KR101585345B1 - Self-assembled albumin nanoparticles for combination therapy of lung cancer and the method for preparing the same - Google Patents

Abstract

The present invention relates to a self-assembling albumin nanoparticle and a manufacturing method of the same. Albumin conjugates having a surface modified with chemical anticancer drugs and fatty acids constitute a self-assembling nanoparticle, and TnF-related apoptosis inducing ligand (TRAIL) is encapsulated inside of the self-assembling nanoparticle to constitute a self-assembling albumin nanoparticle. In addition, the present invention relates to a pharmaceutical composition to prevent or treat cancers, which is composed of the self-assembling albumin nanoparticle. The self-assembling albumin nanoparticle with encapsulated TRAIL and a surface modified with chemical anticancer drugs and fatty acids can be used for pulmonary aspiration and can target lung cancer, while reducing toxicity inside of body and side effects which can be caused by crosslinking agents when a conventional albumin nanoparticle is used as a carrier for drug delivery. In addition, the self-assembling albumin nanoparticle can control a release of drug while reducing side effects by decreasing the administration capacity of drugs through combined administration of both chemical anticancer drugs and TRAIL, thereby being widely used for the purpose of treating and preventing cancers and, in particular, for treating lung cancer.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a self-assembled albumin nanoparticle and a method for preparing the same,

The present invention relates to a self-assembled albumin nanoparticle in which a chemically-chemically-chemically-modified and albumin-conjugated surface modified with a fatty acid constitutes self-assembled nanoparticles and trail (TRAIL) is enclosed within the nanoparticles, and a method for producing the same. The present invention also relates to a pharmaceutical composition for preventing or treating cancer comprising the self-assembled albumin nanoparticle.

Human serum albumin is the most abundant protein in plasma of human body. It is relatively stable in pH change, organic solvent, etc., and is not only highly biocompatible, but also decomposes well in the body. In addition, the compounds are accumulated in cancer sites and inflammation sites, and thus they are attracting interest as drug delivery systems for treating inflammation such as cancer treatment and rheumatoid arthritis.

In particular, albumin has been studied in a variety of drug delivery systems for cancer treatment. For example, it has been reported that methotrexate, one of the anticancer drugs, is directly conjugated to the surface of albumin, thereby increasing drug delivery and anti-cancer effects to the cancer site (Clin. Cancer Res. 5: 753-759, 1999). This can be said to be a study using the advantage that various substances can be modified on the albumin surface due to various residues present on the albumin surface.

In addition, studies have been actively conducted to produce microparticles and nanoparticles using albumin. Since albumin is a biodegradable substance that is toxic and non-immunogenic and degradable in the body, microparticles and nanoparticles formed from albumin are very useful as drug carriers. This is not limited to cancer treatment, but is applied to other diseases and many studies are being carried out. Methods for preparing albumin microparticles and nanoparticles include a method of binding each albumin using a crosslinker such as glutaldehyde (Eur. J. Pharm. Biopharm. 87: 132-141, 2004) And a method of producing albumin by causing denaturation itself (Biomaterials 29: 4022-4028, 2008). The method using a crosslinking agent has a disadvantage in that the crosslinking agent itself can exhibit toxicity in the body. In addition, the method of producing microparticles and nanoparticles using the modification of albumin also has the disadvantage of extinguishing the intrinsic properties of albumin. Therefore, there has been a demand for development of a drug delivery vehicle having a high bioavailability and anticancer effect while minimizing the use of a cross-linking agent to reduce toxicity.

On the other hand, hydrophobic interactions are interactions in which a small number of hydrophilic groups with low affinity to water molecules are trying to converge in aqueous solution, and this interaction occurs thermodynamically spontaneously. It can be understood that non-polar materials are not soluble in the water phase or aggregation with each other is due to such hydrophobic interaction. It can be widely applied in pharmacology because it plays a major role in folding in the three-dimensional structure of proteins or in the formation of micelles. In addition, hydrophobic interactions can also be applied to chromatographic analysis of substances, which is a method of separating hydrophobic interactions using hydrophobic interactions between molecules with a hydrophobic matrix. In the field of pharmacology, in the case of micelles by polymeric polymerization, the hydrophobic part of the polymer forms a center by hydrophobic interaction, and its center can contain a hydrophobic drug, thus acting as a drug delivery system (J. Control Release 145: 116-123, 2010). In another example, a hydrophobic substance is bound to a polymer in the form of a branch, and a polymer is entangled with the polymer by hydrophobic interaction to prepare a hydrogel and encapsulate the drug (J. Control. Release 103 : 235-243, 2005).

The lungs have a very wide surface area (~ 100 m ² ), the thickness of the cells forming the alveolar sac (0.1-0.5 ㎛) is very thin and the density of the cells is lower than other cells. have. It has been known to be effective pathway for local diseases such as asthma / chronic bronchial obstruction because it is fast enough to reach the systemic circulation when the drug is delivered through the lungs and does not undergo liver first pass metabolism.

In addition, due to the above-described characteristics, not only the lung cells exhibit a high permeability to macromolecules but also the amount of bioenzymes present in the lung mucosa is relatively small. Therefore, most of them are injection-dependent Is known to have a great effect on the body's delivery pathway of proteins and peptide drugs (molecular weight of several thousands to tens of thousands Da). Indeed, several reports have reported that these drugs reach a peak blood concentration of up to 30 minutes and bioavailability relative to the subcutaneous route to 50% (Leuprolide). In addition, due to the improvement of patients' convenience that they can mediate themselves, studies on drug delivery systems and delivery media through the respiratory system are actively under way.

The TNF-related apoptosis inducing ligand (TRAIL) is a type of tumor necrosis factor (TNF), and is a cell membrane protein involved in cell apoptosis. It is known that the extracellular portion from the arginine at position 114 to glycine at position 281 affects the self-destruction of the cell (MH Kim, et al., BBRC 2004, 321 , 930-935). It is also known that three trail molecules exhibit a structurally bound trimeric structure and that such a trail of trimeric structures binds to a receptor involved in cell death and induces apoptosis (FC Kimberley, et al. Cell Research 2004, 14, 359-372).

