CN112011543A - Preparation method of triptolide aptamer conjugate - Google Patents
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- CN112011543A CN112011543A CN201910460762.2A CN201910460762A CN112011543A CN 112011543 A CN112011543 A CN 112011543A CN 201910460762 A CN201910460762 A CN 201910460762A CN 112011543 A CN112011543 A CN 112011543A
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Abstract
The invention relates to the field of organic chemistry, in particular to a preparation method of triptolide aptamer conjugate. The invention provides a preparation method of triptolide aptamer conjugate, which comprises the following steps: 1) reacting triptolide with p-nitrophenyl chloroformate to prepare p-nitrophenyl formate triptolide; 2) reacting the p-nitrophenyl formate triptolide with a nucleic acid aptamer to prepare the triptolide nucleic acid aptamer conjugate. The preparation method of the triptolide aptamer conjugate aims at the conjugation of triptolide and the aptamer, and the triptolide and p-nitrophenyl chloroformate are reacted and activated in one step and then efficiently and quickly react with the amino aptamer, so that the reaction time is greatly shortened, the coupling process is simplified, the utilization rate of the triptolide is improved, the overall yield can reach over 90 percent, and the triptolide aptamer conjugate has good industrialization prospect.
Description
Technical Field
The invention relates to the field of organic chemistry, in particular to a preparation method of triptolide aptamer conjugate.
Background
Triptolide is an epoxidized diterpene lactone natural product with various biological activities extracted and separated from tripterygium wilfordii which is a traditional Chinese medicine. Triptolide is an effective immunosuppressive and anti-inflammatory agent, which can effectively inhibit mitogen-induced lymphocyte proliferation and allogeneic mixed lymphocyte reaction; inhibit the expression of various cytokines and show good curative effect in treating leukemia and other cancers.
An aptamer is an antibody-like oligonucleotide sequence that specifically binds to a target protein. The aptamer can be chemically synthesized, is easier to modify and more stable than an antibody, has smaller size and organism immunoreaction, and therefore has wide application in diagnosis and treatment of tumors. The coupling of the drug molecules and the aptamer can improve the water solubility of the drug molecules and the enrichment of the drug molecules in tumor cells, reduce the toxic and side effects of the drug molecules on normal cells, and realize the targeted delivery of the drug molecules. Known aptamers such AS Sgc8c and AS1411 have been widely used in tumor diagnosis and treatment. Sgc8c is obtained by screening an exponential enrichment ligand system evolution technology (cell-SELEX) taking a cell as a target, and can be specifically combined with PTK7 protein on the surface of the cell to realize high-affinity recognition on the target cell, so that the target cell is internalized into the cell. AS1411 is a nucleolin-targeting aptamer capable of specific recognition of a variety of tumor cells. Therefore, the aptamer is coupled with the triptolide and the structure modifier thereof to improve the antitumor activity of the triptolide and the structure modifier thereof. CN 106589049 discloses an improved synthesis method of a conjugate of triptolide and aptamer, which forms a conjugate of triptolide and aptamer through a four-step reaction to obtain a 58% synthesis yield. The technical method needs four steps of reaction, and reduces the use efficiency of triptolide.
Disclosure of Invention
In view of the above-mentioned shortcomings of the prior art, the present invention aims to provide a method for preparing triptolide aptamer conjugate, which is used to solve the problems in the prior art.
In order to achieve the above and other related objects, the present invention provides a method for preparing a triptolide aptamer conjugate, comprising:
1) reacting triptolide with p-nitrophenyl chloroformate to prepare p-nitrophenyl formate triptolide;
2) reacting the p-nitrophenyl formate triptolide with a nucleic acid aptamer to prepare the triptolide nucleic acid aptamer conjugate.
In some embodiments of the present invention, in the step 1), the reaction is performed in the presence of an acid-binding agent.
In some embodiments of the invention, in the step 1), the molar ratio of triptolide to p-nitrophenyl chloroformate is 1: 1.2-10, preferably 1: 3 to 6.
In some embodiments of the invention, in step 1), the reaction is carried out in the presence of a reaction solvent.
In some embodiments of the present invention, in the step 1), the reaction temperature is 10 to 40 ℃, preferably 20 to 30 ℃.
In some embodiments of the invention, in step 1), the acid scavenger is selected from organic bases.
