CN103285826A - Blood purification adsorbing agent for restraining grown and transfer of tumor and application thereof - Google Patents
Blood purification adsorbing agent for restraining grown and transfer of tumor and application thereof Download PDFInfo
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- CN103285826A CN103285826A CN2013102658139A CN201310265813A CN103285826A CN 103285826 A CN103285826 A CN 103285826A CN 2013102658139 A CN2013102658139 A CN 2013102658139A CN 201310265813 A CN201310265813 A CN 201310265813A CN 103285826 A CN103285826 A CN 103285826A
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Abstract
The invention discloses a blood purification adsorbing agent for restraining grown and transfer of tumor and application of the blood purification adsorbing agent. The adsorbing agent takes natural macromolecule or synthetic macromolecule as a carrier, a biological cross-linking agent or a chemical crosslinking agent as an arm, natural small molecules or large molecules with polyhydric group and acidity or basic group with an affine adsorptive action with cell factors (such VEGF, bFGF, MMP-9,TGF,IL and the like) in the tumor microenvironment, or artificially synthesized small molecules or macromolecule substances as a genin; the carrier, the arm and the genin are combined through a biological or chemical synthesizing method, and the adsorbing agent for selectively removing cell factors promoting tumor grown or transfer in patient blood is prepared. The blood purification adsorbing agent provided by the invention selectively removes cell factors promoting tumor grown or transfer in blood by adopting a blood perfusion method, so that the suitable microenvironment for tumor growth is eliminated, the cell factors are effectively antagonized so as not to promote tumor tumorigenesis and progression, meanwhile, an immune system is also protected, and cancer growth or cancer metastasis is prevented.
Description
Technical field
The invention belongs to the biological medicine technology field, relate to a kind ofly for the extracorporeal blood perfusion, remove the adsorbent and the application thereof that promote the cell factor of tumor growth and transfer in the tumor microenvironment.
Background technology
Cancer is main one of the disease that causes death that threatens human health.There is millions of New Development cases of cancers every year in China.Shifting then is the most fearful feature of tumour, and 90% patient is dead because of the relapse and metastasis of tumour.Therefore metastases is key subjects and the research difficult point for the treatment of and prevention of tumour all the time.
Operation, radiotherapy, chemotherapy are the tradition three big means of oncotherapy, and operation can be excised the cancer cell that has formed lump, and radiotherapy, chemotherapy can be killed all or part of adult cancer cell.And the side effect of chemicotherapy has greatly damaged patient's immune system, and then induces the childhood cancer cell to be activated from resting state, fission, sudden change rapidly, forms new cancer cell, the basic reason of metastasis of cancer that Here it is recurrence.Numerous results of study show, before the immunity of tumor patient was not recovered, all thought merely to be not only by the methods for the treatment of of direct kill cancer cell futile, and are harmful to, and have consequently accelerated patient's death! World Health Organization's statistics, contrast the decennary tumor patient death rate of nineteen ninety and 2000, not only do not descend, 17% real data has on the contrary risen, drawn why the medical science and technology level is more and more advanced, oncotherapy technology and medicine are also more and more, and the conclusion that the death rate of tumor patient goes up not down: because we also do not have really to be familiar with generation, the development of tumour and shift track, can't find the key of opening effective oncotherapy!
Tumor migration is the process of a complexity, tumour cell after former position comes off through local infiltration, blood vessel invade, existence the blood circulation, at a distance organ go back to the nest and implantation, essence are soaked into, the growth of new environmental adaptation and secondary tumor.In this process, the residing microenvironment of tumour cell is being brought into play key effect to the transfer of tumour.Tumor microenvironment generally refers in the tumor growth process, the local Stable State Environment that is constituted jointly by tumour cell, stroma cell and extracellular matrix etc., when local cell during by tumour cell transformation, produce a large amount of growth factors, cell chemotactic factor, inflammatory factor and the substrate degradation factor around it, for the generation of tumour, development, invasion and attack, transfer etc. provide necessary material base.
Along with the development of Protocols in Molecular Biology, the target tumor microenvironment becomes the new way for the treatment of tumour.The eighties in last century, scientist finds out tumour immunotherapy and claims biotherapy (Biotherapy, treatment tumour new ideas BT) again.This is the 4th kind of tumor therapy after operation, chemotherapy, radiotherapy, and this new treatment also begins to be applied to clinical treatment the eighties.The biological immune therapy can be used different materials and method at different types of tumors.As anti-VEGF (VEGF) treatment that generates at tumor vessel, the immunotherapy of tumors that increases at immunosuppressive factor etc.These biological therapy methods have been brought into play important effect in oncotherapy.But biotherapy also has the defective of self, and the expense of immune response, non-specific damage, costliness etc. all is the major issue that urgent need will solve.
