Albumin nanoplatform for boron neutron capture therapy and composition for boron neutron capture therapy comprising same

Abstract

The present invention relates to an albumin nanoplatform for boron neutron capture therapy and a composition for boron neutron capture therapy comprising same. The albumin-based nanoplatform according to the present disclosure can effectively deliver boron to specific tumor tissues by simultaneously conjugating a boron compound containing an excessive amount of boron and a targeting molecule as a carrier for specifically targeting tumor tissues via click chemistry functional groups introduced into the albumin surface. In particular, the nanoplatform enables sufficient delivery of boron molecules to target tumor tissues even at doses less than one-tenth of those required by conventional boron neutron capture therapy (BNCT) drugs, and thus can be applied as a composition for boron neutron capture therapy and as an anticancer therapeutic agent.

Description

Albumin nanoplatform for boron neutron capture therapy and composition for boron neutron capture therapy comprising the same

[0001] The present invention relates to an albumin nanoplatform for boron neutron capture therapy and a composition for boron neutron capture therapy comprising the same, and more particularly, to a technology for providing an albumin-based boron neutron capture therapy composition that selectively targets cancer tissue for boron neutron capture therapy and stably and effectively delivers boron to cancer tissue with only a small dose, thereby improving the effectiveness of anticancer treatment.

[0002] Neutron Capture Therapy (NCT) is a treatment technique that deposits neutron-capture drugs in tumors and then irradiates them with thermal neutrons to selectively destroy tumor cells. Because low-energy thermal neutrons selectively induce nuclear fission reactions in drug-loaded cancer cells while leaving normal cells largely unharmed, neutron capture therapy is a highly effective cancer treatment compared to conventional radiotherapy. Therefore, to prevent damage to normal tissue, a major drawback of radiotherapy, the strategy of concentrating the drug in tumor cells is central to neutron capture therapy.

[0003] Boron Neutron Capture Therapy (BNCT), which uses boron as a neutron capture drug, 10 B(n,a) 7 It is based on the principle of Li nuclear fission reaction. It is an isotope of boron. 10B atoms capture thermal neutrons, undergoing a fission reaction and decaying, producing alpha and lithium particles and gamma rays. The range of these emitted alpha and lithium particles is approximately 5 to 7 micrometers, which corresponds to the diameter of a cell (approximately 10 micrometers), ultimately damaging only cells adjacent to the boron drug. Therefore, BNCT was expected to be a treatment method that could selectively treat tumor cells while minimizing damage to normal cells by utilizing boron drugs that selectively accumulate in tumor cells.

[0004] This boron neutron capture therapy has been used in the treatment and research of various cancers such as brain tumors, head and neck cancer, melanoma, and colon cancer, but has not been widely commercialized due to the problem of the cancer cell concentration rate of boron compounds and the limitation of requiring a nuclear reactor facility.

[0005] Boron drugs currently in clinical use include BPA (p-boronophenylalanine) and BSH (sulfhydryl borane, Na2B 12 H 11 SH) is a representative example. BPA is an amino acid derivative and is known to accumulate in cancer tissues with active protein synthesis. However, due to the nature of amino acids, it is rapidly excreted from the body through the kidneys, so it must be administered over approximately 2 hours to perform BNCT. In addition, BSH is structurally very hydrophobic, so when administered into the body, most of it is initially taken up by the liver and then excreted through the intestines. Therefore, there is a problem that the drug must be administered in excessive doses to achieve the desired therapeutic effect.

[0006] Korean Patent Publication No. 10-2008-0107309 is known as a method for producing a nanohybrid by introducing a boron compound into a layer of a metal double layer hydroxide and a technology for maintaining a high concentration of boron in cells by providing the nanohybrid produced by the method as a composition for boron neutron capture therapy. However, it only discloses a nanohybrid using a metal ion and does not disclose a nanocarrier that can deliver a sufficient amount of boron molecules to target cancer tissues with only a smaller dose than conventional boron neutron capture therapy (BNCT) drugs using a bio-derived material.

[0007] Therefore, continued research is needed to develop satisfactory radiopharmaceuticals for neutron capture therapy that exhibit high biocompatibility and stability while solving the problem of excessive boron compound administration.

[0008] The purpose of the present invention is to provide an albumin-based nanoplatform for boron neutron capture therapy (BNCT).

[0009] Another object of the present invention is to provide a composition for boron neutron capture therapy comprising an albumin-based nanoplatform.

[0010] Another object of the present invention is to provide a composition for preventing or treating cancer, which contains an albumin-based nanoplatform for boron neutron capture therapy as an active ingredient.

[0011] The present invention provides an albumin-based nanoplatform for boron neutron capture therapy (BNCT), which is obtained by a click chemistry reaction between albumin having an azide (N3) or cyclooctyne functional group bound thereto and a carrier having an azide (N3) or cyclooctyne functional group bound thereto, wherein the carrier comprises at least one selected from the group consisting of boron compounds and target molecules, and when the albumin is bound to the azide functional group, the carrier is bound to the cyclooctyne functional group, and when the albumin is bound to the cyclooctyne functional group, the carrier is bound to the azide functional group.

[0012] In the present invention, the number of azide or cyclooctane functional groups introduced into the albumin may be 1 to 30.

[0013] In the present invention, the nanoplatform may contain 1 to 15 boron compounds.

[0014] In the present invention, the boron compound may be at least one selected from the group consisting of carborane and sodium borocaptate (BSH).

[0015] In the present invention, the nanoplatform may contain 1 to 10 target molecules.

[0016] In the present invention, the target molecule may include at least one selected from the group consisting of folic acid or a derivative thereof, a sugar compound, and RGD (arginyl-glycyl-aspartic acid) or a derivative thereof.

[0017] In the present invention, the transmitting material further comprises a radioactive isotope, and the radioactive isotope is 3 H, 11 C,18 F, 14 Cl, 32 P, 35 S, 36 Cl, 45 Ca, 51 Cr, 57 Co, 58 Co, 59 F, 64 Cu, 67 Ga, 68 Ga, 89 Zr, 90 Y, 99 Mo, 99m Tc, 111 In, 131 I, 125 I, 124 I, 123 I, 186 Re, 188 Re, 225 Ac, 212 Pb, 117m Sn, and 177 It may be one or more selected from the group consisting of Lu.

[0018] In the present invention, the radioactive isotope is labeled with a chelating agent, and the chelating agent is NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid), DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DFO (3-[6,17-dihyroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxylamino)-6,11,17,22-tetraazaheptaeicosane]thiourea), DTPA (diethylenetriaminepentaacetic acid), N2S2(diaminedithiol), p-SCN-Bn-NOTA (2-(4'-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid), NODAGA (1,4,7-triazacyclononane,1-glutaric acid-4,7-acetic acid),p-SCN-Bn-DOTA (2-(4'-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane1,4,7,10-tetraacetic acid), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid),p-SCN-Bn-DTPA (2-(4-isothiocyanatobenzyl)-diethylenetriaminepentaacetic acid),p-SCN-Bn-DFO It may be at least one selected from the group consisting of (1-(4-Isothiocyanatophenyl)-3-[6,17-dihyroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxylamino)-6,11,17,22-tetraazaheptaeicosane]thiourea) and HYNIC (hydrazinonicotinic acid).

[0019] In the present invention, the transmitting material further includes a fluorescent material, and the fluorescent material is FNR (Ferrodoxin NADP(+) reductase), cyanine-based fluorescent material, TAMRA (tetramethylrhodamine-5-maleimide), Flamma ® It may be at least one selected from the group consisting of fluorescent substances and ICG (indocyanine green).

