CN1951495B - Nano target medicine for magnetothermal therapy of malignant tumor - Google Patents

Nano target medicine for magnetothermal therapy of malignant tumor Download PDF

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CN1951495B
CN1951495B CN2006101319946A CN200610131994A CN1951495B CN 1951495 B CN1951495 B CN 1951495B CN 2006101319946 A CN2006101319946 A CN 2006101319946A CN 200610131994 A CN200610131994 A CN 200610131994A CN 1951495 B CN1951495 B CN 1951495B
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magnetic powder
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target medicine
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CN1951495A (en
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朱宏
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Abstract

The invention relates to a method for preparing the nanometer target drug used to treat cancer. Wherein, it is a nanometer magnetic-antibody/ligand drug and nanometer magnetic target drug used in cancer magnetic thermal treatment, with high specific thermal power; it comprises 1:0.0001-0.20 effective molecule and guide molecule, while the effective molecule is magnetic powder whose diameter is lower than 1000nm, and the specific thermal power SAR is 10-7000W/gFe; the guide molecule comprises antibody or ligand or magnetic powder; the diameter of target drug is 2-1000nm; the preparation comprises that coupling the magnetic powder and guide molecule in water, organic or inorganic mixture. The inventive drug can effective kill cancer cell and treat cancer.

Description

Nano target medicine for magnetothermal therapy of malignant tumor
Technical field
The present invention relates to a kind of nano target medicine for magnetothermal therapy of malignant tumor and preparation method thereof and application, nano-magnetic powder-antibody that promptly a kind of magnetothermal therapy of malignant tumor of high yield ratio of specific heat power is used/part targeted drug and nano-magnetic powder magnetic targeted drug.
Background technology
Global New Development malignant tumor case in 2000 is about 1,000 ten thousand, and is dead 6,200,000, existing ill example 2,200 ten thousand.Estimate that the year two thousand twenty malignant tumor new cases will reach 1,500 ten thousand, dead 1,000 ten thousand, existing ill example 3,000 ten thousand.Malignant tumor is becoming human first killer of new century, will bring about great losses to global social economy, carries out the malignant tumor newtype drug and treat Study on new method to have very significant meaning.
Traditional treating malignant tumor method mainly is operation, radiation and chemotherapy.Usually have only the malignant tumor patient of 10%-20% to have an opportunity to perform the operation, and perform the operation and easily bring bigger misery to patient, postoperative recurs easily and shifts; And the radiation and chemotherapy targeting is indeterminate, also destroys normal cell when killing tumor cell, and toxic and side effects is bigger.In recent years, owing to have the appearance of the targeted drug of characteristics such as efficient, low toxicity, side effect are little, the targeted drug of effect molecules such as antibody and ligand coupling active nucleus, toxin, chemotherapeutics, prodrug has become the antineoplastic novel drugs of broad research and application and the new tool for the treatment of malignant tumor, and the research of targeted drugs such as antibody and part provides new opportunity and effect with the treatment and the prevention that are applied as malignant tumor.But, effect molecules such as link coupled active nucleus such as antibody or part, toxin, chemicals, prodrug still have stronger toxic and side effects and defective such as limitation significantly, the novel effector molecule that treating malignant tumor need have no side effect with targeted drug are arranged in clinical practice in the above-mentioned targeted drug.
Magnetothermal therapy of malignant tumor is that a kind of following four kinds of physics and bioequivalence of utilizing answered, and promptly (1) magnetic heating seed (being magnetic-particle or material, as nano-magnetic powder) can produce magnetothermal effect by the outer alternating magnetic field of inductor; (2) blood flow of malignant tumor has only 10%～15% of normal structure, after the heating tumor acceleration of tumor normal surrounding tissue blood flow, rapid heat dissipation, temperature rise little, and heat radiation difficulty in the tumor, temperature can projecting normal structures 5 ℃～10 ℃; (3) normal cell is dead more than 48 ℃, and malignant cell is greater than 42 ℃ of death, tumor cell poor heat resistance; (4) alternating magnetic field of lower frequency penetrance in tissue good, unattenuated, do not damage normal structure, can arrive the inside tumor of deep and large volume safely and effectively; Making the magnetic heating seed that imports tumor focus produce magnetic hysteresis under the action of alternating magnetic field of magnetic thermotherapy treatment instrument gives birth to heat effect, heats malignant tumor to 42 ℃～48 ℃, and normal structure does not heat up substantially, realizes accurately locating and the thermotherapy new method of the selectively killing malignant tumor that has no side effect.In recent years, a large amount of animal experiments and clinical experimental study achievement proved, used magnetic thermotherapy treatment malignant tumor and not only can effectively kill tumor, tumor is disappeared fully, and do not have toxic and side effects.The magnetic thermotherapy is a kind of oncotherapy new method with broad prospect of application, and its safety that appears as malignant tumor is cured provides hope.Great deal of research results proves that the magnetic heating seed in the magnetic thermotherapy is a kind of targeted drug physical effect molecule that has no side effect.But up to now, magnetic heating seed still need import tumor focus by modes such as heeling-in, local injections in the magnetic thermotherapy, and Shang Weijian makes the magnetic thermotherapy with targeted drug initiatively with utilize the magnetic behavior of magnetic heating seed to make the magnetic thermotherapy with magnetic targeted drug and utilize active, the magnetic target function of medicine to make drug targeting import tumor focus, be used for the bibliographical information of magnetothermal therapy of malignant tumor treatment with antibody or part and magnetic heating seed (being nano-magnetic powder) coupling.
Summary of the invention
The objective of the invention is provides a kind of nano target medicine at the deficiencies in the prior art part, be a kind of magnetothermal therapy of malignant tumor nano-magnetic powder-antibody target medicine, nano-magnetic powder-part targeted drug and nano-magnetic powder magnetic targeted drug of high calorific capacity, just have antibody-mediated, the receptor-mediated nano-magnetic powder targeted drug that the magnetothermal therapy of malignant tumor of high yield ratio of specific heat power or S. E. A. (SAR, specific absorption rate) uses and the nano-magnetic powder magnetic targeted drug of magnetic guiding.
Nano-magnetic powder nano target medicine provided by the invention (comprising nano-magnetic powder-antibody nano target medicine and nano-magnetic powder-part targeted drug) is meant the active targeted drug that guide molecule (as antibody or part) and nano-magnetic powder (being magnetic heating seed) coupling are made, the nano-magnetic powder nano target medicine also is meant the magnetic thermotherapy magnetic targeted drug that nano-magnetic powder process surface modification is modified or bag is wrapped and is made into, and used nano-magnetic powder SAR is 10～7000W/gFe, particularly 500～7000W/gFe.
The present invention takes following technical scheme to realize:
The invention provides a kind of nano target medicine for magnetothermal therapy of malignant tumor, comprising the quality proportioning 1: 0.0001～0.20 effector molecule and guide molecule, described effector molecule is a nano-magnetic powder, this magnetic powder particle diameter is no more than 1000nm, heat absorption ratio SAR is 10～7000W/gFe, described guide molecule comprises antibody and part, and the particle diameter of this targeted drug is 2～1000nm.
Above-mentioned nano target medicine for magnetothermal therapy of malignant tumor, its conjugate of making by solution mixing coupling with described nano-magnetic powder and antibody or part constitutes, nano-magnetic powder wherein is the magnetic-particle through surface modification is modified or bag is wrapped and makes, it also can be untreated naked nano-magnetic powder, the quality proportioning of nano-magnetic powder and antibody or part is 1: 0.0001～0.20 in the conjugate, but the nano-magnetic powder surface of conjugate 1 antibody of coupling or part, also a plurality of antibody of coupling or part at least; The conjugate particle grain size is 2～1000nm, and the biological activity that it remains with coupling antibody or part has antibody-mediated or receptor-mediated active target function, magnetic target function and nano-particle passive target function.
Magnetothermal therapy of malignant tumor of the present invention mainly is to be made of the magnetic-particle that nano-magnetic powder is made through surface modification, modification or bag quilt, parcel or undressed naked nano-magnetic powder with the nano-magnetic powder magnetic targeted drug.Repairing, adorn or wrap by the quality proportioning of nano-magnetic powder and surface modification trim in the magnetic-particle of, parcel through surface modification is 1: 0.0001～0.20, the quality proportioning of nano-magnetic powder and pan coating, wrappage is 1: 0.01～5.0, modification, modification or bag quilt, coated magnetic particle grain size are 2～1000nm, and it has magnetic target function and nano-particle passive target function.
The preferred 2-400nm of targeted drug particle diameter of the present invention.When diameter of aspirin particle during less than 100nm, nano target medicine has the long stable circulation performance of good body inner blood and can effectively utilize the EPR effect (enhancing permeability andretention effects) of tumor, and the targeted drug particle diameter is to be advisable less than 100nm usually.
Above-mentioned conjugate or magnetic-particle become nano target medicine with solution mixing system, and this solution comprises double solvent and/or water (preferred deionized water).Described nano target medicine is based on injection, in injection, can make medicine keep good dispersibility and stability, have good penetrance and with malignant cell antigen or good specificity and the affinity of receptor.It is used in vivo and in vitro all has good targeting, and with the normal cell no cross reaction.
Nano target medicine of the present invention is by administrations such as arteriovenous injection, lumbar injection or direct (comprising: Wicresoft, intervention, injection etc.) importings, but dense poly-being positioned at antibody or part of its high power has malignant tumor focus position than high specific and affinity, also but dense gathering of high power is positioned at malignant tumor focus position interior or that put Magnet outward, shows initiatively target function and magnetic target function; Under the effect of magnetic thermotherapy treatment instrument alternating magnetic field, nano-magnetic powder in the medicine of the dense poly-positioning tumor lesions position of high power is because magnetothermal effect is heated to more than 42～48 ℃ and 48 ℃ tumor, tumor is implemented warm and high pyromagnetic thermotherapy treatment of targeting and prevention, can kill cancerous cell effectively, cure malignant tumor.For the ease of storing, nano target medicine of the present invention also can be made into the form of solid or powdery medicine powder body.
Because the targeted drug particle diameter is during less than 100nm, nano target medicine has the long stable circulation performance of good body inner blood and can effectively utilize the EPR effect of tumor, the grain diameter of the nano-magnetic powder that the present invention is used is preferably 1～400nm, more preferably to be advisable less than 100nm.Nano-magnetic powder comprises ferromagnetism, ferrimagnetism and super-paramagnetism nano magnetic powder, as Fe 2O 3, Fe 3O 4, ferromagnetism, ferrimagnetism nano-magnetic powder such as Fe, and particle diameter is less than the Fe of critical dimension 2O 3, Fe 3O 4, super-paramagnetism nano magnetic powder such as Fe.