At the deficiency that above therapy faces, be necessary to develop a kind of new technology of auxiliary for treating cancer.MMP(metal matrix protease promotion tumor growth or the transitional cell factor such as VEGF, the FGF(fibroblast growth factor of blood purification technology in can selective clearing blood)) etc., thereby reach the purpose of regulating the tumor microenvironment balance, will be expected to become the 5th kind of therapy in the treatment of cancer.
Summary of the invention
The present invention seeks to the deficiency that exists at the oncotherapy prior art, provide a kind of for removing the cell factor that blood promotes tumor growth or transfer
Blood-purifying adsorbing agent and application thereofThereby, reach the purpose of regulating tumor microenvironment.
At the design feature that promotes the cell factor of growth of cancers and transfer in the microenvironment of pathogenesis of cancer, the synthetic a kind of chemical aglucon of design, and it is coupled on the adsorbing agent carrier, prepare and have the blood-purifying adsorbing agent that promotes the cell factor of tumor growth and transfer in the selective clearing tumor microenvironment.
Provided by the invention
Be used for suppressing the blood-purifying adsorbing agent of tumor growth and transfer,Relate to a kind of for external whole blood or plasma perfusion, remove the adsorbent that promotes the cell factor of tumor growth or transfer in the blood, this adsorbent is carrier with natural polymer or synthetic high polymer, be arm with biological crosslinking agent or chemical cross-linking agent, with derive from tumor microenvironment in cell factor to have the affine adsorbing synthetic or natural molecule that has polyhydroxy and acidity or basic group be aglucon, synthetic method by biological or chemical is carrier-arm-aglucon combination, prepares to have the blood-purifying adsorbing agent that promotes the cell factor of tumor growth or transfer in the selective clearing blood.
Described natural polymer carrier material is cellulose, shitosan or collagen; The synthetic high polymer carrier material is polyvinyl alcohol, polystyrene or polyacrylic acid lipid (Tianjin Ourui Biology Technology Co., Ltd. provides); Because they have better biocompatibility, make spheroid carrier easily, it contains a large amount of for reactive activity site (hydroxyl or amido etc.).
Described biological crosslinking agent is the crosslinking agent with special-shaped dual-functional group; Chemical cross-linking agent is glutaraldehyde, ethylenediamine or epoxychloropropane;
Promote in the described blood that the cell factor of tumor growth or transfer is VEGF, bFGF, TGF, IL or MMP-9 etc.
It is to be carrier with water-fast polystyrene microsphere, collagen microballoon, esters of acrylic acid microballoon, chitosan microball, cellulose microsphere or polyvinyl alcohol microballoon, be aglucon to design the synthetic affinity molecule to respective fine intracellular cytokine in the tumor microenvironment, a kind of selective cytokine adsorbent for preparing.The adsorbent that obtains by this method preparation has the effect of cytokines that promotes tumor growth or transfer in the selective clearing blood samples of patients.
Described adsorbing agent carrier is spherical, and particle diameter is at 50-800um, with 100-500um scope the best.
Described biological crosslinking agent is the crosslinking agent with special-shaped dual-functional group, specifically be DPS(3,3'-dithio dipropyl acid two (N-maloyl imines ester), DCC(dicyclohexyl carbodiimide) or PMPI(N-[p-Maleimidophenyl] isocyanate) etc.; Chemical cross-linking agent is glutaraldehyde, ethylenediamine or epoxychloropropane etc.
Described aglucon is natural little molecule or big molecule, or artificial synthetic little molecule or polymer substance.
Specifically be derive from tumor microenvironment in cell factor (as VEGF, bFGF, MMP-9, TGF, IL etc.) have affine adsorbing natural little molecule or the big molecule that has polyhydroxy and acidity or basic group, or artificial synthetic little molecule or polymer substance, as each seed amino acid, vitamin, sulfonic group, amido, heparin, chondroitin sulfate etc.
The synthetic method of described biological or chemical refers at present by document all synthetic methods and reaction condition as can be known.
Adsorbent provided by the invention can be used for whole blood or plasma perfusion, removes the cell factor (as VEGF, bFGF, MMP-9, TGF, IL etc.) that promotes tumor growth or transfer that has in the blood.
Adsorbent provided by the invention not only can be used as preoperative prophylactic treatment, also can be used for postoperative prevention cancer metastasis.Not only be used for treatment for cancer.Also can be generalized to the treatment field of other diseases.