[0020] In the present invention, the nanoplatform can selectively target cancer cells or cancer tissues in which folate receptors are overexpressed.

[0021]

[0022] The present invention also provides a composition for boron neutron capture therapy comprising the albumin-based nanoplatform for boron neutron capture therapy.

[0023] The present invention also provides a pharmaceutical composition for preventing or treating cancer, comprising the albumin-based nanoplatform for boron neutron capture therapy.

[0024] In the present invention, the cancer disease may include cancer cells or cancer tissues in which a folate receptor is overexpressed.

[0025] In the present invention, the cancer disease may be selected from head and neck cancer, ovarian cancer, lung cancer, cervical cancer, bone cancer, breast cancer, central or peripheral nervous system cancer, digestive cancer, germ cell cancer, adenocarcinoma, blood cancer, renal-urinary tract cancer, liver cancer, pleural cancer, prostate cancer, sarcoma, and skin cancer.

[0026] The albumin-based nanoplatform according to the present invention can effectively deliver boron to specific cancer tissues by combining a boron compound containing an excess of boron with a target molecule that specifically targets cancer tissues as a delivery material using a click chemical functional group introduced to the albumin surface, and in particular, the albumin-based nanoplatform of the present invention can deliver a sufficient amount of boron molecules to target cancer tissues even with a dose that is 10 times less than that of a conventional boron neutron capture therapy (BNCT) drug, and thus can be utilized as a composition for boron neutron capture therapy and an anticancer therapeutic agent.

[0027] Figure 1 is a schematic diagram of a process for determining the pharmacodynamic characteristics of a drug in a mouse model using a drug labeled with a radioactive isotope according to one embodiment of the present invention.

[0028] FIG. 2 is a schematic diagram of a nanoplatform for albumin-based boron neutron capture therapy targeting cancer according to one embodiment of the present invention.

[0029] Figure 3 shows a reaction scheme for synthesizing an azide-bonded carborane according to one embodiment of the present invention.

[0030] Figures 4a and 4b are the results of fluorescence analysis confirming the cancer cell targeting and infiltration ability of the albumin nanoplatform according to the number of folic acid bonds according to one embodiment of the present invention.

[0031] Figure 5 shows, according to one embodiment of the present invention, folic acid and [ 10 B] This is the result of a fluorescence analysis confirming the cancer cell targeting ability of albumin nanoparticles bound to carborane (FA-ANP-FNR648) and albumin nanoparticles not bound to folic acid (ANP-FNR648).

[0032] Figure 6a shows an Alb-ADIBO cancer mouse model constructed using a head and neck cancer cell line according to one embodiment of the present invention. 11 -[ 10 B] 6 -FA 5-FNR648 or Alb-ADIBO 11 -FA 3 -[ 10 B] 6 -These are the results of a fluorescence analysis that confirmed the targeting ability of each nanoparticle to cancer tissue after injecting FNR648.

[0033] Figure 6b compares the albumin nanoplatform uptake of KB cell lines and FaDu cell lines according to one embodiment of the present invention.

[0034] FIG. 7 is a standard graph of intensity versus concentration of boron used when labeling an albumin nanoplatform with a fluorescent material according to one embodiment of the present invention.

[0035] Figure 8a shows the results of confirming the efficiency of radioisotope labeling on an albumin nanoplatform according to one embodiment of the present invention using Radio-TLC.

[0036] Figures 8b and 8c are cancer mouse models constructed using head and neck cancer cell lines according to one embodiment of the present invention. 64 Cu-NOTA-Alb-ADIBO 11 -B 10 -FA or 64 Cu-NOTA-Alb-ADIBO 11 -FA-B 10 This is a PET image result that confirmed the targeting ability of each nanoparticle to tumor tissue.

[0037] FIG. 9 is a standard graph of intensity versus concentration of boron used when labeling an albumin nanoplatform with a radioactive isotope according to one embodiment of the present invention.

[0038] FIG. 10 is a graph showing the correlation between a PET signal and boron concentration according to one embodiment of the present invention.

[0039] Hereinafter, specific implementations of the present invention will be described in more detail. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one skilled in the art to which the present invention pertains. In general, the nomenclature used herein is well known and commonly used in the art.

[0040]

[0041] The present invention relates to an albumin-based nanoplatform for boron neutron capture therapy (BNCT) for selectively targeting cancer tissues and delivering boron into cancer tissues, a composition for boron neutron capture therapy including the same, and a composition for preventing or treating cancer diseases.

[0042] Targeted drug delivery platforms are diversifying with technological advancements and the use of convergent new materials. However, their utility is limited due to the time required for infusion into the body or rapid uptake by the spleen. In particular, concerns have been raised regarding increased dosage and immune responses, and existing nanoparticles suffer from inherent toxicity due to their material composition. In the present invention, we utilize albumin, a biocompatible material, and demonstrate that albumin-based nanocarriers, which bind an excess of boron compounds to a single albumin particle, effectively target cancer tissues and deliver high concentrations of boron compounds with only a small dose. Furthermore, we demonstrate that the albumin-based nanocarriers themselves can be used to treat cancer.

[0043] More specifically, one aspect of the present invention is a nanoplatform obtained by a click chemistry reaction of albumin having an azide (N3) or cyclooctyne functional group attached thereto and a carrier having an azide (N3) or cyclooctyne functional group attached thereto,

[0044] The above-mentioned transmitting agent comprises at least one selected from the group consisting of boron compounds and target molecules,

[0045] An albumin-based nanoplatform for boron neutron capture therapy is provided, wherein when the albumin is combined with an azide functional group, the carrier is combined with a cyclooctyne functional group, and when the albumin is combined with a cyclooctyne functional group, the carrier is combined with an azide functional group.

[0046] In the present invention, albumin refers to a protein that constitutes the basic substance of cells, is found in high concentrations in the blood, and is produced in the liver. Albumin has the lowest molecular weight among simple proteins found in nature. Serum albumin in the blood maintains and restores plasma volume, preventing shock caused by excessive bleeding and being used in surgery and burn treatment. It is also known to have an oxygen-carrying capacity similar to hemoglobin.

[0047] The albumin described above may include any albumin that can be formulated, but is preferably derived from human plasma or recombinant human serum albumin produced through genetic engineering, but is not limited thereto. Genetic information regarding the albumin of the present invention can be obtained from known databases such as NCBI GenBank.

[0048] In the present invention, the number of amino groups (-NH2) exposed on the surface of albumin may be 15 to 30.

[0049] In a specific embodiment of the present invention, a nanoplatform capable of exhibiting a selective anticancer therapeutic effect on target cancer tissues by effectively delivering a boron compound to target cancer tissues based on biocompatible albumin was manufactured, and a representative human serum albumin (HSA) was used, and a cyclooctane or azide (N3) group, which is one functional group of click chemistry, was introduced to the surface of human serum albumin under reaction conditions that minimized the denaturation of HSA.

[0050] In the present invention, azide (N3) is a reactive group composed of three nitrogen atoms, has high reactivity, and is known to play a role as an electron donor in the 1,3-dipolar cycloaddition reaction, which is a type of Cu-free click chemistry, to form a triaza-5-membered ring.

[0051] The cyclooctyne functional group is an eight-membered aliphatic ring containing a triple bond under ring strain. It is known to act as an electron acceptor, particularly in the 1,3-dipolar cycloaddition reaction, a type of Cu-free click chemistry, to form a triaza-5-membered ring. The structural characteristics of cyclooctyne, namely the triple bond structure under ring strain, enable click chemistry even without a Cu(I) catalyst.

