Patent Application
20170209373
This application claims the benefit under 35 U.S.C. §119, the priority of Korean Patent Application No. 10-2016-0115881, filed Jan, 27, 2016,in the Korean Intellectual Property Office, the disclosures of which are hereby incorporated by the references.
The present invention relates to a drug delivery system (DDS), which crosses the blood-brain barrier (BBB) formed by self-assembly of an amphiphilic block copolymer, comprising a self-assembled structure having an average diameter of 5 nm to 20 nm of a core-shell structure with a hydrophobic core and a hydrophilic shell; and a hydrophobic drug supported in the hydrophobic core of the self-assembled structure, a pharmaceutical composition for preventing or treating cerebral diseases comprising the drug delivery system as an active ingredient, and a method for preparing the drug delivery system.
The blood-brain barrier (BBB) is a cellular barrier composed of tight junctions having a high magnitude of electrical resistance of at least 0.1 Ωm between vascular endothelial cells that are adjacent to related pericytes and astrocytes, and it acts as a barrier having highly selective permeability to separate circulating blood from brain extracellular fluids in the central nervous system (CNS), thereby playing a role of a gateway to protect the central nervous system by regulating the entry and exit of nutrients and other substances to the brain. Normally, the BBB not only selectively transfers molecules such as glucose and amino acids that are essential for brain function, but also passes water, some gases, and fat-soluble molecules by passive diffusion. On the other hand, the BBB blocks the entry and exit of lipophilic, potential neurotoxins by an active transport mechanism mediated by permeable p-glycoprotein. The BBB is formed along all capillaries and is composed of tight junctions around capillaries that do not exist in normal circulation. As such, although the BBB serves to prevent the transport of bacteria, pathogens, and potential hazardous substances in blood to the brain that can be carried through the blood, due to the barriers of such blood vessels, most drugs for the central nervous system show low efficiency of transcranial delivery, and therefore, in order to compensate for this, such drugs are administered at a high dose, which may cause serious side effects in surrounding organs.
Therefore, there is a need to find an efficient drug delivery system that can cross the BBB in order to ensure the therapeutic effects of chemodrugs, at the same time preventing negative systemic effects.
Using the self-assembling properties of amphiphilic block copolymers, the present inventors undertook diligent research efforts in order to discover a drug delivery system that can cross the BBB and deliver a drug specifically to the brain even when the drug is administered systematically, and as a result, the present invention has been completed by confirming that, a drug delivery system, which comprises a self-assembled structure having an average diameter of 5 nm to 20 nm of a core-shell structure with a hydrophobic core and a hydrophilic shell, formed by self-assembly of an amphiphilic block copolymer by mixing an amphiphilic block copolymer comprising a hydrophilic block of poly(ethylene oxide) (PEO) and a hydrophobic block of poly(propylene oxide) (PPO); and a hydrophobic drug at a predetermined ratio, in which the hydrophobic drug to be delivered is supported in the hydrophobic core, can deliver the hydrophobic drug specifically to the brain and release the drug, even when systematically administered.
The first aspect of the present invention provides a drug delivery system (DDS), which crosses the blood-brain barrier (BBB), comprising: a self-assembled structure having an average diameter of 5 nm to 20 nm of a core-shell structure with a hydrophobic core and a hydrophilic shell, in which a hydrophobic block of an amphiphilic block copolymer is positioned inside the structure and a hydrophilic block of the amphiphilic block copolymer is positioned out of the structure, respectively, and the amphiphilic block copolymer comprises the hydrophilic block of poly(ethylene oxide) (PEO) and the hydrophobic block of poly(propylene oxide) (PPO); and a hydrophobic drug supported in the hydrophobic core of the self-assembled structure, wherein the ratio of the amphiphilic block copolymer to the hydrophobic drug is adjusted for crossing the blood-brain barrier.
The second aspect of the present invention provides a pharmaceutical composition for preventing or treating cerebral diseases, comprising the drug delivery system according to the first aspect of the present invention as an active ingredient.
The third aspect of the present invention provides a method for preparing a drug delivery system, which crosses the blood-brain barrier, having an average diameter of 5 nm to 20 nm of a core-shell structure with a hydrophilic shell, a hydrophobic core and a hydrophobic drug supported therein, comprising: a first step of airblowing a solution containing a hydrophobic drug, and a amphiphilic block copolymer comprising a hydrophilic block of poly(ethylene oxide) (PEO) and a hydrophobic block of poly(propylene oxide) (PPO) in an organic solvent, in which the ratio of the amphiphilic block copolymer to the hydrophobic drug is adjusted for crossing the blood-brain barrier, to evaporate the organic solvent; and a second step of adding a water-containing solvent to the product obtained from the previous step followed by dispersion by an ultrasonic disperser.
