CN106267228B - A kind of albumen and its application loading chemotherapeutics and radiotherapeutic drug altogether - Google Patents

A kind of albumen and its application loading chemotherapeutics and radiotherapeutic drug altogether Download PDF

Info

Publication number
CN106267228B
CN106267228B CN201610668501.6A CN201610668501A CN106267228B CN 106267228 B CN106267228 B CN 106267228B CN 201610668501 A CN201610668501 A CN 201610668501A CN 106267228 B CN106267228 B CN 106267228B
Authority
CN
China
Prior art keywords
hsa
ptx
chemotherapeutics
albumen
loading
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201610668501.6A
Other languages
Chinese (zh)
Other versions
CN106267228A (en
Inventor
杨凯
刘庄
田龙龙
陈倩
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Suzhou Baimai Biomedical Co ltd
Original Assignee
Suzhou University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Suzhou University filed Critical Suzhou University
Priority to CN201610668501.6A priority Critical patent/CN106267228B/en
Publication of CN106267228A publication Critical patent/CN106267228A/en
Application granted granted Critical
Publication of CN106267228B publication Critical patent/CN106267228B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/081—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins the protein being an albumin, e.g. human serum albumin [HSA], bovine serum albumin [BSA], ovalbumin

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Animal Behavior & Ethology (AREA)
  • Optics & Photonics (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicinal Preparation (AREA)

Abstract

The present invention relates to pharmaceutical technology field more particularly to a kind of albumen and its applications for loading chemotherapeutics and radiotherapeutic drug altogether.In some embodiments, I-131 and PTX is loaded on albumin.The chemotherapeutics and the albumen of radiotherapeutic drug provided by the invention of loading altogether has good water-soluble and biocompatibility, it can be realized the combination therapy of chemotherapy and radiotherapy, it is effectively reduced between capable of effectively reducing tumor tissues and presses, improve the weary oxygen of tumour, material is improved in the homogeneous distribution of tumour, and then improves the therapeutic effect of radiotherapy.And preparation method is very simple.