[0052] The cyclooctyn functional group is 4-cyclooctyn-1-yl( ), 3-cyclooctyn-1-yl( ), 2-cyclooctyn-1-yl( ), Monofluorinated cyclooctyne (MOFO) ( ), Difluorinated cyclooctyne (DIFO) ( ), Dimethoxyazacyclooctyne (DIMAC) ( ), Dibenzocyclooctyne(DIBO)( ), Azadibenzocyclooctyne (ADIBO) group ( ) and biarylazacyclooctynone (BARAC) ( ) may be one or more selected from the group consisting of, but is not necessarily limited thereto.

[0053] Albumin having an azide (N3) or cyclooctyne functional group attached thereto can be obtained by the steps of (a) dissolving albumin in phosphate-buffered saline (PBS), (b) dissolving azide-NHS or cyclooctyne-NHS in DMSO, and (c) mixing the obtained solutions and reacting them at 20 to 37°C for 30 minutes to 1 hour.

[0054] (a) In step, the phosphate buffer solution (PBS) may have a pH of 6.8 to 7.6, and preferably a pH of 7.0 to 7.4.

[0055] (b) The amount of DMSO used to prepare the azide-NHS solution or the cyclooctane-NHS solution in step may be 2% (v / v) or less of the total reaction solution.

[0056] (c) The mixing molar ratio of albumin and azide-NHS or cyclooctane-NHS in step may be 1:1 to 1:25.

[0057] (c) When mixing an albumin solution and an azide-NHS solution in step (c), the functional group bound to the albumin may be an azide functional group, and when mixing an albumin solution and a cyclooctane-NHS solution in step (c), the functional group bound to the albumin may be a cyclooctane functional group.

[0058] In one embodiment of the present invention, a human serum albumin (HSA) solution and an ADIBO-NHS solution were mixed and reacted at 37°C for 30 minutes to produce HSA-ADIBO.

[0059] The number of click-reactive functional groups (azide or cyclooctane functional groups) introduced onto the albumin surface is preferably 1 to 30, more preferably 6 to 20. In this case, the drug can remain in the blood for a long time after injection into the body, increasing the likelihood of uptake into the target site. Conversely, if the number of click-reactive functional groups exceeds the above range, the drug may be immediately uptaken into the liver upon injection, limiting uptake into other target disease sites.

[0060] The number of click-reactive functional groups introduced on the albumin surface can be controlled by the reaction ratio of albumin and azide-NHS or cyclooctane-NHS.

[0061] An albumin-based nanoplatform for boron neutron capture therapy, which is one embodiment of the present invention, is obtained by mixing a solution containing albumin having an azide (N3) or cyclooctyne functional group bonded thereto and a solution containing a carrier having an azide (N3) or cyclooctyne functional group bonded thereto, and performing a click chemistry reaction, wherein the click chemistry reaction may be a copper-free click chemistry reaction.

[0062] In one embodiment of the present invention, the azide functional group used as the click chemistry functional group is an electron donor, and the cyclooctane functional group is an electron acceptor. Therefore, when the functional group bonded to albumin is an azide functional group, the functional group bonded to the carrier is preferably a cyclooctane functional group, and when the functional group bonded to albumin is a cyclooctane functional group, the functional group bonded to the carrier is preferably an azide functional group.

[0063] Since the albumin-based nanoplatform for boron neutron capture therapy of the present invention has multiple click-reactive functional groups, various carrier substances can be bound to multiple reactive groups.

[0064] The above-mentioned delivery substance means a substance that is combined with albumin and delivered into a living body, and in the present invention, the delivery substance includes at least one of a boron compound and a target molecule.

[0065] The boron compound is bound to the albumin nanoplatform of the present invention and delivered to target cancer cells to enable effective boron neutron capture therapy.

[0066] The boron compounds used in the present invention include carborane and sodium borocaptate (BSH), and carborane is preferred because it can bind more boron to albumin. In an embodiment of the present invention, [containing 10 boron atoms per molecule] 10 [B] We confirmed that more boron can be delivered to cancer cells at a lower dose using carborane.

[0067] Carboranes are cluster compounds composed of boron, carbon, and hydrogen atoms, essentially large boron compounds in which boron hydride fragments are linked together to form boron cages.

[0068] In the present invention, 10 per mole 10 [ having B10 B] The azide (N3) is attached so that the carborane compound can be combined with the click chemical functional group ADIBO of albumin. 10 B]Carborane-N3 was synthesized, and its structure is as shown in Equation 1 below.

[0069]

[0070] [Formula 1]

[0071]

[0072]

[0073] The albumin-based nanoplatform for boron neutron capture therapy of the present invention may contain 1 to 15 boron compounds, preferably 2 to 12, and more preferably 4 to 9.

[0074] The nanoplatform of the present invention may also include a targeting molecule to precisely deliver boron to cancer cells.

[0075] The term "target molecule" of the present invention refers to a molecule that targets a specific environment in a living body, and specifically refers to a molecule that targets a substance existing in a specific environment in a living body and guides the albumin nanoplatform of the present invention to the corresponding specific environment. The target molecule of the present invention can guide the albumin nanoplatform of the present invention to a specific environment, or confirm the presence or absence of a specific environment, the location of the specific environment, the degree of the specific environment, etc. In the present invention, the specific environment in a living body targeted by the target molecule may be a tissue-specific specific environment or a disease-specific specific environment. In other words, the target molecule may target a substance existing tissue-specifically or a disease-specific substance. A tissue-specific substance may be a protein, RNA, a compound accumulated or produced, etc. existing tissue-specifically, and a disease-specific substance may be a protein, RNA, a compound accumulated or produced, etc. existing depending on the type of disease.

[0076] The albumin-based nanoplatform of the present invention may contain 1 to 10 target molecules, more preferably 1 to 8. In this case, the nanoplatform's targeting potential to the target site upon injection into the human body can be increased, thereby increasing the likelihood of uptake into the target site.

[0077] In particular, in the present invention, the target molecule may be folate or a derivative thereof, a sugar compound, or RGD (arginyl-glycyl-aspartic acid) or a derivative thereof that targets a folate receptor present in large numbers in cancer tissue.

[0078] In a preferred embodiment of the present invention, the target molecule comprises folic acid.

[0079] Folic acid is a vitamin, also known as vitamin B9 or vitamin M. Folic acid binds specifically to the folate receptor. The folate receptor is a tumor-associated glycosylphosphatidylinositol anchor protein that allows bound folate or folate-conjugated substances to be uptaken through receptor-mediated endocytosis.

[0080] In the present invention, it was confirmed that the albumin nanoplatform bound to folic acid can accurately deliver boron compounds to cancer cells by targeting folic acid receptors overexpressed in head and neck cancer, ovarian cancer, lung cancer, cervical cancer, etc.

[0081] The folic acid can be bound to the albumin nanoplatform in numbers of 1 to 10, and it is preferable that it is bound in numbers of 1 to 6.

[0082] In the present invention, the target molecule may include a sugar compound. The sugar compound may be a mannosyl group, a galactosyl group, or a glucosyl group. The structures of the mannosyl group, galactosyl group, and glucosyl group are as shown in Formulas 2 to 4 below, respectively.