Above-mentioned nano-magnetic powder preferably passes through surface modification, modify or the bag quilt, parcel, refer in particular to the nano-magnetic powder surface through surfactant/surface modifier modification, modify or by organic substance/inorganic substances bag quilt, parcel, the nano-magnetic powder surface can be through 1 time or surfactant repeatedly, the surface modifier modification, modify or organic substance bag quilt, parcel, purpose is to make nano-magnetic powder have the surface of functionalization, as the anionization surface, the cationization surface, the nonionic surface, or amination surface, the sulfhydrylation surface, the hydroxylating surface, carboxylated surface, or the polymer surfaces of surface property blind date, the biomacromolecule surface, the inorganic substances surface, also can with antibody, part passes through micromolecule, macromole or directly combination, utilize electrostatic force, hydrophobic interaction, covalence key, Van der Waals force, effect such as short range repulsive force and absorption forms link coupled functionalized surface, wherein surfactant and surface modification comprise sodium lauryl sulphate, dodecylbenzene sodium sulfonate, enuatrol, n-capric acid, Polyethylene Glycol (PEG), glucosan, polyglutamic acid, Sensor Chip CM 5 or glycosaminoglycan, dextran, ferrum ammonium (DF), ethylenediamine, diethylenetriamine pentaacetic acid (DTPA), diazanyl niacinamide usp (HYNIC), nucleotide, polypeptide, coupling agent comprises 3-aminopropyl-trimethyl TMOS, the aminopropyl trimethyl silane, arbitrary or combination in γ-methacryloxypropyl trimethoxy silane etc., also can select other nano-magnetic powder surface modification dressing agent for use, the quality proportioning of nano-magnetic powder and surfactant or surface modifier is 1: 0.0001～0.20; Organic substance comprises the arbitrary or combination in liposome, polylactic acid, polycaprolactone, chitosan or the protein etc., inorganic substances are the arbitrary of silicon dioxide, aluminium oxide or Jin Dengzhong or combination, or organic substance and inorganic substances combination, the mass ratio of nano-magnetic powder and organic/electrodeless material is 1: 0.01～5.0; Also can select other organic substance or inorganic substances for use, and organic substance and inorganic substances are arbitrary or the combination.
Described antibody is at tumor specific antigen and tumor associated antigen, idiotypic determinant, the receptor of some cytokine, the product of hormone and some tumor genes is the various antibody of relevant target molecule, comprise the allos polyclonal antibody, immunity blood combine with the cell engineering human monoclonal antibody (McAb) of generation of the Mus resource monoclonal antibody (McAb) that produces and/or immunology that combines with cell engineering, as single-chain antibody (ScFv), the multivalence miniantibody, single domain antibodies, humanized antibody and human antibody etc., wherein molecular weight is less, antibody such as monoclonal antibody fragment that specificity and affinity are higher or genetic engineering antibody are in immunogenicity, the resistance aspect of penetrance and various hydrolytic enzyme all is better than conventional monoclonal antibody, the easier arrival tumor locus that penetrates of its monoclonal antibody than routine.Antibody is selected from anti-CEA (carcinoembryonic antigen), AFP (alpha-fetoprotein), CA19-9 (carbohydrate antigen 19-9), CA125 (cancer antigen 125), PSA (prostate soecificantigen), hCG (hunan chorionic gonadotropin), Trigen, TheraCIMh-R3, Smart ID10, Vitaxin, 4B5, SS1 (dsFv) PE38, ABX-EGF, BrevaRex, CA15-3, PAP, CA50, SCC, β 2M (β 2-microglobulin), arbitrary or its combination among Rituxan, anti-VEGF, epratuzmab, NSE (neuro-specific enolase), HER2 and TPA (tissue polypeptide antigen) antibody and the human liver cancer monoclonal antibody HAb18 (humanizedantibody 18), also can select other antibody for use.
Described part comprises: folic acid part, transferrins, saccharide part, polypeptide (as antibody fragment), nucleic acid, hormone and other endogenous molecule etc., and wherein the folic acid part comprises: folic acid, folinic acid, dihydrofoilic acid, tetrahydrofolic acid, tetrahydrochysene pterin, the many glutaric acid of pterin acyl, 2-deaminize-hydroxyl folic acid, 1-deaminizes-hydroxyl folic acid, 1 denitrification folic acid, 3-denitrification folic acid, 8-denitrification folic acid, 5-methyl tetrahydrofolate (5-CH 3H 4Folate), 5-formoxyl tetrahydrofolic acid (5-CHOH 4Folate), (methotrexate, MTX) and 5, (5,10-CHH folate, also can select other part for use at DDATHF) arbitrary or its combination among a kind of in waiting to the 10-methylene tetrahydrofolate to the antifol methotrexate.
The coupled direct cross-linking method of antibody that is meant of described antibody and nano-magnetic powder, the strong connection method of the glutaraldehyde method that medicine is connected with antibody by micromolecule, cis-aconitic acid anhydride method, active ester method, N-succinoamino-3-(2-pyridine radicals two sulfur) propanoic acid methods (SPDP method), succinic anhydrides, maleic anhydride, glutaric anhydride, hydrazine, oligopeptide, hydazone derivative method and peptide etc., macromole such as glucosan, polyglutamic acid, Sensor Chip CM 5, glycosaminoglycan and polymerization peptide carrier etc. do carrier indirectly in conjunction with cross-linking method.
Described part is meant that with the nano-magnetic powder coupling part passes through micromolecule, macromole or directly combines absorbent-type coupling and the covalent coupling that forms, as avidin-biotin reaction coupling, with the strong link coupled part of acyl ammonia-nano-particle coupling, with the strong link coupled part of thioether-nano-particle coupling, with the strong link coupled part of Schiff-nano-particle coupling.
Described folacin coupled aglucon ferrum ammonium (DF), diethylenetriamine pentaacetic acid (DTPA) and the diazanyl niacinamide usp (HYNIC) of needing usually, these aglucons itself have amino or carboxyl; Coupling is to utilize amino in the aglucon with the carboxyl reaction in the folic acid, and aglucon plays the bifunctional chelating agent effect, and folic acid and other are coupled together; Do not have aminoly in the aglucon, need to connect ethylenediamine or diamine arm in the folate molecule.
Described is coupling in water or organic and inorganic matter (as ethanol, acetic acid, toluene, phosphoric acid, two methylene sulfones) and mixed solution thereof by solution mixing coupling.
Described targeted drug for the nano-particle state is in order to make targeted drug have good penetrance, and has specificity and the affinity good with the related neoplasms cell in injection.This targeted drug makes its initiatively dense poly-being positioned at around the malignant tumor focus of targeting and high power by injection, it in the concentration of this focus 1～1000 times of normal structure blood Chinese medicine concentration, its nano-magnetic powder SAR under the action of alternating magnetic field of magnetic thermotherapy treatment instrument is 10-7000W/gFe, and wherein nano-magnetic powder SAR is advisable with 501～7000W/gFe.
Nano-magnetic powder targeted drug injection of the present invention comprises the injection of making after the targeted drug coupling and faces the injection of preparing with targeted drug drying solid powder body with preceding.
The targeted drug injection of described double solvent or water (preferred deionized water) is that targeted drug is dispersed in normal saline, phosphate buffer (PBS), Polyethylene Glycol, nucleotide etc. preparation or the preparation of medicine in water, deionized water in the double solvent of inorganic, organic, biological, abitotic substance are arranged, the weight ratio of targeted drug and double solvent or water is 1: 0.5～100, and solute and water weight ratio are no more than 1: 5 in the solvent.
The inventor has proposed the new ideas of nano-magnetic powder as a kind of physical effect molecule that has no side effect of antibody target medicine, proposed simultaneously with the nano-magnetic powder is effector molecule, with antibody or part is that the coupling of targeting guide molecule prepares nano-magnetic powder-antibody or part nano target medicine, the New Policy that is used for malignant tumor targeting magnetic thermotherapy, be a kind of nano-magnetic powder and antibody coupling to be made the magnetothermal therapy of malignant tumor targeted drug, be used for making the initiatively dense poly-tumor focus that is positioned at of targeting of targeted drug by quiet tremulous pulse or lumbar injection, and under the effect of external magnetic thermotherapy treatment instrument alternating magnetic field, dense poly-focus nano-magnetic powder produce magnetic hysteresis and give birth to heat effect tumor is heated to more than 42～48 ℃ and 48 ℃, and normal structure does not heat up substantially, the selectively killing malignant tumor, the realization malignant tumor is efficiently killed, the targeting magnetic thermotherapy treatment of safe without toxic side effect and the New Policy of prevention.Simultaneously, developed the nano-magnetic powder technology of preparing that SAR can reach 7000W/gFe.Zoopery proves, uses nano target medicine targeting magnetic thermotherapy of the present invention and can cure malignant tumor fully, and have no side effect.
Targeted drug of the present invention is the nano-particle medicine that the nano-magnetic powder coupling by antibody and different-grain diameter constitutes, because medicine has been realized nanorize, it has good penetrance, biocompatibility in vivo, has long cyclical stability in blood, make it not only have antibody targeted active target function, and have magnetic target function of carrying the magnetic medicine and the particulate passive target function of Nano medication of utilizing the EPR effect of tumor, be a kind of compound targeted drug with active target function and multiple other target function.
Monoclonal antibody fragment in the targeted drug of the present invention, genetic engineering antibody are because its molecular weight is less, than the easier arrival tumor locus that penetrates of the monoclonal antibody of routine.The magnetic thermotherapy that adopt that molecular weight is less, specificity and the higher antibody of affinity constitutes has with nano-magnetic powder-antibody target medicine and better penetrates function and target function.
Targeted drug of the present invention take the nano-magnetic powder as magnetic thermotherapy magnetic heating seed, antibody and part are guide molecule, nano-magnetic powder wherein can pass through arteriovenous or route of administration such as lumbar injection or importing, under target functions such as the guiding of antibody and part and magnetic target function, realize the drug main moving-target to the dense poly-lesions position that is positioned at tumor of magnetic target, under the effect of magnetic thermotherapy treatment instrument alternating magnetic field, the nano-magnetic powder of patient's focus can produce significant magnetic hysteresis and give birth to heat effect, make tumor focus be warming up to 42～95 ℃ rapidly, tumor is carried out 42～48 ℃ magnetic thermotherapy treatment, also can be higher than 48 ℃ magnetic thermotherapy treatment.
Magnetic thermotherapy of the present invention can effectively be killed with antibody by 1 magnetic thermotherapy more than 1 hour and 1 hour usually with the application of targeted drug has malignant tumor than high specific and affinity, even tumor is disappeared fully; Also can effectively kill with antibody and have malignant tumor, even tumor is disappeared fully than high specific and affinity by the magnetic thermotherapy more than 1 hour and 1 hour repeatedly, general therapeutic with drug dose less than 100mg.
