CN110917122A - Preparation method of platelet drug-loading system carrying anti-tumor drug - Google Patents
Preparation method of platelet drug-loading system carrying anti-tumor drug Download PDFInfo
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- CN110917122A CN110917122A CN201911330170.5A CN201911330170A CN110917122A CN 110917122 A CN110917122 A CN 110917122A CN 201911330170 A CN201911330170 A CN 201911330170A CN 110917122 A CN110917122 A CN 110917122A
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Abstract
The invention belongs to the technical field of medical treatment, and particularly relates to a preparation method of a platelet drug-loading system carrying an anti-tumor drug, which comprises the following steps: the method comprises the following steps: preparation of platelets: adding a sodium citrate solution into the whole blood at the ambient temperature of 15-30 ℃, and centrifuging by using a centrifugal machine to obtain platelet plasma; step two: mixing the medicines: adding the platelets prepared in the step one into an anti-tumor drug, and then carrying out oscillation reaction for 1.5-2.5 hours to obtain a mixture of the platelets and the anti-tumor drug; step three: and D, separating the mixture of the platelets and the antitumor drugs in the step two through an agarose gel 2B separation column, and centrifugally washing to obtain a platelet drug-loading system carrying the antitumor drugs. The invention realizes the comprehensive effect of effectively treating the tumor by taking the platelet as a drug-carrying system.
Description
Technical Field
The invention relates to the technical field of medical treatment, in particular to a preparation method of a platelet drug-loading system carrying an anti-tumor drug.
Background
Tumor is a large disease threatening human health and life, the incidence rate tends to increase year by year, the current main treatment methods comprise operation treatment, chemotherapy and radiotherapy, but the operation treatment can not be generally carried out on some patients with advanced stage, and in addition, the chemotherapy is often supplemented before and after the operation treatment on some patients with tumors with higher malignancy, so the chemotherapy still plays an important role in the treatment of the tumors at present. However, chemotherapy has significant side effects including bone marrow suppression, low immune function, digestive tract reaction, etc., so that it is of great importance to research and develop drug carriers capable of delivering antitumor drugs into or around tumor cells in order to reduce side effects.
At present, drug carriers mainly comprise magnetic nano drug carriers, liposome carriers, particle carriers and the like, but the magnetic nano drug carriers, the liposome carriers, the particle carriers and the like have various defects such as low drug loading rate, poor stability, strong toxic effect and the like, so the biological drug carriers are introduced in a plurality of drug loading systems by virtue of the characteristics of good biocompatibility, slow-controlled drug release and the like.
Platelets are fragments shed from bone marrow mature megakaryocytes, are about 2um in diameter, are about (1-3) x 1011/L in adult blood, have a lifespan of about 7-14 days, and on average 9 days, and are mostly cleared by Kupfer cells in the liver, so that they have good biocompatibility, safety, and long blood circulation time. Platelets play an important role in hemostasis, wound healing, thrombosis and other processes, and have close relation with the occurrence and development of tumors. The tumor cells activate the platelets through a series of reactions, the activated platelets are gathered around the tumor cells through the action of platelet aggregation induced by the Tumor Cells (TCIPA) to form a platelet-tumor cell compound, the adhesion of the platelets and the tumor cells is promoted just through the action, the activated platelets release the ingested drugs, the effect of the platelets for ingesting the drugs and small particles is different from the phagocytosis of a macrophage system, i.e., the endocytosed substances are not killed and eliminated, so that the original structure and activity of the drugs are ensured, and the drug effect is retained. In addition, researches show that the drug loading rate of the platelets is far higher than that of the nanoparticles, and the platelets and the cytotoxic drugs have a synergistic effect and can increase the killing effect of the platelets. Therefore, the platelet is used as a drug carrier, and has the advantages of rich content, high biocompatibility, high drug loading rate and increase of the cytotoxic drug effect.