Hereinbelow, the present invention will be described in more detail.
The present invention is based on a discovery that when a drug delivery system, a self-assembled structure of a core-shell structure with a hydrophobic core and a hydrophilic shell formed by self-assembly, in which a hydrophobic block of an amphiphilic block copolymer is positioned inside the structure and a hydrophilic block of the amphiphilic block copolymer is positioned out of the structure, respectively, in an aqueous solvent and the amphiphilic block copolymer comprises the hydrophilic block of poly(ethylene oxide) (PEO) and the hydrophobic block of poly(propylene oxide) (PPO), is prepared by supporting a hydrophobic drug in its hydrophobic core, it is mainly accumulated in the brain by crossing the blood-brain barrier even when it is systemically administered by intravenous injection if the size of the drug delivery system is adjusted to have an average diameter of 5 nm to 20 nm. Specifically, by adjusting the ratio of the amphiphilic block copolymer to the internally supported hydrophobic drug, the present inventors showed that it is possible to provide a drug delivery system having the above-mentioned size and/or effect.
The present invention can provide a drug delivery system (DDS), which crosses the blood-brain barrier (BBB), comprising: a self-assembled structure having an average diameter of 5 nm to 20 nm of a core-shell structure with a hydrophobic core and a hydrophilic shell, in which a hydrophobic block of an amphiphilic block copolymer is positioned inside the structure and a hydrophilic block of the amphiphilic block copolymer is positioned out of the structure, respectively, and the amphiphilic block copolymer comprises the hydrophilic block of poly(ethylene oxide) (PEO) and the hydrophobic block of poly(propylene oxide) (PPO); and a hydrophobic drug supported in the hydrophobic core of the self-assembled structure.
For example, the drug delivery system may comprise the drug delivery system comprises the hydrophobic drug and the amphiphilic block copolymer at a mass ratio (w/w) of (0.025 to 2):20. Alternatively, the drug delivery system may comprise the hydrophobic drug and the amphiphilic block copolymer at a mass ratio (w/w) of (0.05 to 1):20 or (0.1 to 0.5):20. If the ratio of the hydrophobic drug to the amphiphilic block copolymer is less than 0.025 mg/20 mg, the drug supporting efficiency may be low, and if the ratio is over 5 mg/20 mg, as the size of the formed drug delivery system becomes larger, it is difficult to cross the blood-brain barrier, and the delivery efficiency to the brain may be lowered.
For example, the amphiphilic block copolymer is a triblock copolymer having an average molecular weight of 1,500 Da to 20,000 Da. Alternatively, it may be a triblock copolymer having an average molecular weight of 8,000 Da to 15,000 Da, but is not limited thereto. For example, the amphiphilic block copolymer may be a block copolymer comprising PEO and PPO blocks at a ratio of (2.5 to 6):1 based on the numbers of EO and PO units constituting the hydrophilic PEO block and hydrophobic PPO block, respectively. The amphiphilic block copolymer can be synthesized and used so as to comprise the hydrophilic and hydrophobic blocks at the above-mentioned ratio and to have a molecular weight in the above-mentioned range, or commercially available products such as Pluronic F-127 or F-68 corresponding thereto can be purchased and used.
For example, the hydrophobic drug is an anticancer agent, antioxidant agent, anti-inflammatory agent, contrast agent, or combination thereof, but is not limited thereto. As the anticancer, antioxidant, and anti-inflammatory agents, a drug capable of being supported in a hydrophobic core of a particle formed by an amphiphilic polymer can be used as the hydrophobic drug without limitation.
Non-limiting examples of the anticancer agent may include curcumin, Doxorubicin (Dox), Paclitaxel (Taxol), and Cisplatin.
Non-limiting examples of the antioxidant agent may include vitamin E, alpha-carotene, and anthocyanine.
Non-limiting examples of the anti-inflammatory agent may include Methotrexate (MTX), Leflunomide, and Tacrolimus.