In some embodiments of the invention, in the step 1), the molar ratio of triptolide to the acid-binding agent is 1: 0.002 to 0.25, preferably 1: 0.05 to 0.08.
In some embodiments of the invention, in step 1), the reaction solvent is selected from haloalkane solvents.
In some embodiments of the present invention, the acid scavenger is selected from organic bases, preferably, the acid scavenger is selected from one or more of pyridine and DMAP in combination.
In some embodiments of the present invention, in step 1), the reaction solvent is selected from one or more of dichloromethane, chloroform, and 1, 2-dichloroethane.
In some embodiments of the present invention, the step 1) further includes: adding water for quenching, extracting by an organic solvent, and desolventizing an organic phase.
In some embodiments of the invention, in step 2), the reaction is carried out in the presence of a base.
In some embodiments of the invention, in the step 2), the molar ratio of the triptolide p-nitrophenylformate to the aptamer is 1: 0.001 to 0.5, preferably 1: 0.005-0.1.
In some embodiments of the invention, in step 2), the reaction is carried out in the presence of a reaction solvent.
In some embodiments of the present invention, in the step 2), the reaction temperature is 10 to 50 ℃, preferably 30 to 40 ℃.
In some embodiments of the invention, in the step 2), the aptamer comprises an amino group, and preferably, the 5' end of the aptamer comprises an amino group.
In some embodiments of the present invention, in the step 2), the reaction time is less than or equal to 4 hours, preferably 15min to 1 hour.
In some embodiments of the invention, in step 2), the reaction is carried out under protection from light.
In some embodiments of the invention, in step 2), the base is selected from organic bases.
In some embodiments of the present invention, in the step 2), the reaction solvent is selected from water and organic solvents.
In some embodiments of the invention, in step 2), the aptamer is selected from the group consisting of Sgc8c aptamer, AS1411 aptamer in combination with one or more of these.
In some embodiments of the invention, in step 2), the base is selected from one or more of triethylamine, N-diisopropylethylamine, or a combination thereof.
In some embodiments of the present invention, in the step 2), the organic solvent used as the reaction solvent is selected from aprotic polar solvents, and more preferably from one or more of N, N-dimethylformamide, dimethylsulfoxide and acetonitrile.
In some embodiments of the present invention, the step 2) further includes: adding water for quenching, extracting by an organic solvent, and desolventizing an organic phase.
The invention also provides the triptolide aptamer conjugate prepared by the preparation method.
Drawings
FIG. 1 shows the mass spectrum of the triptolide-AS 1411 aptamer conjugate in example 1 of the present invention.
FIG. 2 is a schematic diagram showing the inhibitory effect of triptolide and the triptolide aptamer conjugates in hepatoma carcinoma cells HepG2 according to example 4 of the present invention.
FIG. 3 is a schematic diagram showing the inhibitory effect of triptolide and the triptolide aptamer conjugate in breast cancer cells MCF-7 according to example 5 of the present invention.
FIG. 4 is a schematic diagram showing the inhibitory effect of triptolide and the triptolide aptamer conjugate in breast cancer cells MDA-MB-231 according to embodiment 6 of the invention.
Detailed Description
The inventor of the invention unexpectedly discovers a novel preparation method of the triptolide aptamer conjugate through a large amount of practical researches, and compared with the prior art, the preparation method greatly improves the reaction efficiency and the reaction yield, so that the process is simpler and more efficient, and the invention is completed on the basis.
The invention provides a preparation method of triptolide aptamer conjugate, which comprises the following steps:
1) reacting triptolide with p-nitrophenyl chloroformate to prepare p-nitrophenyl formate triptolide;
2) reacting the p-nitrophenyl formate triptolide with a nucleic acid aptamer to prepare the triptolide nucleic acid aptamer conjugate, wherein the nucleic acid aptamer comprises a terminal amino group.
The preparation method of the triptolide aptamer conjugate provided by the invention can comprise the following steps: reacting the p-nitrophenyl formate triptolide with a nucleic acid aptamer to prepare the triptolide nucleic acid aptamer conjugate, wherein the reaction equation is as follows:
wherein the amino group-containing band represents a nucleic acid aptamer.