Description

A kind of albumen and its application loading chemotherapeutics and radiotherapeutic drug altogether
Technical field
The present invention relates to pharmaceutical technology field more particularly to a kind of albumen for loading chemotherapeutics and radiotherapeutic drug altogether and its Using.
Background technique
Cancer is one of several great malignant diseases for 21 century threatening human health.Although from the fifties in last century, A large amount of human and material resources, financial resources are put in the prevention and treatment of cancer in many decades, but in this respect acquired by the mankind It is in progress still extremely limited.
Operation, radiation and chemotherapy together and claim three big treatment means of cancer.Chemotherapy is the abbreviation of chemotherapy, is led to It crosses and reaches therapeutic purposes using chemotherapeutic agent killing cancer cell.Chemotherapy is a kind of means of systemic therapy, no matter using assorted Approach administration (oral, vein and Cavity administration etc.), chemotherapeutics all can be as blood circulation be throughout the overwhelming majority of whole body Organ and tissue.Therefore, to some tumours for having whole body to sow tendency and transferred Advanced cancers, chemotherapy is all main Treatment means.Radiotherapy, including Inner irradiation and outer radiotherapy are the radiation exposure tumours using various different-energies, with suppression System and killing cancer cell.Wherein Inner irradiation is α, β, the gamma-rays generated using radioactive isotope.About 70% cancer is suffered from Person needs to use radiotherapy during treating cancer, and there are about 40% cancers can be eradicated with radiotherapy.By radiotherapy and chemotherapy Combination therapy can effectively improve therapeutic effect.But classic chemotherapy and Inner irradiation are low in tumour accumulation rate, drug resistance of tumor By force, treatment side effect is big.In recent years, with the development of nanosecond medical science, using multifunctional nano carrier can dramatically enhancing Small molecule and radiotherapy nucleic are treated in the enrichment of tumour, and then enhances the effect of oncotherapy.But tumor hypoxia microenvironment causes Tumour all shows resistance to radiation and chemotherapy, and the weary oxygen microenvironment for improving tumour has important meaning for improving curative effect.
But existing some nano-carrier poor biocompatibilities, understand long-term delay in vivo, and material Preparation process is complicated, and at high cost, these all limit further application.It is total using a kind of simple method nano materials The combination therapy and imaging of load radiotherapeutic drug and chemotherapeutics for tumour were not reported also.
Summary of the invention
In view of this, a kind of loading chemotherapeutics and radiotherapeutic drug altogether the technical problem to be solved in the present invention is that providing Albumen and its application, its significant effect is better than individually loading radiotherapeutic drug or independent dress after loading chemotherapeutics and radiotherapeutic drug altogether Carry radiotherapeutic drug.
The present invention provides a kind of albumen for loading chemotherapeutics and radiotherapeutic drug altogether.
Radiotherapeutic drug and chemotherapeutics are loaded on albumen by the present invention jointly using albumen as carrier, so that being made Product the combination therapy of radiation and chemotherapy can be achieved, and can be used for the imaging of tumour.Since albumen is a large amount of in blood In the presence of, there is good biocompatibility so that it is produced by the present invention be mounted with radiotherapy, the albumen of chemotherapeutics can be There is good dispersibility in organism, and then improve prevention and treatment and/or the imaging effect of tumour.
Albumen is albumin in the albumen provided by the invention for loading chemotherapeutics and radiotherapeutic drug altogether；Chemotherapeutics is purple China fir alcohol；Radiotherapeutic drug is I-131.
Albumin (also known as albumin, albumin, Alb), for the single chain polypeptide containing 585 amino acid residues, molecular weight is 67kDa contains 17 disulfide bond in molecule, the component without containing sugar.Albumin is the protein that content is most in blood plasma, accounts for blood plasma The 40%~60% of total protein.Taxol (Paclitaxel, PTX) molecular weight is 854Da, molecular formula C47H51NO14, it is one The classical small molecule chemotherapeutic drug of kind, has been widely used in clinic.I-131 is denoted as131I, iodine -131 is a kind of Common radiotherapy nucleic, especially has good therapeutic effect to thyroid disease.
The present invention adsorbs small molecule chemotherapeutic drug taxol using albumin HSA as substrate, by hydrophobic forces, and by I- 131 labels are on the tyrosine of albumen, to obtain the albumen provided by the invention for loading chemotherapeutics and radiotherapeutic drug altogether, mark It is denoted as131I-HSA-PTX。
The molar ratio of albumen and chemotherapeutics is in the albumen provided by the invention for loading chemotherapeutics and radiotherapeutic drug altogether 1:5。
The molar ratio of albumen and I-131 can be any in the albumen provided by the invention for loading chemotherapeutics and radiotherapeutic drug altogether Regulation.