Meanwhile, a contrast agent is a substance that functions to provide clear images so that a disease occurring at a site which cannot be confirmed by simple imaging can be diagnosed early and treated, and contrast agents for magnetic resonance imaging (MRI), contrast agents for computed tomography (CT), contrast agents for positron emission tomography (PET), ultrasound contrast agents, and fluorescent contrast agents, etc., may be used. Non-limiting examples of the contrast agent may include contrast agents for magnetic resonance imaging (MRI), which are paramagnetic or superparamagnetic substances of a transition metal ion including gadolinium (Gd), manganese (Mn), copper (Cu), and chromium (Cr), of hydrophobic complexes of the transition metal ion including gadopentetate dimeglumine (Gd-DTPA) and gadoterate meglumine (Gd-DOTA), of fluorine-containing compounds including perfluorocarbons and perfluoropropane, of iron oxide-based, manganese-based, copper-based, and chromium-based nanoparticles, and compounds obtained by modifying the surface of the nanoparticles with a hydrophobic substance, etc.; contrast agents for computed tomography (CT) of iodinated hydrophobic substances derived from iodinated poppy seed oil, and nanoparticles composed of a metal element including bismuth (Bi), gold (Au), and silver (Ag), etc.; contrast agents for positron emission tomography (PET) of a radioactive isotope including 99mTc, 123I, 166Ho, 111In, 90Y, 153Sm, 186Re, 188Re, 68Ga, and 177Lu, and of hydrophobic complexes of the radioactive isotope prepared using diethylenetriaminepentaacetate (DTPA), etc.; ultrasonic contrast agents which are hydrophobic compounds of perfluoropropane, perfluorohexane, sulfur hexafluoride, perfluoropentane, and decafluorobutane, etc.; and fluorescent contrast agents such as fluorescein, rhodamine, Nile red, Cy-3, and Cy-5, etc., and near-infrared fluorescent substances described below can be included without limitation.
For example, the drug delivery system may further comprise a near-infrared fluorescent substance in the hydrophobic core of the self-assembled structure. In particular, the near-infrared fluorescent substance may be comprised at a mass ratio (w/w) of (0.025 to 0.5):20, (0.025 to 0.3):20, or (0.05 to 0.25):20 relative to the amphiphilic block copolymer, wherein the sum of the masses of the hydrophobic drug and the near-infrared fluorescent substance may be adjusted to have a mass ratio (w/w) of (0.05 to 2.5):20, (0.1 to 1):20, or (0.1 to 0.3):20 relative to the mass of the amphiphilic block copolymer. For example, the near-infrared fluorescent substance may be selected from fluorescent substances exhibiting fluorescence in a range of 640 nm to 1,000 nm. For example, it may be a series of indane derivatives or a series of 1,3-indandione-based derivatives comprising a carbazole ring or aniline ring, but is not limited thereto. Specifically, non-limiting examples of the near-infrared fluorescent substance may include 2,2′-(2-((9-ethyl-9H-carbazol-3-yl)methylene)-1H-indene-1,3(2H)-diylidene)dimalononitrile, (Z)-2-(2-((9-ethyl-9H-carbazol-3-yl)methylene)-3-oxo-2,3-dihydro-1H-inden-1-ylidene)malononitrile, (Z)-2-(2-(4-(dimethylamino)benzylidene)-3-oxo-2,3-dihydro-1H-inden-1-ylidene)malononitrile, 2,2′-(2-(4-(dimethylamino)benzylidene)-1H-indene-1,3(2H)-diylidene)dimalononitrile, 2,2′-(1H-indene-1,3(2H)-diylidene)dimalononitrile, (2,2′-(1H-indene-1,3(2H)-diylidene)dimalonic acid, (2E,2′Z)-diethyl 2,2′-(1H-indene-1,3(2H)-diylidene)bis(2-cyanoacetate), (2E,2′Z)-2,2′-(1H-indene-1,3(2H) -diylidene)bis(2-cyanoacetic acid), (E)-ethyl 2-cyano-2-(3-(dicyanomethylene)-2,3-dihydro-1H-inden-1-ylidene)acetate, 2,2′,2″-([1,2′-biindenylidene]-1′,3,3′(2H)-triylidene)trimalonic acid, 2,2′,2″-((1Z,1′Z,3E,3′E)-[1,2′-biindenylidene]-1′,3,3′(2H)-triylidene)tris(2-cyanoacetic acid), triethyl 2,2′,2″-((1Z, 1′Z,3E,3′E)-[1,2′-biindenylidene]-1′,3,3′(2H)-triylidene)tris(2-cyanoacetate), and 2,2′,2″-([1,2′-biindenylidene]-1′,3,3′(2H)-triylidene)trimalononitrile.