[0083]

[0084] [Formula 2]

[0085]

[0086] [Formula 3]

[0087]

[0088] [Formula 4]

[0089]

[0090]

[0091] Specifically, an albumin nanoplatform containing a mannosyl group or galactosyl group as a delivery agent has targeting ability for macrophages, thereby enabling drug delivery to macrophages, and as a result, can function as a platform for preemptive diagnosis and treatment of metastatic cancer.

[0092] Mannose receptors are known to be abundant on cells involved in defense mechanisms, with Kupffer cells being a prime example of such immune cells. When the transmitter is mannosyl, it targets Kupffer cells, so it typically acts in the liver, but it can also affect the blood, muscles, spleen, and lungs.

[0093] In addition, when the transmitter is a galactosyl group, it can exhibit hepatobiliary excretion through the gall bladder. In addition, the albumin-based nanoplatform of the present invention, in which a glucosyl group is bound as a transmitter, has a targeting ability for M1 type macrophages, and through this, it can be selectively taken up by inflammatory macrophages in damaged tissues, suppress the inflammatory response and ROS production induced by inflammatory macrophages, restore the function of damaged mitochondria, and suppress cell death in damaged tissues, so that it can be used as a nanoplatform that can prevent or treat progression to serious diseases.

[0094] In the present invention, the albumin nanoplatform may contain 4 to 8 sugar compounds, more preferably 5 to 7 sugar compounds.

[0095] Additionally, in the present invention, the target molecule may include RGB.

[0096] RGD is a tripeptide composed of three amino acids—arginine (L-arginine), glycine (glycine), and aspartic acid (L-aspartic acid). It was first discovered in the cell adhesion domain of fibronectin. Because it has a strong affinity for integrin receptors, which are primarily expressed on the endothelial cell layer of tumor blood vessels, it is widely used as a universal tool in research, including drug-targeted therapies and simple reagents.

[0097] The nanoplatform of the present invention may also further include one or more carriers in addition to the boron compound and the target molecule, in which case, multiple substances may be simultaneously subjected to a click chemical reaction with albumin having an azide or cyclooctane functional group attached thereto, and multiple substances may be sequentially subjected to a click chemical reaction, respectively.

[0098] In the present invention, the delivery material may further include a radioisotope, and the radioisotope may be labeled with a chelating agent.

[0099] Radioisotopes are elements with the same atomic number but different atomic masses. Among these isotopes, those that are radioactive are called radioisotopes. These radioisotopes can be used as important markers for diagnosing diseases or checking pharmacodynamics by utilizing the characteristic of attenuating radioactivity by emitting gamma rays or other subatomic particles. The radioisotopes that can be used as markers in the present invention are those known in the art and can be used without limitation. 3 H, 11 C, 18 F,14 Cl, 32 P, 35 S, 36 Cl, 45 Ca, 51 Cr, 57 Co, 58 Co, 59 F, 64 Cu, 67 Ga, 68 Ga, 89 Zr, 90 Y, 99 Mo, 99m Tc, 111 In, 131 I, 125 I, 124 I, 123 I, 186 Re, 188 Re, 225 Ac, 212 Pb, 117m Sn, and 177 It may be at least one selected from the group consisting of Lu, and preferably 11 C, 18 F, 64 Cu, 67 Ga, 68 Ga, 89 Zr, 99m Tc, 111 In and 123 It may be I, but is not necessarily limited to this.

[0100] Chelating agents are those that bind radioactive isotopes to albumin, for example, NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid), DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DFO (3-[6,17-dihyroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxylamino)-6,11,17,22-tetraazaheptaeicosane]thiourea), DTPA (diethylenetriaminepentaacetic acid), N2S2 (diaminedithiol), p-SCN-Bn-NOTA (2-(4'-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid), NODAGA (1,4,7-triazacyclononane,1-glutaric acid-4,7-acetic acid),p-SCN-Bn-DOTA (2-(4'-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid),p-SCN-Bn-DTPA (2-(4-isothiocyanatobenzyl)-diethylenetriaminepentaacetic acid),p-SCN-Bn-DFO It may be at least one selected from the group consisting of (1-(4-Isothiocyanatophenyl)-3-[6,17-dihyroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxylamino)-6,11,17,22-tetraazaheptaeicosane]thiourea) and HYNIC (hydrazinonicotinic acid), but is not necessarily limited thereto.

[0101] The delivery material can be radiolabeled by including a radioisotope, and the radiolabeled material can be used in research to determine the correlation between the results of quantitative evaluation based on nuclear medicine imaging and the mass results of the radiolabeled material.

[0102] In one embodiment of the present invention, an albumin nanoplatform containing a boron compound is a radioactive isotope. 64 By labeling with Cu, obtaining PET images and measuring PET signals, it was confirmed that there was a correlation between boron concentration and PET signal.

[0103] In addition, unlike the existing BPA, it is possible to predict the drug intake in advance through quantitative analysis based on isotopes, and to evaluate at what point in time neutron beam irradiation is most effective in treating diseases.

[0104] In this regard, as shown in Fig. 1, after establishing a head and neck cancer mouse model, the drug uptake in the tumor, blood, and liver is compared based on nuclear medicine imaging, and the pharmacodynamic characteristics related to the drug's absorption, distribution, and excretion in the body are identified by confirming the residual amount of the drug in the blood, so that it is possible to evaluate at what point after actual drug injection the neutron beam should be irradiated to enable the most effective cancer treatment.

[0105] A fluorescent material refers to a material that emits a specific wavelength of visible light for a specific wavelength, and in the present invention, all fluorescent materials that can be combined with an albumin-based nanoplatform for boron neutron capture therapy and used to identify the location of the nanoplatform are included.

[0106] Specifically, in the present invention, fluorescent materials that can be used as markers are those known in the art without limitation, and include, but are not limited to, rhodamine-based fluorescent materials including rhodamine, TAMRA, etc.; fluorescein-based fluorescent materials including fluorescein isothiocyanate (FITC) and fluoreceinamidite (FAM), etc.; bodipy-based fluorescent materials (borondipyrromethene); alexa fluor-based fluorescent materials; and cyanine-based fluorescent materials including Cy3, Cy5, Cy7, and indocyanine green.

[0107] In the present invention, the transmitting material may further include a fluorescent material.

[0108] A fluorescent material refers to a material that emits a specific wavelength of visible light for a specific wavelength, and in the present invention, all fluorescent materials that can be combined with an albumin-based nanoplatform for boron neutron capture therapy and used to identify the location of the nanoplatform are included.

[0109] Specifically, in the present invention, fluorescent materials that can be used as markers are those known in the art without limitation, and include, but are not limited to, rhodamine-based fluorescent materials including rhodamine, TAMRA, etc.; fluorescein-based fluorescent materials including fluorescein isothiocyanate (FITC) and fluorescein amidite (FAM), etc.; bodipy-based fluorescent materials (boron-dipyrromethene); alexa fluor-based fluorescent materials; and cyanine-based fluorescent materials including Cy3, Cy5, Cy7, and indocyanine green.

[0110] In the present invention, the albumin-based nanoplatform for boron neutron capture can precisely deliver boron compounds to target cancer cells by selectively targeting cancer cells or cancer tissues in which folate receptors are overexpressed.

[0111]

[0112] Another aspect of the present invention provides a composition for boron neutron capture therapy comprising the albumin-based nanoplatform.