In the embodiment of the present invention, the molar ratio of taxol and albumin is in the albumin of paclitaxel loaded and I-131 altogether 1:5。
The present invention is not construed as limiting the ratio of I-131 and albumin, can arbitrarily regulate and control.In the embodiment of the present invention, fill altogether Carrying the ratio of I-131 and albumin in the albumin of taxol and I-131 is 200 μ Ci:3mg.
The preparation method of the albumen provided by the invention for loading chemotherapeutics and radiotherapeutic drug altogether, comprising: chemotherapeutics dress After being loaded onto albumen, radiotherapeutic drug is loaded；Alternatively, radiotherapeutic drug is loaded to albumen, chemotherapeutics is loaded.
In the present invention, the solvent for loading the chemotherapeutics is ethyl alcohol, and the condition of the loading is stirring.
In the present invention, the solution for loading the radiotherapeutic drug is water, and catalyst is toluene-sodium-sulfonchloramide；The condition of the loading is Concussion.
In certain embodiments, the preparation side of the albumen provided by the invention for loading chemotherapeutics and radiotherapeutic drug altogether Method are as follows: radiotherapy nucleic I-131 is tagged on albumin, molten chemotherapeutics PTX in ethanol is then added, is stirred overnight, Centrifugation removal precipitating, material are dispersed in supernatant.
In this embodiment, the source of I-131 is Na131I, Na131The ratio of I and HSA is 100 μ Ci:2mg.
The concentration of HSA is 2mg/mL in HSA aqueous solution.
The catalyst of I-131 is loaded as toluene-sodium-sulfonchloramide, the mass ratio of toluene-sodium-sulfonchloramide and HSA are 0.3:2.
Toluene-sodium-sulfonchloramide is dissolved in PB buffer, concentration 10mg/mL.
By Na131I, the loading of I-131 is carried out after HSA aqueous solution, the mixing of toluene-sodium-sulfonchloramide solution.
The condition for loading I-131 is concussion, and time 20min, reaction temperature is room temperature.
In order to remove unreacted Na131I and toluene-sodium-sulfonchloramide further include ultrafiltration step after loading I-131.The ultrafiltration uses 10kDa super filter tube.
After loading I-131, obtain131The HSA of I label, is denoted as131I-HSA。
?131PTX is loaded on I-HSA, PTX is dissolved in ethyl alcohol, concentration 20mg/mL.
In order to make the mass ratio of the material 1: 5 of HSA and PTX in products therefrom, when loading PTX, the mass ratio of PTX and HSA For 1:10.
It will be by ultrafiltration131I-HSA solution is mixed with PTX ethanol solution, carries out PTX loading.
The condition for loading PTX is stirring, and the time is 8h~12h, and reaction temperature is room temperature.
After stirring, through centrifugation removal precipitating, product131I-HSA-PTX is dispersed in supernatant.
The condition of centrifugation is 14800 revs/min, 5 minutes.
Preparation method in another embodiment is, the albumen provided by the invention for loading chemotherapeutics and radiotherapeutic drug altogether The preparation method comprises the following steps: PTX is molten in ethanol, it is added in albumin aqueous solution, is stirred overnight, centrifugation removal precipitating, intermediate It is dispersed in supernatant.Then, radiotherapy nucleic I-131 is tagged on intermediate, obtains final material.
In this embodiment, in order to make the mass ratio of the material 1:5 of HSA and PTX in products therefrom, when loading PTX, PTX Mass ratio with HSA is 1:13.
HSA aqueous solution is mixed with PTX ethanol solution, carries out PTX loading.
In the ethanol solution of PTX, the concentration of PTX is 20mg/mL.
The concentration of HSA is 2mg/ml in HSA aqueous solution.
The condition for loading PTX is stirring, and the time is 8h~12h, and reaction temperature is room temperature.
After stirring, through centrifugation removal precipitating, product HSA-PTX is dispersed in supernatant.
The condition of centrifugation is 14800 revs/min, 5 minutes.
The source of I-131 is Na131I, Na131The ratio of I and HSA is 100 μ Ci:2mg.
The catalyst of I-131 is loaded as toluene-sodium-sulfonchloramide, the mass ratio of toluene-sodium-sulfonchloramide and HSA are 0.3:2.
Toluene-sodium-sulfonchloramide is dissolved in PB buffer, concentration 10mg/mL.
By Na131I, the supernatant containing HSA-PTX, toluene-sodium-sulfonchloramide solution carry out the loading of I-131 after mixing.
The condition for loading I-131 is concussion, and time 20min, reaction temperature is room temperature.
In order to remove unreacted Na131I and toluene-sodium-sulfonchloramide further include ultrafiltration step after loading I-131.The ultrafiltration uses 10kDa super filter tube.
After loading I-131, obtain131The HSA of I label, is denoted as131I-HSA。
?131PTX is loaded on I-HSA,
The albumen of loading chemotherapeutics and radiotherapeutic drug altogether provided by the invention is preparing the application in tumor imaging agent.
The albumen provided by the invention for loading chemotherapeutics and radiotherapeutic drug altogether, wherein the ray of radiotherapeutic drug transmitting is available In the imaging of tumour.Experiments indicate that using I-131 as radiotherapeutic drug, using PTX as chemotherapeutics, using albumin as carrier, It is obtained131I-HSA-PTX is injected into model mice body, the image after the image of CT imaging and gamma imaging merges, table The position of bright gamma ray is concentrated mainly on tumor locus, the readability of imaging to be significantly better than free I-131 or131I- HSA.Illustrate, it is provided by the invention131I-HSA-PTX can have very high enrichment at the position of tumour, and load PTX does not have Have an impact the imaging effect of I-131, plays positive influence to imaging effect instead.