Targeted drug of the present invention not only can be used for having targeting magnetic thermotherapy treatment than the malignant tumor of high specific and affinity with antibody, and can be used for having than the prevention of the malignant tumor targeting magnetic thermotherapy of high specific and affinity with antibody, the active targeting magnetic thermotherapy treatment that promptly can be used for detected and undetected malignant tumor, the active targeting magnetic thermotherapy treatment that minimum malignant tumor and imaging equipment can't detected early stage malignant tumor is only arranged, the active targeting magnetic thermotherapy treatment of the early stage malignant tumor when several cell is only arranged, the active targeting magnetic thermotherapy of transfering malignant tumor cell and cancer embolus is killed treatment, and the normal person regularly and aperiodically carries out the prophylactic treatment that malignant cell active targeting magnetic thermotherapy is killed.
The present invention also provides the preparation method of nano-magnetic powder targeted drug, comprises with the quality proportioning being that 1: 0.0001～0.20 magnetic powder and guide molecule coupling in the solution that comprises water or organic substance ethanol, acetic acid, toluene, two methylene sulfones or inorganic substances phosphoric acid or its mixed solution obtain; Preferred earlier described magnetic powder the processing with 1: 0.0001～0.20 part by weight with above-mentioned surfactant and/or surface modifier mixed coupling with guide molecule again in above-mentioned solution.
The present invention also provides the application of above-mentioned nano-magnetic powder targeted drug in the medicine of preparation treatment and prophylaxis of tumours.
Nano target medicine of the present invention has following characteristics: (1) nano-magnetic powder-antibody and part initiatively targeted drug are to be the antibody-mediated or receptor-mediated active targeting magnetic thermotherapy Nano medication of effector molecule with the nano-magnetic powder, have target function initiatively, magnetic and medicated magnetic target function and the passive target function of nano-carrier medicine, wherein said antibody or part are guide molecule of the present invention; (2) the nano-magnetic powder magnetic targeted drug is to be the magnetic target magnetic thermotherapy Nano medication of the magnetic guiding of effector molecule with the nano-magnetic powder with biocompatibility in the good body; (3) not only selecting SAR in the targeted drug for use is the nano-magnetic powder of 10～500W/gFe, and the magnetic powder SAR that selects for use can be up to 500～7000W/gFe, although being the targeted drug that the nano-magnetic powder of 10～500W/gFe and 500～7000W/gFe is made, SAR may be used to tumor magnetic thermotherapy nano target medicine, but if need to be used to 5 * 6 * 7cm cerebroma magnetic thermotherapy treatment of 12W heat energy heating, then tumor focus needs 10,500 and magnetic powder 1.2,0.024, the 0.0017g of 7000W/gFe respectively in theory, and the density of considering ferrum and ferrum oxide is about 7.9 and 5.25g/cm respectively 3The magnetic powder volume that cerebroma magnetic thermotherapy needs in theory is not about 150,3,0.22 μ l and 230,4.6,0.32 μ l with ferrum and ferrum oxide score, the nano-magnetic powder of needs or nano target medicine still less had higher clinical practice performance and economic worth when the nano target medicine that the nano-magnetic powder of 500～7000W/gFe is made was used for the treatment of; (4) nano-magnetic powder in the medicine has no side effect to human body or animal body, can be used for malignant tumor targeting magnetic thermotherapy treatment and prevention safe without toxic side effect at people or animal vivo degradation; (5) nano-magnetic powder-antibody of nano-scale, part targeted drug and nano-magnetic powder magnetic targeted drug have good penetrance, can by arteriovenous injection, lumbar injection or directly route of administration such as (comprising: Wicresoft, intervention, injection etc.) importings make the drug main moving-target to or dense gathering of magnetic target be positioned at the malignant tumor focus; (6) nano target medicine for magnetothermal therapy of malignant tumor of the present invention is applied to malignant tumor initiatively targeting or magnetic target magnetic thermotherapy treatment and prevention, can kill malignant tumor and cancerous cell fully, a kind of targeting magnetic thermotherapy is cured fully, the new tool of safe without toxic side effect for the treatment of malignant tumor and prevention provide.
The specific embodiment
For more clear explanation goal of the invention and technical scheme, be described in further detail by following embodiment.
Embodiment 1
With particle size distribution range is the Fe of 1～5nm, 10～20nm and 20～40nm, 50～100nm, 350～390nm or 950nm 2O 3Or Fe 3O 4Or the Fe nano-magnetic powder adds nano-magnetic powder and enuatrol quality proportioning respectively and is respectively in 1: 0.0001,1: 0.05,1: 0.20 the aqueous solution sodium oleate, ultrasonic mixing 10min, inserting in 85 ℃ of water-baths 600r/min stirred 8 hours, through centrifugalize, except that supernatant, use the deionized water thorough washing, make the nano-magnetic powder of enuatrol modification.
Embodiment 2
With particle size distribution range is the Fe of 5～10nm, 50～100nm, 200～300nm or 950nm 2O 3Or Fe 3O 4Or the Fe nano-magnetic powder adds nano-magnetic powder and n-capric acid quality proportioning respectively and is respectively in 1: 0.0001,1: 0.01,1: 0.20 the n-capric acid aqueous solution, ultrasonic mixing 10min, inserting in 70 ℃ of water-baths 1000r/min stirred 6 hours, through the centrifugal supernatant that removes, use the deionized water thorough washing, make the nano-magnetic powder of n-capric acid modification.
Embodiment 3
With particle size distribution range is the Fe of 30～50nm, 40～80nm, 50～100nm or 250～350nm 2O 3Or Fe 3O 4Or the Fe nano-magnetic powder adds nano-magnetic powder and sodium lauryl sulphate quality proportioning respectively and is respectively in 1: 0.0001,1: 0.01,1: 0.20 the lauryl sodium sulfate aqueous solution, ultrasonic mixing 10min, inserting in 80 ℃ of water-baths 800r/min stirred 1 hour, through centrifugalize, except that supernatant, use the deionized water thorough washing, make the nano-magnetic powder of sodium lauryl sulphate modification.
Embodiment 4
With particle size distribution range is the Fe of 1～5nm, 10～20nm, 20～40nm, 40～80nm or 150～200nm 2O 3X or Fe 3O 4Or the Fe nano-magnetic powder adds nano-magnetic powder and dodecylbenzene sodium sulfonate quality proportioning respectively and is respectively in 1: 0.0001,1: 0.01,1: 0.20 the sodium dodecyl benzene sulfonate aqueous solution, ultrasonic mixing 10min, inserting in 85 ℃ of water-baths 500r/min stirred 1 hour, through centrifugalize, except that supernatant, use the deionized water thorough washing, make the nano-magnetic powder of dodecylbenzene sodium sulfonate modification.
Embodiment 5
Modified Nano magnetic powder in the foregoing description 1,2,3 is added nano-magnetic powder and dodecylbenzene sodium sulfonate quality proportioning respectively to be respectively in 1: 0.01,1: 0.10,1: 0.20 the sodium dodecyl benzene sulfonate aqueous solution, ultrasonic mixing 10min, inserting in 85 ℃ of water-baths 1000r/min stirred 1 hour, through centrifugalize, except that supernatant, use the deionized water thorough washing, make the nano-magnetic powder of oleic acid and dodecylbenzene sodium sulfonate, n-capric acid and dodecylbenzene sodium sulfonate, sodium lauryl sulphate and the modification of dodecylbenzene sodium sulfonate layer bilayer.
Embodiment 6
With particle size distribution range is the Fe of 1～5nm, 10～20nm, 20～40nm, 40～80nm, 350～390nm or 950nm 2O 3Or Fe 3O 4Or the Fe nano-magnetic powder adds nano-magnetic powder and dextran (WM10000) quality proportioning respectively and is respectively in 1: 0.01,1: 0.50,1: 1.00,1: 2.00,1: 5.0 the dextran aqueous solution, ultrasonic mixing 10min, inserting in 70 ℃ of water-baths 1000r/min stirred 1 hour, through centrifugalize, except that supernatant, use the deionized water thorough washing, make the nano-magnetic powder of dextran bag quilt.
Embodiment 7
With particle size distribution range is the Fe of 1～5nm, 10～20nm, 20～40nm, 40～80nm or 350～390nm 2O 3Or Fe 3O 4Or the Fe nano-magnetic powder adds nano-magnetic powder and Gentran 40 000 or 20000 quality proportionings respectively and is respectively in 1: 0.01,1: 0.50,1: 1.00,1: 2.00,1: 5.0 the glucan aqueous solution, ultrasonic mixing 10min, inserting in 70 ℃ of water-baths 1000r/min stirred 1 hour, through centrifugalize, except that supernatant, use the deionized water thorough washing, make the nano-magnetic powder of glucosan bag quilt.
Embodiment 8
With particle size distribution range is the Fe of 1～5nm, 10～20nm, 20～40nm, 40～80nm, 350～390nm or 950nm 2O 3Or Fe 3O 4Or the Fe nano-magnetic powder adds nano-magnetic powder and human serum albumin's quality proportioning respectively and is respectively in 1: 0.05,1: 0.20,1: 1.00 the human albumin aqueous solution, ultrasonic mixing 10min, through centrifugalize, except that supernatant, use the deionized water thorough washing, make the nano-magnetic powder that the human serum albumin wraps quilt.
Embodiment 9
With particle size distribution range is the Fe of 1～5nm, 10～20nm, 20～40nm, 40～80nm, 350～390nm or 950nm 2O 3Or Fe 3O 4Or the Fe nano-magnetic powder adds nano-magnetic powder and chitosan mass proportioning respectively and is respectively in 1: 0.05,1: 0.20,1: 1.00 the chitosan aqueous solution, ultrasonic mixing 10min, inserting in 70 ℃ of water-baths 800r/min stirred 1 hour, through centrifugalize, except that supernatant, use the deionized water thorough washing, make the nano-magnetic powder of chitosan bag quilt.
Embodiment 10
With particle size distribution range is the Fe of 1～5nm, 10～20nm, 20～40nm, 40～80nm or 350～390nm 2O 3Or Fe 3O 4Or the Fe nano-magnetic powder to add nano-magnetic powder and Macrogol 2000,4000,6000,20000 quality proportionings respectively be in 1: 0.05,1: 0.20,1: 0.50,1: 1.00 the Polyethylene Glycol aqueous solution, ultrasonic mixing 10min, inserting in 80 ℃ of water-baths 500r/min stirred 10 hours, through centrifugalize, except that supernatant, use the deionized water thorough washing, make polyethyleneglycol modified nano-magnetic powder.