The most striking difference between the other TNF superfamilies, TNF and CD95L, is that they do not induce the death of normal tissue cells. Since TNF and CD95L also induce cell death, a variety of pharmaceutical applications have been attempted. These proteins such as TNF and CD95L induce the death of cancer cells and hyperactivated immune cells as well as affect normal cells. On the other hand, it is known that trail induces auto-apoptosis of various kinds of cancer cells and hyperactivated immune cells and does not affect general cells. This is known to be due to differences in the expression of trail receptor in each cell.

There are five types of tracer receptors so far. Representative apoptosis-related receptors include DR4 (TRAIL-R1) and DR5 (TRAIL-R2). When the trail binds to these receptors, the cell self-destroys through the activation of the death-related domain in the cell and various signal transduction systems. In addition, there are three receptors, DcR1, DcR2, and OPG (osteoprotegerin). These three receptors are known not to be involved in cell death. The difference in the degree of expression of DR4 and DR5 involved in apoptosis in normal cells and cancer cells is known to be insignificant. On the other hand, the other three receptors that are not related to apoptosis are well expressed in normal cells, but the expression level is low or not expressed in cancer cells. Therefore, in the case of trail, the binding of DcR1, DcR2 and OPG, which do not bind to the death-related domain, predominantly occurs in normal cells and does not induce apoptosis of cells. On the other hand, in cancer cells and hyperactivated immune cells, DR4 and DR5 It is known that self-apoptosis of the cell is induced by binding with Such selective cell death is considered to be a very useful property in the clinical application of trail.

Cancer cells, glioma cancer, lung cancer cells, prostate cancer cells, brain tumor and multiple myeloma have been reported as cancer cells which are observed by the trail-like cell death, and they exhibit excellent anticancer activity in animal experiments .

However, when only the protein of the natural trail is used for treatment, the cell can not bind properly to the receptor according to the low trimer formation ratio, and the cell death effect is delayed. In addition, there is a problem that not only toxicity can be shown to normal cells by using only the trail itself but also the half life is short. In the case of the trail, it has been reported that it has different half-life depending on the animal species used in the experiment, and it has been reported to have a half-life of about 30 minutes in rodents and in apes (H. Xiang, et al. Drug Metabolism and Disposition 2004, 32, 1230-1238).

Chemotherapeutic drugs are drugs that act on various metabolic pathways of cancer cells and exhibit cytotoxic or growth inhibitory effects. In particular, various anticancer drugs and compounds, including doxorubicin, which are used in chemotherapy for lung cancer, are injected mainly by intravenous injection. However, it has been reported that various side effects, which are represented by cardiovascular diseases, appear in chemotherapy using intravenous injection.

Under these circumstances, the inventors of the present invention have found that, while being able to be used for lung inhalation, they have a target orientation to lung cancer, and also, when using conventional albumin nanoparticles as drug delivery materials, they can reduce toxicity and side effects, As a result of diligent efforts to develop the particles, we have developed albumin nanoparticles with chemically modified anticancer agents and fatty acids surface-modified and trail-encapsulated. In addition, the albumin nanoparticles have high biocompatibility and are confirmed to be effective in prevention or treatment of lung cancer because it is possible to reduce adverse effects and to control the release of drugs by reducing administration dose through the combination of chemo-chemotherapeutic agent and trail, Thereby completing the invention.

An object of the present invention is to provide a method for producing self-assembled albumin nanoparticles in which a chemical anticancer agent and a fatty acid are surface-modified and trail (TRAIL) is enclosed.

Another object of the present invention is to provide a self-assembled albumin nanoparticle in which the chemical anticancer agent and the fatty acid produced by the above-described method are surface-modified and TRAIL is encapsulated.

It is still another object of the present invention to provide a pharmaceutical composition for preventing or treating cancer, which comprises the self-assembled albumin nanoparticle.

In one aspect, the present invention provides a method for producing self-assembled albumin nanoparticles in which a chemical anticancer agent and a fatty acid are surface-modified and trail (TRAIL) is encapsulated.

Specifically, the method of the present invention comprises the steps of (a) modifying albumin with a chemotherapeutic agent; (b) mixing and stirring albumin with a reducing agent and a fatty acid in the step (a); (c) adjusting the pH of the mixed solution stirred in step (b) and centrifuging to obtain an albumin conjugate wherein the chemical anticancer agent and the fatty acid are surface-modified; (d) adding the albumin conjugate obtained in step (c) to a sample containing TRAIL (TNF-related apoptosis inducing ligand) protein and ultrasonically pulverizing the same; And (e) obtaining the self-assembled albumin nanoparticle from the reaction solution after the step (d).

The step (a) is a step of modifying albumin with a chemo-chemotherapeutic agent, wherein albumin and a chemo-chemotherapeutic agent are modified with a choloric acid group and a maleimide group, or a maleimide group and a cholol group, respectively, But the present invention is not limited thereto.

That is, in step (a), the albumin may be modified with a choloric or maleimide group, and the chemo-chemotherapeutic agent may be modified with a maleimide group or a chiral group. In the step (a), the chelate or maleimide group may be modified to an amine group present on the surface of albumin or a chemo-chemotherapeutic agent.

Specifically, 2-iminothiolane or NHS-PEG-SH may be used for modifying the surface of the albumin of step (a) with a chelating agent. In particular, 2-iminoconazole can be used , But is not limited thereto. The cross-linker for modifying the surface of the albumin of step (a) with a maleimide group and joining it with a chelating agent is α- [3- (3-Maleimido-1-oxopropyl) amino] propyl-ω- succinimidyl-oxycarboxy), polyoxyethylene (MAL-PEG-NHS), succinimidyl 4- [N-maleimidomethyl] cyclohexane-1-carboxylate or sulfosuccinimidyl 4- [N-maleimidomethyl] cyclohexane- Sulfo-SMCC may be used, but is not limited thereto.

Specifically, 2-iminothiolane or NHS-PEG-SH may be used for modifying the chemical anticancer agent of the step (a) with a chelating agent. In particular, 2-iminothiourea can be used But is not limited thereto. The cross-linker, which modifies the chemical anticancer agent of step (a) with a maleimide group and bonds it to the cholol, is a mixture of α- [3- (3-Maleimido-1-oxopropyl) amino] propyl-ω- (succinimidyl -oxycarboxy, polyoxyethylene (MAL-PEG-NHS), succinimidyl 4- [N-maleimidomethyl] cyclohexane-1-carboxylate or sulfosuccinimidyl 4- [N-maleimidomethyl] cyclohexane-1-carboxylate Sulfo-SMCC can be used, but it is not limited thereto.