The albumen of loading chemotherapeutics and radiotherapeutic drug altogether provided by the invention is in the drug that preparation inhibits tumour growth Application.
Tumor imaging not only may be implemented in radiotherapy nucleic I-131, but also oncotherapy may be implemented；Small-molecule drug PTX It is not only chemotherapeutics, kills cancer cell, and tumour oxygen content can be improved.According to the prior art,131I-HSA or HSA- The effect and use that PTX is used in combination131The effect of I-HSA-PTX is compared,131Killing ability of the I-HSA-PTX to tumour cell Than by individual HSA-PTX and131The united effect of I-HSA is more significant (p < 0.05).
The albumen provided by the invention for loading chemotherapeutics and radiotherapeutic drug altogether is in preparation detection tumour and/or treatment tumour Product in application.
The albumen of loading chemotherapeutics and radiotherapeutic drug altogether provided by the present invention can be used in the detection or treatment of tumour. It is obtained using albumin as carrier using PTX as chemotherapeutics using I-131 as radiotherapeutic drug131I-HSA-PTX can be to more Kind tumour plays the role of good treatment and detection.In the embodiment of the present invention,131I-HSA-PTX is to breast cancer (breast cancer cell Good inhibiting effect 4T1) is played, it can be in the good enrichment of tumor locus.In this embodiment, tumour is breast cancer.
The present invention also provides detection tumour and/or the method for treating tumour, this method includes being provided using the present invention Total loading chemotherapeutics and radiotherapeutic drug albumen.
The method of detection tumour is after giving the albumen provided by the invention for loading chemotherapeutics and radiotherapeutic drug altogether, into The imaging of row gamma.
The method for treating tumour are as follows: give the albumen provided by the invention for loading chemotherapeutics and radiotherapeutic drug altogether.
The present invention provides the total albumen for loading chemotherapeutics and radiotherapeutic drug, in some embodiments, fill on albumin Carry I-131 and PTX.The chemotherapeutics and the albumen of radiotherapeutic drug provided by the invention of loading altogether has good water-soluble and biology Compatibility can be realized the combination therapy of chemotherapy and radiotherapy, is effectively reduced between capable of effectively reducing tumor tissues and presses, and improve swollen The weary oxygen of tumor improves material in the homogeneous distribution of tumour, and then improves the therapeutic effect of radiotherapy.And preparation method is very simple It is single.
Detailed description of the invention
Fig. 1 synthesizes the albumen for loading chemotherapeutics and radiotherapy nucleic131The schematic diagram of I-HSA-PTX, wherein Fig. 1-a shows The synthesis path of embodiment 1, Fig. 1-b show the synthesis path of embodiment 2；
Fig. 2 show HSA,131I-HSA, HSA-PTX and131I-HSA-PTX is hydrated particle diameter distribution；
Fig. 3 shows the fluorescence co-focusing photo that 4T1 cell is incubated for the HSA-PTX of fluorescent marker and HSA respectively；Wherein, scheme 3-a shows that laser co-focusing proves the cell endocytic of the HSA and HSA-PTX of fluorescent marker；Fig. 3-b shows that flow cytometer proves fluorescence The cell endocytic of the HSA and HSA-PTX of label；Fig. 3-c shows HSA the and HSA-PTX cell of flow cytometer test fluorescent marker The fluorescence intensity of endocytosis；
Fig. 4 shows131I-HSA-PTX、131Treatment of the I-HSA and HSA-PTX in cellular level, wherein * * show with131I-HSA Effect, which is compared, has extremely significant sex differernce, p < 0.01；* * shows has extremely significant sex differernce compared with HSA-PTX effect, and p < 0.001, p value is calculated with ANOVA；
Fig. 5 shows131I-HSA-PTX、131I-HSA and HSA-PTX passes through different time points in tail vein injection to Mice Body Gamma imaging；
Fig. 6 shows131In point of each tissue when I-HSA-PTX is by tail vein injection to Mice Body in different time points Cloth；
After Fig. 7 shows tail vein injection HSA-PTX, the photoacoustic imaging of mouse tumor in different time points；Wherein, Fig. 7-a shows The photoacoustic imaging blood oxygen saturation of tumour different time points after tail vein injection HSA-PTX；Fig. 7-b shows tumour under different frame numbers Blood oxygen saturation；Tumour blood oxygen saturation when Fig. 7-c shows different time points；
Fig. 8 show mouse distinguish tail vein injection HSA-PTX and HSA24 hours after tumour immunity fluorescence；Wherein, Fig. 8-a is small Weary oxygen immunofluorescence slice after tail vein injection HSA and HSA-PTX；After Fig. 8-b mouse tail vein injection HSA and HSA-PTX Hypoxia-inducible factor lα immunofluorescence slice；Weary oxygen immunofluorescence after Fig. 8-c mouse tail vein injection HSA and HSA-PTX；Figure Hypoxia-inducible factor lα immunofluorescence after 8-d mouse tail vein injection HSA and HSA-PTX；