Although clinical experiments of antiplatelet drugs in tumor patients are not reported so far in the prior art, with the deep research on tumorigenesis, development and metastasis and interaction mechanism of platelets and tumors, the antiplatelet drugs are very likely to become one of the comprehensive tumor treatment means, cancer biology always centers on tumors, and with the increasing importance on the role of stromal cells in tumorigenesis, development and metastasis, cancer biologists are reevaluating the role of non-tumor cells in the cancerization process, including the role of platelets, so that the adjuvant treatment of cancer through the antiplatelet drugs is more and more regarded, and therefore, the development of a preparation method of a platelet drug loading system carrying antitumor drugs is urgently needed.
Disclosure of Invention
This section is for the purpose of summarizing some aspects of embodiments of the invention and to briefly introduce some preferred embodiments. In this section, as well as in the abstract and the title of the invention of this application, simplifications or omissions may be made to avoid obscuring the purpose of the section, the abstract and the title, and such simplifications or omissions are not intended to limit the scope of the invention.
The invention is provided in view of the above and/or the problems in the existing preparation method of the platelet drug-carrying system carrying the anti-tumor drug.
Therefore, the invention aims to provide a preparation method of a platelet drug carrier system carrying an anti-tumor drug, which can realize effective treatment of tumors by platelets by taking the platelets as the drug carrier system.
To solve the above technical problem, according to an aspect of the present invention, the present invention provides the following technical solutions:
a preparation method of a platelet drug-loaded system carrying an anti-tumor drug comprises the following steps:
the method comprises the following steps: preparation of platelets: adding a sodium citrate solution into the whole blood at the ambient temperature of 15-30 ℃, and centrifuging by using a centrifugal machine to obtain platelet plasma;
step two: mixing the medicines: adding the platelets prepared in the step one into an anti-tumor drug, and then carrying out oscillation reaction for 1.5-2.5 hours to obtain a mixture of the platelets and the anti-tumor drug;
step three: and D, separating the mixture of the platelets and the antitumor drugs in the step two through an agarose gel 2B separation column, and centrifugally washing to obtain a platelet drug-loading system carrying the antitumor drugs.
As a preferable scheme of the preparation method of the platelet drug-loaded system carrying the antitumor drug, the preparation method comprises the following steps: the rotating speed of the centrifuge is 1500-2000rpm, and the centrifugation time is 10-15 minutes.
As a preferable scheme of the preparation method of the platelet drug-loaded system carrying the antitumor drug, the preparation method comprises the following steps: and the mixing condition in the second step is a dark environment with the mixing temperature of 36-38 ℃.
As a preferable scheme of the preparation method of the platelet drug-loaded system carrying the antitumor drug, the preparation method comprises the following steps: the anticancer drug in the step two is one or a mixture of prostacyclin, aspirin, emopamil, fluoxetine, maprotiline, ketanserin, sarpogrelate, abciximab, eptifibatide and tirofiban.
As a preferable scheme of the preparation method of the platelet drug-loaded system carrying the antitumor drug, the preparation method comprises the following steps: the volume ratio of the whole blood to the sodium citrate solution with the weight percentage of 2-4% in the step one is 7:1-10: 1.
As a preferable scheme of the preparation method of the platelet drug-loaded system carrying the antitumor drug, the preparation method comprises the following steps: the number of platelets in the first step is controlled to be (2.5-3.0) multiplied by 105/ul。
Compared with the prior art: the preparation method of the platelet drug-loading system carrying the antitumor drug realizes that the platelets are used as the drug-loading system, and the platelets can effectively treat tumors.
Detailed Description
The present invention will be described in detail with reference to the following embodiments in order to make the aforementioned objects, features and advantages of the invention more comprehensible.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways than those specifically described herein, and it will be apparent to those of ordinary skill in the art that the present invention may be practiced without departing from the spirit and scope of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.
In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention will be described in further detail below.