In the present invention, when 2-aminocyclohexene is used to modify a chemical anticancer agent or albumin by a chelating agent, a chemical anticancer agent or albumin: 2-aminocyclohexane can be reacted in a ratio of 1: 1 to 1: 10 equivalent, 1: 1 to 1: 5 equivalent.

In a specific embodiment of the present invention, doxorubicin, which is a representative chemotherapeutic agent, was reacted with 2-aminocyclohexane in the ratio of 1: 2 equivalents for up to 3 hours in order to modify doxorubicin into a chelate.

The chemical anticancer agent may be prepared in the form of a dispersion using a solution containing deionized water, but is not limited thereto.

In the present invention, a chemical anticancer agent or a cross-linker for modifying the albumin surface with a maleimide group and conjugating with a chelate group is a [3- (3-Maleimido-1-oxopropyl) amino] propyl-ω- (succinimidyl-oxycarboxy ), polyoxyethylene (MAL-PEG-NHS), succinimidyl 4- [N-maleimidomethyl] cyclohexane-1-carboxylate or sulfosuccinimidyl 4- [N-maleimidomethyl] cyclohexane- , Especially sulfo-SMCC, may be used, but the present invention is not limited thereto. But is not limited thereto. In the above reaction, albumin and sulfo-SMCC can be reacted at a ratio of 1: 1 to 1: 50, particularly 1:10 to 1:30.

In one specific embodiment of the present invention, sulfo-SMCC was used as a cross-linking agent for modifying the maleimide group on the albumin surface in order to bond the chemotherapeutic agent modified with the chelating agent and albumin. Albumin and sulfo-SMCC were added in the ratio of 1: 20 and reacted for up to 2 hours.

The albumin may be in the form of a dispersion using a solution containing deionized water, but is not limited thereto.

For reference, an amine group present in albumin or a chemical anticancer agent can be modified with a thiol group or a maleimide group, and when a thiol group is modified in one substance, a maleimide group can be modified in another substance to enable conjugation of two polypeptides .

In the present invention, the surface-modified albumin and the chemical anticancer agent may be prepared in the form of a dispersion using a solution containing deionized water, and the two dispersions may be mixed and stirred to induce a bonding reaction. Stirring of the two dispersions may be carried out at 10 to 40 캜 for 1 to 24 hours, particularly at 15 to 25 캜 for 2 to 6 hours.

In a specific embodiment of the present invention, the surface-modified albumin and the chemotherapeutic agent were prepared in the form of a dispersion using a solution containing deionized water, and the two dispersions were mixed and stirred at 20 ° C for 3 hours. As a result, albumin whose surface was modified with a chemotherapeutic agent was obtained.

Since the albumin nanoparticles of the present invention enter the living body through inhalation, the albumin used in the preparation should have biocompatibility and biodegradability. That is, it is preferable that the above-mentioned albumin is safe for living cells and does not harm the human body when the trail protein and the chemical anticancer drug are completely decomposed at the end of the release.

The albumin that may be used in the method of the present invention may include human serum albumin, bovine serum albumin, ovalbumin, or a combination thereof, particularly human serum albumin , But is not limited thereto.

In the present invention, the step (b) is a step of mixing the albumin modified with the chemical anticancer agent of the step (a) with a reducing agent and a fatty acid and stirring the mixture. By this step, an albumin conjugate in which a fatty acid is modified on the surface of the albumin is prepared.

In the present invention, the reducing agent may be used without limitation, which is a reducing agent for promoting the bonding between fatty acid and albumin, and in particular may be sodium cyanoborohydride.

The term "fatty acid" in the present invention refers to a chain-like carboxylic acid, which is used in the present invention as an aldehyde form. In the present invention, the fatty acid may be selected from the group consisting of octanal, nonanal, decanal, undecanal, dodecanal, tridecanal, tetradecanal, May be selected from the group consisting of pentadecanal, hexadecanal, octadecanal, icosanal and docosanal but may be conjugated to albumin to cause hydrophobic interactions As long as it is not limited to that kind.

In the step (b) of the present invention, a reducing agent and a fatty acid are added to albumin modified with a chemotherapeutic agent and stirred. In this step, stirring may be carried out at 1 to 40 ° C for 1 to 96 hours, preferably at 1 to 10 ° C for 24 to 60 hours.

In one embodiment of the present invention, sodium cyanoborohydride and octanal are used as a typical reducing agent for the albumin dispersion modified with the chemical anticancer agent obtained in the step (a), octanal as a representative fatty acid, For 48 hours to prepare an albumin conjugate.

The step (c) of the present invention is a step of obtaining the albumin conjugate surface modified with a chemotherapeutic agent and the fatty acid prepared in the step (b). Specifically, in step (c), the pH of the mixed solution stirred in step (b) is adjusted and centrifuged to obtain an albumin conjugate in which a chemotherapeutic agent and a fatty acid are surface-modified.

In the present invention, the pH control in step (c) is to further increase the formation of nanoparticles by self-assembly of the albumin-modified albumin surface by hydrophobic interactions through the change of the pH to the vicinity of the isoelectric point of albumin to be. Specifically, the pH of the step (c) may be adjusted to pH 4 to 6, preferably pH 5 to 6 since the isoelectric point of human serum albumin is pH 4 to pH 6 have. In a specific embodiment of the present invention, the pH was adjusted to 5.5 in step (c).

In the present invention, the solution containing the albumin nanoparticles is centrifuged to remove the supernatant, and the precipitated albumin conjugate is obtained.

In the step (d), the albumin conjugate obtained in step (c) is added to a sample containing TRAIL protein and ultrasonically pulverized, and the ultrasonic treatment is applied to a mixture solution containing the albumin conjugate and the TRAIL protein And the step of enclosing the TRAIL protein into the albumin nanoparticles.

In the present invention, the ultrasonic pulverization in the step (d) may be performed at a frequency of 10 to 30 kHz, an output amplitude of 5 to 25%, preferably a frequency of 15 to 25 kHz, a frequency of 10 to 20% Output amplitude. The experimenter can arbitrarily modify the frequency and the amplitude as long as it corresponds to the ultrasonic wave that can be generated at the above-mentioned amplitude at the frequency.