Fig. 9 shows vivo tumor combination therapy experimental result picture；Fig. 9-a shows combination therapy tumor growth curve；Fig. 9-b shows The volume of mouse tumour after various processing 19 days, Predicted additive effect are that calculated estimated tumour is independent Radiotherapy and independent chemotherapy combined treatment (131I-HSA+HSA-PTX effect)；* show with significant difference, p < 0.05；* shows tool There are extremely significant sex differernce, p < 0.01；* * shows that with extremely significant sex differernce, p < 0.001, p value is calculated with ANOVA；
Figure 10 shows mouse after various processing, and tumor tissue section H&E and TUNEL are dyed.
Specific embodiment
The present invention provides a kind of albumen and its application for loading chemotherapeutics and radiotherapeutic drug altogether, those skilled in the art Present disclosure can be used for reference, realization of process parameters is suitably modified.In particular, it should be pointed out that all similar substitutions and modifications pair It is it will be apparent that they are considered as being included in the present invention for those skilled in the art.Method and application of the invention is Through being described by preferred embodiment, related personnel obviously can not depart from the content of present invention, in spirit and scope to this The methods and applications of text are modified or appropriate changes and combinations, carry out implementation and application the technology of the present invention.
Refer to body under the effect of the various tumorigenesis factors for tumour (tumour), local organization hyperplasia is formed by newly Biological (neogrowth).The classification of tumour usually occurs as foundation with tissue, each classification press again its differentiation and maturation degree and its Benign and malignant two major classes are divided on the difference that body influences.Common tumour betides: rectum, colon, mammary gland, uterus, Stomach, liver, lung, bone, neck etc..
Chemotherapeutics refers mainly to chemicals, can act on growth of tumour cell breeding different links on, inhibit or Tumour cell is killed, is one of the main means of current treatment tumour.
Radiotherapeutic drug is mainly radionuclide, is controlled using α, β, gamma-rays and all kinds of X-rays that radionuclide generates Kill cancer cell in the irradiation such as X-ray, electron beam, neutron beam, proton beam and other particles beams that treatment machine or accelerator generate affected part.
Tumor imaging, which refers to by detecting radioactive method, observes distribution of the radioactive isotope in human internal organ, To diagnose internal organs with the presence or absence of the position where lesion and determining lesion.
Reagent that the present invention uses, animal, cell are all common commercially available product, can all be bought in market.
Below with reference to embodiment, the present invention is further explained:
The preparation of embodiment 1 loads the albumen of chemotherapy and radiation drug altogether131I-HSA-PTX
Preparation process is as shown in Fig. 1-a, using albumin HSA as substrate, by reprinting chemotherapeutics and radiotherapy nucleic.
Take the Na of 100 μ Ci131The HSA aqueous solution of the 2mg/ml of 1ml is added in I, adds the toluene-sodium-sulfonchloramide of the 10mg/ml of 30 μ L Solution (is dissolved in PB buffer), and concussion shakes up 20 minutes, with the super filter tube ultrafiltration of 10kDa, removes unreacted Na131I and chlorine Amine T, obtains131The HSA solution of I label.Then PTX (molten in ethanol) (additional amount meeting of PTX of the 20mg/ml of 13 μ L is added Change the size of nano particle, the amount of PTX is more, and nano particle diameter is bigger), it is stirred overnight, 14800 revs/min are centrifuged 5 points Clock removal precipitating, product131I-HSA-PTX is dispersed in supernatant
The preparation of embodiment 2 loads the albumen of chemotherapy and radiation drug altogether131I-HSA-PTX
Preparation process is as shown in Fig. 1-b, using albumin HSA as substrate, by reprinting chemotherapeutics and radiotherapy nucleic.
By the PTX of the 20mg/ml of 13ul (molten in ethanol), the HSA aqueous solution of the 2mg/ml of 1ml is added, is stirred overnight, 14800 revs/min of centrifugation removal in 5 minutes precipitatings, intermediate HSA-PTX are dispersed in supernatant.Then it is added in supernatant The Na of 100uCi131The toluene-sodium-sulfonchloramide solution (being dissolved in PB buffer) of the 10mg/ml of I and 30ul, concussion shake up 20 minutes, use The super filter tube ultrafiltration of 10KDa, removes unreacted Na131I and toluene-sodium-sulfonchloramide, obtain final product131I-HSA-PTX。
Embodiment 3131The characterization of I-HSA-PTX
To made from embodiment 1 or 2131I-HSA-PTX is characterized, embodiment 1 or 2 gained131I-HSA-PTX's swashs Photokinesis light scatters result as shown in Fig. 2, the hydration radius of original HSA and radiolabeled HSA are about 8nm, radioactivity Mark the size without changing HSA；Intermediate HSA-PTX and final product131The hydration radius of I-HSA-PTX may each be about 110nm, radioactive label do not change scantling and the loading of PTX.
The cell endocytic of 4 material of embodiment
By the HSA of fluorescent marker and HSA-PTX respectively with 4T1 cell incubation 6 hours, free material is washed away with PBS, carefully Born of the same parents fix, DAPI is dyed, endocytosis of the confocal laser scanning microscope cell to material.Then with further with flow cytometer Endocytosis of the quantitative test cell to material.
As a result as shown in figure 3, apparent red fluorescence shows that material by cell endocytic, and does not enter nucleus in cell. Flow cytometry data further demonstrates cell to the endocytosis of material.