Embodiment 11
The alcoholic solution of 2: 1 soybean lecithin of preparation mol ratio, cholesterol is injected into alcoholic solution the Fe that contains 1～5nm, 10～20nm, 20～40nm, 40～80nm or 350～390nm fast with syringe 2O 3Or Fe 3O 4Or in the phosphate buffered solution of Fe nano-magnetic powder, respectively 300,400,600rpm and 40 ℃ stir down and removed ethanol in 30 minutes, the centrifugal liposome of removing the magnetic powder that is not wrapped and not wrapping up magnetic powder makes the nano-magnetic powder of liposome.If select the nano-magnetic powder of particle diameter for use, then can make the liposome nano-magnetic powder of mean diameter less than 50nm less than 40nm.
Embodiment 12
It is in 1: 1 the solution that the nanometer magnetic of 20～40nm is added 400ml ethanol/water ratio, and powerful ultra-sonic dispersion 10 minutes adds volume ratio and be γ-methacryloxypropyl trimethoxy silane (CH of 2% again 2C (CH 3) C (O) OC 3H 6Si (OCH 3) 3, silicone couplet KH-570), keep system lock, avoiding solvent evaporates, 50 ℃ of following strong agitation 5 hours, cooling back magnetic separated, washing with alcohol, deionized water wash is then by nanometer SiO 2: the quality proportioning of nano-magnetic powder is respectively 5.0,1.0,0.5, adds nanometer SiO at 0.1: 1 2, powerful ultra-sonic dispersion mixed 10 minutes, and magnetic separates, removes absolute ethanol washing 3 times, removes supernatant, adds 1%PBS, makes parcel different-thickness SiO 2Nano-magnetic powder.
Embodiment 13
Get 0.6%HAuCl4 aqueous solution 2.5ml, add distilled water 120ml, regulate pH to 7.2 with 0.2mol/lK2CO3, down payment and magnetic powder quality proportioning are 5,3, add nano-magnetic powder at 0.01: 1, add white phosphorus diethyl ether solution 1ml (1 part of saturated diethyl ether solution of white phosphorus adds 4 parts of ether) again, room temperature vibration 15 minutes, solution is bronzing, be heated to the wine redness, make the nano-magnetic powder of parcel different-thickness nanometer gold.
Embodiment 14
The nano-magnetic powder of the liposome that the foregoing description 11 is made and the quality of equal volume are that the PBS solution solution of 2%PEG5000 mixes, under 10 ℃, leave standstill 60min, the PBS solution that adds equal volume again, stirred 1 hour at 10 ℃ of following 800r/min, through centrifugalize, except that supernatant, use the deionized water thorough washing, make the nano-magnetic powder of the liposome of long stable circulation in the body that PEG modifies.
Embodiment 15
With iron-holder in the foregoing description 6 is the nano-magnetic powder aqueous solution and the 500mgNaIO of 20mg dextran bag quilt 4Mix, the lucifuge oxidation is 60 minutes under the room temperature, again with 1000ml 20mM sodium borate (pH8.5) dialysis 20 hours, get its 1/10 with the 0.5ml 20mMNaIO of 1.0mg monoclonal antibody HAb18 4Buffer mixes, and effect is 4 hours under the room temperature, reuse 50 μ l 0.25MNaBH 44 ℃ were reduced 30 minutes down, and this moment, the dextran glycosyl on nano-magnetic powder surface changed aldehyde radical into, with the amine groups generation covalent bond of monoclonal antibody molecule, separate through magnetic then, deionized water wash removes supernatant, add 1%PBS, make nano-magnetic powder-anti-human liver cancer monoclonal antibody HAb18 nano target medicine.
Embodiment 16
With iron-holder in the foregoing description 7 is the nano-magnetic powder aqueous solution and the 500mgNaIO of 20mg glucosan bag quilt 4Mix, the lucifuge oxidation is 60 minutes under the room temperature, again with 1000ml20mM sodium borate (pH8.5) dialysis 20 hours; Get its 1/10 with 0.25mg monoclonal antibody HAb18 fragment HAb18F (ab ') 20.5ml 20mMNaIO 4Buffer mixes, and effect is 4 hours under the room temperature, reuse 50 μ l0.25MNaBH 44 ℃ of down reduction 30 minutes separate through magnetic then, and nano-magnetic powder-anti-human liver cancer monoclonal antibody HAb18 fragment HAb18F (ab ') is made in the 1%PBS washing 2Nano target medicine
Embodiment 17
It is in 1: 1 the solution that nanometer magnetic is added 400ml ethanol/water ratio, and powerful ultra-sonic dispersion 10 minutes adds volume ratio and be γ-methacryloxypropyl trimethoxy silane (CH of 2% again 2C (CH 3) C (O) OC 3H 6Si (OCH 3) 3, silicone couplet KH-570), keep system lock, avoiding solvent evaporates, 50 ℃ of following strong agitation 5 hours, cooling back magnetic separated, washing with alcohol, deionized water wash, then by magnetic powder: the quality proportioning of antibody is respectively and added ABX-EGF antibody in 20: 0.01,1,5,10, and powerful ultra-sonic dispersion mixed 10 minutes, magnetic separation, deionized water wash 3 times, remove supernatant, add 1%PBS, make nano-magnetic powder-ABX-EGF antibody nano target medicine.
Embodiment 18
With the anti-CEA antibody process of 3mg PBS dialysed overnight, volume is adjusted into 2ml, special-shaped bi-functional cross-linking agent N-succinimido-3-(the 2-pyridine two sulfur) propionic ester (SPDP) of the 20mmol/l 10 μ l that add the dehydrated alcohol preparation, at room temperature slowly stirred 60 minutes, and, used the PBS eluting through sephadex G25 chromatographic column, collect first peak, be condensed into 2ml, filtration sterilization makes the anti-CEA antibody-PDP of purification; Learn from else's experience again 60The iron-holder that the foregoing description 8 human serum albumins that the Co radiation sterilization is handled wrap quilt evenly spreads among the 3ml for the 30mg nano-magnetic powder, adds 10～20 μ lSPDP, stirs 60 minutes, and the centrifugal residue SPDP that removes makes bag by the PDP of magnetic powder; 3ml, concentration are in the hac buffer of 0.05mmol/l, pH4.5 again with its ultra-sonic dispersion, adding dithiothreitol, DTT (DTT) to concentration is 50mmol/l, stirred 30 minutes under the room temperature, with reduction human serum albumin-PDP, the centrifugal residue DTT that removes, use the PBS washing precipitation, get human serum albumin-PDP-SH sulfydryl; It is suspended among the 2mlPBS, mixes with anti-CEA antibody-PDP immediately, stirred 24 hours under the room temperature or powerful ultrasonic 10 minutes, centrifugal, with 1% normal saline washing precipitation, make nano-magnetic powder-anti-CEA antibody nano target medicine.
Embodiment 19
The nano-magnetic powder aqueous solution of the chitosan bag quilt of the foregoing description 9 iron content 10mg is dropwise added in the toluene solution that is dissolved with surfactant succinic acid diethyl monooctyl ester sodium sulfonate mechanical agitation emulsifying.Splash into 25% glutaraldehyde solution, reaction is 1 hour under the 2000r/min mechanical agitation.Surfactant and organic solvent are removed with ethanol, acetone, distilled water cyclic washing in reaction back, isolate magnetic component with the magnetic separator frame, wash, and are dispersed in 4 ℃ of distilled water, make contain abundant aldehyde radical in the surface the chitosan bag by nano-magnetic powder.This nano-magnetic powder is dispersed in 5% the glutaraldehyde solution room temperature vibration 2 hours.With PBS buffer washing, by magnetic powder: the quality proportioning of antibody is respectively and added Anti-HER 2s in 20: 0.01,0.1,1.0,10,4 ℃ of reactions 12 hours, usefulness PBS and 0.2% NaN 3The 1%PBS buffer respectively wash 3 times, be dispersed among the 2mlPBS, make nano-magnetic powder-Anti-HER 2 nano target medicine.
Embodiment 20
With particle size distribution range be 1～5nm, 10～20nm, 20～40nm, 50～100nm or 950nm nano-magnetic powder (its SAR be 501～7000W/gFe) in deionized water powerful ultra-sonic dispersion 10 minutes, be 0.001,0.1,1, add Rituxan antibody at 2: 10 by antibody and nano-magnetic powder mass ratio then, mix rapidly, powerful ultra-sonic dispersion mixed 10 minutes, adding concentration is 1% BPS, centrifugal, with 1%PBS washing 3 times, make nano-magnetic powder-Rituxan antibody target medicine.
Embodiment 21
The surfactant-modified nano-magnetic powder that the foregoing description 1～5 is made was the powerful ultra-sonic dispersion of deionized water 10 minutes, be respectively 0.001,0.1,1,2. by the quality proportioning of antibody and nano-magnetic powder then: 10 add anti-CEA antibody, mix rapidly, powerful ultra-sonic dispersion 10 minutes, the BPS that adds concentration 1%, centrifugal, nano-magnetic powder-anti-CEA antibody nano target medicine is made in 1%PBS washing 3 times.
Embodiment 22
The iron-holder that the foregoing description 8 human serum albumins that the 60Co radiation sterilization of learning from else's experience is handled wrap quilt evenly spreads among the 3ml for the 30mg nano-magnetic powder, adds 10～20 μ lSPDP, stirs 60 minutes, and the centrifugal residue SPDP that removes makes bag by the PDP of magnetic powder; With 0.1,1.0,5.0, the 10mg transferrins is dissolved in the phosphate buffer solution, be mixed with the solution of mass ratio 0.1～50%, adding is wherein fast to stir following SPDP coupled to Nano magnetic at-20～30 ℃ and 800～1200r/min, powerful ultra-sonic dispersion mixes, magnetic separates, adds 1% normal saline washing 3 times, makes nano-magnetic powder-transferrins nano target medicine.
Embodiment 23
The solution that under-20～30 ℃ and 800～1200r/min stir, the foregoing description 10 polyethyleneglycol modified nano-magnetic powders are added fast 0.1,1.0,5.0, the 10mg transferrins is dissolved in the mass ratio 0.1～50% that is mixed with in the phosphate buffer solution, powerful ultra-sonic dispersion mixes, magnetic separates, goes 1%PBS washing 3 times, makes nano-magnetic powder-transferrins nano target medicine.