[0113] According to one embodiment of the present invention, a cancer mouse model constructed by transplanting a head and neck cancer cell line is injected with a boron compound as shown in FIG. 2. 10 B]Albumin nanoplatform (Alb-ADIBO) in which carborane and target molecule folic acid are combined through click chemistry 11 -[ 10 B] 6 -FA 5 -FNR648 or Alb-ADIBO 11 -FA 3 -[ 10 B] 6 -FNR648) was injected, and in vivo fluorescence expression was confirmed. As a result of extracting cancer tissue from each experimental animal and quantifying it, it was confirmed that fluorescence expression increased by about 60% in cancer tissue with overexpression of folate receptors compared to cancer tissue with underexpression of folate receptors. Accordingly, it was confirmed that the albumin nanoplatform of the present invention can selectively target cancer tissues, and through this, it was confirmed that boron is effectively delivered to cancer tissues.

[0114] In addition, in the case of BPA, a conventional BNCT drug, to confirm the BNCT treatment effect 10It has been reported that the cancer tissue accumulation concentration of B is approximately 4 to 6 μg (0.4 to 0.6 μmol) per g of cancer tissue, and approximately 250 mg / kg of BPA is required to secure the cancer tissue concentration at this level. In addition, in a recent animal experiment using BPA, it was reported that when 5 mg was injected per mouse, 30 μg / g was taken up in cancer tissue, which indicates a 1.2% delivery ability to cancer per mouse when the cancer weight is 2 g (60 μg / 5000 μg).

[0115] However, according to one embodiment of the present invention, 16 μg of albumin-bound per mouse is established by transplanting the head and neck cancer cell line. 10 B] Carborane was injected and the uptake was confirmed through fluorescence analysis, confirming that 1 to 1.5 μg / g was taken up by cancer tissue. At this time, the weight of the cancer isolated from the mouse was 1.3 g. Based on this, the delivery efficiency of the albumin nanoplatform to the cancer per mouse was evaluated, and the delivery efficacy was confirmed to be 8 to 12% compared to the injected amount.

[0116] Additionally, 2.3 μg of albumin-bound [ per mouse was established by transplanting head and neck cancer cell lines. 10 B] As a result of injecting carbolane and confirming the uptake amount through PET imaging, it was confirmed that approximately 290 ng was uptaken into cancer tissue per mouse, and based on this, the delivery efficacy of the albumin nanoplatform was confirmed to be approximately 12.6%.

[0117] This means that the albumin nanoplatform of the present invention can effectively deliver boron molecules for BNCT treatment by selectively targeting cancer tissues at a dose more than 10 times lower than that of the conventional BNCT drug, BPA.

[0118] Furthermore, existing boron neutron capture therapy compositions had the problem of requiring long-term injections because they were excreted through the kidneys or liver, but the boron neutron capture therapy composition according to the present invention still remains in cancer cells even after 24 hours, so a single injection can sufficiently maintain the boron concentration for a necessary period of time, thereby causing a change in the cancer treatment mechanism.

[0119]

[0120] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating cancer, comprising the albumin-based nanoplatform for boron neutron capture therapy as an active ingredient.

[0121] The cancer disease may include cancer cells or cancer tissues in which folate receptors are overexpressed, and may be, for example, one or more cancer diseases selected from the group consisting of head and neck cancer, ovarian cancer, lung cancer, cervical cancer, bone cancer, breast cancer, central or peripheral nervous system cancer, digestive cancer, germ cell cancer, adenocarcinoma, blood cancer, renal-urinary tract cancer, liver cancer, pleural cancer, prostate cancer, sarcoma, and skin cancer.

[0122] The pharmaceutical composition may further comprise an appropriate pharmaceutically acceptable carrier, excipient or diluent according to a conventional method. The pharmaceutically acceptable carrier is one commonly used in the preparation of a formulation, and includes, but is not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

[0123] The pharmaceutical composition of the present invention may further include, in addition to the above ingredients, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. With regard to suitable pharmaceutically acceptable carriers and formulations, each ingredient can be preferably formulated according to the method disclosed in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0124] The pharmaceutical composition of the present invention can be administered either orally or parenterally, and parenteral administration includes intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc.

[0125] Oral dosage forms include, for example, tablets, pills, hard and soft capsules, solutions, suspensions, emulsions, syrups, and granules. These dosage forms may further contain, in addition to the active ingredient, diluents (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycine), lubricants (e.g., silica, talc, stearic acid and its magnesium or calcium salts, and / or polyethylene glycol). In addition, the tablets may contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidine, and, if desired, disintegrants or effervescent mixtures such as starch, agar, alginic acid or its sodium salt, and / or absorbents, coloring agents, flavoring agents, and sweetening agents. The above formulation can be prepared by conventional mixing, granulating or coating methods.

[0126] In addition, a representative example of a formulation for parenteral administration is an injectable preparation, and solvents for the injectable preparation include water, Ringer's solution, isotonic saline solution, or a suspension.

[0127] The sterile fixed oil of the above injectable preparation can be used as a solvent or suspending medium, and any non-irritating fixed oil, including mono- and di-glycerides, can be used for this purpose. The above injectable preparation can also use a fatty acid such as oleic acid.

[0128] The composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment. The effective dosage level may be determined based on the type and severity of the patient's disease, the activity and sensitivity of the drug to the drug, the time of administration, the route of administration and excretion rate, the duration of treatment, factors including concurrently used drugs, and other factors well known in the medical field. The composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or in multiple doses. It is important to take all of the above factors into consideration and administer an amount that achieves the maximum effect with the minimum amount without causing side effects, and this can be easily determined by a skilled artisan.

[0129] Specifically, the effective dose of the composition according to the present invention may vary depending on the patient's age, sex, and weight, and is generally 0.001 to 150 mg per kg of body weight, preferably 0.01 to 100 mg, administered daily or every other day, or divided into 1 to 3 times a day. However, since the dosage may increase or decrease depending on the route of administration, sex, body weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

[0130]

[0131] Example

[0132]

[0133] The present invention is described in more detail through the following examples. However, these examples are intended to illustrate some experimental methods and configurations of the present invention, and the scope of the present invention is not limited to these examples.

[0134]

[0135] Manufacturing Example 1: Manufacturing of an Albumin-Based Nanoplatform for Boron Neutron Capture Therapy

[0136]

[0137] 1-1. Preparation of albumin with ADIBO functional group

[0138]

[0139] Human serum albumin (HSA) was dissolved in phosphate-buffered saline (PBS, pH 7.4) at a concentration of 40 mg / mL, and the albumin solution was dispensed at 20 mg / 0.5 mL per vial. ADIBO-NHS was dissolved in DMSO (10 mg / 50 μL) and added to the albumin solution to prepare a volume of 500 μL.

[0140] Albumin and ADIBO-NHS were reacted at a ratio of 1:14 at room temperature for 6 hours, and purified using an amicon Ultra-0.5 centrifugal filter tube to obtain Alb-ADIBO.

[0141]

[0142] 1-2. Preparation of carboranes with azide functional groups

[0143]

[0144] By the same process as in Fig. 3 10 A carborane compound is prepared by combining B with an azide (N3) that can be combined with the ADIBO functional group. 10 B] Carborane-N3 compound was synthesized.

[0145] First, 4-o-carboranylnitrobenzene (3) was obtained from o-carborane (1). 4-o-carboranylnitrobenzene was hydrogenated using Pd / C / H2 to synthesize 4-o-carboranylaniline (4) in a yield of 95%.