Embodiment 5131I-HSA-PTX is in cell-based therapies
By various concentration131I-HSA-PTX、131I-HSA and HSA-PTX and MTT is tested after 4T1 cell incubation 72 hours.
Wherein,131The concentration of I-HSA-PTX is followed successively by 0,6.25 μ Ci/mL131I and 0.15 μ g/mL PTX, 12.5 μ Ci/ mL 131I and 0.31 μ g/mL PTX, 25 μ Ci/mL131I and 0.62 μ g/mL PTX, 50 μ Ci/mL131I and 1.25 μ g/mL PTX、100μCi/mL 131I and 2.5 μ g/mL PTX, 200 μ Ci/mL131I and 5 μ g/mL PTX；
131In I-HSA131The concentration of I is followed successively by 0,6.25 μ Ci/mL, 12.5 μ Ci/mL, 25 μ Ci/mL, 50 μ Ci/mL, 100 μCi/mL,200μCi/mL；
The concentration of PTX is 0,0.15 μ g/mL, 0.31 μ g/mL, 0.62 μ g/mL, 1.25 μ g/mL, 2.5 μ g/ in HSA-PTX mL、5μg/mL。
As a result as shown in figure 4, statistics indicate that131The cell activity of I-HSA-PTX is significantly lower than other control groups, shows131I-HSA-PTX has stronger radiotherapy and chemotherapy combined therapeutic effect.Its significant effect is better than131I-HSA (p < 0.001) or HSA-PTX(p<0.01)。
Embodiment 6131I-HSA-PTX passes through tail vein injection to intravital gamma imaging experiment
It randomly chooses nude mice of the back with 4T1 tumour and carries out imaging test.
By what is dissociated131I(131I 200μCi)、131I-HSA(131200 μ Ci of I) and131I-HSA-PTX(131I200 μ Ci, PTX 9mg/kg) by carrying out point in different times in toy gamma imaging system in tail vein injection to Mice Body Acquisition picture in real time is carried out, the enriching quantity of tumor locus is observed.
As a result free as shown in figure 5, over time131I is metabolized away from bladder quickly, in tumor region It is not enriched with significantly；131I-HSA and131I-HSA-PTX is gradually increased in the gamma ray signal of tumor region, other tissues Gamma ray signal gradually die down, show131I-HSA and131I-HSA-PTX is gradually decomposed and is metabolized.Fig. 5 is by mouse CT imaging and gamma imaging image merge after image, show that the position of gamma ray is concentrated mainly on tumor locus.Fig. 5 The result shows that,131The imaging effect of I-HSA-PTX is significantly better than131I-HSA or free131I。
Embodiment 7131The distribution experiments of I-HSA-PTX in vivo
It randomly chooses nude mice of the back with 4T1 tumour and carries out imaging test.
It will131By in tail vein injection to Mice Body, (dosage is I-HSA-PTX131200 μ Ci, PTX 9mg/kg of I), 24 Or after 72 hours, experiment mice sacrifice takes out important organ weighing and is placed in gamma counter.
As a result as shown in fig. 6,131I-HSA-PTX is woven with higher enrichment in tumor group.After 72 hours, other groups are enriched in Knitting becomes very low with the material content of organ, is gradually decomposed and is metabolized；But it is still with higher in tumor tissues Enrichment.
Embodiment 8HSA-PTX improves tumor hypoxia in living body level
Influence of the HSA-PTX to tumour oxygen content is directly tested first with photoacoustic imaging, is had containing 1 back is measured The mouse of 4T1 tumour is from tail vein injection HSA-PTX (PTX 9mg/kg), respectively 0,12,24 and 48 hour test tumor area The blood oxygen saturation in domain.As a result as shown in fig. 7, the blood oxygen saturation at mouse tumor position is significantly raised after injection HSA-PTX, 48 Blood oxygen saturation shows the sustainable oxygen content for improving tumor tissues of HSA-PTX still in higher level after hour.
Then illustrate that HSA-PTX improves the weary oxygen of mouse tumor by immunofluorescence dyeing.It is swollen with 4T1 to choose two backs The mouse distribution of tumor is from tail vein injection HSA-PTX and HSA, the Pimonidazole of injection 60mg/kg at 22.5 hours, and 24 hours The tumour of mouse is taken afterwards, and frozen section, carries out photoacoustic imaging measurement tumour blood oxygen saturation (Fig. 7) and determination of immunofluorescence method is weary Oxygen and hypoxia-inducible factor lα expression (Fig. 8).The result shows that tumour after relative injection HSA, mouse injection HSA-PTX Weary oxygen region significantly reduces.Hypoxia-inducible factor lα expression is significantly reduced as shown in Fig. 7-b.To injection131I-HSA- Tumor hypoxia is detected with hypoxia-inducible factor lα expression after PTX, and result is similar to HSA-PTX, and no conspicuousness is poor Different (p > 0.05).
Embodiment 9HSA-PTX oncotherapy in living body level
The nude mice that 20 backs have 4T1 tumour is chosen, is randomly divided into four groups, every group five, respectively at interval of three days tails Intravenous injection physiological saline, HSA-PTX (PTX 9mg/kg),131I-HAS(131200 μ Ci of I) and131I-HSA-PTX(131I 200 μ Ci, PTX 9mg/kg), co-injection four times.Tumor growth curve result as shown in Fig. 9~10, comparison HSA-PTX and131I-HSA Group,131I-HSA-PTX inhibits the growth of tumour more significantly；And by calculating discovery,131I-HSA-PTX ratio will be independent HSA-PTX and131The united effect of I-HSA is more significant.After four injections treatment, tumor tissues are removed.Pass through H&E Dyeing and TUNEL dyeing discovery, pass through131I-HSA-PTX processing, most of tumour cell are dead.
The above is only the preferred embodiment of the present invention, it is noted that those skilled in the art are come It says, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications also should be regarded as Protection scope of the present invention.