Embodiment 24
13.2mg2-is deaminized-hydroxyl folic acid FA is dissolved in 1ml dimethyl sulfoxine DMSO, stirs to add a certain amount of two hexamethylene carbodiimide DCC and N-hydroxy succinic acid imines NHS down, adds 100mg Polyethylene Glycol diamino PEG-NH 2, reaction is 4 hours under the room temperature, adds the 5ml deionized water, removes by filter insoluble matter, the supernatant lyophilization, and ether washs dry thing, makes yellow solid FA-PEG-NH 2Get the foregoing description 9 contain 10 or the nano-magnetic powder of 100mg chitosan bag quilt dissolve in the 1ml deionized water, ultra-sonic dispersion mixed 10 minutes, added 0.1mg FA-PEG-NH 2, by amino and granule coupling, to educate altogether under the room temperature 3 hours, the magnetic separating, washing removes supernatant, adds normal saline, makes nano-magnetic powder-folic acid nano target medicine.
Embodiment 25
13.2mg5-methyl tetrahydrofolate FA is dissolved in 1ml dimethyl sulfoxine DMSO, stirs a certain amount of two hexamethylene carbodiimide DCC of adding and N-hydroxy succinic acid imines NHS down, add 100mg Polyethylene Glycol diamino PEG-NH 2, reaction is 4 hours under the room temperature, adds the 5ml deionized water, removes by filter insoluble matter, the supernatant lyophilization, and ether washs dry thing, makes yellow solid FA-PEG-NH 2Get the foregoing description 7 contain 10 or the nano-magnetic powder of 100mg glucosan bag quilt dissolve in the 1ml deionized water, ultra-sonic dispersion mixed 10 minutes, added 0.1mg FA-PEG-NH 2, by amino and granule coupling, to educate altogether under the room temperature 3 hours, the magnetic separating, washing removes the last cleer and peaceful nano-magnetic powder that does not wrap coupling folic acid, adds 1%PBS, makes nano-magnetic powder-folic acid targeted drug.
Embodiment 26
Under agitation an amount of dicyclohexylcarbodiimide and N-hydroxy succinic acid imines NHS adding are dissolved in the 50mg folic acid of 1ml dimethyl sulfoxine (triethylamine that wherein adds 0.025ml), room temperature reaction spends the night, remove by filter by-product of dicyclohexylurea in the reaction, be evaporated to dried, yellow solid powder folic acid active ester.With the foregoing description 8 contain 100,200 or the human serum albumin of the 500mg ferrum colloid water preparation that wraps the nano-magnetic powder of quilt to transfer pH be 9, add the dimethylsulfone solution of an amount of folic acid active ester, stirring at room.The magnetic separated and collected, nano-magnetic powder-folic acid targeted drug is made in the washing of 1% normal saline.
Embodiment 27
According to the Mathias method (referring to 1998.Nuclear Medicine and Biology, 25:585～587) synthetic diethylenetriamine pentaacetic acid-folic acid (being DTPA-Folate), 10mgDTPA-Folate is dissolved in the 1.5ml water, adding the 100mg particle size distribution range is the nano-magnetic powder of 1～5nm, 5～10nm, 10～20nm, 20～40nm or 50～100nm, powerful ultra-sonic dispersion 10 minutes, 400r/min stirred 1 hour under the room temperature, magnetic separates, 1%PBS washs 3 times, add 1%PBS, make nano-magnetic powder-folic acid targeted drug.
Embodiment 28
Bark fetching is inoculated 16 of colon cancer cell LS174T nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, two groups of every nude mices are the nano-magnetic powder deionized water solution 0.2ml of the iron-holder 40mg that makes of intravenous injection the foregoing description 7, glucosan bag quilt that particle diameter is about 300～400nm all, but one group applies the magnetic field 30 minutes of 0.6T at external tumor locus, and the phenanthridines algoscopy of using ferrous ion records external tumor locus and applies the particle content of tumor locus in magnetic field than not adding high about 7 times of magnetic field.
Embodiment 29
Bark fetching is inoculated 16 of colon cancer cell LS174T nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, two groups of every nude mices are the iron-holder 40mg that makes of intravenous injection the foregoing description 10 all, particle diameter is about the nano-magnetic powder deionized water solution 0.2ml of the interior long stable circulation of polyethyleneglycol modified body of 3～20nm, but one group applies the magnetic field 30 minutes of 0.8T at external tumor locus, and the phenanthridines algoscopy of using ferrous ion records the nano-magnetic powder more than 50% that external tumor locus applies magnetic field and accumulates in tumor locus.
Embodiment 30
Bark fetching is inoculated 16 of colon cancer cell LS174T nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, two groups of every nude mices are the iron-holder 40mg that makes of intravenous injection the foregoing description 14 all, particle diameter is about the nano-magnetic powder deionized water solution 0.2ml of the liposome of long stable circulation in the body that the PEG of 50～100nm modifies, but one group in the external magnetic field that applies 0.6T 30 minutes, the phenanthridines algoscopy of using ferrous ion records external tumor locus and applies the particle content of tumor locus in magnetic field than not adding high about 9 times of magnetic field.
Embodiment 31
Bark fetching is inoculated 16 of hepatoma carcinoma cell HHCC nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, two groups of every nude mices are the iron-holder 30mg that makes of intravenous injection the foregoing description 15 all, particle diameter is about nano-magnetic powder-anti-human liver cancer monoclonal antibody HAb18 nano target medicine 1%PBS aqueous solution 0.2ml of 20～40nm, but one group in the external magnetic field that applies 0.3T 60 minutes, the phenanthridines algoscopy of using ferrous ion records external tumor locus and applies the particle content of tumor locus in magnetic field than not adding high about 138 times of magnetic field.
Embodiment 32
Bark fetching is inoculated 16 of sarcoma S-180 Kunming mouses down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, two groups of every mices all iron-holder 40mg, the particle diameter made of intravenous injection the foregoing description 23 are about nano-magnetic powder-transferrins nano target medicine 1%PBS aqueous solution 0.2ml of 10～30nm, in the external magnetic field that applies 0.3T 30 minutes, the phenanthridines algoscopy of using ferrous ion recorded external tumor locus and applies the particle content of tumor locus in magnetic field than not adding high about 43 times of magnetic field.
Embodiment 33
Bark fetching is inoculated 16 of hepatoma carcinoma cell BEL7404 nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, two groups of every nude mices all iron-holder 40mg, the particle diameter made of intravenous injection the foregoing description 26 are about nano-magnetic powder-folic acid nano target medicine normal saline solution 0.2ml of 5～30nm, but one group in the external magnetic field that applies 0.3T 30 minutes, the phenanthridines algoscopy of using ferrous ion records external tumor locus and applies the particle content of tumor locus in magnetic field than not adding high about 31 times of magnetic field.
Embodiment 34
Using the be wrapped grain diameter and the surface of nano-magnetic powder granule, nano-magnetic powder-antibody or ligand conjugates of Zetasizer3000 laser particle size analyzer and JEM-2010UHR high-resolution transmission electron microscope observation the foregoing description modifying and decorating or bag changes, find that (1) the foregoing description 1,2,3 and 4 forms the surfactant layers less than 1nm usually, embodiment 5 then formation connects the surfactant layer of about 1nm; (2) the foregoing description 6,7,8,9,10 and 14 can form the organic material layer of about 10-20nm, and embodiment 11 then forms the liposome layer of about 8-10nm, and embodiment 12,13 forms the inorganic material layer of about 5-50nm; (3) the foregoing description 15,16,17,18,19,20 and nano-magnetic powder-antibody coupling matter particle diameter of 21 only can reach increases about 6～20nm than nano-magnetic powder modifying and decorating thing, the enveloped and naked nano-magnetic powder of bag before the coupling; (4) the foregoing description 22 and 23, nano-magnetic powder-transferrins conjugate particle diameter only can reach than nano-magnetic powder modifying and decorating thing, the enveloped and naked nano-magnetic powder of bag before the coupling and increase about 8～20nm; (5) the foregoing description 24,25,26 and nano-magnetic powder-folate conjugate particle diameter of 27 only can reach increases about 5～10nm than nano-magnetic powder modifying and decorating thing, the enveloped and naked nano-magnetic powder of bag before the coupling.Select for use above-mentioned nano-magnetic powder-antibody nano target medicine, nano-magnetic powder-part nano target medicine stable dispersion or the nano-magnetic powder magnetic Nano targeted drug of the nano-magnetic powder preparation of 1～40nm to have good dispersibility and reach dispersion stabilization more than several months to 4 year.
Embodiment 35
Show through JEM-2010UHR high-resolution transmission electron microscope analysis, in the foregoing description by the super-paramagnetism nano magnetic powder of the about 10nm of grain diameter and anti-CEA antibody, human liver cancer monoclonal antibody HAb18 fragment HAb18F (ab ') 2Be about 20nm with the grain diameter of the nano target medicine (conjugate) of Anti-HER 2 etc., in the medicine nano-magnetic powder with can form good coupling by antibody, nano-magnetic powder surface 1 antibody of coupling at least in the medicine.
Embodiment 36
With the preparation targeted drug in the foregoing description and nano-magnetic powder modifying and decorating thing, bag is enveloped can make the solid lyophilized powder through concentration through lyophilization, so that long term storage and convenient the use.During actual the use, only needing the mass ratio that above-mentioned targeted drug lyophilized powder is mixed with targeted drug and deionized water, PBS aqueous solution, normal saline solution, nucleotide aqueous solution with the PBS aqueous solution of deionized water or 1% or 1% normal saline solution or nucleotide aqueous solution (wherein the part by weight of deionized water and nucleotide is 1: 0.20 in the injection solvent) respectively is 1 :≤0.5 targeted drug injection, can use.Equally, only needing that above-mentioned nano-magnetic powder modifying and decorating thing, the enveloped lyophilized powder of bag are added an amount of aqueous solution promptly can be used for further processing using.Usually, also can directly to make the mass ratio of targeted drug and deionized water, PBS aqueous solution, normal saline solution or nucleotide aqueous solution be that 1 :≤0.5 targeted drug injection uses to the targeted drug for preparing in the foregoing description.
Embodiment 37
(1) to record the immunocompetence of the nano-magnetic powder that grain diameter is 20～70nm-anti-CEA antibody nano target medicine be 63% for ELISA (enzyme-linked immunosorbent assay) experiment.(2) the vitro stability experiment shows, grain diameter is that nano-magnetic powder-anti-CEA antibody nano target medicine injection of 20～70nm was hatched 24 hours through 37 ℃, no matter still be among the human serum albumin in normal saline, nano-magnetic powder comes off all less than 5%, proves that this nano target medicine is external stable.