In the preparation method provided by the invention, the aptamer generally comprises an amino group, and preferably, the 5' end of the aptamer comprises an amino group. The aptamer may be a polynucleotide strand modified with an amino group at the 5 ' end, and the method of modifying the amino group at the 5 ' end of the polynucleotide strand will be well known to those skilled in the art, and a polynucleotide strand modified with an amino group at the 5 ' end can also be purchased from a publicly available commercial source. For example, the nucleic acid fragment may be modified with a C6 phosphoramidite monomer to obtain a-NH-substituted 5' terminus2A modified nucleic acid fragment. In a preferred embodiment of the present invention, the C6 phosphoramidite monomer can be modified at the 5' end using a DNA solid phase synthesis method, which is described in detail with reference to Wurtz, n.r.; turner, j.m.; baird, e.e.; dervan, P.B., Fmoc solid phase synthesis of polyamides stabilizing and imidazole amino acids Org Lett 2001,3(8), 1201-:
the hydroxyl on the sugar ring at the 5' end of the nucleic acid fragment is crosslinked with a phosphoramidite monomer, and the reaction equation is as follows:
wherein the-O-linked curve part represents the remainder of the nucleic acid fragment and the filled circle represents the solid support CPG. Amino group modification is carried out on a DNA phosphate skeleton, and the protective group of the prepared product is removed, thus obtaining the 5' end-NH2The modified nucleic acid fragment is further connected with a connecting group through the amino group, so that the aptamer fragment shown in the structural formula as follows can be obtained:
in a specific embodiment of the present invention, the aptamer may be a combination including, but not limited to, one or more of Sgc8c aptamer, AS1411 aptamer, and the like. The dosage of the p-nitrophenyl formate triptolide is usually excessive relative to the amount of the aptamer, so that the aptamer can be fully reacted to improve the reaction efficiency. For example, the molar ratio of triptolide p-nitrophenylformate to aptamer may be 1: 0.001-0.1, 1: 0.001 to 0.005, 1: 0.005-0.01, 1: 0.01-0.05, 1: 0.05-0.1, 1: 0.1 to 0.3, or 1: 0.3 to 0.5.
In the preparation method provided by the invention, the reaction of the triptolide p-nitrophenylformate and the aptamer is usually carried out in the presence of alkali. The base may typically be an organic base, for example, a combination including, but not limited to, one or more of triethylamine, N-diisopropylethylamine, and the like. One skilled in the art can select an appropriate amount of base, for example, the molar ratio of triptolide p-nitrophenylformate to base can be 1: 10-500, 1: 10-20, 1: 20-50, 1: 50-100, 1: 100 to 200, or 1: 200 to 500.
In the preparation method provided by the invention, the reaction of the p-nitrophenyl formate triptolide and the aptamer is usually carried out in the presence of a reaction solvent. The reaction solvent is generally a good solvent for the reaction system, and may be, for example, water and/or an organic solvent, which may be generally an aprotic polar solvent, so that a good reaction system may be formed with water. For example, the organic solvent may be a combination including, but not limited to, one or more of N, N-dimethylformamide, dimethylsulfoxide, acetonitrile, and the like. The amount of solvent used should be selected by one skilled in the art, for example, 0.5-5 mL of water per 1mg of aptamer and 0.1-10 mL of organic solvent per 1mg of triptolide p-nitrophenylformate can be used.
In the preparation method provided by the invention, the reaction of the triptolide p-nitrophenylformate and the aptamer can be carried out at room temperature or under a heating condition, for example, the reaction temperature can be 10-50 ℃, 10-20 ℃, 20-30 ℃, 30-40 ℃, or 40-50 ℃. The reaction time can be adjusted by those skilled in the art according to the reaction progress, for example, the reaction progress can be judged by TLC, chromatography, etc., and for example, the reaction time of the step can be 4 hours or less, 15-30 min, 30-45 min, 45 min-1 h, 1-2 h, 2-4 h.
In the preparation method provided by the present invention, a person skilled in the art may select an appropriate method to perform post-treatment on the reaction product of the reaction between triptolide p-nitrophenylformate and the aptamer, for example, the method may further include: adding water for quenching, extracting by an organic solvent, and desolventizing an organic phase. The product obtained by organic phase desolventization may be further purified, for example, by column chromatography or the like.