Claims (4)

1. a kind of albumen for loading chemotherapeutics and radiotherapeutic drug altogether；
The albumen is albumin；The chemotherapeutics is taxol；The radiotherapeutic drug is I-131；
The molar ratio of the albumen and chemotherapeutics is 1: 5；
The preparation method of the albumen for loading chemotherapeutics and radiotherapeutic drug altogether includes: that chemotherapeutics is loaded to albumen, dress Carry radiotherapeutic drug；Alternatively, radiotherapeutic drug is loaded to albumen, chemotherapeutics is loaded；
The solvent for loading the chemotherapeutics is ethyl alcohol, and the condition of the loading is stirring；
The solution for loading the radiotherapeutic drug is water, and catalyst is toluene-sodium-sulfonchloramide；The condition of the loading is concussion.
2. the albumen for loading chemotherapeutics and radiotherapeutic drug described in claim 1 altogether is preparing the application in tumor imaging agent.
3. the albumen of chemotherapeutics and radiotherapeutic drug is loaded described in claim 1 altogether in the drug that preparation inhibits tumour growth Using.
4. the albumen of chemotherapeutics and radiotherapeutic drug is loaded described in claim 1 altogether in preparation detection tumour and/or treatment tumour Product in application.
CN201610668501.6A 2016-08-15 2016-08-15 A kind of albumen and its application loading chemotherapeutics and radiotherapeutic drug altogether Active CN106267228B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610668501.6A CN106267228B (en) 2016-08-15 2016-08-15 A kind of albumen and its application loading chemotherapeutics and radiotherapeutic drug altogether