Advantage of the present invention and beneficial effect:
The present invention is by being coupled to affinity ligand on the adsorbing agent carrier; see through the mode of blood perfusion, specificity is removed the cell factor of pathogenic or promotion growth of cancers and transfer in the blood, has protected immune system; suppress growth of tumor, prevented cancer metastasis.
The specific embodiment
The invention process process comprises that adsorbing agent carrier is selected, selection, aglucon and the carrier coupling of carrier activation, aglucon and to the adsorption experiment of the total cell factor of tumor patient blood (VEGF, MMP-9, bFGF etc.).Adsorbing agent carrier provides by Tianjin Ourui Biology Technology Co., Ltd., and the method that the detection of cell factor adopts ELLSA kit (U.S. company BD) to provide in the blood plasma detects, and patient's blood plasma is provided by the patient of Tianjin Tumour Hospital aspiration.
Embodiment 1:VEGF preparation of adsorbent
Carrier: polyvinyl alcohol microballoon (available from Tianjin Ourui Biology Technology Co., Ltd.)
Arm: epoxychloropropane (available from Tianjin chemical reagent factory)
Aglucon: heparin (available from sigma company)
Preparation process:
Polyvinyl alcohol microballoon activation: add the NaOH of 100ml3mol/L in the 100ml polyvinyl alcohol microballoon, 600ml epoxychloropropane (EPI), 40 ℃ down concussion carry out 3 hours priming reactions, it is standby fully to wash the back with distilled water.
Aglucon is fixed: get above-mentioned activation polyvinyl alcohol carrier 100ml, add 200ml PBS(phosphate buffer, pH7.2) with swollen microsphere.The heparin 200ml of the 10mg/ml for preparing in advance of excessive adding then in 45 ℃ of concussion reactions 4 hours down, inhales and abandons reactant liquor, and PBS cleans 3 times, each 5 minutes.Namely obtain having the adsorbent of affine absorption VEGF.
Adsorption experiment: get 10 parts of cancer patient's blood plasma, every part of 10ml places the 15ml test tube respectively.Each test tube adds the adsorbent 2ml of above-mentioned preparation respectively then, and 37 ℃ of concussions were adsorbed 2 hours, surveyed VEGF concentration in the blood plasma, and concrete data are seen following table 1.
Embodiment 2:MMP-9 preparation of adsorbent
Carrier: chitosan microball (available from Tianjin Ourui Biology Technology Co., Ltd.)
Arm:DPS (3,3'-dithio dipropyl acid two (N-maloyl imines ester)
(available from Sigma company)
Aglucon: taurine (available from sigma company)
Preparation process:
Chitin carrier activation: get chitosan microball 100ml, add 200ml PBS(phosphate buffer, pH7.2) with the swelling chitosan microball.Then the DPS of the 1mg/ml concentration for preparing in advance of excessive adding (3,3'-dithio dipropyl acid two (N-maloyl imines ester) 10ml, room temperature reaction 30 minutes are inhaled and are abandoned reactant liquor, and PBS cleans 3 times, each 5 minutes, the adsorbing agent carrier that obtains activating.
Aglucon is fixed: get above-mentioned activation chitin carrier 100ml, add 200ml PBS(phosphate buffer, pH7.2) with swollen microsphere.The taurine solution 300ml of the 5mg/ml for preparing in advance of excessive adding then in 45 ℃ of concussion reactions 3 hours down, inhales and abandons reactant liquor, and PBS cleans 3 times, each 5 minutes.Namely obtain having the adsorbent of affine absorption MMP-9.
Adsorption experiment: get 10 parts of cancer patient's blood plasma, every part of 10ml places the 15ml test tube respectively.Each test tube adds the adsorbent 2ml of above-mentioned preparation respectively then, and 37 ℃ of concussions were adsorbed 2 hours, surveyed MMP-9 concentration in the blood plasma, and concrete data are seen following table 1.
Embodiment 3:bFGF preparation of adsorbent
Carrier: cellulose microsphere (available from Tianjin Ourui Biology Technology Co., Ltd.)
Arm: epoxychloropropane (available from Tianjin chemical reagent factory)
Aglucon: phosphopyridoxal pyridoxal phosphate (available from Beijing Braun Aktiengesellschaft)
Preparation process:Cellulose microsphere activation: extracting cellulose microballoon 100ml, the NaOH that adds 100ml3mol/L, 600ml epoxychloropropane (EPI), 3 hours priming reactions are carried out in concussion under 40 ℃, after fully washing with distilled water, add 100ml 2mol/L NaOH, the ethylenediamine of 500ml, the concussion reaction is 12 hours under 75 ℃, and the distilled water washing is standby.