The drug delivery system of the present invention can be used as a pharmaceutical composition for preventing or treating cerebral diseases comprising the drug delivery system as an active ingredient. For example, the pharmaceutical composition may further comprise a carrier and may be provided in an injectable form, but is not limited thereto.
In a specific exemplary embodiment of the present invention, it was confirmed that even when the drug delivery system comprising curcumin as a drug was intravenously injected in the form of an aqueous dispersion, it was efficiently delivered to the brain by crossing the blood-brain barrier, and furthermore, in the brain tumor model, as it was selectively delivered to the tumor site of the brain and released the drug, it was confirmed that the drug delivery system suppressed the growth of tumor and exhibited a therapeutic effect of making the tumor size significantly smaller, even when used in an amount as low as 1/2,000 relative to a simple solution of the same drug.
Further, the drug delivery system of the present invention can be prepared via a first step of airblowing a solution containing a hydrophobic drug, and a amphiphilic block copolymer comprising a hydrophilic block of poly(ethylene oxide) (PEO) and a hydrophobic block of poly(propylene oxide) (PPO) in an organic solvent, to evaporate the organic solvent; and a second step of adding a water-containing solvent to the product obtained from the previous step followed by dispersion by an ultrasonic disperser.
For example, in the second step, the water-containing solvent may be used in which the final concentration of the amphiphilic block copolymer used in the solution is set to be in a range of 5 mg/mL to 50 mg/mL. More specifically, the concentration of the amphiphilic block copolymer used in the solution may be in a range of 10 mg/mL to 30 mg/mL, but is not limited thereto.
For example, as the organic solvent, a volatile organic solvent capable of simultaneously dissolving a hydrophobic drug and a polymeric surfactant, that is, an amphiphilic polymer, can be used without limitation. Specifically, the organic solvent may be dichloromethane, tetrahydrofuran, chloroform, or a mixture thereof, but is not limited thereto.
For example, the solution of the first step may further comprise a near-infrared fluorescent substance, but is not limited thereto, and the content thereof is as described above. For example, the near-infrared fluorescent substance is preferably a fluorescent substance which does not cause self-quenching of fluorescence, even if it is densely concentrated.
For example, if the hydrophobic drug is a molecule which exhibits fluorescence, the near-infrared fluorescent substance may be selected to have an absorption spectrum in the fluorescence wavelength range of the hydrophobic drug. When combined as above, in the case of exciting at the absorption wavelength of the hydrophobic drug, by a phenomenon of fluorescence energy transfer, fluorescence generated therefrom is absorbed by adjacent near-infrared fluorescent substances supported therewith, thereby generating fluorescence thereof, and by detecting the fluorescence of the fluorescent substances, it can be confirmed whether the drug delivery system was delivered to a lesion by monitoring the position thereof, and at the same time, it can be confirmed from changes in the intensity thereof whether the drug was released.
Since the drug delivery system of the present invention i) is formed by self-assembly of an amphiphilic block copolymer comprising a hydrophilic block of poly(ethylene oxide) (PEO) and a hydrophobic block of poly(propylene oxide) (PPO), ii) has a core-shell structure with a hydrophobic core and a hydrophilic shell so that it is capable of supporting a hydrophobic drug in the hydrophobic core thereof, and iii) has a small size of an average diameter of 5 nm to 20 nm, even when administered systematically by intravenous injection, it does not become accumulated in other organs such as liver, pancreas, and kidney, and is able to cross the blood-brain barrier and be efficiently delivered to the brain, enabling to release the drug specifically to the lesion, and therefore the drug delivery system can be valuably used for preventing or treating cerebral diseases by supporting various drugs inside.
Hereinbelow, the present invention will be described in detail with accompanying exemplary embodiments. However, the exemplary embodiments disclosed herein are only for describing the invention more specifically and should not be construed as limiting the scope of the present invention.
The nanoparticles containing the drug and/or fluorescent dye according to the present invention were prepared by, for example, dispersing in water curcumin, which is an anticancer agent, as the drug, and/or a carbazole derivative of an arylvinyl compound which is a fluorescent dye absorbing near-infrared ray, with an amphiphilic polymer as a homogeneous mixture, thereby inducing to spontaneously and uniformly form a colloid. No precipitates were observed in such dispersion, and it indicates that the drug and/or fluorescent dye molecules, which are water-insoluble, were successfully loaded in the hydrophobic interior space of the self-assembled nanostructure of the amphiphilic polymer. The composition of such particles and a specific preparation method thereof are as follows.