Further, the ultrasonic pulverization of the present invention may be performed for 1 minute to 5 minutes under the above output conditions, and preferably for 1 minute to 3 minutes. In the present invention, it is preferable to perform ultrasonic treatment in a vessel containing ice to prevent denaturation of the albumin nanoparticle due to high temperature.

In the present invention, the albumin conjugate refers to a substance in which a chemotherapeutic agent and a fatty acid are conjugated to albumin, and refers to a chemotherapeutic agent and albumin in which fatty acid is surface-modified. As the albumin conjugate of the present invention includes a fatty acid structure, a hydrophobic interaction occurs between the same molecules in a hydrophilic environment, and the nanoparticles can be formed by self-assembly.

The term "TNF-related apoptosis inducing ligand (TRAIL)" in the present invention is a kind of tumor necrosis factor (TNF), which refers to a cell membrane protein involved in cell apoptosis. For the purpose of the invention, the trail protein is a protein that is enclosed in albumin nanoparticles, and is a protein capable of exhibiting an anti-cancer effect by being enclosed in albumin nanoparticles through hydrophobic interaction, but is not limited thereto.

In addition, in the present invention, the trail protein may be a natural or recombinant trail obtained by genetic modification, and may include a zipper amino acid sequence leading to the formation of a trimer of trail or / and a terminal group which facilitates separation and purification Trail. The trail protein may be a human trail having an amino acid sequence of 281 amino acids, and in particular, the trail protein may be a human trail having an amino acid sequence from amino acid 114 to 281 glycine, preferably an amino acid sequence of SEQ ID NO: 1 Lt; / RTI >

In addition, the trail protein can be obtained by attaching an isoleucine zipper to the N-terminal. It is easy to form a trail protein in the form of a trimer capable of effectively recognizing a trail protein having an isoleucine zipper at its N-terminus, so that it can exhibit an excellent anti-cancer effect as compared with a natural type trail.

In the present invention, the step (e) may be a step of obtaining self-assembled albumin nanoparticles containing a TRAIL protein from the ultrasonic pulverized mixture in step (d), which may be performed by centrifugation. The centrifugation may be performed at 5000 RPM to 15000 RPM, preferably at 8000 to 12000 RPM. Specifically, it may be centrifugation for 1 minute to 10 minutes, in particular centrifugation for 3 to 7 minutes. In a specific embodiment of the present invention, the ultrasonically pulverized mixture in step (d) is centrifuged at 10,000 rpm for 5 minutes to obtain self-assembled albumin nanoparticles of the present invention.

In one embodiment of the present invention, the doxorubicin is modified to albumin by using doxorubicin as a representative chemical anticancer agent. Octanal and cyanoborohydride are mixed with dobutrovirin-modified albumin and then agitated at low temperature to form nanoparticles. Then, it is added to the solution containing the trail, and the ultrasonic wave is applied by an ultrasonic grinder to enclose the trail in the albumin nanoparticle. In this way, albumin nanoparticles having chemically chemotherapeutic agents and fatty acids surface-modified and embedded with trails were prepared (Example 1 and Fig. 1A, Fig. 1B).

Since the self-assembled albumin nanoparticles of the present invention enter the body through respiration and the like, the protein drugs and albumin used in the preparation must have biocompatibility and biodegradability, and protein substances such as albumin are denatured or aggregated in an organic solvent It is preferred that all reactions carried out in the preparation process of the present invention are carried out in a hydrophilic solvent or aqueous solution, since they may lose their inherent function and activity.

In order to efficiently bind the thiol group or maleimide group-modified albumin with a fatty acid or a chemotherapeutic agent, it is preferable to conduct the reaction in a concentrated state at a high concentration.

In another aspect, the present invention provides a self-assembled albumin nanoparticle prepared by the above-described method, wherein the chemical anticancer agent and the fatty acid are surface-modified and the TRAIL is encapsulated.

The chemical anticancer agent, fatty acid and trail are as described above.

The term "albumin nanoparticle" in the present invention refers to nanoparticle-sized particles composed of albumin and can be produced through the production method of the present invention, but is not limited thereto. The term "self-assembled albumin nanoparticle" in the present invention means a nanoparticle formed by self-assembly of a hydrophobic interaction between the same molecules in a hydrophilic environment by including a fatty acid structure in an albumin conjugate.

In the present invention, the self-assembled albumin nanoparticle comprises a chemical anticancer agent and an albumin conjugate surface modified with a fatty acid, wherein the albumin conjugate constitutes self-assembled nanoparticles by hydrophobic interaction, and TRAIL ) May be enclosed.

In another aspect, the present invention provides a pharmaceutical composition for preventing or treating cancer, wherein the chemical anticancer agent of the present invention and the self-assembled albumin nanoparticle in which the fatty acid is surface-modified and the TRAIL is encapsulated.

Since such a pharmaceutical composition is surface-modified with a chemical anticancer agent and includes albumin nanoparticles including trail therein, it can be effectively used for the combined administration of cancer, and self-assembled albumin nanoparticles having biocompatibility and biodegradability As a drug delivery system, it was possible to reduce toxicity and side effects caused by crosslinking agents.

The albumin nanoparticle of the present invention has high biocompatibility and it is confirmed that administration dose of chemo-chemotherapeutic agent and trail can be decreased by decreasing the dose of the drug and controlling the release of the drug, and thus it is effective for the prevention or treatment of cancer.

The composition of the present invention is characterized in that it is used for prevention or treatment of lung cancer through anticancer activity against cancer cells and targeted targeting through lung inhalation by the combined administration of trail and chemotherapeutic agent.

The composition may comprise an effective amount of the chemotherapeutic agent of the present invention, surface-modified and trail-enclosed albumin nanoparticles, and pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, and other adjuvants and / or carriers. The composition of the present invention can be formulated in the form of an inhalation mixture and a (freeze) dry powder in a conventional manner, and various inhalable formulations can be prepared according to techniques well known in the art or known in the art Can be manufactured. The composition comprising the chemical anticancer agent and the fatty acid thus prepared and the albumin nanoparticle encapsulated with the trail is delivered to the alveoli through the lungs to continuously release the drug, and the increased anticancer effect and side effect reduction effect .