Embodiment mode 1
The invention provides a preparation method of a platelet drug-loading system carrying an anti-tumor drug, which comprises the following steps:
the method comprises the following steps: preparation of platelets: adding a sodium citrate solution into whole blood at the ambient temperature of 15 ℃, wherein the volume ratio of the whole blood to the sodium citrate solution with the weight percentage of 4 percent is 7:1, centrifuging by using a centrifugal machine to obtain platelet plasma, and controlling the number of platelets to be 3.0 multiplied by 105The rotation speed of the centrifugal machine is 1500rpm, and the centrifugation time is 15 minutes;
step two: mixing the medicines: adding an anti-tumor drug into the platelets prepared in the step one, and then carrying out oscillation reaction for 1.5 hours in a dark environment at the temperature of 36-38 ℃ to obtain a mixture of the platelets and the anti-tumor drug, wherein the anti-tumor drug is one or more of prostacyclin, aspirin, emopamil, fluoxetine, maprotiline, ketanserin, sarpogrelate, abciximab, eptifibatide and tirofiban;
step three: and D, separating the mixture of the platelets and the antitumor drugs in the step two through an agarose gel 2B separation column, and centrifugally washing to obtain a platelet drug-loading system carrying the antitumor drugs.
Embodiment mode 2
The invention provides a preparation method of a platelet drug-loading system carrying an anti-tumor drug, which comprises the following steps:
the method comprises the following steps: preparation of platelets: adding a sodium citrate solution into whole blood at the ambient temperature of 30 ℃, wherein the volume ratio of the whole blood to the sodium citrate solution with the weight percentage of 2 percent is 10:1, centrifuging by utilizing a centrifugal machine to obtain platelet plasma, and controlling the number of the platelets to be 2.5 multiplied by 105The rotation speed of the centrifugal machine is 2000rpm, and the centrifugation time is 10 minutes;
step two: mixing the medicines: adding an anti-tumor drug into the platelets prepared in the step one, and then carrying out oscillation reaction for 2.5 hours in a dark environment at the temperature of 36-38 ℃ to obtain a mixture of the platelets and the anti-tumor drug, wherein the anti-tumor drug is one or more of prostacyclin, aspirin, emopamil, fluoxetine, maprotiline, ketanserin, sarpogrelate, abciximab, eptifibatide and tirofiban;
step three: and D, separating the mixture of the platelets and the antitumor drugs in the step two through an agarose gel 2B separation column, and centrifugally washing to obtain a platelet drug-loading system carrying the antitumor drugs.
Embodiment 3
The invention provides a preparation method of a platelet drug-loading system carrying an anti-tumor drug, which comprises the following steps:
the method comprises the following steps: preparation of platelets: adding sodium citrate solution into whole blood at the ambient temperature of 22.5 ℃, wherein the volume ratio of the whole blood to the sodium citrate solution with the weight percentage of 3 percent is 8.5:1, centrifuging by utilizing a centrifugal machine to obtain platelet plasma, and controlling the number of the platelets to be 2.55 multiplied by 105The rotation speed of the centrifuge is 1750rpm, and the centrifugation time is 12 minutes;
step two: mixing the medicines: adding an anti-tumor drug into the platelets prepared in the step one, and then carrying out oscillation reaction for 2 hours in a dark environment at 36-38 ℃ to obtain a mixture of the platelets and the anti-tumor drug, wherein the anti-tumor drug is one or more of prostacyclin, aspirin, emopamil, fluoxetine, maprotiline, ketanserin, sarpogrelate, abciximab, eptifibatide and tirofiban;
step three: and D, separating the mixture of the platelets and the antitumor drugs in the step two through an agarose gel 2B separation column, and centrifugally washing to obtain a platelet drug-loading system carrying the antitumor drugs.