[0146] Specifically, 4-fluoronitrobenzene (0.98 g, 7.0 mmol, (2)) dissolved in 10 mL of dry DMF was added via a cannula to a stirred solution of o-carborane (1.0 g, 7.2 mmol, (1)) and NaH (0.24 g, 1.2 mmol) dissolved in 20 mL of dry DMF at 0 °C for 30 min. The reaction mixture was maintained at 0 °C for 10 min and then slowly heated to room temperature. The mixture was stirred for an additional hour, and 15 mL of 10% HCl aqueous solution was added to the reaction mixture to terminate the reaction. The crude 4-o-carbonylnitrobenzene was extracted with AcOEt (20 mL × 2), and the organic layer was washed with distilled water (20 mL × 2) and brine, dried over MgSO4, and concentrated. The residue was then purified by flash column chromatography (EA-n-hexane, 1:4, Rf = 0.53) to obtain 1.6 g (90%) of 4-o-carboranylnitrobenzene (3).

[0147] 4-o-Carboranylnitrobenzene was dissolved in EtOH (100 mL), 10% Pd / C was added, and the mixture was stirred under H2 until a large amount of hydrogen was consumed. The mixture was then filtered, the solvent was evaporated using a rotary evaporator, and the residue was purified by flash column chromatography (EA-n-hexane, 1:4, Rf = 0.34) to obtain 1.2 g (95% yield) of 4-o-carbonylaniline (4).

[0148] Next, 11-azido-3,6,9-trioxaundecanoic acid was added in DMF solvent to obtain azide-bonded carborane (6).

[0149]

[0150] 1-3. Manufacturing of albumin nanoplatforms combined with carbolane and folic acid

[0151]

[0152] First, folate-N3 was prepared by introducing an azide functional group to the amino acid side chain of the N-terminus of the folic acid derivative.

[0153] Dissolve Alb-ADIBO in PBS [ 10 B]Carborane-N3 was mixed in a molar ratio of 1:8 and reacted at 37°C for 1 hour.

[0154] Next, folic acid-N3 was mixed in a molar ratio of 1:6 and reacted at 37°C for 1 hour to sequentially introduce carborane and folic acid.

[0155] In addition, the order of introduction of carbolane and folic acid was reversed to form Alb-ADIBO, respectively. 11 -B 10 -FA and Alb-ADIBO 11 -FA-B 10 The number of bonds for each compound was confirmed through molecular weight confirmation using MALDI-TOF, as shown in Table 1 below.

[0156]

[0157] MALDI-TOF DOF Albumin (Alb) Boron (B) 10 )Folic acid (FA)Alb-ADIBO 11 -B 10 -FA16.595.64Alb-ADIBO 11 -FA-B 10 16.433.88

[0158]

[0159] 1-4. Manufacturing of albumin nanoplatforms conjugated with fluorescent materials

[0160]

[0161] Fluorescence-conjugated Alb-ADIBO of FNR648-N3 using HSA-ADIBO 11 -B 10 -FA-FNR648 and Alb-ADIBO 11 -FA-B 10 -FNR648 complex was prepared. 100 nmol / 10 ㎕ of FNR648-N3 was prepared, and the corresponding volume (㎕) of fluorescent material was taken as the molar number of HSA-ADIBO and added to the HSA-ADIBO solution. After reacting at 37℃ for 60 minutes, Alb-ADIBO bound to FNR648-N3 was detected through a PD-10 (desalting) column. 11 -B 10 -FA-FNR648 and Alb-ADIBO 11 -FA-B 10 - Only the FNR648 complex was isolated and obtained.

[0162]

[0163] 1-5. Manufacturing of isotope-labeled albumin nanoplatforms

[0164]

[0165] In HSA-ADIBO and NOTA-N3 64 Cu labeled 64 Using Cu-NOTA-N3 64 Cu-NOTA-Alb-ADIBO 11 -B 10 -FA and 64 Cu-NOTA-Alb-ADIBO 11 -FA-B 10 The complex was prepared. 18 nmol / 100 μl labeled with specific activity 64 Prepare Cu-NOTA-N3 and add the same amount of moles of HSA-ADIBO. 64 Take the corresponding volume (㎕) of Cu-NOTA-N3 and add Alb-ADIBO 11 -B 10 -FA or Alb-ADIBO 11 -FA-B 10After adding to the solution and reacting at 37℃ for 30 minutes, purification is performed. 64 Cu-NOTA-N3 combined 64 Cu-NOTA-Alb-ADIBO 11 -B 10 -FA and 64 Cu-NOTA-Alb-ADIBO 11 -FA-B 10 Only the complex was isolated and obtained.

[0166]

[0167] Experimental Example 1: Confirmation of targeting ability of head and neck cancer cell lines according to folic acid binding number

[0168]

[0169] The targeting ability of albumin nanoparticles to tumor cells was confirmed using the KB cell line overexpressing the folate receptor (FR) to target head and neck cancer.

[0170] First, FA-ANP was constructed by conjugating 1, 3, and 5 azide (N3)-introduced folic acid compounds (FA-N3) that can bind to the ADIBO functional group of albumin nanoparticles (ANP), respectively. To confirm cell uptake ability, a fluorescent substance (FNR-648-N3) with azide (N3) was additionally introduced to the albumin surface, thereby preparing FA-ANP-FNR648. This was treated with KB cells to confirm the cellular uptake level of albumin nanoparticles conjugated with folic acid as a target substance.

[0171] Place cover glasses in a 12-well plate and seed KB cells at 1 × 10 per well. 5 Cells were seeded at 1 cell / ml and cultured at 37°C for 24 hours. After replacing the medium, the cells were treated with FA-ANP-FNR648 conjugated with 1, 3, and 5 folic acids, respectively, and cultured at 37°C in the dark for 1 hour. The cells were then washed and fixed, stained with DAPI, and the fluorescence expression and intensity were confirmed, which are shown in Figures 4a and 4b.

[0172] As can be seen in Figures 4a and 4b, all of the albumin nanoparticles conjugated with 1, 3, and 5 folates were found to be uptaken at very high levels by targeting KB cells compared to the ADIBO-albumin nanoparticles without folate conjugation.

[0173]

[0174] Experimental Example 2: Confirmation of the Cellular Invasion Ability of the Carborane-Folic Acid-Albumin Nanoplatform

[0175]

[0176] [ 10 B] FA-ANP was constructed using a folic acid compound (FA-N3) introduced with an azide (N3) that can be combined with the ADIBO functional group of albumin nanoparticles (ANP) combined with carborane, and a fluorescent substance (FNR-648-N3) introduced with an azide (N3) was additionally introduced to the albumin surface to confirm cell penetration ability, thereby preparing FA-ANP-FNR648.

[0177] Using the KB cell line of Experimental Example 1 as the experimental subject, a cell line in which the receptor was blocked by pre-treating the cells with an excessive amount of folic acid was prepared as a control group, and ANP-FNR648, which does not bind folic acid, was used as a control group, and the cells were treated and fluorescence expression was confirmed.

[0178] Referring to Figure 5, it was confirmed through fluorescence that FA-ANP-FNR648 was bound to tumor cells, but in the case of cells that had been previously treated with excessive amounts of folic acid to block the receptor and when ANP that was not bound to folic acid was used, it was confirmed that it was not targeted at all to the KB cell line.

[0179]

[0180] Experimental Example 3: Confirmation of targeting ability in an animal model of head and neck cancer

[0181]

[0182] Among head and neck cancer cell lines, the KB cell line with overexpression of the folate receptor and the FaDu cell line with underexpression were each inoculated into nude mice to establish a cancer animal model.

[0183] Using 5 animal models each, Alb-ADIBO was administered at a concentration of 0.46 mg / 0.1 mL of albumin and 0.0165 mg / 0.1 mL of carborane. 11 -[ 10 B] 6 -FA 5 -FNR648 or Alb-ADIBO 11 -FA 3 -[ 10 B] 6 -0.1 mL of FNR648 was injected, and fluorescence images were simultaneously acquired using IVIS equipment.

