice plate bed boards, 5000/hole (the RPMI-1640 culture solution 90ul containing 10% fetal calf serum)；NTH cell strain is control group
A group, FRO cell strain are experimental group B group, are divided into A1 group (AP-1-M-Dox processing group) and A2 according to the difference of cell processing mode
Group (Dox processing group), B1 group (AP-1-M-Dox processing group) and B2 group (Dox processing group)；Subsequently in each group difference hole
The adriamycin (Dox) or AP-1-M-Dox (hole 10uL/) of various concentration is added in cell, and totally 9 group, first group of cell are not made to locate
Reason, as negative control, 8 groups of concentration of remaininging successively increase 2 times, and minimum concentration is about 0.016umol/ml, maximum concentration 50umol/
mL；After being cultivated 3 hours, 3 hours in the saturation vapour carbon dioxide incubator of 37 DEG C of 5%CO2 later to all hole inner cells into
Row changes liquid processing (the RPMI-1640 culture solution 90ul containing 10% fetal calf serum)；Finally cultivated 48 hours in incubator, it is last every
Hole adds 10 μ L CCK, after cultivating 3h in incubator, measures 450nm OD value with microplate reader.
As a result illustrate that AP-1-M-Dox has good cytotoxic effect, but its toxic effect outline for ATC cell FRO
It is weaker than Dox, and it has weaker cytotoxic effect for the normal thyroid cell NTH for not expressing CD133, toxic effect
It is markedly less than Dox.
Sequence table
<110>Zhejiang Prov. Tumor Hospital
<120>a kind of targeting combines the aptamer drug conjugates and application thereof of CD133 albumen
<130> CN-CN-2018-1031-1
<141> 2018-12-06
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 46
<212> DNA
<213> Artificial Sequence
<400> 1
taccagccgt ttccccggag ggtcacccct gacgcattcg gttgac 46
Claims (9)
1. the aptamer that a kind of targeting combines CD133 albumen, which is characterized in that the targeting combines the core of CD133 albumen
SEQ ID NO.1 in sour aptamers sequence such as sequence table.
2. the application that a kind of targeting combines the aptamer of CD133 albumen, which is characterized in that aptamers described in claim 1
The application of chemotherapy of tumors target medicine carrier as the CD133 positive.
3. the aptamer drug conjugates that a kind of targeting combines CD133 albumen, which is characterized in that the targeting combines
The aptamer drug conjugates of CD133 albumen are the aptamer and cytotoxicity of the sequence such as SEQ ID NO.1
The conjugate of drug.
4. a kind of targeting according to claim 3 combines the aptamer drug conjugates of CD133 albumen, feature exists
In the coupling mode of the aptamer and cytotoxic drug is by being rich in the connection of the aptamer 5 ' end
GC base-pair is to cytotoxic drug.
5. a kind of targeting according to claim 3 combines the aptamer drug conjugates of CD133 albumen, feature exists
In the end of conjugate aptamer 5 ' connects 8 pairs of GC base sequences.
6. a kind of targeting according to claim 3 combines the aptamer drug conjugates of CD133 albumen, feature exists
In the cytotoxic drug is adriamycin.
7. the purposes that a kind of targeting combines the aptamer drug conjugates of CD133 albumen, which is characterized in that claim 3
Combine the aptamer drug conjugates of CD133 albumen in preparation for treating CD133 sun to any one targeting described in 6
Application in the antineoplastic agents of property.
8. a kind of tumor drug formulation of the targeted therapy CD133 positive, which is characterized in that the preparation contains claim 3 to 6
Targeting described in any one combines acceptable carrier or tax on the aptamer drug conjugates and pharmacy of CD133 albumen
Shape agent.
9. the preparation method that a kind of targeting combines the aptamer drug conjugates of CD133 albumen, which is characterized in that exist first
The long GC base sequence of 5 ' end connection the preceding paragraphs of aptamer, then again by the aptamer and cytotoxic drug
It is incubated for, 1:10 concentration proportioning, and aptamers drug conjugates i.e. AP-1-M-Dox is obtained with this.
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| CN114574495A (en) * | 2020-12-01 | 2022-06-03 | 上海交通大学医学院附属仁济医院 | Aptamer R50 modified by nucleoside derivative |
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| CN111721932A (en) * | 2019-03-20 | 2020-09-29 | 复旦大学 | Screening method of small molecule compound taking CD133 as target spot and application of small molecule compound in pharmacy |
| WO2022000805A1 (en) * | 2020-06-29 | 2022-01-06 | 浙江大学 | Nucleic acid aptamer targeting cell surface antigen mfi2 protein, and use thereof |
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