Embodiment 4
The invention provides a preparation method of a platelet drug-loading system carrying an anti-tumor drug, which comprises the following steps:
the method comprises the following steps: preparation of platelets: adding sodium citrate solution into whole blood at the ambient temperature of 20 ℃, wherein the volume ratio of the whole blood to the 3 percent by weight of the sodium citrate solution is 9:1, centrifuging by using a centrifugal machine to obtain platelet plasma, and controlling the number of the platelets to be 3.0 multiplied by 105The rotation speed of the centrifuge is 1800rpm, and the centrifugation time is 12 minutes;
step two: mixing the medicines: adding an anti-tumor drug into the platelets prepared in the step one, and then carrying out oscillation reaction for 2 hours in a dark environment at 36-38 ℃ to obtain a mixture of the platelets and the anti-tumor drug, wherein the anti-tumor drug is one or more of prostacyclin, aspirin, emopamil, fluoxetine, maprotiline, ketanserin, sarpogrelate, abciximab, eptifibatide and tirofiban;
step three: and D, separating the mixture of the platelets and the antitumor drugs in the step two through an agarose gel 2B separation column, and centrifugally washing to obtain a platelet drug-loading system carrying the antitumor drugs.
By combining the above embodiments, the preparation method of the platelet drug-loading system carrying the antitumor drug can effectively realize that the platelets are used as the drug-loading system, and the platelets can effectively treat tumors, so that the preparation method has good economic benefits.
While the invention has been described above with reference to an embodiment, various modifications may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. In particular, the various features of the disclosed embodiments of the invention may be used in any combination, provided that no structural conflict exists, and the combinations are not exhaustively described in this specification merely for the sake of brevity and resource conservation. Therefore, it is intended that the invention not be limited to the particular embodiments disclosed, but that the invention will include all embodiments falling within the scope of the appended claims.
Claims (6)
1. A preparation method of a platelet drug-loaded system carrying an anti-tumor drug is characterized by comprising the following steps: the preparation method of the platelet drug-loading system carrying the antitumor drug comprises the following steps:
the method comprises the following steps: preparation of platelets: adding a sodium citrate solution into the whole blood at the ambient temperature of 15-30 ℃, and centrifuging by using a centrifugal machine to obtain platelet plasma;
step two: mixing the medicines: adding the platelets prepared in the step one into an anti-tumor drug, and then carrying out oscillation reaction for 1.5-2.5 hours to obtain a mixture of the platelets and the anti-tumor drug;
step three: and D, separating the mixture of the platelets and the antitumor drugs in the step two through an agarose gel 2B separation column, and centrifugally washing to obtain a platelet drug-loading system carrying the antitumor drugs.
2. The method for preparing a platelet drug carrier system carrying an antitumor drug according to claim 1, wherein the method comprises the following steps: the rotating speed of the centrifuge is 1500-2000rpm, and the centrifugation time is 10-15 minutes.
3. The method for preparing a platelet drug carrier system carrying an antitumor drug according to claim 1, wherein the method comprises the following steps: and the mixing condition in the second step is a dark environment with the mixing temperature of 36-38 ℃.
4. The method for preparing a platelet drug carrier system carrying an antitumor drug according to claim 1, wherein the method comprises the following steps: the anticancer drug in the step two is one or a mixture of prostacyclin, aspirin, emopamil, fluoxetine, maprotiline, ketanserin, sarpogrelate, abciximab, eptifibatide and tirofiban.
5. The method for preparing a platelet drug carrier system carrying an antitumor drug according to claim 1, wherein the method comprises the following steps: the volume ratio of the whole blood to the sodium citrate solution with the weight percentage of 2-4% in the step one is 7:1-10: 1.
6. The method for preparing a platelet drug carrier system carrying an antitumor drug according to claim 1, wherein the method comprises the following steps: the number of platelets in the first step is controlled to be (2.5-3.0) multiplied by 105/ul。
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| CN112891317A (en) * | 2021-02-05 | 2021-06-04 | 东南大学 | Preparation method of platelet drug delivery system |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104771762A (en) * | 2015-03-23 | 2015-07-15 | 南京大学医学院附属鼓楼医院 | Preparation method of antitumor drug carrying platelet drug loading system |
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| CN104771762A (en) * | 2015-03-23 | 2015-07-15 | 南京大学医学院附属鼓楼医院 | Preparation method of antitumor drug carrying platelet drug loading system |
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| CN112891317A (en) * | 2021-02-05 | 2021-06-04 | 东南大学 | Preparation method of platelet drug delivery system |
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Application publication date: 20200327 |