In a specific embodiment of the present invention, the chemotherapeutic agent of the present invention and the self-assembled albumin nanoparticle in which the fatty acid is surface-modified and trailed (TRAIL) are administered to an animal model inducing lung cancer, and the cancer treatment effect is confirmed. As a remedy, animals without H226 cells were injected into control, non-drug group (Group 1), albumin nanoparticle dosage group (doxorubicin-modified) (Group 2), albumin nanoparticle drug loaded with trail (Group 3), and the albumin nanoparticle administration group (Group 4) of the present invention, in which doxorubicin was modified and the trail was sealed, was administered three times a week for 4 weeks. The effect of the anticancer activity on the control group and Groups 1 to 4 was confirmed by comparative observation of lung tissue after dissection and tissue staining using TUNEL reagent.

As a result, the cross-section of the lungs of the mice treated with the albumin nanoparticles encapsulating doxorubicin and trail showed that lung cancer cells were significantly less distributed than the cross-sections of the lungs of the other groups (Figs. 5A and 5B) It was observed that cells treated with doxorubicin and trail treatment showed more apoptotic cells in the lung than in the other groups (Fig. 6). That is, it was confirmed that the composition comprising the chemical anticancer agent of the present invention and the albumin nanoparticle surface-modified with the fatty acid and encapsulated in the trail was administered to the lung cancer-induced mice, ).

As used herein, the term "cancer" refers to a cancer selected from the group consisting of lung cancer, non-small cell lung cancer, colon cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, , Colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small bowel cancer, endocrine cancer, thyroid cancer, pituitary cancer, adenocarcinoma, soft tissue sarcoma, urethra (CNS) tumors, primary central nervous system lymphoma, spinal cord tumor, prostate cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, kidney cell carcinoma, renal pelvic carcinoma, , Brainstem glioma, and pituitary adenoma, and may be, but not limited to, lung cancer.

As used herein, the term "prophylactic" refers to any act that inhibits or delays the onset of cancer by administration of a composition according to the present invention. As used herein, the term "treatment" refers to any act that inhibits or delays the deterioration of cancer by the administration of the composition according to the invention and restores it from the cancer.

The pharmaceutical composition of the present invention can be used as a single preparation, and can be used as a combination preparation by further containing a drug known to have an approved cancer treatment effect.

By "pharmaceutically acceptable" as used herein is meant not significantly irritating the organism and not interfering with the biological activity and properties of the administered active substance.

The pharmaceutical compositions of the present invention comprising a pharmaceutically acceptable carrier may be of various oral or parenteral formulations. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain one or more excipients, such as starch, calcium carbonate, sucrose or lactose lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The pharmaceutical composition may be in the form of tablets, pills, powders, granules, capsules, suspensions, solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze- It can have one formulation.

In addition, the pharmaceutical composition of the present invention may contain a pharmaceutically effective amount of a chemotherapeutic agent and self-assembled albumin nanoparticles in which fatty acid is surface-modified and TRAIL is encapsulated. The term "pharmaceutically effective amount " as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will vary depending on the species and severity, age, sex, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer an amount that can achieve the maximum effect in a minimal amount without causing side effects, and can be readily determined by those skilled in the art. Preferably, the pharmaceutical composition according to the present invention may contain 0.001 to 1500 μg / ml of the chemotherapeutic agent and the self-assembled albumin nanoparticle in which the fatty acid is surface-modified and the TRAIL is encapsulated, in the pharmaceutical composition, May be included in the range of 0.001 to 1000 占 퐂 / ml. Alternatively, the composition according to the present invention may contain 1 to 10% by weight, particularly 5 to 10% by weight, of a chemo-chemotherapeutic agent and self-assembled albumin nanoparticles surface-modified with a fatty acid and containing TRAIL have.

In another aspect, the present invention provides a method for preventing or treating cancer by administering a pharmaceutical composition comprising a chemotherapeutic agent, a self-assembled albumin nanoparticle surface-modified with a fatty acid and a TRAIL, . ≪ / RTI >

The above-described self-assembled albumin nanoparticles and cancers in which the chemical anticancer agent and the fatty acid are surface-modified and the trail (TRAIL) is encapsulated are as described above.

In the present invention, the term " individual " refers to all animals, including humans, from which a cancer has developed or may develop. By administering a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt thereof to a suspected cancer- The individual can be treated efficiently. Specifically, the individual is a mammal such as a cow, a horse, a sheep, a pig, a goat, a camel, a nutrient, a dog, or a cat that requires treatment of similar symptoms as well as a human, But is not limited thereto.

As used herein, the term "administering " means introducing a pharmaceutical composition of the present invention to a patient in any suitable manner, and the route of administration of the composition of the present invention is not particularly limited as long as it can reach the target tissue, It can be administered via various routes of parenteral administration.

The method of treatment of cancer of the present invention includes administering a pharmaceutically effective amount of a chemotherapeutic agent and a self-assembled albumin nanoparticle surface-modified with a fatty acid and containing TRAIL. That is, the method for treating cancer of the present invention includes administering a pharmaceutical composition of the present invention in which the chemical anticancer agent and the self-assembled albumin nanoparticle surface-modified with TRAIL are surface-modified and the pharmaceutically effective amount do.

The term "pharmaceutically effective amount" as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will vary depending on the species and severity, age, sex, The type of drug, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of the treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention may be administered as an individual therapeutic agent or in combination with another therapeutic agent, and may be administered sequentially or simultaneously with a conventional therapeutic agent. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art. The preferred dosage of the composition of the present invention will depend on the condition and the weight of the patient, the severity of the disease, the type of drug, the route of administration and the period of time, and the appropriate total daily dose may be determined by treatment within the proper medical judgment, Generally, an amount of 0.001 to 1000 mg / kg, preferably 0.05 to 200 mg / kg, more preferably 0.1 to 100 mg / kg, can be administered in a single dose, divided into several times a day. The composition is not particularly limited as long as it is an object for prevention or treatment of rheumatoid arthritis, and any object can be applied. For example, it can be applied to any non-human animal such as a monkey, a dog, a cat, a rabbit, a guinea pig, a rat, a mouse, a cattle, a sheep, a pig, Including but not limited to, For example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

The composition of the present invention can be formulated in a conventional manner by inhalation, dry powder or injection. Any additive that may be included in the composition may be any of those conventionally used in dry powders. For example, the base may comprise a mixture of inorganic salts such as sodium chloride or a mixture thereof, a mixture of saccharides such as mannitol, lactose, dextran, glucose, and mixtures of amino acids such as arginine. In addition, preservatives such as methylhydroxybenzoate and propylhydroxybenzoate or surfactants such as lecithin and nonionic surfactants can be added. The injectable formulations are preferably isotonic aqueous solutions and may contain sterilized and / or adjuvants such as preservatives, stabilizers, wettable or emulsifying accelerators, salts for the control of osmotic pressure and / or buffers and other therapeutically useful substances .