The preparation method of the triptolide aptamer conjugate provided by the invention can also comprise the following steps: the triptolide is reacted with p-nitrophenyl chloroformate to prepare the p-nitrophenyl formate triptolide, and the reaction equation is as follows:
in the preparation method provided by the invention, the reaction of triptolide and p-nitrophenyl chloroformate is usually carried out in the presence of an acid-binding agent. The acid-binding agent can generally play a catalytic role in the reaction, accelerate the acylation reaction and generally has good stability in the reaction system. The acid scavenger may typically be an organic base, for example, may be a combination including, but not limited to, one or more of pyridine, DMAP, and the like. The acid-binding agent is generally used in a catalytic amount, for example, the molar ratio of triptolide to acid-binding agent may be 1: 0.002-0.01, 1: 0.01 to 0.05, 0.05 to 0.08, or 1: 0.08 to 0.25.
In the preparation method provided by the invention, the using amount of the p-nitrophenyl chloroformate is usually excessive relative to the triptolide, so that the full reaction of the triptolide serving as a substrate can be ensured. For example, the molar ratio of triptolide to p-nitrophenyl chloroformate may be 1: 1.2-10, 1: 1.2-1.5, 1: 1.5-3, 1: 3-6, 1: 6 to 10. During the reaction, triptolide and p-nitrophenyl chloroformate may be mixed, and an acid-binding agent may be added gradually to avoid a large amount of heat release in a short time, and a suitable temperature reduction means (e.g., ice bath, etc.) may be adopted when the acid-binding agent is added, which should be known to those skilled in the art.
In the preparation method provided by the invention, the reaction of triptolide and p-nitrophenyl chloroformate is usually carried out in the presence of a reaction solvent, wherein the reaction solvent is usually a good solvent of a reaction system. For example, in this step, the reaction solvent may be generally a haloalkane type solvent, and more specifically may be a combination including, but not limited to, one or more of dichloromethane, chloroform, 1, 2-dichloroethane, and the like. The amount of the solvent to be used should be selected by those skilled in the art, and for example, in this step, 5mL to 50mL of the reaction solvent can be used per 1g of the starting material.
In the preparation method provided by the invention, the reaction of triptolide and p-nitrophenyl chloroformate can be carried out at room temperature, for example, the reaction temperature can be 10-40 ℃, 10-20 ℃, 20-30 ℃ or 30-40 ℃. The reaction time can be adjusted by those skilled in the art according to the reaction progress, for example, the reaction progress can be judged by TLC, chromatography, etc., and the reaction time of the step can be 1-8 h, 1-2 h, 2-4 h, 4-6 h, or 6-8 h.
In the preparation method provided by the present invention, a person skilled in the art may select an appropriate method to perform post-treatment on the reaction product of the reaction between triptolide and p-nitrophenyl chloroformate, for example, the method may further include: adding water for quenching, extracting by an organic solvent, and desolventizing an organic phase. The product obtained by organic phase desolventization may be further purified, for example, by column chromatography or the like.
The preparation method of the triptolide aptamer conjugate aims at the conjugation of triptolide and the aptamer, and the triptolide and p-nitrophenyl chloroformate are reacted and activated in one step and then efficiently and quickly react with the amino aptamer, so that the reaction time is greatly shortened, the coupling process is simplified, the utilization rate of the triptolide is improved, the overall yield can reach over 90 percent, and the triptolide aptamer conjugate has good industrialization prospect.
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
It is to be understood that the processing equipment or apparatus not specifically identified in the following examples is conventional in the art.
Furthermore, it is to be understood that one or more method steps mentioned in the present invention does not exclude that other method steps may also be present before or after the combined steps or that other method steps may also be inserted between these explicitly mentioned steps, unless otherwise indicated; it is also to be understood that a combined connection between one or more devices/apparatus as referred to in the present application does not exclude that further devices/apparatus may be present before or after the combined device/apparatus or that further devices/apparatus may be interposed between two devices/apparatus explicitly referred to, unless otherwise indicated. Moreover, unless otherwise indicated, the numbering of the various method steps is merely a convenient tool for identifying the various method steps, and is not intended to limit the order in which the method steps are arranged or the scope of the invention in which the invention may be practiced, and changes or modifications in the relative relationship may be made without substantially changing the technical content.