In order to prepare curcumin-encapsulated amphiphilic polymeric nanoparticles (NP-cur), 0.1 mg of curcumin (Sigma-Aldrich) and 20 mg of Pluronic F-127, which is an amphiphilic polymer, were completely dissolved in 0.5 mL of dichloromethane (Junsei), followed by airblowing to selectively evaporate only the solvent. 2 mL of water was added to the dried mixture, and it was dispersed uniformly with an ultrasonic disperser. The concentration of curcumin contained in the finally prepared curcumin-encapsulated amphiphilic polymeric nanoparticle dispersion was 50 μg/mL.
Transmission electron microscopy (TEM) images of the NP-cur prepared as described above were measured, and the result is shown in
Except that 0.2 mg of a series of 1,3-indandione-based derivatives containing a carbazole ring or an aniline ring, for example, 2,2′-(2-((9-ethyl-9H-carbazol-3-yl)methylene)-1H-indene-1,3(2H)-diylidene)dimalononitrile (CbV10), (Z)-2-(2-((9-ethyl-9H-carbazol-3-yl)methylene)-3-oxo-2,3-dihydro-1H-inden-1-ylidene)malononitrile (CbV9), (Z)-2-(2-(4-(dimethylamino)benzylidene)-3-oxo-2,3-dihydro-1H-inden-1-ylidene)malononitrile (AnV9), and 2,2′-(2-(4-(dimethylamino)benzylidene)-1H-indene-1,3(2H)-diylidene)dimalononitrile (AnV10) was used as the near-infrared fluorescent dye instead of curcumin, CbVn or AnVm-encapsulated amphiphilic polymeric nanoparticles (NP-CbVn and NP-AnVm) were prepared in the same manner as in Example 1-1. The concentration of CbVn or AnVm contained in the finally prepared CbVn or AnVm-encapsulated amphiphilic polymeric nanoparticle dispersion was about 100 μg/mL. For reference, CbVn or AnVm was synthesized by referencing the methods disclosed in ACS Appl. Mater. Interfaces, 2013, 5: 8881.
TEM images of the NP-CbV10 prepared as described above were measured, and the result is shown in
Except that a series of indane derivatives were used as the near-infrared fluorescent dye instead of a series of 1,3-indandione derivatives containing a carbazole ring or an aniline ring, the nanoparticles loading the near-infrared fluorescent dye were prepared in the same manner as in Example 1-2. As the indane derivatives, a total of 9 types of commercially available compounds were purchased from the company TCI, etc. and used, and the types are as follows:
Except that 0.2 mg of CbV was additionally used in addition to 0.1 mg of curcumin, the amphiphilic polymeric nanoparticles (NP-cur/CbV) containing curcumin and CbV were prepared in the same manner as in Example 1-1. The concentrations of curcumin and CbV contained in the finally prepared amphiphilic polymeric nanoparticle dispersion containing curcumin and CbV were 50 μg/mL and 100 μg/mL, respectively.
TEM images of the NP-cur/CbV as prepared above were measured, and the result is shown in
As described above, the optical properties of the amphiphilic polymeric nanoparticles loading an optically active drug and/or fluorescent dye molecule prepared according to Examples 1-1 to 1-4 were consistent with the intrinsic optical properties which were displayed when the optically active drug loaded therein was dissolved in an organic solvent. It was shown therefrom that the optically active drug and/or fluorescent dye was successfully loaded in the interior space of the amphiphilic polymeric nanoparticle, and thus it was confirmed that even when concentrated at a high density in the interior space of the nanoparticle, it exhibited effective fluorescence without self-quenching by adjacent molecules.
Meanwhile, it was confirmed that, when irradiating excitation light at a wavelength of 450 nm, at which the absorption of curcumin is high and the absorption by CbV is hardly displayed, to NP-cur/CbV simultaneously loading both curcumin and CbV, the fluorescence of curcumin due to the 450 nm excitation light was quenched and the fluorescence of CbV increased at the same time. It can be explained by the fluorescence energy transfer which occurs because the fluorescence of curcumin occurring under excitation at 450 nm is spectrally overlapped with the absorption wavelength range of CbV, and such molecules are highly densely packed inside the nanoparticles thereby placing within a distance of several nanometers. Therefore, in subsequent experiments, drug release from the particles was non-invasively confirmed using the fluorescence energy transfer phenomenon between curcumin and CbV.