In a specific embodiment of the present invention, self-assembled albumin nanoparticles with chemically-modified chemotherapeutic agent and fatty acid surface-modified and trailed (TRAIL) were treated with a lung cancer-induced mouse model and compared with lung sections of other comparative groups , It was confirmed that lung cancer cells were significantly less distributed and that many apoptotic cells were present (Fig. 6).

In another aspect, the present invention provides a use of a chemotherapeutic agent, a self-assembled albumin nanoparticle in which a fatty acid is surface-modified and a TRAIL is encapsulated, in the prevention or treatment of cancer.

The self-assembled albumin nanoparticles of the present invention can be used for lung inhalation by surface-modifying a chemical anticancer agent and a fatty acid and encapsulating a trail (TRAIL), so that they have target orientation to lung cancer, and when conventional albumin nanoparticles are used as a drug delivery vehicle Can reduce the toxicity and side effects that may be caused by the cross-linking agents, and can reduce the side effects and reduce the drug release by decreasing the dose through the combined administration of the chemo-chemotherapeutic agent and trail, and thus can be widely used for cancer treatment and prevention Especially lung cancer.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a diagram schematically showing a chemical structure of a chemical anticancer agent modified on the surface of albumin and a process (a) for modifying the chemical agent and a process (b) for encapsulating a TRAIL protein in albumin nanoparticles formed through hydrophobic interaction to be.
2 is an electron micrograph of albumin nanoparticles prepared by the method of the present invention. a) is a photograph of several particles in one shot, and b) is a photograph of an enlarged shot of several particles.
Fig. 3 shows whether the albumin surface was modified with a chemotherapeutic agent by peak movement through high performance liquid chromatography.
FIG. 4 is a view comparing and comparing the cytotoxic effect of doxorubicin and trail (TRAIL) protein in the lung cancer cell line (H226) and the apoptosis effect when coadministered.
FIG. 5A shows the anticancer activity of the albumin nanoparticles of the present invention against lung cancer, which is a comparative diagram of apparent morphology and size of dissected mouse lungs.
FIG. 5B shows the anticancer activity of the albumin nanoparticles of the present invention against lung cancer, which shows the weight of the extracted lung of the dissected mouse.
FIG. 6 is a graph showing the anticancer activity and the cross-section of lung tissue of doxorubicin and trail (TRAIL) in the lung tissue of mice in which lung cancer was induced by a lung cancer cell line (H226).

Hereinafter, the present invention will be described in more detail with reference to examples. It is to be noted, however, that these examples are illustrative of some experimental methods and compositions for illustrative purposes of the present invention, and the scope of the present invention is not limited to these examples.

Example  1: chemical chemotherapeutic agent and fatty acid surface-modified Trail Enclosed  Production of albumin nanoparticles

1-1. Preparation of albumin conjugates with surface chemically modified chemotherapeutic agents and fatty acids

Albumin nanoparticles modified fatty acids and chemotherapeutic agents on albumin surface. Representatively, in the present invention, doxorubicin was used as a chemical anticancer agent modifying the surface of albumin.

More specifically, 0.8 mg of doxorubicin and 0.4 mg of 2-iminosoylene were dissolved in 800 μl of a PBS (pH 7.4) solution. Dissolved in 200 μl, and the two solutions were mixed to make 1 ml. After stirring for 3 hours, the amine groups on the surface of doxorubicin were modified with a chelate group.

Meanwhile, 5 mg of human serum albumin was added to 1 ml of PBS (pH 7.4) to dissolve gently. After adding 0.7 mg of sulfo-SMCC, sulfo-SMCC was bound to the albumin surface by stirring at room temperature for 2 hours. The solution was centrifuged (Mw cut off 10,000) to remove unreacted sulfo-SMCC and concentrated to a volume of 0.5 ml.

The albumin solution bound with sulfo-SMCC concentrated to 0.5 ml was added to 1 ml of PBS (pH 7.4) solution containing doxorubicin modified with an amine group on the surface of the chitosan, and doxorubicin was added to the albumin surface with the crosslinking agent sulfo-SMCC, Lt; / RTI > for 3 hours.

Sodium cyanoborohydride was added to 20 ml of PBS (pH 7.4) solution in which 8 mg of albumin modified with doxorubicin was dissolved. Then, 6 쨉 l of octanol was added to the solution and stirred at 4 째 C for 48 hours. After the reaction was completed, the mixture was adjusted to pH 5.5 and centrifuged at 10,000 RPM for 5 minutes to obtain an albumin conjugate. The obtained albumin conjugate was washed three times by redispersing in PBS (pH 7.4) solution.

1-2: Preparation of albumin nanoparticles chemically chemically chemically and octane-aliquotted and trail-filled

Using the albumin conjugate prepared in Example 1-1, trail-filled albumin nanoparticles were prepared by self-assembly by hydrophobic interaction generated by surface-modified octanoic acid.

Specifically, precipitated particles were collected by centrifuging the obtained albumin conjugate, redispersed in 2 ml of trail-containing solution, and then subjected to ultrasonic treatment using an ultrasonic disintegrator (Sonic & Materials Inc. Newtown, CT, USA) Amplitude) 15% by ultrasonication for 2 minutes to enclose the trail in the albumin nanoparticles. At this time, the ultrasonic pulverization process was carried out in a container containing ice (on ice) in order to prevent denaturation of proteins (trail and / or albumin) contained in the mixture as high heat was generated.

Thereafter, centrifugation was performed at 10,000 rpm for 5 minutes to obtain final self-assembled albumin nanoparticles having the trail therein.

Figs. 1A and 1B show a brief schematic diagram of Examples 1-1 and 1-2.

Example  2: Formation of albumin nanoparticles by scanning electron microscopy ( Morphology ) Confirm

In order to confirm the morphology of the albumin nanoparticles prepared in Example 1, sample observation was performed using a scanning electron microscope (FE-SEM; LEOSUPRA 55 GENESIS 2000).