Example 1
Preparing triptolide p-nitrophenylformate:
adding 360mg (1mmol) of triptolide into a 100mL two-neck round-bottom flask, adding 40mL of dichloromethane for dissolving, adding 1.007g (5mmol) of p-nitrophenyl chloroformate, stirring, adding 0.5mL of pyridine under ice bath, returning to room temperature, monitoring the reaction by TLC after 4 hours of reaction, stopping the reaction after the triptolide completely disappears, adding 50mL of water, extracting with 50mL of dichloromethane for three times, combining organic phases, washing with 0.1M hydrochloric acid solution and saturated saline water for three times respectively, drying with anhydrous sodium sulfate, and carrying out reduced pressure rotary evaporation to remove the dichloromethane. The separation by chromatography column gave 450mg of a white solid as the desired product in 85% yield.
Example 2
Preparation of triptolide AS1411 aptamer conjugate:
15.8mg (30. mu. mol) of triptolide p-nitrophenylformate (prepared in example 1) was dissolved in 3mL of N, N-dimethylformamide, and 2.99mg (0.3. mu. mol) of an aptamer (amino-modified aptamer, available from Biotech, Inc., Beijing Populaceae, sequence: 5'-TTTTTG GTG GTG GTG GTT GTG GTG GTG GTG G-3', SEQ ID No.1) was dissolved in 3mL of ultrapure water (dd H)2O), mixing the two solutions, adding 0.5mL of triethylamine, keeping out of the light, and carrying out oscillation reaction at 37 ℃ for 1 hour. Purifying with reversed-phase preparative column after 1 hr, and freeze-drying the collected solution containing target product to obtain 2.82mg triptolide aptamer conjugate with mass spectrum detection result shown in FIG. 1, yield 90.8%, MS: calibrated 10360.0(Found: 10359.5).
Example 3
Preparing triptolide Sgc8c aptamer conjugate:
referring to example 2, the feed amount of each raw material was the same on a molar basis, and only the aptamer was replaced with Sgc8c aptamer from AS1411 aptamer (amino-modified aptamer was purchased from Biotech, Inc., Beijing Strongraceae, sequence: 5'-ATCTAACTGCTGCGCCGCCGGGAAAATACTGTACGGTTAGA-3', SEQ ID No.2), the yield of the product was 91.2%, and MS: sized: 13199.4(Found: 13199.9).
Example 4
HepG2 cells were seeded into 96-well plates (each5000 cells per well), culturing at 37 degrees under 5% carbon dioxide for 24 hours, and adding culture media containing triptolide and triptolide aptamer conjugates (prepared in example 2) at different concentrations. The culture was incubated at 37 ℃ under 5% carbon dioxide for 72 hours. The inhibitory effect of the drug was measured after 72 hours using the CCK-8 kit, and the results are shown in FIG. 2. As shown in FIG. 2, the triptolide and the triptolide aptamer conjugate have good inhibition effect on liver cancer HepG2 cells, and the IC of the triptolide and the triptolide aptamer conjugate50Values were 40nM and 165nM, respectively.
Example 5
Experimental methods referring to example 4, HepG2 cells were changed to MCF-7 cells, and the results of the experiments are shown in FIG. 3. As can be seen from FIG. 3, IC of triptolide and triptolide aptamer conjugates50Values were 35nM and 102nM, respectively.
Example 6
Experimental methods referring to example 4, HepG2 cells were changed to MDA-MB-231 cells, and the results of the experiments are shown in FIG. 4. As can be seen from FIG. 4, IC of triptolide and triptolide aptamer conjugates50Values were 42nM and 151nM, respectively.
In conclusion, the present invention effectively overcomes various disadvantages of the prior art and has high industrial utilization value.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Sequence listing
<110> university of Hunan
<120> preparation method of triptolide aptamer conjugate
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 31
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
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tttttggtgg tggtggttgt ggtggtggtg g 31
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<212> DNA
<213> Artificial Sequence (Artificial Sequence)
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atctaactgc tgcgccgccg ggaaaatact gtacggttag a 41
Claims (10)
1. A method for preparing a triptolide aptamer conjugate, comprising:
1) reacting triptolide with p-nitrophenyl chloroformate to prepare p-nitrophenyl formate triptolide;
2) reacting the p-nitrophenyl formate triptolide with a nucleic acid aptamer to prepare the triptolide nucleic acid aptamer conjugate.