200 μL of the NP-cur or NP-CbV aqueous dispersion prepared according to Examples 1-1 and 1-2 was injected in tail veins of male mice (CAnN.Cg-Foxnlnu/Crl, 5 weeks old, Orient Bio, Korea). At certain time intervals before the intravenous injection and thereafter for four hours, fluorescence signals were tracked and observed using a fluorescence imaging device (IVIS-Spectrum; Perkin-Elmer, USA), and the results are shown in
Further, as described above, after intravenous injection in mice using the aqueous dispersion of the nanoparticles loading 2,2′-(1H-indene-1,3(2H)-diylidene)dimalononitrile prepared according to Example 1-3, the fluorescence signals were tracked and observed using the fluorescence imaging device, and the results are shown in
200 μL of the NP-cur/CbV aqueous dispersion prepared according to Example 1-4 was injected in the tail veins of male mice. At certain time intervals before the intravenous injection and thereafter for four hours, fluorescence signals were tracked and observed using the fluorescence imaging device, and the results are shown in
As described above, through Examples 2-1 and 2-2, the nanoparticles loading the drug and/or fluorescent dye according to the present invention were systemically administered to the normal mouse model via intravenous injection, and the behavior of the particles in vivo was monitored by fluorescence imaging to determine whether they could cross the blood-brain barrier (BBB). As shown in
However, the fluorescence signals in the brain after administration of NP-cur or NP-CbV showed a tendency to increase over time and then decrease again, the increase in the fluorescence signals in the brain over time was due to the fact that after the particles loading CbV or curcumin were systemically administered, they were circulating through the blood and were accumulated in the brain, and the decrease in the signals was due to the release and dispersion of CbV or curcumin molecules from the nanoparticles delivered to the brain.
In order to confirm the release of curcumin from the NP-cur delivered to the brain, according to Example 2-2, using the nanoparticles (NP-cur/CbV) simultaneously loading both curcumin and CbV, which is a fluorescent dye capable of fluorescence energy transfer therewith, the energy transfer property between these optically active molecules was observed. As shown in
In order to non-invasively confirm the accumulation specific to the brain, particularly to tumor lesions of the nanoparticles loading the drug according to the present invention and the tumor treating effect due to the release of the drug to the lesions according to the same, a bioluminescent glioblastoma multiforme model was used. Since the bioluminescent glioblastoma multiforme model exhibits bioluminescence in brain tumor lesions upon administration of luciferin, the position and/or size of formed lesions can be confirmed therefrom. Therefore, it became possible to monitor whether the lesion was targeted by comparison of the position of a lesion confirmed by the bioluminescence and the position at which the nanoparticles of the present invention were accumulated, which was obtained through fluorescence images.
In order to identify the accumulation behavior of the drug-containing nanoparticles of the present invention to cancerous lesions after crossing the blood-brain barrier, bioluminescence and fluorescence signals were observed using the bioluminescent glioblastoma multiforme (GBM) model. After drilling a fine hole 3.5 mm deep with a 26G needle (Hamilton Company, USA) at a point 0.2 mm posterior to and 2.2 mm lateral from the cranial bregma of a male mouse (CAnN.Cg-Foxnlnu/Crl, 5 weeks old, Orient Bio, Korea), 1.5 μL of physiological saline in which 1×105 luciferase-expressing U-87 MG cells were stably dispersed was administered through the hole.
7 days after transplantation of cancer cells, luciferin dissolved in physiological saline was intraperitoneally injected at a dosage of 150 mg/kg, then after 20 minutes, optical images were collected to observe bioluminescence signals. Thereafter, 200 μL of the NP-CbV aqueous dispersion prepared according to Example 1-2 was intravenously injected into the same mouse, and optical images were collected at certain time intervals to observe near-infrared fluorescence signals. The bioluminescence signals and near-infrared fluorescence signals collected therefrom were compared and analyzed, and are shown in
The NP-cur aqueous dispersion prepared according to Example 1-1 was intravenously injected daily at a dose of 200 μL from day 3 of the transplantation of the cancer cells in the brain tumor model prepared according to Example 3-1. Luciferin was administered on days 7, 14, and 21 after the administration of the NP-cur aqueous dispersion was initiated, and bioluminescence signals were observed in the same manner as in Example 3-1. For the control groups to which the drug was not administered, a group (control of
As shown in
| Number | Date | Country | Kind |
|---|---|---|---|
| 10-2016-0115881 | Jan 2016 | KR | national |