The final albumin nanoparticles prepared in Example 1 were lyophilized and attached to a sample magnet, coated with argon gas, and observed with a scanning electron microscope (FIG. 2).

As a result, as shown in FIG. 2, it was confirmed that the albumin nanoparticle of the present invention had a diameter of 200 nm to 400 nm.

Example  3: Determination of albumin surface expression of chemical anticancer drug by high performance liquid chromatography

Albumin in which the chemotherapeutic agent-modified albumin prepared in Example 1 was identified through high performance liquid chromatography (PLRP-S 300A 8 μM 150 x 4.6 mm, manufactured by Agilent Technologies, USA). The mobile phase used was deionized purified water and 0.1% trifluoroacetic acid (TFA) was added to acetonitrile (CH ₃ CN). The ratio of mobile solvent deionized water to acetonitrile was linearly changed from 80:20 to 20:80 (%) for 8 minutes and linearly changed from 20:80 to 0: 100 for 2 minutes. Thereafter, the sample was linearly changed from 0: 100 to 80:20 for 1 minute and then maintained at 80:20 for 3 minutes. The column temperature was 40 ° C and the flow rate was 1 ml / min. The chromatogram was obtained at a wavelength of 220 nm of the detector UV.

As shown in Fig. 3, there were four peaks of albumin, sulfo-SMCC-modified albumin, albumin in which sulfo-SMCC was modified with chemical anticancer drug, and albumin in which both chemotherapeutic agent and fatty acid were all modified. It was confirmed that the peak of the albumin in which the anticancer agent and the fatty acid were modified shifted to the right side more than the peak of the albumin in which the chemical anticancer agent and the fatty acid were not modified.

Example  4: Confocal  Using a laser scanning microscope in vitro  Confirmation of anti-cancer effect

The anticancer effect of the albumin nanoparticles prepared in Example 1, that is, the combined chemotherapeutic effect of trail and doxorubicin released from albumin nanoparticles, was observed through a confocal laser scanning microscope (Carl Zeiss, Meta LSM510, Germany).

Specifically, lung cancer cells H226 were dispensed at 1 × 10 ⁴ cells per well and cultured for 24 hours to induce cell stabilization. Then, each of the cells is treated with albumin nanoparticles containing 200 μg of the final albumin and cultured for 24 hours. After the cells are fixed with paraformaldehyde, 50 μl of TUNEL (Transferase (TdT) -mediated dUTP nick end labeling) reagent is added and the light is blocked for 1 hour. It was then treated with DAPI (4 ', 6-diamidino-2-phenylindole) reagent and observed through a confocal laser scanning microscope, the results of which are shown in FIG. Here, doxorubicin inside the cell is red due to its own fluorescence, and apoptotic cells are green.

As a result, as shown in FIG. 4, it can be seen that the albumin nanoparticles prepared using the method of the present invention show an increased drug efficacy due to the combined administration of doxorubicin and trail.

Example  5: Albumin nanoparticles in vivo  Lung cancer chemotherapeutic effect confirmation

The anti-cancer effect of the albumin nanoparticles of the present invention prepared in Example 1 was confirmed and evaluated using 5-week-old male BALB / c nu / nu mice.

Specifically, the adaptive environment of mice was maintained for 2 weeks under constant conditions such as temperature (22 ± 3 ° C), humidity (55 ± 5%) and light (repeated bright and dark for 12 hours). Four weeks prior to dosing, ⁵ × 10 ⁵ H226 cells per mouse were injected intravenously via the tail vein to induce lung cancer. When induction of lung cancer by injected H226 cells was stabilized for 4 weeks, group 1, group 2, albumin nanoparticle-modified group 2, group 3, trail-enclosed albumin nanoparticle group 3, , And the albumin nanoparticle administration group (Group 4) of the present invention, in which doxorubicin was modified and the trail was enclosed, was administered three times a week for 4 weeks. Here, mice not injected with H226 cells were set as a control. As a result, the anticancer activity effects of Groups 1 to 4 were confirmed by comparative observation of lung tissues after dissection and tissue staining, and the results are shown in FIG.

As a result, as shown in FIG. 5, in the case of Group 4 treated with albumin nanoparticles in which the chemical anticancer agent of the present invention and the fatty acid were expressed and trailed, the apparent appearance of the lungs was significantly smaller than the other groups , And showed a lung size similar to that of the control group (Fig. 5A). In addition, the weight of the lungs was significantly lower than that of the other groups, indicating that the albumin nanoparticles of the present invention had an excellent anti-cancer effect (FIG. 5B).

Example  6: In lung tissue Doxorubicin Trail  Confirm antitumor activity

The anticancer effects of trail and doxorubicin through the lungs of the mice treated with the albumin nanoparticles prepared in Example 1 were observed with an optical microscope and a confocal laser scanning microscope.

Specifically, the adaptive environment of mice was maintained for 2 weeks under constant conditions such as temperature (22 ± 3 ° C), humidity (55 ± 5%) and light (repeated bright and dark for 12 hours). Four weeks prior to dosing, ⁵ × 10 ⁵ H226 cells per mouse were injected intravenously via the tail vein to induce lung cancer. When induction of lung cancer by injected H226 cells was stabilized for 4 weeks, group 1, group 2, albumin nanoparticle-modified group 2, group 3, trail-enclosed albumin nanoparticle group 3, , And the albumin nanoparticle administration group (Group 4) of the present invention, in which doxorubicin was modified and the trail was enclosed, was administered three times a week for 4 weeks. Here, mice not injected with H226 cells were set as a control.

As a result, the anticancer activity effects of Groups 1 to 4 were confirmed by comparative observation of lung tissue after dissection and tissue staining using TUNEL reagent. After dissection, add 50 μl of TUNEL (Transferase (TdT) -mediated dUTP nick end labeling) reagent to the section of the slice obtained from the lung tissue and cut off the light for 1 hour. Then, it was treated with DAPI (4 ', 6-diamidino-2-phenylindole) reagent, and the results were observed with an optical microscope and a confocal laser scanning microscope. Here, apoptotic cells were green.

As a result, as shown in the first column of Fig. 6, the cross-section of the lungs of the mice treated with the albumin nanoparticles encapsulated with doxorubicin and trail showed that the lung cancer cells were significantly less distributed Can be confirmed. In addition, as can be seen from the results from the second column, it can be seen that apoptotic cells were significantly more abundant in the lungs of the treatment group in which doxorubicin and trail were used than in the other groups, indicating that they exhibited an increased efficacy .