Aglucon is fixed: get the microsphere supported 100ml of above-mentioned activated cellulose, add 200ml PBS(phosphate buffer, pH7.2) with swollen microsphere.2% the phosphopyridoxal pyridoxal phosphate 100ml for preparing in advance of excessive adding then, room temperature reaction 30 minutes is inhaled and is abandoned reactant liquor, PBS cleans 3 times, each 5 minutes, namely obtain having can with the adsorbent of bFGF selective absorption.
Adsorption experiment: get 10 parts of cancer patient's blood plasma, every part of 10ml places the 15ml test tube respectively.Each test tube adds the adsorbent 2ml of above-mentioned preparation respectively then, and 37 ℃ of concussions were adsorbed 2 hours, surveyed bFGF concentration in the blood plasma, and concrete data are seen following table 1.
VEGF, MMP-9 and bFGF level are relatively in table 1, the absorption front and back blood plasma
| Group | Sample number (n) | Before the absorption (pg/ml) | Absorption back (pg/ml) | The p value |
| VEGF | 10 | 868.5±148.2 | 260.7±36.2 | ＜0.01 |
| MMP-9 | 10 | 281.9±20.7 | 151.1±17.8 | ＜0.01 |
| bFGF | 10 | 18.3±12.9 | 4.4±1.2 | ＜0.01 |
Claims (5)
1. blood-purifying adsorbing agent that be used for to suppress tumor growth and transfer, it is characterized in that, this adsorbent is carrier with natural polymer or synthetic high polymer, be arm with biological crosslinking agent or chemical cross-linking agent, with derive from tumor microenvironment in cell factor to have the affine adsorbing synthetic or natural molecule that has polyhydroxy and acidity or basic group be aglucon, synthetic method by biological or chemical is carrier-arm-aglucon combination, prepares to have the blood-purifying adsorbing agent that promotes the cell factor of tumor growth or transfer in the selective clearing blood;
Described natural polymer carrier material is cellulose, shitosan or collagen; The synthetic high polymer carrier material is polyvinyl alcohol, polystyrene or polyacrylic acid lipid;
Described biological crosslinking agent is the crosslinking agent with special-shaped dual-functional group; Chemical cross-linking agent is glutaraldehyde, ethylenediamine or epoxychloropropane;
Described aglucon is natural little molecule or big molecule, or artificial synthetic little molecule or polymer substance;
Promote in the described blood that the cell factor of tumor growth or transfer is VEGF, bFGF, TGF, IL or MMP-9.
2. adsorbent according to claim 1 is characterized in that referring to the synthetic method of described biological or chemical at present by document all synthetic methods and reaction condition as can be known.
3. adsorbent according to claim 1 and 2 is characterized in that described biological crosslinking agent specifically is DPS, DCC or PMPI.
4. adsorbent according to claim 1 and 2 is characterized in that described adsorbing agent carrier for spherical, and particle diameter is at 50-800um.
5. the application of the described adsorbent of claim 1 is used for whole blood or plasma perfusion, removes the cell factor that promotes tumor growth or transfer that has in the blood, specifically is VEGF, bFGF, TGF, IL or MMP-9.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103585977A (en) * | 2013-11-22 | 2014-02-19 | 珠海健帆生物科技股份有限公司 | Cytokine adsorbent for hemoperfusion and preparation method thereof |
| CN111686704A (en) * | 2020-07-02 | 2020-09-22 | 苏州仝康医疗科技有限公司 | Blood purification adsorbent and preparation method and application thereof |
| CN111701581A (en) * | 2020-06-29 | 2020-09-25 | 南开大学 | Low-density lipoprotein, cholesterol and triglyceride adsorbent and preparation method and application thereof |
| CN113926435A (en) * | 2021-11-26 | 2022-01-14 | 华中科技大学 | Preparation method and application of collagen microsphere adsorbent |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103585977A (en) * | 2013-11-22 | 2014-02-19 | 珠海健帆生物科技股份有限公司 | Cytokine adsorbent for hemoperfusion and preparation method thereof |
| CN111701581A (en) * | 2020-06-29 | 2020-09-25 | 南开大学 | Low-density lipoprotein, cholesterol and triglyceride adsorbent and preparation method and application thereof |
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| CN113926435A (en) * | 2021-11-26 | 2022-01-14 | 华中科技大学 | Preparation method and application of collagen microsphere adsorbent |
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Application publication date: 20130911 |