[0184] Additionally, fluorescence images were checked 24 hours after injection, and cancer tissue was removed from each animal model and quantified.

[0185] Referring to Figure 6a, it can be confirmed that cancer tissues were selectively targeted in all animal models regardless of the order of introduction of boron and folic acid.

[0186] Additionally, it was confirmed from Figure 6b that both the KB cell line and the FaDu cell line were targeted, and it was confirmed that the KB receptor with overexpressed folate receptor showed an increase in signal of approximately 60% compared to the FaDu receptor with underexpressed folate receptor.

[0187]

[0188] Experimental Example 4: Determination of intratumoral boron uptake in an animal model of head and neck cancer.

[0189]

[0190] Mass analysis was performed to confirm the boron concentration in cancer tissue of an animal model of head and neck cancer administered with albumin nanoparticles of Experimental Example 3.

[0191] First, to confirm the boron used in this experiment, a standard graph of intensity versus boron concentration was obtained, as shown in Figure 7. Next, three mice targeted with albumin nanoparticles were selected, and cancer tissue was isolated and mass analyzed.

[0192] Tumor tissue isolated from each animal was ground, diluted 10-fold, and the concentration determined. Furthermore, the final concentration, taking into account the dilution factor, was determined and converted to boron uptake (ng / g) per tumor weight (g), taking tissue size into account, and is shown in Table 2.

[0193]

[0194]

[0195]

[0196] It was confirmed that the uptake into cancer tissue by the KB cell line was much higher than that into cancer tissue by the FaDu cell line, and in particular, the FaDu cell line showed an uptake rate so low that it could not be measured.

[0197] Furthermore, in the case of KB② among the cancer tissues used in the experiment, since only half of the isolated cancer was used, KB②, which was confirmed to be 740 ng / g, can be converted to approximately 1500 ng / g. Therefore, in the case of KB, the uptake within cancer tissue was confirmed to be at the level of 1000 to 1500 ng / g, which, when converted, indicates an uptake per cancer weight of 1 to 1.5 μg / g.

[0198]

[0199] Experimental Example 5: Conversion of intake through mass analysis

[0200]

[0201] The actual intake amount was converted to the head and neck cancer mouse model administered with the albumin nanoparticles of Experimental Example 3 above.

[0202] The albumin nanoplatform used is Alb-ADIBO, in which 11 ADIBOs are introduced on the albumin surface and 3 folic acid and 6 boron compounds are combined. 11 -FA 3 -[ 10 B] 6 Based on the compound. The above albumin nanoplatform was injected into mice at 0.46 mg / 0.1 mL, and the concentration of carborane in the solution was 0.0165 mg / 0.1 mL. Therefore, if the boron injected into the mouse is converted to μg, it is 16 μg per mouse. 10 B] It can be seen that carborane has been injected.

[0203] In the above experimental example 4, it was confirmed that the intake according to cancer mass was 1 to 1.5 μg / g, and considering that the cancer weight per mouse was approximately 1.3 g, it can be converted to 1 to 1.5 μg / g × 1.3 g = 1.3 to 1.95 μg / total tumor.

[0204] Also, the injected [ 10 B] Carborane is 16 μg per mouse, which is 1 to 1.5 μg / 16 μg × 100 = 8 to 12%, and it was confirmed that approximately 10% of the injected amount was taken up by the cancer.

[0205]

[0206] BPA, the most widely used boron compound for BNCT, is known to be taken up by the tumor at 30 μg / g when 5,000 μg is injected per mouse. When the tumor weight is 2 g, 60 μg of BPA is taken up by the tumor per mouse, which represents a very low injection dose of 1.2%.

[0207] In order to compare the uptake of the albumin nanoparticles of the present invention into cancer tissues with the uptake of BPA into cancer tissues, the uptake of the albumin nanoparticles was converted to the same amount as that of BPA.

[0208] When 16 μg of albumin nanoparticles were injected, an uptake per cancer weight of 1.6 μg / g, which is approximately 10%, was confirmed. In order to achieve an uptake per cancer weight of 30 μg / g, as with the BPA, an injection of approximately 18 times the amount is required.

[0209] That is, 288μg of [ 10 B]Carborane should be injected into mice, and 16 μg of [ per 0.46 mg of albumin 10 B] Since carborane is bound, approximately 10 mg of albumin is required.

[0210] However, the amount of 288 μg of boron compound is more than 50 times less than the amount of 5,000 μg of the BAP compound, [ 10 B] In the case of carborane, 10 per mole [ 10 Because B] exists, it can show the same therapeutic effect even if injected at 1 / 10 level, and 1 mg of albumin and 28.8 μg of [ per mouse 10 B]Carborane alone can exhibit the same cancerous intake as 5,000 μg of BPA. If the above mouse injection dose is converted to a drug dose notation, it is approximately 1.4 mg / kg [ 10 B] 50 mg / kg of albumin nanoplatform containing carborane, which can be administered as an injection with a volume of 2 mL.

[0211]

[0212] Experimental Example 6: Confirmation of targeting ability through nuclear medicine imaging evaluation in an animal model of head and neck cancer.

[0213]

[0214] FA (folate-N3) and 10 B ([ 10 B] Caborane) introduced albumin nanoplatform 64 Nuclear medicine distribution evaluation was performed in a head and neck cancer animal model constructed using the same method as Experimental Example 3 by labeling Cu.

[0215] First, it is manufactured in the same manner as described in Manufacturing Example 1-5, but 1 to 6 FA (folate-N3) are added, [ 10 B] Composed of 1 to 9 carboranes 10 B-Albumin-FA 64 NOTA-N3 with Cu introduced was introduced through a click reaction. 64 Cu-NOTA-Alb-ADIBO 11 -B 10 -FA and 64 Cu-NOTA-Alb-ADIBO 11 -FA-B 10 A complex was manufactured.

[0216] The labeling efficiency of the manufactured complex was confirmed by Radio-TLC, and the results are shown in Figure 8a. After final purification, a purity of 95% or higher was confirmed, and then it was used in animal experiments.

[0217] A total of 18 head and neck cancer mouse models were injected with the same amount, and three mice were sacrificed at a time interval to obtain blood and tumor tissues. The size of the obtained tumor tissues and the activity of the radioactive isotope are shown in Table 3 below.

[0218]

[0219]

[0220]

[0221] Additionally, the PET (Positron Emission Tomography) imaging results confirming fluorescence expression by isotopes in 18 and 3 head and neck cancer models are shown in Fig. 8b and Fig. 8c, respectively.

[0222] As can be seen in Table 3, Figures 8b and 8c, cancer tissues were selectively targeted in all animal models. 64 Cu labeled 10B-Albumin-FA was confirmed to circulate in the blood for up to 4 hours and then be slowly taken up by the liver or tumor site, remaining in the tumor site even after 24 hours and being excreted after 48 hours.

[0223]

[0224] Experimental Example 7: Determination and Conversion of Boron Uptake in Blood and Tumors

[0225]

[0226] Mass analysis was performed to confirm the boron concentration in the cancer tissue of the head and neck cancer animal model injected with albumin nanoparticles of Experimental Example 6.

[0227] To confirm the boron used in this experiment, a standard graph of boron intensity versus concentration was obtained, as shown in Figure 9. Next, mass spectrometry was performed on tumor tissue and blood isolated from 18 mice. Tumor tissue and blood isolated from each animal were pre-diluted 100-fold and the concentration measured. Finally, the concentration was determined by considering the dilution factor and tissue size, and converted to ppb to obtain the final concentration, which is shown in Table 4.