From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention without departing from the scope of the present invention as defined by the appended claims.

<110> Research & Business Foundation SUNGKYUNKWAN UNIVERSITY <120> Self-assembled albumin nanoparticles for combination therapy          of lung cancer and the method for preparing same <130> KPA140859-KR <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 167 <212> PRT <213> hTRAIL 114-281 <400> 1 Arg Glu Arg Gly Pro Gln Arg Val Ala Ala His Ile Thr Gly Thr Arg   1 5 10 15 Gly Arg Ser Asn Thr Leu Ser Ser Pro Asn Ser Lys Asn Glu Lys Ala              20 25 30 Leu Gly Arg Lys Ile Asn Ser Trp Glu Ser Ser Arg Ser Gly His Ser          35 40 45 Phe Leu Ser Asn Leu His Leu Arg Asn Gly Glu Leu Val Ile His Glu      50 55 60 Lys Gly Phe Tyr Tyr Ile Tyr Ser Gln Thr Tyr Phe Arg Phe Gln Glu  65 70 75 80 Glu Ile Lys Glu Asn Thr Lys Asn Asp Lys Gln Met Val Gln Tyr Ile                  85 90 95 Tyr Lys Tyr Thr Ser Tyr Pro Asp Pro Ile Leu Leu Met Lys Ser Ala             100 105 110 Arg Asn Ser Cys Trp Ser Lys Asp Ala Glu Tyr Gly Leu Tyr Ser Ile         115 120 125 Tyr Gln Gly Gly Ile Phe Glu Leu Lys Glu Asn Asp Arg Ile Phe Val     130 135 140 Ser Val Thr Asn Glu His Leu Ile Asp Met Asp His Glu Ala Ser Phe 145 150 155 160 Phe Gly Ala Phe Leu Val Gly                 165

Claims (18)

(a) modifying albumin with a chemotherapeutic agent;
(b) mixing and stirring the albumin with a reducing agent and an aldehyde compound in the step (a);
(c) adjusting the pH of the mixed solution stirred in step (b) and centrifuging to obtain an albumin conjugate surface-modified with a chemotherapeutic agent and an aldehyde compound;
(d) adding the albumin conjugate obtained in step (c) to a sample containing TRAIL (TNF-related apoptosis inducing ligand) protein and ultrasonically pulverizing the same; And
(e) obtaining a self-assembled albumin nanoparticle from the reaction solution having been subjected to the step (d), wherein the chemical anticancer agent and the aldehyde compound are surface-modified and the TRAIL is sealed; .
The method of claim 1, wherein the albumin is selected from the group consisting of human serum albumin, bovine serum albumin, ovalbumin, and combinations thereof.
4. The method of claim 1, wherein the chemotherapeutic agent is selected from the group consisting of doxorubicin, cisplatin, mitomycin, 5-fluorouracil, tamoxifen, sorafenib, octreotide, and combinations thereof.
The method according to claim 1, wherein the chemical anticancer agent in step (a) is surface-modified with a chelate group, the albumin in step (a) is surface-modified with maleimide groups, Wherein the maleimide group of the albumin reacts and is carried out in combination.
The method according to claim 1, wherein the chemical anticancer agent in step (a) is surface-modified with maleimide groups, the albumin in step (a) is surface-modified with a chelate group, Wherein the chelating agent of tile and albumin reacts and is carried out in combination.
The method according to claim 4 or 5, wherein the surface modification with the chewing gum is carried out using 2-iminoconazole.
6. The method according to claim 4 or 5, wherein the modification with the maleimide group is carried out using sulfo-SMCC (sulfosuccinimidyl 4- [N-maleimidomethyl] cyclohexane-1-carboxylate).
The method of claim 1, wherein the aldehyde compound of step (b) is selected from the group consisting of octanal, nonanal, decanal, undecanal, dodecanal, tridecanal, Wherein the method is selected from the group consisting of tetradecanal, pentadecanal, hexadecanal, octadecanal, icosanal, and docosanal. .
The method of claim 1, wherein the reducing agent in step (b) is sodium cyanoborohydride.
2. The method according to claim 1, wherein the stirring of step (b) is performed at 1 to 40 DEG C for 1 to 96 hours.
The method according to claim 1, wherein the ultrasonic pulverization in step (d) is performed at a frequency of 10 to 30 kHz and an output amplitude of 5% to 25% for 1 minute to 5 minutes.
The method according to claim 1, wherein the step (e) is performed by centrifuging the reaction solution through the step (d).
A self-assembled albumin nanoparticle prepared by the manufacturing method according to claim 1, wherein the chemical anticancer agent and the aldehyde compound are surface-modified and the TRAIL is encapsulated.
14. The method of claim 13, wherein said chemotherapeutic agent is selected from the group consisting of doxorubicin, cisplatin, mitomycin, 5-fluorouracil, tamoxifen, soraphenib, octreotide, particle.
14. The method of claim 13, wherein the aldehyde compound is selected from the group consisting of octanal, nonanal, decanal, undecanal, dodecanal, tridecanal, wherein the self-assembled albumin nanoparticle is selected from the group consisting of tetradecanal, pentadecanal, hexadecanal, octadecanal, icosanal and docosanal.
14. The nanoparticle of claim 13, wherein the self-assembled albumin nanoparticle comprises a chemical anticancer agent and an albumin conjugate surface modified with an aldehyde compound, wherein the albumin conjugate constitutes self-assembled nanoparticles by hydrophobic interaction, Self-assembled albumin nanoparticles in which TRAIL is encapsulated.
A pharmaceutical composition for preventing or treating cancer, which comprises the self-assembled albumin nanoparticle according to any one of claims 13 to 16.
18. The method of claim 17, wherein the cancer is selected from the group consisting of lung cancer, non-small cell lung cancer, colon cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, gastric cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, soft tissue sarcoma, soft tissue sarcoma, urethral cancer, penile cancer, endometrial carcinoma, endometrial carcinoma, endometrial carcinoma, uterine cancer, vaginal cancer, vulvar carcinoma, Renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer And pituitary adenoma. &Lt; RTI ID = 0.0 &gt; A &lt; / RTI &gt;

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