2. The method according to claim 1, wherein in the step 1), the reaction is carried out in the presence of an acid-binding agent;
and/or, in the step 1), the molar ratio of the triptolide to the p-nitrophenyl chloroformate is 1: 1.2-10, preferably 1: 3-6;
and/or, in the step 1), the reaction is carried out in the presence of a reaction solvent;
and/or in the step 1), the reaction temperature is 10-40 ℃, and preferably 20-30 ℃.
3. The method according to claim 1, wherein in step 1), the acid scavenger is selected from the group consisting of organic bases;
and/or, in the step 1), the molar ratio of the triptolide to the acid-binding agent is 1: 0.002 to 0.25, preferably 1: 0.05 to 0.08;
and/or, in the step 1), the reaction solvent is selected from halogenated alkane solvents.
4. The preparation method of claim 1, wherein the acid-binding agent is selected from organic bases, preferably, the acid-binding agent is selected from one or more of pyridine and DMAP;
and/or, in the step 1), the reaction solvent is selected from one or more of dichloromethane, chloroform and 1, 2-dichloroethane.
5. The method of claim 1, wherein the step 1) further comprises: adding water for quenching, extracting by an organic solvent, and desolventizing an organic phase.
6. The method according to claim 1, wherein in the step 2), the reaction is carried out in the presence of a base;
and/or, in the step 2), the molar ratio of the triptolide p-nitrophenylformate to the aptamer is 1: 0.001 to 0.5, preferably 1: 0.005 to 0.1;
and/or, in the step 2), the reaction is carried out in the presence of a reaction solvent;
and/or in the step 2), the reaction temperature is 10-50 ℃, and preferably 30-40 ℃;
and/or, in the step 2), the aptamer comprises an amino group, preferably, the 5' end of the aptamer comprises an amino group;
and/or, in the step 2), the reaction time is less than or equal to 4 hours, preferably 15 min-1 h;
and/or, in the step 2), the reaction is carried out under the condition of avoiding light.
7. The method according to claim 1, wherein in the step 2), the base is selected from the group consisting of organic bases;
and/or, in the step 2), the reaction solvent is selected from water and organic solvents;
and/or, in the step 2), the aptamer is selected from Sgc8c aptamer and AS1411 aptamer in combination of one or more.
8. The preparation method according to claim 1, wherein in the step 2), the base is selected from one or more of triethylamine, N-diisopropylethylamine;
and/or, in the step 2), the organic solvent used as the reaction solvent is selected from aprotic polar solvents, and is more preferably selected from one or more of N, N-dimethylformamide, dimethyl sulfoxide and acetonitrile.
9. The method of claim 1, wherein the step 2) further comprises: adding water for quenching, extracting by an organic solvent, and desolventizing an organic phase.
10. The triptolide aptamer conjugate prepared by the preparation method according to any one of claims 1-9.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115227829A (en) * | 2022-02-22 | 2022-10-25 | 成都中医药大学 | Novel acid-sensitive aptamer triptolide conjugate and application thereof |
| CN115990264A (en) * | 2022-12-21 | 2023-04-21 | 中国科学院基础医学与肿瘤研究所(筹) | PTK7 targeted aptamer coupled drug |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2015085447A1 (en) * | 2013-12-11 | 2015-06-18 | 香港浸会大学 | New triptolide derivatives and preparation method and use thereof |
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| WO2015085447A1 (en) * | 2013-12-11 | 2015-06-18 | 香港浸会大学 | New triptolide derivatives and preparation method and use thereof |
| CN106589049A (en) * | 2015-10-19 | 2017-04-26 | 中国中医科学院中医临床基础医学研究所 | Synthesis method for modified triptolide-aptamer conjugate |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115227829A (en) * | 2022-02-22 | 2022-10-25 | 成都中医药大学 | Novel acid-sensitive aptamer triptolide conjugate and application thereof |
| WO2023160354A1 (en) * | 2022-02-22 | 2023-08-31 | 成都中医药大学 | Novel acid-sensitive aptamer triptolide conjugate and application |
| CN115227829B (en) * | 2022-02-22 | 2023-10-13 | 成都中医药大学 | Acid-sensitive aptamer triptolide conjugate and application thereof |
| CN115990264A (en) * | 2022-12-21 | 2023-04-21 | 中国科学院基础医学与肿瘤研究所(筹) | PTK7 targeted aptamer coupled drug |
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