Embodiment 38
(1) to record the immunocompetence of the nano-magnetic powder that grain diameter is 40～80nm-human liver cancer monoclonal antibody HAb18 nano target medicine be 57% for ELISA experiment.(2) the vitro stability experiment shows, grain diameter is that nano-magnetic powder-human liver cancer monoclonal antibody HAb18 nano target medicine injection of 40～80nm was hatched 24 hours through 37 ℃, no matter still be among the human serum albumin in normal saline, nano-magnetic powder comes off all less than 5%, and nano-magnetic powder-human liver cancer monoclonal antibody HAb18 nano target medicine is external stable.
Embodiment 39
(1) be that ferromagnetic nano magnetic powder-anti-CEA antibody nano target medicine of 40～80nm mixes with colon cancer cell LS174T with grain diameter, detect through the Zetasizer2000 laser particle size analyzer when ultrasonic power is 1～25W, nano-magnetic powder-anti-CEA antibody nano target medicine and cancerous cell and nano-magnetic powder granule and antibodies do not come off.(2) be that ferrimagnetism nano-magnetic powder-anti-CEA antibody nano target medicine and colon cancer cell LS174T, hemocyte, the normal liver cell of 40～80nm mixes with grain diameter, after magnetic separates, only colon cancer cell LS174T is detained under magnetic field, and hemocyte separates with colon cancer cell LS174T fully with normal liver cell.Experimental result shows, nano-magnetic powder and anti-CEA antibody have stronger coupling adhesion and stability, nano-magnetic powder-anti-CEA antibody nano target medicine and colon cancer cell LS174T have good targeting and stronger affinity, have excellent biological compatibility with blood and liver etc.
Embodiment 40
(1) be that ferromagnetic nano magnetic powder-human liver cancer monoclonal antibody HAb18 nano target medicine of 20～70nm mixes with human liver cancer cell HHCC with grain diameter, detect through the Zetasizer2000 laser particle size analyzer when ultrasonic power 1～25W, nano-magnetic powder-human liver cancer monoclonal antibody HAb18 nano target medicine and cancerous cell and nano-magnetic powder granule and antibodies do not come off.(2) be that nano-magnetic powder-human liver cancer monoclonal antibody HAb18 nano target medicine and human liver cancer cell HCC, hemocyte, the normal liver cell of 20～70nm mixes with grain diameter, after magnetic separates, only have human liver cancer cell HCC to be detained under magnetic field, hemocyte separates with hepatoma carcinoma cell HHCC fully with normal liver cell.Experimental result shows, nano-magnetic powder and human liver cancer monoclonal antibody HAb18 have stronger coupling adhesion and stability, nano-magnetic powder-human liver cancer monoclonal antibody HAb18 nano target medicine and hepatoma carcinoma cell HHCC have good targeting and stronger affinity, and nano-magnetic powder-human liver cancer monoclonal antibody HAb18 nano target medicine and blood and liver etc. have excellent biological compatibility.
Embodiment 41
Be to put to death after 30 minutes behind super-paramagnetism nano magnetic powder-anti-CEA antibody nano target medicine 30mg of 20～50nm for the nude mice tail vein injection grain diameter of subcutaneous lotus colon cancer cell LS174T tumor, get vitals such as the tumor and the heart, brain, spleen, kidney, intestinal, liver and carry out scanning electron microscope study.Electronic Speculum and EDAX results show, (the Fe element is the essential element of nano-magnetic powder to the Fe element, accounts for 70% of nano-magnetic powder quality in the nude mice tumor capillary blood of injection nano target medicine and in the tumor tissues.) account for 10 kinds of essential elements such as Fe in the tissue, Ca, Si, Al, Mg, K, Na, P, O, C part by weight respectively from matched group (not injecting targeted drug) nude mice tumor capillary blood and in the tumor tissues 0.08% and 0% bring up to 23.11% and 0.83%, and also be higher than in 0.14% and other organ in 0.14% in the non-tumor vessel blood of the nude mice of injecting targeted drug, the normal liver tissue below 0.14%.Nano target medicine in nude mice tumor capillary blood and in the tumor tissues than in matched group (not injecting nano target medicine) the nude mice tumor capillary blood and in the tumor tissues and in the nude mice non-tumor vessel blood of injection nano target medicine and increase at least 289 times in the normal liver tissue respectively (owing to do not detect Fe in the matched group tumor, can't calculate) and 165 times and 165 times, be that the content of dispersion of nano target medicine in tumor is higher than 165 times at other position at least, nano-magnetic powder-anti-CEA antibody nano target medicine has significant Targeting Performance in vivo, can make dense gathering of targeted drug targeting be positioned at tumor focus, and not assemble at other positions.
Embodiment 42
Be to put to death after 30 minutes behind super-paramagnetism nano magnetic powder-human liver cancer monoclonal antibody HAb18 nano target medicine 30mg of 20～50nm for the nude mice tail vein injection grain diameter of subcutaneous lotus hepatoma carcinoma cell HHCC tumor, get vitals such as the tumor and the heart, brain, spleen, kidney, intestinal, liver and carry out scanning electron microscope study.Electronic Speculum and EDAX results show, (the Fe element is the essential element of nano-magnetic powder to the Fe element, accounts for 70% of nano-magnetic powder quality in the nude mice tumor capillary blood of injection nano target medicine and in the tumor tissues.) account for 10 kinds of essential elements such as Fe in the tissue, Ca, Si, Al, Mg, K, Na, P, O, C part by weight respectively from matched group (not injecting nano target medicine) nude mice tumor capillary blood and in the tumor tissues 0.08% and 0% bring up to 24.38% and 0.76%, and also be higher than in 0.14% and other organ in 0.14% in the non-tumor vessel blood of the nude mice of injecting nano target medicine, the normal liver tissue below 0.14%.Nano target medicine in nude mice tumor capillary blood and in the tumor tissues than in matched group (not injecting targeted drug) the nude mice tumor capillary blood and in the tumor tissues and injection target nanometer in the non-tumor vessel blood of the nude mice of medicine and in the normal liver tissue, increase at least 304 times respectively (owing to do not detect Fe in the matched group tumor, can't calculate) and 174 times and 174 times, be that the content of dispersion of nano target medicine in tumor is higher than 174 times at other position at least, nano-magnetic powder-human liver cancer monoclonal antibody HAb18 nano target medicine has significant Targeting Performance in vivo, can make dense gathering of nano target medicine targeting be positioned at tumor focus, and not assemble at other positions.
Embodiment 43
Bark fetching is inoculated 16 of gastric carcinoma cells N87 nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus injection deionized water 0.3ml, second group of every nude mice abdominal cavity injection contains 30mg, the injection 0.3ml of 30～50nm Superparamagnetic Iron Oxide nano-magnetic powder (its SAR is 1000W/gFe)-Anti-HER 2 nano target medicine, second group of nude mice inserted under the magnetic thermotherapy treatment instrument alternating magnetic field 42～43 ℃ of treatments 1 hour 2 times, treat and put to death above-mentioned nude mice after 10 days, separate tumor, take by weighing tumor weight.The result shows that first group of tumor weighs 2.36 ± 0.53g, and second group of tumor weighs 0.97 ± 0.29g, and the tumour inhibiting rate of treatment group is 59%, and the P value is less than 0.01.
Embodiment 44
Bark fetching is inoculated 16 of hepatoma carcinoma cell HHCC nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus injection deionized water 0.3ml, second group of every nude mice abdominal cavity injection contains 30mg, the injection 0.3ml of 20～40nm Superparamagnetic Iron Oxide (its SAR is 2000W/gFe)-human liver cancer monoclonal antibody HAb18 nano target medicine, second group of nude mice inserted under the magnetic thermotherapy treatment instrument alternating magnetic field 42～43 ℃ of treatments 65 minutes 2 times, treat and put to death above-mentioned nude mice after 10 days, separate tumor, take by weighing tumor weight.The result shows that first group of tumor weighs 2.97 ± 0.65g, and second group of tumor weighs 1.02 ± 0.31g, and the tumour inhibiting rate of treatment group is 66%, and the P value is less than 0.01.
Embodiment 45
Bark fetching is inoculated 16 of people's pulmonary carcinoma A549 nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus injection deionized water 0.3ml, second group of every nude mice abdominal cavity injection contains 30mg, the injection 0.3ml of 50～100nm Superparamagnetic Iron Oxide nano-magnetic powder (its SAR is 200W/gFe)-anti-CEA antibody nano target medicine, second group of nude mice inserted under the magnetic thermotherapy treatment instrument alternating magnetic field 42～43 ℃ of treatments 65 minutes 2 times, treat and put to death above-mentioned nude mice after 10 days, separate tumor, take by weighing tumor weight.The result shows that first group of tumor weighs 1.97 ± 0.53g, and second group of tumor weighs 0.79 ± 0.37g, and the tumour inhibiting rate of treatment group is 60%, and the P value is less than 0.01.
Embodiment 46
Bark fetching is inoculated 16 of colon cancer cell LS174T nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus injection deionized water 0.3ml, second group every nude mice tail vein injection iron-holder 50mg, the injection 0.3ml of 40～80nm nano-magnetic powder (its SAR is 300W/gFe)-anti-CEA antibody nano target medicine, second group of nude mice inserted under the magnetic thermotherapy treatment instrument alternating magnetic field 42～43 ℃ of treatments 1 hour 1 time, treat and put to death above-mentioned nude mice after 10 days, separate tumor, take by weighing tumor weight.The result shows, first group of tumor weighs 2.97 ± 0.65g, second group of tumor weighs 0.12 ± 0.31g, the tumour inhibiting rate of treatment group is 96%, the P value is less than 0.01, and wherein 2 nude mice tumors disappear fully, and 3 nude mice tumor remnant tissue pathology picture analyzings are found the whole coagulation necrosiss of tumor cell, and 2 nude mice tumor remnant tissue pathology picture analyzings find only to have not downright bad on a small quantity tumor cell.Simultaneously second group of nude mice carried out body weight and untoward reaction observation treatment back ten days, and after nude mice puts to death, core, vitals such as brain, spleen, kidney, intestinal, liver do pathologic finding, show no obvious abnormalities.The zoopery result shows, adopt nano-magnetic powder-anti-CEA antibody nano target medicine carry out the magnetic thermotherapy can realize targeting, efficiently, safely, have no side effect and kill tumor, even tumor is disappeared fully.