[0228]

[0229]

[0230]

[0231]

[0232] Based on the contents of Table 4, the targeting ability of the albumin nanoplatform was evaluated by converting the intake of boron to a head and neck cancer mouse model administered with albumin nanoparticles.

[0233] The targeting ability of the nanoplatform was evaluated based on data on tumors obtained from mice 4 hours after injection of the albumin nanoplatform (tumor, 4 h).

[0234] The average boron concentration considering the tissue size of the tumor obtained from the above mouse is approximately 1834 ((1858 + 1727 + 1918) / 3) ng / g, and when the average boron concentration is multiplied by the average cancer tissue size of approximately 0.157 ((0.140 + 0.212 + 0.122) g, it is approximately 290 ng, so it can be seen that approximately 290 ng of boron is taken up by the tumor per mouse.

[0235] Injected per mouse [ 10 B] Since carborane is about 2.3 μg, it can be seen that the targeting efficiency for tumors is 0.29 μg / 2.3 μg x 100% = 12.6%. This figure is similar to the targeting efficiency of boron for cancer tissues in Experimental Example 5, and it can be seen through this experiment that the albumin nanoplatform of the present invention can effectively target tumor tissues even with a very small injection amount compared to the BAP compound used in the past.

[0236]

[0237] Experimental Example 8: Correlation between Boron Residue in Blood and Nuclear Medicine Signals

[0238]

[0239] In order to confirm the correlation between the boron residue in the blood and PET images, blood from 18 mice obtained in Experimental Example 7, from which relatively uniform sampling was possible, was used to confirm whether there was a correlation between the nuclear medicine signal (PET signal) and the boron residue in the blood.

[0240] The PET signal (%ID / g), i.e., the radiation signal amount and the boron concentration were measured and shown in Table 5. Based on the results in Table 5, a graph of the change in boron concentration according to the PET signal was derived and shown in Fig. 10.

[0241]

[0242] hrPET(%ID / g)SDMass(ppb)SD045.224.581173115.39218.752.8640749.07410.84 2.77515.6739.5487.981.02312.3348.96243.410.721524.34481.220.5118318.78

[0243]

[0244] As can be confirmed from the graph in Figure 10, it was found that there was a correlation between the PET signal and the boron concentration in the blood.

[0245]

[0246] Although some implementation forms of the present invention have been described above, the present invention is not limited to the implementation forms described above, and can be implemented by modifying and changing them within a scope that does not deviate from the gist of the present invention, and it should be understood that forms with such modifications and changes also fall within the technical spirit of the present invention.

Claims

1. A nanoplatform obtained by a click chemistry reaction of albumin with an azide (N3) or cyclooctyne functional group and a carrier with an azide (N3) or cyclooctyne functional group. The above-mentioned transmitting agent comprises at least one selected from the group consisting of boron compounds and target molecules, An albumin-based nanoplatform for boron neutron capture therapy (BNCT), wherein when the albumin is bound to an azide functional group, the carrier is bound to a cyclooctyne functional group, and when the albumin is bound to a cyclooctyne functional group, the carrier is bound to an azide functional group.

2. In paragraph 1, An albumin-based nanoplatform for boron neutron capture therapy, wherein 1 to 30 azide or cyclooctane functional groups are introduced into the albumin.

3. In paragraph 1, An albumin-based nanoplatform for boron neutron capture therapy, wherein the nanoplatform comprises 1 to 15 boron compounds.

4. In paragraph 1, An albumin-based nanoplatform for boron neutron capture therapy, wherein the boron compound is at least one selected from the group consisting of carborane and sodium borocaptate (BSH).

5. In paragraph 1, An albumin-based nanoplatform for boron neutron capture therapy, wherein the nanoplatform comprises 1 to 10 target molecules.

6. In paragraph 1, An albumin-based nanoplatform for boron neutron capture therapy, wherein the target molecule comprises at least one selected from the group consisting of folic acid or a derivative thereof, a sugar compound, and RGD (arginyl-glycyl-aspartic acid) or a derivative thereof.

7. In paragraph 1, The above-mentioned carrier further comprises a radioactive isotope, wherein the radioactive isotope is 3 H, 11 C, 18 F, 14 Cl, 32 P, 35 S, 36 Cl, 45 Ca, 51 Cr, 57 Co, 58 Co, 59 F, 64 Cu, 67 Ga, 68 Ga, 89 Zr, 90 Y, 99 Mo, 99m Tc, 111 In, 131 I, 125 I, 124 I, 123 I, 186 Re, 188 Re, 225 Ac, 212 Pb, 117m Sn, and 177 An albumin-based nanoplatform for boron neutron capture therapy, comprising at least one selected from the group consisting of Lu.

8. In paragraph 7, The above radioactive isotope is labeled with a chelating agent, and the chelating agent is NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid), DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DFO (3-[6,17-dihyroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxylamino)-6,11,17,22-tetraazaheptaeicosane]thiourea), DTPA (diethylenetriaminepentaacetic acid), N2S2(diaminedithiol), p-SCN-Bn-NOTA (2-(4'-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid), NODAGA (1,4,7-triazacyclononane,1-glutaric acid-4,7-acetic acid),p-SCN-Bn-DOTA (2-(4'-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane1,4,7,10-tetraacetic acid), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid),p-SCN-Bn-DTPA (2-(4-isothiocyanatobenzyl)-diethylenetriaminepentaacetic acid),p-SCN-Bn-DFO An albumin-based nanoplatform for boron neutron capture therapy, comprising at least one selected from the group consisting of (1-(4-Isothiocyanatophenyl)-3-[6,17-dihyroxy-7,10,18,21-tetraoxo-27-[N-acetylhydroxylamino)-6,11,17,22-tetraazaheptaeicosane]thiourea) and HYNIC (hydrazinonicotinic acid).

9. In paragraph 1, The above-mentioned transmitter further contains a fluorescent substance, and the fluorescent substance is FNR (Ferrodoxin NADP(+) reductase), cyanine-based fluorescent substance, TAMRA (tetramethylrhodamine-5-maleimide), Flamma ® An albumin-based nanoplatform for boron neutron capture therapy, comprising at least one selected from the group consisting of fluorescent substances and indocyanine green (ICG).

10. In paragraph 1, An albumin-based nanoplatform for boron neutron capture therapy, wherein the nanoplatform selectively targets cancer cells or cancer tissues in which folate receptors are overexpressed.

11. A composition for boron neutron capture therapy comprising an albumin-based nanoplatform for boron neutron capture therapy according to any one of claims 1 to 10.

12. A pharmaceutical composition for preventing or treating cancer, comprising an albumin-based nanoplatform for boron neutron capture therapy according to any one of claims 1 to 10.

13. In paragraph 12, A pharmaceutical composition for preventing or treating a cancer disease, wherein the cancer disease comprises cancer cells or cancer tissues in which a folate receptor is overexpressed.

14. In paragraph 12, A pharmaceutical composition for preventing or treating a cancer disease, wherein the cancer disease is selected from head and neck cancer, ovarian cancer, lung cancer, cervical cancer, bone cancer, breast cancer, central or peripheral nervous system cancer, digestive cancer, germ cell cancer, adenocarcinoma, blood cancer, renal-urinary tract cancer, liver cancer, pleural cancer, prostate cancer, sarcoma, and skin cancer.

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