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610668501.6A CN106267228B (en) 2016-08-15 2016-08-15 A kind of albumen and its application loading chemotherapeutics and radiotherapeutic drug altogether

Publications (2)

Publication Number Publication Date
CN106267228A CN106267228A (en) 2017-01-04
CN106267228B true CN106267228B (en) 2019-08-23

Family

ID=57670317

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610668501.6A Active CN106267228B (en) 2016-08-15 2016-08-15 A kind of albumen and its application loading chemotherapeutics and radiotherapeutic drug altogether

Country Status (1)

Country Link
CN (1) CN106267228B (en)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104043135A (en) * 2014-06-13 2014-09-17 苏州大学 Albumin indocyanine green paclitaxel compound as well as preparation method and application thereof
CN105148293A (en) * 2015-08-04 2015-12-16 山东省医学科学院放射医学研究所 Iodine-131 fibrous protein as well as preparation method and application thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104043135A (en) * 2014-06-13 2014-09-17 苏州大学 Albumin indocyanine green paclitaxel compound as well as preparation method and application thereof
CN105148293A (en) * 2015-08-04 2015-12-16 山东省医学科学院放射医学研究所 Iodine-131 fibrous protein as well as preparation method and application thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
"The use of 131I labelled human serum albumin in the assessment of improved lymph flow following buried dermis flap operation in cases of post-mastectomy lymphoedema of the arm";By R. F. Harvey，et al;《British Journal of Radiology》;19690531;第42卷(第496期);摘要 *

Also Published As

Publication number Publication date
CN106267228A (en) 2017-01-04

Similar Documents

Publication Publication Date Title
CN106572993B (en) Application of the EP4 antagonist in the drug of preparation treating cancer
US20200276230A1 (en) Particles for the treatment of cancer in combination with radiotherapy
CN108377643A (en) The therapy of the cell death of cancer cell is deprived by iron death induction nutrient using extra small nano-particle
CN111888336B (en) Calcium carbonate poly (lactic acid-glycolic acid) composite particles and preparation and application thereof
US7803350B2 (en) Radioactive arsenic-containing compounds and their uses in the treatment of tumors
Sheng et al. Theranostic radioiodine-labelled melanin nanoparticles inspired by clinical brachytherapy seeds
CN107213479B (en) A kind of composition and purposes comprising catalase
Hansen et al. Liposome accumulation in irradiated tumors display important tumor and dose dependent differences
WO2018097335A1 (en) Novel bsh complex for boron neutron capture therapy
CN106267228B (en) A kind of albumen and its application loading chemotherapeutics and radiotherapeutic drug altogether
CN108144074A (en) Melanin nano particle, hydrogel and the preparation method of a kind of radioiodination, application
CA3047152A1 (en) Radiolabelled material for targeted administration
CN106606783B (en) A kind of targeting is passed altogether to be released the drug of photosensitizer and chemotherapeutics and passs release system
Guo et al. Radioiodine based biomedical carriers for cancer theranostics
Zhang et al. Radionuclide 131I-labeled albumin-indocyanine green nanoparticles for synergistic combined radio-photothermal therapy of anaplastic thyroid cancer
Herman et al. Trimodality therapy (drug/hyperthermia/radiation) with BCNU or mitomycin C
CA3178425A1 (en) Gold nanoparticle-containing medicine
CN113456832A (en) Transferrin-modified antibody-entrapped nanoparticle and application thereof
Novell et al. Therapeutic aspects of radio-isotopes in hepatobiliary malignancy
KR20220123260A (en) Chitosan hydrogel-chelator manufacturing method for cancer treatment
KR101942196B1 (en) Composition for multiple imaging agent comprising metal ions contained melanin nanoparticle
US20120134918A1 (en) Gum arabic coated 198gold radioactive nanoparticles for cancer therapy
EP1477174B1 (en) Radioactive arsenic-containing compounds and their uses in the treatment of tumors
US20230390248A1 (en) Composition for enhancing tumor penetration of anticancer drug, comprising losartan as active ingredient, and use thereof
JP6598395B2 (en) Intravenous formulations containing boron amino acids and boron neutron capture therapy

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
TR01 Transfer of patent right
TR01 Transfer of patent right

Effective date of registration: 20201104

Address after: Unit 102-1, building A4, 218 Xinghu street, Suzhou Industrial Park, Suzhou area, China (Jiangsu) pilot Free Trade Zone, Suzhou City, Jiangsu Province

Patentee after: Suzhou Baimai biomedical Co.,Ltd.

Address before: 215123 No. 199 benevolence Road, Suzhou Industrial Park, Jiangsu, China

Patentee before: Suzhou University