Embodiment 47
Bark fetching is inoculated 16 of hepatoma carcinoma cell HHCC nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus injection deionized water 0.3ml, second group every nude mice tail vein injection iron-holder 50mg, 40～80nm nano-magnetic powder (its SAR is 300W/gFe)-human liver cancer monoclonal antibody HAb18 fragment HAb18F (ab ') 2The injection 0.3ml of nano target medicine inserts under the magnetic thermotherapy treatment instrument alternating magnetic field 42～43 ℃ of treatments 1 hour 1 time with second group of nude mice, treat and puts to death above-mentioned nude mice, separation tumor after 10 days, takes by weighing tumor weight.The result shows, first group of tumor weighs 2.97 ± 0.65g, second group of tumor weighs 0.12 ± 0.31g, the tumour inhibiting rate of treatment group is 96%, the P value is less than 0.01, and wherein 2 nude mice tumors disappear fully, and 2 nude mice tumor remnant tissue pathology picture analyzings are found the whole coagulation necrosiss of tumor cell, and 3 nude mice tumor remnant tissue pathology picture analyzings find only to have not downright bad on a small quantity tumor cell.Simultaneously, second group of nude mice carried out body weight and untoward reaction observation, show no obvious abnormalities, after nude mice is put to death, core, vitals such as brain, spleen, kidney, intestinal, liver do pathologic finding, show no obvious abnormalities treatment back ten days.The zoopery result shows, adopt nano-magnetic powder-human liver cancer monoclonal antibody HAb18 nano target medicine carry out the magnetic thermotherapy can realize targeting, efficiently, safely, have no side effect and kill tumor, even tumor is disappeared fully.
Embodiment 48
Bark fetching is inoculated 16 of sarcoma S-180 Kunming mouses down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every mouse tumor position injecting normal saline 0.3ml, second group every the iron-holder 60mg that mouse mainline the foregoing description 22 is made, the nano-magnetic powder of 20～50nm (its SAR is 510W/gFe)-transferrins nano target medicine 1%PBS aqueous solution 0.3ml, in the external magnetic field of 0.3T that applies after 30 minutes, second group of mice inserted under the magnetic thermotherapy treatment instrument alternating magnetic field 42～43 ℃ of treatments 1 hour 2 times, treat and put to death above-mentioned mice after 10 days, separate tumor, take by weighing tumor weight.The result shows that first group of tumor weighs 2.13 ± 0.46g, and second group of tumor weighs 0.64 ± 0.42g, and the tumour inhibiting rate of treatment group is 70%, and the P value is less than 0.01.
Embodiment 49
Bark fetching is inoculated 16 of colon cancer cell LS174T nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus injecting normal saline 0.5ml, second group of every nude mice be the iron-holder 100mg that makes of intravenous injection the foregoing description 7 all, the nano-magnetic powder of the glucosan bag quilt of 20～30nm (its SAR is 110W/gFe) deionized water solution 0.5ml, apply the magnetic field of 0.6T after 30 minutes at external tumor locus, second group of nude mice inserted under the magnetic thermotherapy treatment instrument alternating magnetic field 42～43 ℃ of treatments 2 hours 1 time, treat and put to death above-mentioned nude mice after 10 days, separate tumor, take by weighing tumor weight.The result shows that first group of tumor weighs 1.86 ± 0.39g, and second group of tumor weighs 0.65 ± 0.41g, and the tumour inhibiting rate of treatment group is 65%, and the P value is less than 0.01.
Embodiment 50
Bark fetching is inoculated 16 of colon cancer cell LS174T nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus injecting normal saline 0.3ml, second group of every nude mice be the iron-holder 60mg that makes of intravenous injection the foregoing description 14 all, nano-magnetic powder (its SAR is 210W/gFe) the deionized water solution 0.3ml of long stable circulation in the polyethyleneglycol modified body of 10～20nm, 2 apply the magnetic field of 0.8T after 30 minutes at external tumor locus, second group of nude mice inserted under the magnetic thermotherapy treatment instrument alternating magnetic field 42～43 ℃ of treatments 1 hour 1 time, treat and put to death above-mentioned nude mice after 10 days, separate tumor, take by weighing tumor weight.The result shows that first group of tumor weighs 2.23 ± 0.61g, and second group of tumor weighs 0.61 ± 0.36g, and the tumour inhibiting rate of treatment group is 73%, and the P value is less than 0.01.
Embodiment 51
Bark fetching is inoculated 16 of colon cancer cell LS174T nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus injecting normal saline 0.2ml, second group every the iron-holder 20mg that nude mice intravenous injection the foregoing description 18 is made, 20～40nm makes nano-magnetic powder (its SAR is 7000W/gFe)-anti-CEA antibody nano target medicine normal saline solution 0.2ml, in the external magnetic field of 0.3T that applies after 60 minutes, second group of nude mice inserted under the magnetic thermotherapy treatment instrument alternating magnetic field 45～48 ℃ of treatments 1 hour 1 time, tumor disappears fully in 1 month moon of treatment back, do not see tumor recurrence or transfer in 12 months, and treatment back nude mice body weight and life-span and not plant the nude mice of control group of tumor basic identical.Zoopery is the result show, adopts nano target medicine magnetic thermotherapy treatment of the present invention to kill fully and cures malignant tumor, and do not see toxic and side effects.
Embodiment 52
Bark fetching is inoculated 16 of sarcoma S-180 Kunming mouses down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus injecting normal saline 0.3ml, second group of every mouse peritoneal injection contains the 60mg that embodiment 22 makes, the injection 0.3ml of 30～50nm Superparamagnetic Iron Oxide (its SAR is 5000W/gFe)-transferrins nano target medicine, second group of mice inserted the following 95 ℃ of treatments of magnetic thermotherapy treatment instrument alternating magnetic field 1 minute 1 time, put to death above-mentioned nude mice after 10 days, take by weighing tumor weight.The result shows that first group of tumor weighs 2.09 ± 0.46g, and second group of tumor weighs 0.44 ± 0.31g, and the tumour inhibiting rate of treatment group is 79%, and the P value is less than 0.01.
Embodiment 53
Bark fetching is inoculated 16 of hepatoma carcinoma cell BEL7404 nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus injecting normal saline 0.3ml, second group of every nude mice abdominal cavity injection contains 80mg, the injection 0.3ml of the Superparamagnetic Iron Oxide that 30～50nm the foregoing description 25 is made (its SAR is 1500W/gFe)-folic acid nano target medicine, second group of nude mice inserted under the magnetic thermotherapy treatment instrument alternating magnetic field 42～43 ℃ of treatments 1 hour 1 time, treat and put to death above-mentioned nude mice after 10 days, separate tumor, take by weighing tumor weight.The result shows that first group of tumor weighs 2.03 ± 0.57g, and second group of tumor weighs 0.54 ± 0.36g, and the tumour inhibiting rate of treatment group is 73%, and the P value is less than 0.01.
Embodiment 54
Bark fetching is inoculated 16 of hepatoma carcinoma cell BEL7404 nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus injecting normal saline 0.2ml, second group of every nude mice abdominal cavity injection contains 50mg, the injection 0.2ml of the Superparamagnetic Iron Oxide that 20～40nm the foregoing description 26 is made (its SAR is 6000W/gFe)-folic acid nano target medicine, second group of nude mice inserted 42～43 ℃ of treatments of magnetic thermotherapy treatment instrument alternating magnetic field 1 hour 1 time, treat and put to death above-mentioned nude mice after 10 days, separate tumor, take by weighing tumor weight.The result shows that first group of tumor weighs 2.13 ± 0.65g, and second group of tumor weighs 0.66 ± 0.32g, and the tumour inhibiting rate of treatment group is 69%, and the P value is less than 0.01.
Embodiment 55
Bark fetching is inoculated 16 of stomach cancer cell SGC-7901 nude mices down, male and female half and half, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus injecting normal saline 0.2ml, second group of every nude mice abdominal cavity injection contains 30mg, the injection 0.2ml of the nano target medicine of the nanometer ferromagnetic powder that 20～40nm the foregoing description 27 is made (its SAR is 500W/gFe)-folic acid, second group of nude mice inserted under the magnetic thermotherapy treatment instrument alternating magnetic field 42～43 ℃ of treatments 1 hour 1 time, treat and put to death above-mentioned nude mice after 10 days, separate tumor, take by weighing tumor weight.The result shows that first group of tumor weighs 1.76 ± 0.39g, and second group of tumor weighs 0.58 ± 0.41g, and the tumour inhibiting rate of treatment group is 67%, and the P value is less than 0.01.
Embodiment 56
Bark fetching is inoculated 16 of the female nude mices of colon cancer cell LS174T down, after when the about 5～10mm of tumor, measuring and calculating gross tumor volume, be divided into 2 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus injecting normal saline 0.2ml, second group every nude mice intravenous injection iron-holder 30mg, the nano oxidized ferromagnetic powder that 20～40nm the foregoing description 18 is made (its SAR is 7000W/gFe)-anti-CEA antibody nano target medicine normal saline solution 0.2ml, second group of nude mice inserted under the magnetic thermotherapy treatment instrument alternating magnetic field 42～45 ℃ of treatments 1 hour 3 times, tumor disappears fully in 1 month moon of treatment back, do not see tumor recurrence or transfer in 12 months, and treatment back nude mice body weight and life-span and not plant the nude mice of control group of tumor basic identical.Zoopery is the result show, adopts nano target medicine magnetic thermotherapy treatment of the present invention to kill fully and cures malignant tumor, and do not see toxic and side effects.
Embodiment 57
Bark fetching is inoculated 24 of hepatoma carcinoma cell BEL7404 nude mices down, male and female half and half, after when the about 10mm of tumor, measuring and calculating gross tumor volume, be divided into 3 groups at random by gross tumor volume, first group is matched group, every nude mice tumor locus is through 3 injecting normal saline 1.0ml, second group of every nude mice imports iron-holder 100mg through 3 injections, the magnetic oxide that 400nm the foregoing description 24 is made (the about 350-400nm of magnetic powder particle diameter) (its SAR is 30W/gFe)-folic acid nano target medicine normal saline solution 1.0ml, the 3rd group of every nude mice imports iron-holder 200mg through 3 injections, the magnetic oxide that 1000nm the foregoing description 22 is made (the about 950-1000nm of magnetic powder particle diameter) (its SAR is 10W/gFe)-folic acid nano target medicine normal saline solution 1.0ml, with second, three groups of nude mices are inserted under the magnetic thermotherapy treatment instrument alternating magnetic field 42～43 ℃ of treatments 1 hour 2 times, treat and put to death above-mentioned nude mice after 10 days, separate tumor, take by weighing tumor weight.The result shows that first, second and third group tumor heavily is respectively 2.76 ± 0.69g, 0.24 ± 0.16g, 0.48 ± 0.33g, and the tumour inhibiting rate of second and three treatment groups is respectively 91% and 83%, and the P value is all less than 0.01.
Embodiment 58
Get each 10 of white mice and Cavia porcelluss, male and female half and half, inject 0.5ml in white mice and Cavia porcellus abdominal cavity respectively and contain nano-magnetic powder-anti-CEA antibody nano target medicine that 0.1g and 1.5ml contain 1.0g the foregoing description 18, observe and do not see untoward reaction and weight loss after 7 days and 30 days, after the execution, core, vitals sections such as brain, spleen, kidney, intestinal, liver do pathologic finding, show no obvious abnormalities.Observe after 30 days put to death after, core, vitals such as brain, spleen, kidney, intestinal, liver do the test of magnetic thermotherapy treatment, not seeing that heating occurs.Usually tumor magnetic thermotherapy treatment only needs 5～100mg nano-magnetic powder-anti-CEA antibody nano target medicine, and from the secondary experimental result of above-mentioned preliminary poison as can be known, nano-magnetic powder targeted drug of the present invention has biological degradability and has no side effect.
Embodiment 59
Get each 10 of white mice and Cavia porcelluss, male and female half and half, inject 0.5ml in white mice and Cavia porcellus abdominal cavity respectively and contain the nano-magnetic powder that 0.3g and 1.5ml contain the n-capric acid modification of 1.0g the foregoing description 2, observe and do not see untoward reaction and weight loss after 7 days and 30 days, after the execution, core, vitals sections such as brain, spleen, kidney, intestinal, liver do pathologic finding, show no obvious abnormalities.Observe after 30 days put to death after, core, vitals such as brain, spleen, kidney, intestinal, liver do the test of magnetic thermotherapy treatment, not seeing that heating occurs.Usually tumor magnetic thermotherapy treatment only needs the nano-magnetic powder of 5～100mg n-capric acid modification, and from the secondary experimental result of above-mentioned preliminary poison as can be known, the nano-magnetic powder of n-capric acid modification of the present invention has biological degradability and has no side effect.
Embodiment 60
Get each 10 of white mice and Cavia porcelluss, male and female half and half, inject the nano-magnetic powder that 0.5ml contains the liposome of long stable circulation in the body that PEG that 0.3g and 1.5ml contain 1.0g the foregoing description 14 modifies respectively in white mice and Cavia porcellus abdominal cavity, observe and do not see untoward reaction and weight loss after 7 days and 30 days, after the execution, core, vitals sections such as brain, spleen, kidney, intestinal, liver do pathologic finding, show no obvious abnormalities.Observe after 30 days put to death after, core, vitals such as brain, spleen, kidney, intestinal, liver do the test of magnetic thermotherapy treatment, not seeing that heating occurs.Usually tumor magnetic thermotherapy treatment only needs the nano-magnetic powder of the liposome of long stable circulation in the body that 5～100mg PEG modifies, from the secondary experimental result of above-mentioned preliminary poison as can be known, the nano-magnetic powder of the liposome of long stable circulation has biological degradability and has no side effect in the body of PEG modification of the present invention.
Embodiment 61
Get each 10 of white mice and Cavia porcelluss, male and female half and half, inject 0.5ml in white mice and Cavia porcellus abdominal cavity respectively and contain nano-magnetic powder-anti-human liver cancer monoclonal antibody HAb18 nano target medicine that 0.3g and 1.5ml contain 1.0g the foregoing description 16, observe and do not see untoward reaction and weight loss after 7 days and 30 days, after the execution, core, vitals sections such as brain, spleen, kidney, intestinal, liver do pathologic finding, show no obvious abnormalities.Observe after 30 days put to death after, core, vitals such as brain, spleen, kidney, intestinal, liver do the test of magnetic thermotherapy treatment, not seeing that heating occurs.Usually tumor magnetic thermotherapy treatment only needs 5～100mg nano-magnetic powder-anti-human liver cancer monoclonal antibody HAb18 nano target medicine, from the secondary experimental result of above-mentioned preliminary poison as can be known, nano-magnetic powder of the present invention-anti-human liver cancer monoclonal antibody HAb18 nano target medicine has biological degradability and has no side effect.
Embodiment 62
Get each 10 of white mice and Cavia porcelluss, male and female half and half, inject 0.5ml in white mice and Cavia porcellus abdominal cavity respectively and contain nano-magnetic powder-transferrins nano target medicine that 0.3g and 1.5ml contain 1.0g the foregoing description 22, observe and do not see untoward reaction and weight loss after 7 days and 30 days, after the execution, core, vitals sections such as brain, spleen, kidney, intestinal, liver do pathologic finding, show no obvious abnormalities.Observe after 30 days put to death after, core, vitals such as brain, spleen, kidney, intestinal, liver do the test of magnetic thermotherapy treatment, not seeing that heating occurs.Usually tumor magnetic thermotherapy treatment only needs 5～100mg nano-magnetic powder-transferrins nano target medicine, and from above-mentioned experimental result as can be known, nano-magnetic powder of the present invention-transferrins nano target medicine has biological degradability and has no side effect.
Embodiment 63
Get each 10 of white mice and Cavia porcelluss, male and female half and half, inject 0.5ml in white mice and Cavia porcellus abdominal cavity respectively and contain nano-magnetic powder-folic acid nano target medicine that 0.3g and 1.5ml contain 1.0g the foregoing description 25, observe and do not see untoward reaction and weight loss after 7 days and 30 days, after the execution, core, vitals sections such as brain, spleen, kidney, intestinal, liver do pathologic finding, show no obvious abnormalities.Observe after 30 days put to death after, core, vitals such as brain, spleen, kidney, intestinal, liver do the test of magnetic thermotherapy treatment, not seeing that heating occurs.Usually tumor magnetic thermotherapy treatment only needs 5～100mg nano-magnetic powder-folic acid nano target medicine, and from the secondary experimental result of above-mentioned preliminary poison as can be known, nano-magnetic powder of the present invention-folic acid nano target medicine has biological degradability and has no side effect.

Claims (17)

1. nano target medicine for magnetothermal therapy of malignant tumor, it is characterized in that comprising the quality proportioning being 1: 0.0001～0.20 effector molecule and guide molecule, described effector molecule is a magnetic powder, this magnetic powder particle diameter is no more than 1000nm, heat production specific power SAR is 10-7000W/gFe, described guide molecule comprises antibody or part, also comprises the magnetic powder that has the magnetic target function simultaneously, and the particle diameter of this targeted drug is 2-1000nm.
2. nano target medicine according to claim 1 is characterized in that wherein said magnetic powder heat production specific power SAR is 500-7000W/gFe.
3. nano target medicine according to claim 1, it is characterized in that the magnetic powder of wherein said magnetic powder for handling through surfactant and/or surface modifier modifying and decorating, the mass ratio of itself and surfactant and/or surface modifier is 1: 0.0001～0.20.
4. nano target medicine according to claim 3, it is characterized in that wherein said surfactant and/or surface modification comprise be selected from sodium lauryl sulphate, enuatrol, n-capric acid, organo-silicon coupling agent, dodecylbenzene sodium sulfonate, nucleotide or the polypeptide arbitrary or the combination.
5. nano target medicine according to claim 1 is characterized in that wherein said magnetic powder is ferromagnetism, ferrimagnetism and super-paramagnetism nano magnetic powder.
6. nano target medicine according to claim 1, it is characterized in that wherein said antibody for being the antibody of target molecule at the receptor of tumor specific antigen, tumor associated antigen, idiotypic determinant, cytokine, with the product of hormone and tumor gene, it comprises allos polyclonal antibody, Mus resource monoclonal antibody, human monoclonal antibody.
7. nano target medicine according to claim 1 is characterized in that wherein said part comprises arbitrary or its combination in folic acid part, transferrins, saccharide part, polypeptide, nucleic acid and the hormone.
8. nano target medicine according to claim 7; it is characterized in that described folic acid part comprise folic acid, folinic acid, dihydrofoilic acid, tetrahydrofolic acid, tetrahydrochysene pterin, the many glutaric acid of pterin acyl, 2-deaminize-hydroxyl folic acid, 1-deaminizes-hydroxyl folic acid, 1-denitrification folic acid, 3-denitrification folic acid, 8-denitrification folic acid, 5-methyl tetrahydrofolate, 5-formoxyl tetrahydrofolic acid, antifol methotrexate and 5, arbitrary or its combination in the 10-methylene tetrahydrofolate.
9. nano target medicine according to claim 1 is characterized in that wherein said antibody is selected from anti-CEA, AFP, CA19-9, CA125, PSA, Trigen, TheraCIMh-R3, Smart ID10, Vitaxin, 4B5, SS1 (dsFv) PE38, ABX-EGF, BrevaRex, CA15-3, PAP, CA50, hCG, SCC, β 2Arbitrary or its combination among M, Rituxan, anti-VEGF, epratuzmab, NSE, HER2 and TPA antibody and the human liver cancer monoclonal antibody HAb18.
10. nano target medicine according to claim 1 is characterized in that wherein said magnetic powder is the magnetic powder of surface through organic or inorganic material parcel.
11. nano target medicine according to claim 10, it is characterized in that wherein said organic substance comprises arbitrary or combination in liposome, polylactic acid, polycaprolactone, Polyethylene Glycol, glucosan, chitosan, dextran or the protein, inorganic substances are arbitrary or combination in silicon dioxide, aluminium oxide or the gold, or the combination of organic substance and inorganic substances.
12. nano target medicine according to claim 10, the raw materials quality ratio that it is characterized in that wherein said magnetic powder and organic or inorganic material is 1: 0.01～5.0.
13. nano target medicine according to claim 1 is characterized in that wherein this medicine is injection, solid or powder injection formulation.
14. the preparation method of the described nano target medicine of claim 1 is characterized in that obtaining comprising the magnetic powder and the guide molecule coupling in water or organic substance ethanol, acetic acid, toluene, two methylene sulfones or inorganic substances phosphoric acid or its mixed solution that with the quality proportioning are 1: 0.0001～0.20.
15. preparation method according to the described nano target medicine of claim 14, it is characterized in that comprising earlier described magnetic powder being handled with surfactant that is selected from sodium lauryl sulphate, enuatrol, n-capric acid, dodecylbenzene sodium sulfonate, organo-silicon coupling agent, nucleotide or polypeptide and/or surface modifier, the weight ratio of magnetic powder and surfactant and/or surface modifier is 1: 0.0001～0.20, mixes coupling with guide molecule in water or solution again.
16. preparation method according to the described nano target medicine of claim 14, it is characterized in that comprising earlier described magnetic powder being wrapped up with the organic or inorganic material, this organic substance comprises liposome, Polyethylene Glycol, glucosan, chitosan, dextran, polylactic acid, polycaprolactone or protein, inorganic substances are silicon dioxide, aluminium oxide, gold, the weight ratio of magnetic powder and organic or inorganic material wrappage is 1: 0.01～5.0, mixes coupling with guide molecule in solution or water again.
17. the application of each described nano target medicine of claim 1-16 in the medicine of preparation treatment and prophylaxis of tumours.
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